Develop a Sample Preparation Procedure for HPLC Analysis ...libproject.hkbu.edu.hk/trsimage/hp/02010364.pdf · Develop a Sample Preparation Procedure for HPLC Analysis of Glucosinolates

Develop a Sample Preparation Procedure for HPLC Analysis of Glucosinolates in Traditional Chinese Medicines

By

LEE Kim Chung (02010364)

A thesis submitted in partial fulfillment of the requirements for the degree of

Bachelor of Science (Honours) in Applied Chemistry

(Concentration in Environmental Studies)

at

Hong Kong Baptist University

22042005

II

ACKNOWLEDGEMENT

This is an ongoing project and has been done by graduated students Miss CY Cheung Miss

WM Au and Mr CK Kwong The negative electrospray ionization-quadrupole time-of- flight

mass spectrometry and MSMS analysis of glucosinolate standards vegetable and Traditional

Chinese Medicine (TCM) samples were done by Dr Zongwei Cairsquos M Phil students Mr WT

Ma and Mr W Chan All other experiments described in this thesis were my own original work

and were carried out by myself under the supervision of Dr Zongwei Cai Thank you for his

valuable advice and guidance for me in this project

Thank you for Prof Albert WM Lee as my observer and provide me Rorippa indica (Linn)

Hiern sample [蔊菜]

Thank you for Dr Zhong-zhen Zhao provide me Leaf of Isatis indigotica Fort sample [大青葉]

Thank you for Dr Zongwei Cairsquos M Phil students Mr WT Ma and Mr W Chan as well as his

PhD student Miss Q Luo and laboratory technician Mr John Ng who have helped me a lot in

this project

Signature of Student

Student Name

Department of Chemistry Hong Kong Baptist University

Date

III


By


Department of Chemistry

ABSTRACT

Glucosinolates which are β-D-thioglucoside-N-hydroxysulfates found in the plant family

Cruciferae especially in Brassica A reversed-phase HPLC method using Hypersil BDS C18

column was developed for analyzing twelve intact glucosinolates (glucoiberin glucocheirolin

progoitrin sinigrin epiprogoitrin glucoraphenin sinalbin gluconapin glucosibarin

glucotropaeolin glucoerucin gluconasturtiin) in three vegetable and ten Traditional Chinese

Medicine (TCM) samples The samples were extracted with methanol followed by filtration

and evaporation Interferences in the organic sample extracts were removed by using activated

Florisil solid-phase extraction A gradient program and mobile phases using methanol and

30mM ammonium acetate at pH50 allowed sufficient retention and baseline separation of the

glucosinolates in the sample extracts Individual glucosinolates were detected by a UV detector

at 233nm Further confirmation was done by using liquid chromatography mass spectrometry

The glucosinolate concentrations in the sample extracts were determined by using an external

calibration method Detection limits of the glucosinolates were 252microgg 78microgg 18microgg

57microgg 83microgg 108microgg 242microgg 198microgg 33microgg 147microgg 43microgg and 175microgg

respectively when 5g of dried TCM was analyzed The average recovery (accuracy) of the

method was 998 and the precisions were from 53 to 146 (relative standard derivation

n=3) respectively

IV

Contents

1 Introduction P1-P6

2 Experimental preparations and procedures

21 Chemicals and reagents P7

21 Vegetable and Traditional Chinese Medicine (TCM) samples

211 Vegetable samples P7

212 Traditional Chinese Medicine (TCM) samples P8

23 Preparation of individual intact glucosinolate standard solutions P9

24 Preparation of intact glucosinolate standard mixture solutions P9

25 Preparation of vegetable and Traditional Chinese Medicine (TCM) samples

251 Sample grinding and extraction P9-P10

252 Clean-up process P10

26 Preparation of buffer solution P11

27 HPLC analysis P11-P12

28 Mass spectrometry analysis P12-P13

3 Experimental results and analysis

31 Qualitative analysis

311 Determination of the retention times for each intact glucosinolate standard P13-P14

312 Identification of the glucosinolates in vegetable and Traditional Chinese Medicine (TCM) samples by using reversed-phase HPLC analysis P15-P16

313 Identification of the glucosinolates in vegetable and Traditional Chinese Medicine (TCM) samples by using ESI-QTOF-MS and MSMS analysis P16-32

32 Quantitative analysis

321 Calibration curves for each individual glucosinolate standard P33-P34

V

322 Detection limits of each glucosinolate P35

323 Method recoveries of each glucosinolate P36-37

324 Glucosinolate concentrations in vegetable and Traditional Chinese Medicine (TCM) samples P38-39

4 Discussion

41 Extraction P40

42 Clean-up process P40-P42

43 Optimization of buffer system P42

44 Gradient program P42-P43

45 Optimization of glucosinolate concentrations in the sample extracts P43-P44

5 Conclusion P45

6 Future plan P45-P46

7 References P47-P48

1

1 Introduction

Glucosinolates are β-D-thioglucoside-N-hydroxysulfates[1] More than 120 individual

glucosinolates differing from each other in their structures of their glycon mioties have been

identified these generally classified as alkyl aliphatic alkenyl hydroxyalkenyl aromatic or

indole[2] The diversity of the R Group leads to a wide variation in the polarity and biological

activity of the natural products[3] The glucosinolates generally occur in the form of the

sodium or potassium salt and the general structure is shown in Figure 1[45]

ΟΟΗ

OH

OH

CH2OH

S C

R

NOSO3

Figure 1 General structure of glucosinolates

Glucosinolates are a class of approximately 100 plant secondary metabolites which contained

in the seeds roots stems and leaves of plants belonging to 11 families of dicotyledonous

angiosperms of which the crucifers are certainly the most important[6] Structural types and

individual concentrations differ according to various factors for example species tissue type

physiological age and plant health and nutrition[2] Glucosinolate concentrations in the

reproductive tissues (florets flowers and seeds) are often as much as 10-40 times higher than

in vegetative tissues [2] Plant myrosinase is widespread in seeds and tissues of the family

Cruciferae and catalyzes the hydrolysis of glucosinolates which are also contained in plant

vacuoles of the cruciferous plants[7] This reaction produces goitrogenic and potentially

hepatoxic compounds eg isothiocyanates thiocyanates nitriles and thiones[7] depending on

reaction conditions such as pH temperature metal ions protein cofactors and the properties

2

of the side chain [2]

R C

N

S

OSO3

β⎯D⎯glucose

R C

S

N

ΟΟ Η

O H

O H

C H 2 O H

O H

HSO4 unstable aglucon Glucose hydrogen suphate ion

R S C N R S C N R C N S

Thiocyanate Isothiocyanate Nitrile Sulfur

Figure 2 Degradation of glucosinolate by myrosinase in the presence of water[8]

The great number of the individual glucosinolates produces a large range of flavours as well as

toxic effect upon consumption[910] Glucosinolates have long been known for the fungicidal

bacteriodical nematocidal and allelopathic properties[1] The activity of isothiocyanates

such as sulforaphane against numerous human pathogens for example Escherichia coli

Salmonella typhimurium and Candida spp could even contribute to the medicinal properties

ascribed to cruciferous vegetables such as cabbage and mustard[1]

Glucosinolate hydrolysis products especially the isothiocyanates were demonstrated that

these molecules affect human health either beneficially or adversely[2] Several mechanism

have been proposed to the cancer prevention by breakdown products from cruciferous

vegetables And they have been proposed to act as blocking agents against carcinogenesis by

+ +

gt pH7 pH3 Fe2+

Myrosinase H2O

3

quinone reductase activity[11] Moreover glucosinolates and derived products would prevent

carcinogen molecules from reaching the target site or interacting with the reactive

carcinogenic molecules or activating the important hepatic enzymes for the protection against

several carcinogens[11] while some glucosinolates and their breakdown products are found to

have anti-nutritional effect in cattle[12]

For the determination of glucosinolates it is performed either by indirectly measuring the

enzymatic degradation products or by directly determining the intact glucosinolates[13]

However enzymatically or chemically released products such as isothiocyanates

oxazolidinethiones thiocyanate ion sulfate nitrile or glucose are also measured in the direct

analysis[13] Since the direct analysis of the intact glucosinolates can reflect the specificity

for the analysis of each individual glucosinolate Therefore the direct analysis of intact

glucosinolates is also used [13]

Product analyzed Method of dramatization and identification Main references

1Total glucosinolates I Palladium chloride for tetrachloropallidate assay Moller et al(1985) Bennert and Pauling (1988)

II Thymol assay Tholen et al(1989) Bennert and Pauling(1988)

III Glucose-release enzyme-coupled assay Heaney et al (1988)

IV Sulphate-release assay Schung (1987 1988)

V ELISA Van Doorn et al (1998)

VI Near infra-red reflectance(NIR) spectroscopy Velasco and Becker (1998)

VII Alkaline degradation and thioglucose detection Jesek et al (1999)

2 Individual intact I Reverse phase HPLC-MS Moller et al (1985) Bjerg and Sorenson (1987)

glucosinolates Hogge et al (1988) Kokkonen et al (1991)

Prestera et al (1996) Lewkw et al (1996)

Zrybko et al (1997) Schutze et al (1999)

Kaushik and Agnihotri (1999)

II Thermospray LC with tandem MS Heeremans et al (1989)

III High performance capillary electrophoresis Arguello et al (1999)

IV Capillary GC-MS GC-MS GC-MS-MS Shaw et al (1987 1989)

3 Desulphoglucosinolates I Reverse phase HPLC Fenwick et al (1983) Quinsac et al (1991)

4

Heaney and Fenwick (1993) Bjergegaard et al

(1995) Hrncirik and Velisek (1997) Robertson

and Botting (1999) Griffiths et al (2000)

II X-ray fluorescence spectroscopy (XRF) Schung and Hane Klaus (1990)

4 Degradation Products I GC or GC-MS Velisek et al (1990) Daxenbichler et al (1991)

II HPLC (all degradation products) Matthaus and Fiebig (1996)

II HPLC (fluorescent labeled products) Karcher and EI Rassi (1998)

III HPLC ( 12-benzenedithiol derivatives of Jiao et al (1998)

isothiocyanates

Table 1 Some of the commonly used methods for the quantitative and qualitative

analysis of the intact glucosinolates desulphoglucosinolates and their breakdown products [14]

Various alternative methods eg GC analysis of the trimethsilyl (TMS) derivatization of

glucosinolates and high-performance liquid chromatography (HPLC) of the

desulfoglucosinolate have been used for direct or indirect determination of total glucosinolate

and individual glucosinolates GC-MS analysis of the glucosinolate breakdown products is

often used [15] After a simple clean-up process the hydrolysis products were determined

qualitatively and quantitatively by GC-FID [16] However some side chains of the

glucosinolates are non-volatile or breakdown products are unstable for the determination [15]

Therefore the most suitable technique is to use the HPLC analysis of the enzymatically

desulfated glucosinolates Desulfated glucosinolate gives a better separation However

desulfated glucosinolates are often subject to the difficulties in interpreting results of the

individual glucosinolates due to concerns over the effect of pH value time and enzyme

concentration on desulfation products[19] Therefore the direct analysis of the intact

glucosinolates is needed for more specific and accurate determination for better interpretation

of analytical results for the reduction of analytical time [13]

It is an ongoing project three more glucosinolate standards would be analyzed and clean-up

process is also studied Twelve intact glucosinolate standards including epiprogoitrin

5

glucocheirolin glucoerucin glucoiberin gluconapin gluconasturtiin glucoraphenin

glucosibarin glucotropaeolin progoitrin sinalbin sinigrin are determined qualitatively and

quantitatively by using the reversed-phase high-performance liquid chromatography (HPLC)

The trivial and chemical names chemical formulas and chemical structures of side-chains and

molecular weights of intact glucosinolate standards used for analysis were shown in Table 2

The retention times of each of the intact glucosinolates in the HPLC column depend on the

polarities of the glucosinolates in the differences by the R groups Longer side chain or

containing of the aromatic ring in the R group will make it relative non-polar On the other

hand shorter side chain or containing the polar R group will make it relative polar

Therefore the HPLC can be used for analysis

6

Trivial Name and Chemical formula of glucosinolate (GS)

Chemical Name of glucosinolate (GS)

Chemical structure of R Group a

Molecular weight b gmol

Glucoiberin (C11H20NO10S3)

3-(methylsulfinyl)propyl-GS

4230327

Glucocheirolin (C11H20NO11S3)

3-(Methylsulfonyl)propyl-GS

4390277

Progoitrin (C11H18NO10S2)

(2R)-2-Hydroxybut-3-enyl-GS

3890450

Sinigrin (C10H16NO9S2)

Prop-2-enyl-GS

3590345

Epiprogoitrin (C11H18NO10S2)

(2S)-2-Hydroxybut-3-enyl-GS

3890450

Glucoraphenin (C12H20NO10S3)

4-(methylsulfinyl)but-3-enyl-GS

4350327

Sinalbin (C14H18NO10S2)

p-Hydroxybenzyl-GS

4250450

Gluconapin (C11H18NO9S2)

But-3-enyl-GS 3730501

Glucosibarin (C15H20NO10S2)

(2R)-2-Hydroxy-2-phenethyl-GS

4390607

Glucotropaeolin (C14H18NO9S2)

Benzyl-GS

4090501

Glucoerucin (C12H22NO9S3)

4-(Methylthio)butyl-GS

4210535

Gluconasturtiin (C15H20NO9S2)

Phenethyl-GS

4230658

Remarks a = the side-chain R in the general structures shown in Figure 1 b = Molecular weights of intact glucosinolates Table 2 The trivial and chemical names chemical formulas and chemical structures of

side-chains and molecular weights of intact glucosinolate standards used for analysis

H2C

H2C

H2CS

O

H3C

H2C

H2C

H2CS

O

H3C

O

H 2CC

O H

H

CH

H 2C

(2R)

H2CC

HH2C

(2S)

H2CC

H

H2CC

HS

O

H3C

H2CHO

H2C

H2CC

HH2C

H2CC

OH

H

H2C

H2C

H2C

H2C

H2CSH3C

H2C

H2C

H 2CC

H

O H

CH

H 2 C

7


21 Chemicals and reagents

Epiprogoitrin glucocheirolin glucoerucin glucoiberin gluconapin gluconasturtiin

glucoraphenin glucosibarin glucotropaeolin progoitrin sinalbin were obtained from KVL

(Frederiksberg C Denmark) Sinigrin was obtained from Sigma (St Louis USA)

HPLC-grade hexane and methanol were obtained from Riedel-de Haeumlnreg (Hanover Germany)

HPLC-grade dichloromethane and ethyl acetate were obtained from Tedia (Fairfield USA)

Ammonium acetate was obtained from Panreac (Barcelona Spain) and formic acid was from

Merck (Darmstadt Germany) Milli-Q water was produced by using a Milli-Qreg Ultrapure

Water Purification Academic System from Millipore (Billerica USA)


221 Vegetable samples

Three vegetable samples were analyzed and shown in Table 3 They were purchased from

Wellcome supermarket in Hong Kong

Local name Scientific name Family name

I Chinese Radish [蘿蔔] Raphanus sativus Cruciferae [十字花科]

II Cherry Tomato [車厘茄] Lycopersicon esculentum Solanaceae [茄科]

III Tomato [番茄] Lycopersicon esculentum Solanaceae [茄科]

Table 3 Local Scientific and Family names of the vegetable samples used for analysis

8

222 Traditional Chinese Medicine (TCM) samples

Ten TCM samples were analyzed and shown in Table 4 Leaf of Isatis indigotica Fort [大青

葉] was a gift from Dr Zhong-zhen Zhao School of Chinese Medicine Hong Kong Baptist

University Rorippa indica (Linn) Hiern [蔊菜] was a gift from Prof Albert WM Lee

Department of Chemistry Hong Kong Baptist University The other TCM samples were

purchased from Mr amp Mrs Chan Hon Yin Chinese Medicine Specialty Clinic amp Good

Clinical Practice Centre Hong Kong Baptist University


1 北板藍根 Root of Isatis indigotica Fort Cruciferae [十字花科]

2 南板藍根 Root of Baphicacanthus cusia (Nees) Bremek Acanthaceae [爵床科]

3 敗醬草 Patrinia scabiosaefolia Fisch ex Trev Valerianaceae [敗醬草科]

4 菥蓂 Thlaspi arvense L Cruciferae [十字花科]

5 大青葉 Leaf of Isatis indigotica Fort Cruciferae [十字花科]

6 廣東大青葉 Leaf of Baphicacanthus cusia (Nees) Bremek Acanthaceae [爵床科]

7 蔊菜 Rorippa indica (Linn) Hiern Cruciferae [十字花科]

8 白芥子 Seed of Sinapis alba L Cruciferae [十字花科]

9 萊菔子 Seed of Raphanus sativus L Cruciferae [十字花科]

10 葶藶子 Seed of Lepidium apetalum Willd Cruciferae [十字花科]

Remarks 1 is the original TCM and commonly confused by 2 in Hong Kong 3 is the original TCM and commonly confused by 4 in Hong Kong

5 is the original TCM and commonly confused by 6 in Hong Kong Table 4 Local Scientific and Family names of the Traditional Chinese Medicine (TCM)

samples used for analysis

9

23 Preparation of individual intact glucosinolate standard solutions

1000ppm of individual intact glucosinolate standard solutions were prepared by dissolving

1mg each of intact glucosinolates (glucoiberin glucocheirolin progoitrin sinigrin

epiprogoitrin glucoraphenin sinalbin gluconapin glucosibarin glucotropaeolin glucoerucin

gluconasturtiin) in 1mL of Milli-Q water respectively

24 Preparation of intact glucosinolate standard mixture solutions



gluconasturtiin) was weighed by precision weighing balance (Sartorius Bradford Germany)

and was dissolved in 1mL of Milli-Q water to prepare stock standard mixture solution with a

concentration of 1000ppm This stock solution was diluted to prepare standard mixture

solutions at 500ppm 400ppm 300ppm 200ppm 100ppm 50ppm and 5ppm


251 Sample grinding and extraction

Fresh vegetable or dried TCM sample was submitted to an initial grinding in an Extra Fine

Blade Blender (Hitachi Ibaraki Japan) for 2 minutes to form vegetable paste or dried TCM

powder 50g of the vegetable paste or 5g of the dried TCM powder was weighed and

blended with 100mL methanol The sample mixture was heated with stirring at 70oC for 15

minutes After cooling to room temperature the sample mixture was filtered through a

Whatman No 40 filter paper (Maidstone England) by suction filtration using water vacuum

pump The sample extract residue was washed twice with 50mL methanol The collected

sample extract was evaporated to dryness under vacuum using a Rotovap (Caframo Germany)

at 55oC with 60 rpm The solid sample extract was then dissolved in 10mL methanol and

10

was centrifuged by cyclone centrifuge (Alltech Deerfield USA) for 15 minutes at

13000rpm The supernate was collected for clean-up process

252 Clean-up process

Non-glucosinolate interferences were eliminated from the organic sample extract by using

activated Florisil solid-phase extraction (SPE) column Florisil sorbent (Fisher Certified

ACS 60-100mesh) (Sigma St Louis USA) was activated overnight at 200oC before using

for solid-phase extraction procedure A 5mL polypropylene syringe barrel was filled with

08g of the activated Florisil sorbent in between two 20microm polypropylene frits

All solvents were kept at a flow rate of 1-2mLmin in a vacuum manifold (Alltech Deerfield

USA) by using water vacuum pump during the clean-up process The activated Florisil

column was rinsed with 5mL of 30 (vv) dichloromethane in hexane 300microL of the organic

sample extract was mixed with 5mL of 30 (vv) dichloromethane in hexane and then

transferred onto the column 5mL of 30 (vv) dichloromethane in hexane was added to

wash the non-polar interferences from the column The glucosinolates in the sample extract

were eluted from the column by using 5mL of 30 (vv) ethyl acetate in methanol The

fraction was evaporated to dryness using a TurboVapreg LV Evaporator (Zymark Hopkinton

USA) under a slow stream of nitrogen The solid sample extract was then dissolved in

300microL of Milli-Q water The aqueous sample extract was centrifuged by the cyclone

centrifuge for 15 minutes at 13000rpm The supernate was collected for HPLC analysis

11

26 Preparation of buffer solution

30mM ammonium acetate buffer solution at pH50 was prepared by dissolving 231g of

ammonium acetate in 1L of Milli-Q water followed by adding a certain amount of 100

formic acid until a calibrated Orion Model 420 pH meter (Delhi India) showed pH50 value

The buffer solution was then filtered through 02microm cellulose acetate filter paper (Alltech

Deerfield USA) by suction filtration using water vacuum pump The buffer solution was

degassed ultrasonically by a Branson 2510 series Ultrasonic degasser (Danbury USA) for 10

minutes and was ready for HPLC analysis

27 HPLC analysis

High-performance liquid chromatography (HPLC) experiments were performed on a Hewlett

Packard HP1100 series HPLC instrument with a diode array detector (DAD) (San Francisco

USA) A reversed-phase Hypersil BDS C18 column (250mm x 46mm id 5micro particle size)

(Alltech Deerfield USA) was used for separation of the glucosinolates in three vegetable

and ten TCM sample extracts 20microL of the aqueous sample extract was injected into the

HPLC system by 100microL HPLC-syringe (Alltech Deerfield USA) Individual intact

glucosinolates were detected by the DAD detector at a UV wavelength of 233nm HP1100

series degasser was used for the degas process HP chemstation was used to control the

operation of the system and performed data analysis A gradient program was used for

sufficient retention and baseline separation of the glucosinolates in which mobile phase A

consisted of 30mM ammonium acetate containing formic acid at pH50 and mobile phase B

consisted of pure methanol The gradient program was shown in Figure 3

12

Gradient program

0

5

10

15

20

25

30

35

0 5 10 15 20 25 30

Time(min)

B

1

Figure 3 Gradient program for separation of the glucosinolates

The flow rate was kept at 1mLmin during the HPLC analysis 100 mobile phase A and 0

mobile phase B were kept for the first 5 minutes Then 0 mobile phase B was gradually

increased to 30 mobile phase B from 5 minutes to 17 minutes 30 mobile phase B was

then kept until the end of the separation The twelve glucosinolate standards were separated

completely under these conditions and their corresponding retention times were recorded

By comparing the retention times of the twelve glucosinolate standards with those of the

sample extracts the presence of the glucosinlates in the sample extracts could be identified

The peak areas of the identified glucosinoates in the sample extracts were recorded and used

for quantitative analysis

28 Mass spectrometry analysis

HPLC fractions of the twelve intact glucosinolates detected in the vegetable and TCM sample

extracts were collected and analyzed by using electrospray ionzation-quadrupole time-of-flight

mass spectrometry (ESI-QTOF-MS) in negative mode and MSMS analysis for the

confirmation of the glucosinolates in the sample extracts

13

The confirmation depended on the masses of the molecular ions and their corresponding

fragment ions The QTOF mass spectrometer was equipped with a turbo ionspray source

(Sciex Q-Star Pulsar i Applied Biosystem Canada) The parameters of the turbo ionspray

were shown in Table 5

Ion source gas 1 25 Declustering potential 1 -650V

Ion source gas 2 8 Focusing potential -1650V

Curtain gas 15 Declustering potential 2 -150V

Ionspray voltage -4000V Collision gas 3

Temperature of Ion source gas 2

200 oC Scan mass mode 50-600 amu

Table 5 Experimental conditions for ESI-QTOF-MS analysis of the glucosinolates



311 Determination of the retention times for each intact glucosinolate standard Figure 4 Chromatogram of 300ppm glucosinolate standard mixture solution

min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800

1000

1200

DAD1 A Sig=2334 Ref=550100 (CHUNG2KCLEE108D)

47

72

51

50

58

37 6

244

70

42

84

92

11

234

11

899

15

084

15

811

16

160

19

101

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

14

In the HPLC analysis the twelve intact glucosinolate standards were baseline separated as

shown in Figure 4 The most polar glucosinolate standard glucoiberin was first eluted out

When the composition of mobile phase B was increased by the gradient program the

relatively non-polar glucosinolates were eluted out in an order of descending the polarities of

the glucosinolate standards The most non-polar glucosinolate standard gluconasturtiin was

the last one to be eluted out

Under the chromatographic conditions described in Chapter 27 the average and relative

retention times for each glucosinolate standard were determined and shown in Table 6

Intact Glucosinolate standard Retention time (min)

Glucoiberin 477 plusmn 002

Glucocheirolin 514 plusmn 002

Progoitrin 583 plusmn 003

Sinigrin 625 plusmn 003

Epiprogoitrin 704 plusmn 003

Glucoraphenin 848 plusmn 004

Sinalbin 1123 plusmn 003

Gluconapin 1190 plusmn 004

Glucosibarin 1508 plusmn 003

Glucotropaeolin 1582 plusmn 003

Glucoerucin 1616 plusmn 003

Gluconasturtiin 1911 plusmn 003

Table 6 The average and relative retention times for each glucosinolate standard

15

312 Identification of the glucosinolates in vegetable and Traditional Chinese Medicine (TCM) samples by using reversed-phase HPLC analysis

The glucosinolates in three vegetable and ten TCM samples were analyzed By comparing

the retention times of the sample extracts with those of the glucosinolate standards the

presence of the glucosinolates in the sample extracts could be identified However the

retention times of the glucosinolates in the sample extracts varied a little bit due to the

complicated martices in the sample extracts Therefore a small volume of 1000ppm

glucosinolate mixture standard solution was spiked into the sample extracts for the

identification of the glucosinolates in the sample extracts By comparing the peak areas of

the corresponding retention times in the chromatogram of original sample extract with the

spiked one the peak areas of the spiked one were increased It identified that the sample

extract contained the glucosinolates being spiked

By repeating the spiked standard method described above it identified that Root of Isatis

indigotica Fort [北板藍根 ] contained glucoiberin glucocheirolin progoitrin sinigrin

epiprogoitrin glucoraphenin sinalbin gluconapin glucotropaeolin glucoerucin and

gluconasturtiin

The chromatograms of Root of Isatis indigotica Fort [北板藍根] extract and Root of Isatis

indigotica Fort [北板藍根] extract with standards spiked were shown in Figures 5a and 5b

respectively

16

Figure 5a Chromatogram of Root of Isatis indigotica Fort [北板藍根] extract

Figure 5b Chromatogram of Root of Isatis indigotica Fort [北板藍根] extract with standards spiked

313 Identification of the glucosinolates in vegetable and Traditional Chinese Medicine

(TCM) samples by using ESI-QTOF-MS and MSMS analysis

By using the spiked standard method described in Chapter 312 the glucosinolates in the

sample extracts can be detected However the retention times of the spiked glucosinolate

standards might be same as those of the interferences in the sample extracts

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin

GlucocheirolinProgoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


48

05

52

12

58

34

62

76

69

71

85

48

11

306

11

883

15

799

16

026

19

255

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

Glucoerucin

17

To pinpoint the co-elution problem mentioned above the electrospray ionzation-quadrupole

time-of-flight mass spectrometry in negative mode was used for further confirmation of the

HPLC fractions of the glucosinolates detected in the sample extracts The identification was

based on the molecular ion mass and the pattern of their corresponding fragment ions It was

a more powerful method than the spiked standard method and capable of providing the

information on the elemental compositions and the structures of the molecules

HPLC fractions of the glucosinolates detected in vegetable and TCM sample extract were

collected The collected fractions were diluted with methanol followed by negative

ESI-QTOF-MS analysis

The deprotoned molecular ion [M-H]- of the HPLC fraction in ESI-QTOF-MS spectrum was

compared with that of the corresponding glucosinolate standard For example the

deprotonated molecular ion [M-H]- of gluconapin standard was found to be mz 3720536 in

ESI-QTOF-MS spectrum By comparing the mass of the deprotonated molecular ion [M-H]-

of the gluconapin standard with that of the Root of Isatis indigotica Fort [北板藍根] extract at

11883min mz 3720233 was found It showed a positive result for the further confirmation

of the gluconapin in the HPLC fraction collected from the Root of Isatis indigotica Fort [北板

藍根] extract at 11883min The ESI-QTOF-MS spectrums of gluconapin standard and Root

of Isatis indigotica Fort [北板藍根] extract at 11883min were shown in Figure 6a and 6b

respectively

18

Figure 6a ESI-QTOF-MS spectrum of gluconapin standard

Figure 6b ESI-QTOF-MS spectrum of Root of Isatis indigotica Fort [北板藍根]

extract at 11883min

-TOF MS 30 MCA scans from Sample 10 (Rt11955-1) of Chung291104wiffa=355978894933761710e-004 t0=566641529975095180e+001

Max 39820 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3000

3200

3400

3600

3800

3982 3720233

2641464 3740192

315054126721773830960 44000993410834 4559759 484019639399712972460

-TOF MS 30 MCA scans from Sample 5 (gluconapin) of Chungwiffa=356036418804506270e-004 t0=568918465398965050e+001

Max 44870 counts

200 210 220 230 240 250 260 270 280 290 300 310 320 330 340 350 360 370 380 390 400mz amu

0

500

1000

1500

2000

2500

3000

3500

4000

4487 3720536

3740497

25523932201537 2272081 2832709 388065535803532931867 32518753392078

[M-H]- Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2

Gluconapin standard MW = 3730501

[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2

Gluconapin in Root of Isatis indigotica Fort [北板藍根] extract at 11883min MW = 3730501

19

However interferences in the sample extracts might have the similar molecular mass For

further confirmation of the glucosinolates present in the sample extracts the MSMS analysis

with resolution of 10000 was done It provides further confirmation of the glucosinolates

detected and structural elucidation The pattern of the fragment ions of the glucosinolates is

different for different compounds even they have similar molecular mass

The MSMS spectrum of the gluconapin standard was shown in Figure 7a the peak at mz

3720600 corresponded to the deprotoned molecular ion [M-H]- of the gluconapin The

observed fragment ion at mz 2920796 resulted from the loss of SO3 from the [M-H]- ion

The peak at mz 2749975 corresponded to the molecular ion with the loss of HSO4 from the

[M-H]- ion The peak of mz 1950337 corresponded to the fragment ion of the glucose

group in gluconapin The peaks of mz 969584 and mz 799501 represented the fragment

ions of HSO4- and SO3

- respectively By comparing the MSMS spectrum of the gluconapin

with that of HPLC fraction collected from the Root of Isatis indigotica Fort [北板藍根]

extract at 11883min similar fragment ion pattern in the sample extract was shown in Figure

7b Therefore the gluconapin was identified in the HPLC fraction collected from the Root of

Isatis indigotica Fort [北板藍根] at 11883min

20

Figure 7a MSMS spectrum of gluconapin standard

Figure 7b MSMS spectrum of Root of Isatis indigotica Fort [北板藍根] at 11883min

-TOF Product (3720) 59 MCA scans from Sample 14 (Rt11955-1(MSMS)-2) of Chung291104wiffa=355978894933761710e-004 t0=566641529975095180e+001

Max 1240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120124 3720156

969548

749854

1950168

25899591300262

274971617897167995151289233

850283 29205781450374 2270103 2409984 3009815564612

-TOF Product (3720) 30 MCA scans from Sample 7 (gluconapinMS2) of Chungwiffa=356036418804506270e-004 t0=568918465398965050e+001

Max 1180 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

118 749866

969584

37206001300304799501 25901761950337 27499751789801850244 1289378 2920796590074 17599271630819

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S

[M-HSO4-H]-[M-SO3-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2

Root of Isatis indigotica Fort [北板藍根] extract at 11883min MW = 3730501

HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S[M-H]-

[M-HSO4-H]-

[M-SO3-H]-

21

By using similar confirmation procedures described as above glucoiberin glucocheirolin

progoitrin sinigrin epiprogoitrin glucoraphenin sinalbin gluconapin glucotropaeolin

glucoerucin and gluconasturtiin detected in the Root of Isatis indigotica Fort [北板藍根]

extract were analyzed and all gave the positive results expect glucoiberin and glucoerucin

Therefore the Root of Isatis indigotica Fort [北板藍根] contained glucocheirolin progoitrin

sinigrin epiprogoitrin glucoraphenin sinalbin gluconapin glucotropaeolin and

gluconasturtiin The other sample extracts were analyzed by the similar methods and the

chromatograms of the sample extracts were shown in the following

Figure 8a Chromatogram of Raphanus sativus [蘿蔔(Chinese Radish)] extract

Figure 8b Chromatogram of Raphanus sativus [蘿蔔(Chinese Radish)] extract with

standards spiked

Progoitrin G

lucocheirolin

Glucoiberin

Glucoerucin

Glucosibarin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


47

60

51

78

58

40

62

42

70

83

86

16

11

271

11

861

15

077

15

834

16

103

19

110

Glucoiberin

Glucocheirolin

Sinigrin Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Progoitrin

Glucoraphenin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


46

75

51

48

61

86

87

09

15

122

16

142

22

Figure 9a Chromatogram of Lycopersicon esculentum [車厘茄(Cherry Tomato)] extract

Figure 9b Chromatogram of Lycopersicon esculentum [車厘茄(Cherry Tomato)] extract

with standards spiked

Figure 10a Chromatogram of Lycopersicon esculentum [番茄(Tomato)] extract

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

11

49

71

56

31

60

64

68

38

82

40

11

051

11

718

14

935

15

656

15

991 1

890

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


82

67

14

950

18

873

Glucoraphenin

Glucosibarin

Gluconasturtiin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


58

53

87

29

16

168

19

153

Progoitrin

Glucoraphenin

Glucoerucin

Gluconasturtiin

23

Figure 10b Chromatogram of Lycopersicon esculentum [番茄(Tomato)] extract with standards spiked


Figure 11b Chromatogram of Root of Isatis indigotica Fort [北板藍根] extract with

standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

78

51

58

58

18

62

57

70

91

86

68

11

291

11

881

15

111

15

847

16

159

19

138

Glucoiberin

Glucocheirolin

Progoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Epiprogoitrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


52

12

58

34

62

76

69

71

85

48

11

306

118

83

15

799

19

255

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

24

Figure 12a Chromatogram of Root of Baphicacanthus cusia (Nees) Bremek [南板藍根] extract

Figure 12b Chromatogram of Root of Baphicacanthus cusia (Nees) Bremek [南板藍根] extract with standards spiked

Figure 13a Chromatogram of Patrinia scabiosaefolia Fisch ex Trev [敗醬草] extract

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

61

50

60

56

98

61

03

69

18

84

01

11

169

11

741

14

944 1

569

2 1

601

1

18

963

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


25

Figure 13b Chromatogram of Patrinia scabiosaefolia Fisch ex Trev [敗醬草] extract with standards spiked

Figure 14a Chromatogram of Thlaspi arvense L [菥蓂] extract

Figure 14b Chromatogram of Thlaspi arvense L [菥蓂] extract with standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


49

78

55

68

60

05

14

842

835

0

Glucosibarin

Glucoraphenin

Sinigrin

Progoitrin G

lucocheirolin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


46

57

50

54 56

93

60

99

69

01

83

67

11

161

11

743 14

941

15

688

16

006

18

908

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

-50 0

50 100 150 200 250 300 350


46

43

50

21

56

67

60

04

68

75

83

47

11

137

11

724

14

931

15

684

16

005

18

962

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

26

Figure 15a Chromatogram of Leaf of Isatis indigotica Fort [大青葉] extract

Figure 15b Chromatogram of Leaf of Isatis indigotica Fort [大青葉] extract with standards spiked

Figure 16a Chromatogram of Leaf of Baphicacanthus cusia (Nees) Bremek [廣東大青葉]

extract

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


47

63

51

69 5

823

6

238

70

65

86

68

11

293

11

873

15

081 1

583

2 1

614

1

19

126

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150

200 250 300 350


55

88

84

78

14

918

Progoitrin

Glucoraphenin

Glucosibarin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


27

Figure 16b Chromatogram of Leaf of Baphicacanthus cusia (Nees) Bremek [廣東大青葉]

extract with standards spiked

Figure 17a Chromatogram of Rorippa indica (Linn) Hiern [蔊菜] extract

Figure 17b Chromatogram of Rorippa indica (Linn) Hiern [蔊菜] extract with standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

59

50

50

56

94 6

094

6

901

83

64

11

141

11

736 1

494

0 1

570

8 1

601

0 18

967

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


62

09

19

122

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

04

49

88

56

02

59

96

67

77

82

20

11

064

11

629

14

850 1

561

1 1

594

1

18

920

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin Sinigrin

Gluconasturtiin

28

Figure 18a Chromatogram of Seed of Sinapis alba L [白芥子] extract

Figure 18b Chromatogram of Seed of Sinapis alba L [白芥子] extract with standards spiked

Figure 18c Chromatogram of five-fold dilution of Seed of Sinapis alba L [白芥子] extract

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

69

51

46 5

819

6

006

70

49

85

26

11

128

11

722

15

166

15

899

16

228 1

919

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

72

58

21

59

99

70

50

11

138

11

723

15

177

16

204 1

919

2

Glucoiberin Progoitrin

Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

53

58

18

61

76

70

27

11

212

11

833

15

119

16

145

19

093


Sinigrin

Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

29

Figure 18d Chromatogram of five-fold dilution of Seed of Sinapis alba L [白芥子]


Figure 19a Chromatogram of Seed of Raphanus sativus L [萊菔子] extract

Figure 19b Chromatogram of Seed of Raphanus sativus L [萊菔子] extract with

standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150

175


47

47

51

30

58

12

617

4

702

3

84

67

11

208

11

834

15

077

158

18 1

614

0

19

084

Glucoiberin

Glucocheirolin Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


57

50 60

92

69

29

79

53

11

809

Progoitrin

Epiprogoitrin

Glucoraphenin

Gluconapin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

33

50

10

56

77 60

70

68

57

79

24

11

128

11

732

14

924 1

567

7 1

599

2

18

929

Glucoiberin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Glucocheirolin

30

Figure 19c Chromatogram of ten-fold dilution of Seed of Raphanus sativus L [萊菔子] extract

Figure 19d Chromatogram of ten-fold dilution of Seed of Raphanus sativus L [萊菔子] extract with standards spiked

Figure 20a Chromatogram of Seed of Lepidium apetalum Willd [葶藶子] extract

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31

Figure 20b Chromatogram of Seed of Lepidium apetalum Willd [葶藶子] extract with standards spiked

Figure 20c Chromatogram of twenty-fold dilution of Seed of Lepidium apetalum Willd [葶藶子] extract

Figure 20d Chromatogram of twenty-fold dilution of Seed of Lepidium apetalum Willd [葶藶子] extract with standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32

By similar confirmations described in Chapter 31 the glucosinolates in the vegetable and

TCM sample extracts were identified and shown in Table 7

Vegetable and Traditional Chinese Medicine Samples

Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Raphanus sativus [蘿蔔(Chinese Radish)] O O X O X X X X O X O X

Lycopersicon esculentum [車厘茄(Cherry Tomato)] X X X X X O X X O X X O

Lycopersicon esculentum [番茄(Tomato)] X X O X X O X X X X O O

Root of Isatis indigotica Fort [北板藍根] X O O O O O` O O X O X O

Root of Baphicacanthus cusia (Nees) Bremek

[南板藍根] X X X X X X X X X X X X

Patrinia scabiosaefolia Fischex Trev

[敗醬草] X X X X X X X X X X X X

Thlaspi arvense L [菥蓂] X O O O X O X X O X X X

Leaf of Isatis indigotica Fort [大青葉] X X O X X O X X O X X X

Baphicacanthus cusia (Nees) Bremek

[廣東大青葉] X X X X X X X X X X X X

Rorippa indica (Linn) Hiern [蔊菜] X X X O X X X X X X X O

Seed of Sinapis alba L [白芥子] O X O O O X O O O X O O

Seed of Raphanus sativus L [萊菔子] X X O O O O X O X X X X

Seed of Lepidium apetalum Willd [葶藶子] X O X O X X O O X X O X

Remarks O = Detected X = Not Detected

Table 7 Identified glucosinolates in vegetable and Traditional Chinese Medicine samples

33


321 Calibration curves for each individual glucosinolate standard

Concentration of standards Peak Areas

Glucosinolates 5ppm 50ppm 100ppm 200ppm 300ppm 400ppm 500ppm Glucoiberin 5750 59528 128474 233979 354886 477439 587806

Glucocheirolin 5042 53342 116418 211301 320047 427474 530813 Progoitrin 6655 69325 151260 274352 414470 552809 686626 Sinigrin 10199 104688 226854 413544 623107 834519 1029754

Epiprogoitrin 5853 60221 130575 237371 360446 480189 597922 Glucoraphenin 4608 46658 101219 184887 278648 373958 466701

Sinalbin 12342 136218 297099 569420 862251 1141440 1404644 Gluconapin 5857 64987 141632 257745 391187 522012 649376

Glucosibarin 6763 69012 149650 271896 411267 549541 680996 Glucotropaeolin 10451 108015 235541 427270 642164 854262 1045704

Glucoerucin 6038 61448 134529 244527 366038 485557 596982 Gluconasturtiin 6797 70973 153600 278357 422178 563459 701529

Table 8 Peak areas of the glucosinolates at 5ppm 50ppm 100ppm 200ppm 300ppm 400ppm and

500ppm

Calibration curves for each glucosinolate standard were obtained by plotting the peak areas

against concentrations for each glucosinolate standard respectively The graph of the

calibration curve for epiprogoitrin was shown in Figure 21 The summary of the equations

and their corresponding correlation coefficients (R2 values) of the calibration curves for each

glucosinolate were shown in Table 9 According to the R2 values of the calibration curves for

each glucosinolate the linearity of all calibration curves were acceptable

dddddddddddddddddddddddddddddddd

34

Calibration curve for Epiprogoitrin

y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600

Concentration of Epiprogoitrin (ppm)

Are

a

Figure 21 Calibration curve for epiprogoitrin

Glucosinolates Equations of the calibration curves R2 values

Glucoiberin y = 11758x + 27812 R2 = 09996

Glucocheirolin y = 10588x + 25809 R2 = 09996

Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996

Epiprogoitrin y = 11916x + 28152 R2 = 09997

Glucoraphenin y = 9290x + 17229 R2 = 09997

Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996

Glucosibarin y = 13585x+ 38123 R2 = 09996

Glucotropaeolin y = 20928x + 98804 R2 = 09993

Glucoerucin y = 11927x + 57822 R2 = 09994

Gluconasturtiin y = 13974x + 34193 R2 = 09996

Table 9 The summary of the equations and their corresponding R2 values of the calibration curves for each glucosinolate

35

322 Detection limits of each glucosinolate

Both instrument detection limits and method detection limits of each glucosinolate can be

calculated by applying the following equation

CL=kSBb

Remarks CL = Concentration related to the smallest measure of response can be detected

k = 3(based on the confidence interval) SB = Standard derivation of the blank of the method b = Slope of the calibration curve for corresponding glucosinolates

By using the the equations of calibration curves for each glucosinolate and equation above the

detection limits for each glucosinolate were calculated and shown in Table 10

Glucosinolates Slope b Standard deviation SB

Instrument Detection Limit ngmicroL

Method Detection Limit microgg a

Method Detection Limit microgg b

Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033

Glucotropaeolin 20928 5113 733 1466 147

Glucoerucin 11927 862 217 434 043

Gluconasturtiin 13974 4080 876 1752 175 Remarks a = Method Detection Limit when 5g of dried TCM was analyzed

b = Method Detection Limit when 50g of fresh vegetable was analyzed

Table 10 Detection limits of each glucosinolate

36

323 Method recoveries of each glucosinolate

In the sample preparation some of the glucosinolates might lose in heating transfer of the

sample extract filtration evaporation and clean-up process Therefore recovery tests were

done to determine both accuracies of the glucosinolates extracted from those sample

preparation procedures described in Chapter 25 and the effects of the interferences on the

glucosinolates in the sample extracts

A recovery test was carried out by using a dried TCM sample Root of Belamcanda chinensis

(L) DC [射干] that had been repeatedly analyzed and showed no existing glucosinolates

50microL of 10000ppm of the glucosinolate mixture standard was spiked into 5 g Root of

Belamcanda chinensis (L) DC [射干] in 100mL methanol solution Then the spiked

sample was prepared by repeating the same experimental procedures described in Chapter 25

Theoretically the final concentration of the glucosinolates would be 50ppm The peak areas

of the glucosinolates determined in the sample extract were compared with those of standard

mixture solutions at 50ppm to obtain the recovery data The recovery test was repeated three

times in order to get the average recoveries (accuracies) and precisions [relative standard

derivations (RSD n=3)] of each standard

Method recoveries of the glucosinolates were calculated by applying the following equation

Peak area of the glucosinolates in the testing solution Recoveries () = X 100

Peak area of the glucosinolates in the 50ppm glucosinolate standard mixture solution

37

By using the similar calculation both the method recoveries and RSD values for each

glucosinolate were obtained and shown in Table 11 The method recoveries of each

glucosinolate were over 8610 with average 9980 The precisions were from 527 to

1460 (RSD n=3) respectively

Glucosinolates Peak areas of testing solution 1

Peak areas of testing solution 2


Peak areas of 50ppm standard mixture solution

Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209

Glucotropaeolin 112400 110650 111962 108015 10338 744

Glucoerucin 60414 58620 58489 61448 9630 878

Gluconasturtiin 68813 68323 69656 70973 9712 551

Table 11 Summary of method recoveries and RSD values for each glucosinolate

Average 9980 928

38

324 Glucosinolate concentrations in vegetable and Traditional Chinese Medicine (TCM) samples

For sinigrin in Thlaspi arvense L [菥蓂] extract

y Peak area of sinigrin in Thlaspi arvense L [菥蓂] extract = 962809

y = 20569x + 63207 for sinigrin

By substituting the peak area of sinigrin in Thlaspi arvense L [菥蓂] extract into the equation

of the calibration curve for sinigrin the concentration of sinigrin in 10mL sample extract was

found

Therefore x = 4650145ppm (mgL-1)

Weight of sinigrin in 5g Thlaspi arvense L [菥蓂] extract

= Concentration of sinigrin x diluted factor recovery of sinigrin

= 4650145ppm x 10 x 10-3L 9803

= 47436mg

Concentration of sinigrin in Thlaspi arvense L [菥蓂]

= Weight of sinigrin weight of Thlaspi arvense L [菥蓂]

= 47436mg 5g

=09487mgg

=94870microgg

Therefore the concentration of sinigrin in Thlaspi arvense L [菥蓂] was found to be

94870microgg

39

By similar calculation and compare the concentrations of the identified glucosinolates in the

sample extract with detection limit the glucosinolate concentrations in three vegetable and ten

TCM samples were obtained and shown in Table 12

Concentration (microgg)


Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Total glucosinolates

Raphanus sativus [蘿蔔(Chinese Radish)] 1302 100 X 265 X X X X 631 X 6116 X 8414

Lycopersicon esculentum [車厘茄(Cherry Tomato)] X X X X X 2663 X X 513 X X 683 3859

Lycopersicon esculentum [番茄(Tomato)] X X 340 X X 3041 X X X X 413 996 4790

Root of Isatis indigotica Fort

[北板藍根] X 955 79532 2891 209966 12454 2432 63609 X 1858 X 956 374653Root of Baphicacanthus

cusia (Nees) Bremek [南板藍根] X X X X X X X X X X X X 000

Patrinia scabiosaefolia Fisch ex Trev

[敗醬草] X X X X X X X X X X X X 000

Thlaspi arvense L [菥蓂] X 4287 1645 98470 X 28105 X X 2895 X X X 135402

Leaf of Isatis indigotica Fort [大青葉] X X 1432 X X 48877 X X 4232 X X X 54541

Leaf of Baphicacanthus cusia (Nees) Bremek

[廣東大青葉] X X X X X X X X X X X X 000

Rorippa indica (Linn) Hiern [蔊菜] X X X 2069 X X X X X X X 6063 8132

Seed of Sinapis Alba L [白芥子] 6631 X 834 431382 4143 X 159108 296687 2675 X 1701 2156 905317

Seed of Raphanus sativus L [萊菔子] X X 2364 1367 3768 1091908 X 2067 X X X X 1101474

Seed of Lepidium apetalum Willd

[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982

Remark X = Not Detected

Table 12 Glucosinolate concentrations in vegetable and Traditional Chinese Medicine (TCM) samples

40

4 Discussion

41 Extraction

To inactivate myrosinase to catalyze the hydrolysis of the glucosinolates into glucose and

unstable products that described in Figure 2 100 methanol was used for extraction of the

glucosinolates from the vegetable and Traditional Chinese Medicine (TCM) samples

Moreover less pigment was extracted and all glucosinolates could be extracted with high

recoveries above 85 Heating the sample extract at 70oC for 15minutes in extraction

process could denature the myrosinase and increase dissolution of the glucosinolates into

methanol After cooling to room temperature the proteins in the organic sample extract were

precipitated out due to the low solubility of proteins in methanol

42 Clean-up process

Both normal-phase solid-phase extraction using activated Florisil column and reversed-phase

solid-phase extraction using C18 column were tried for the clean-up process The activated

Florisil clean-up process was described in Chapter 25 And both C18 and activated Florisil

clean-up processes could achieve more than 88 of method recoveries for all glucosinolates

in testing conditions They differed from each other by the ability of removal of the

interferences in the sample extracts

In C18 clean-up process C18 cartridge (Alltech Deerfield USA) was rinsed with 5mL of

methanol followed by 5mL of deionized water The 5mL aqueous sample extract was

transferred onto the column followed by collection of the glucosinolates due to their poor

retentions on C18 column 5mL of the 10 methanol in deionized water was added to elute

the remaining glucosinolates The final volume of the sample extracts was 10mL

41

Glucosinolates were very polar that they had poor retention on the C18 column And C18

clean-up process was done by retaining of the non-polar interferences on the sorbent

However the interferences with the co-retention time of the glucotropaeolin in the Root of

Isatis indigotica Fort [ 北板藍根 ] were eluted together with the glucosinolates The

chromatograms of the original Root of Isatis indigotica Fort [北板藍根] and Root of Isatis

indigotica Fort [北板藍根] after C18 clean-up process were shown in Figure 21a and 21b

respectively

Activated Florisil clean-up method could remove the interferences with the co-retention time

of the glucotropaeolin in the Root of Isatis indigotica Fort [北板藍根] as shown in Figure 11a

Florisil was activated at 200oC for overnight that could remove volatile organic compounds in

the sorbent And therefore the activated Florisil could then be used as adsorption sorbent for

adsorption of slightly polar interferences By washing the column with 30 dichoromethane

in hexane the non-polar interferences were removed from the sample extracts Organic sample

extracts were prepared to load onto the column as water in the aqueous sample extract was too

polar that could inactivate the activated Florisil column

Figure 21a Chromatogram of the original Root of Isatis indigotica Fort [北板藍根]

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42

Figure 21b Chromatogram of the original Root of Isatis indigotica Fort [北板藍根] after

C18 clean-up process

43 Optimization of buffer system

Glucosinolates were present in ionic states in aqueous medium The retention of the

glucosinolates on the reversed-phase HPLC column was usually poor because of high polarity

of the glucosinolates Therefore 30mM ammonium acetate containing formic acid at pH50

was used for HPLC analysis The buffer solution was used to achieve better retention and

good peak shape for the glucosinolates Formic acid was a strong acid and easily protonate

the glucosinolates in aqueous medium Ammonium acetate was completely ionize in the

aqueous medium and suppressed the protonated glucosinolates to ionize The protonated

glucosinolates were relatively non-polar and had sufficient retention on the reversed-phase

C18 HPLC column for baseline separation of them

44 Gradient program

When 100 buffer solution was kept at a flow rate of 1mLmin in isocratic condition some

(relatively non-polar) peaks of the glucosinolates eluted from HPLC column more than one

hour and became flattened in peak shape When 100 methanol was kept at a flow rate of

1mLmin in isocratic condition the glucosinolate standards were eluted out together

Therefore gradient program was used The gradient program was used in HPLC system to

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350

DAD1 A Sig=2334 Ref=550100 (CHUNG2SUB00006D)

Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43

increase the resolutions and lower the detection limits of the glucosinolates To get a better

retention of the glucosinolates on the HPLC column a 100 aqueous phase was used as the

initial condition Methanol was used as the organic phase and was increased to 30 from 5

minutes to 17 minutes using the gradient program in order to separate the glucosinolates

completely and elute the relatively non-polar glucosinolates

45 Optimization of glucosinolate concentrationsin the sample extracts

The glucosinolate concentrations in some TCM samples especially the seed TCM samples

were found to be very high For example seed of Lepidium apetalum Willd [葶藶子]

contained 1264894microgg of the gluconapin and seed of Raphanus sativus L[萊菔子] contained

1091908microgg of the glucoraphenin Therefore the pre-concentration process was necessary

for quantitative analysis Otherwise the glucosinolate concentration in the sample extracts

might be out of the range of the calibration curve or even the HPLC column might be

overloaded The column overloading could be seen with back tailing of the peak in the

chromatogram For example the gluconapin in seed of Lepidium apetalum Willd [葶藶子]

extract overloaded the column as shown in Figure 22a And its corresponding peak area

7178070 was above the range of the calibration curve from 5857 to 649376 After

twenty-fold dilution of the seed of Lepidium apetalum Willd [葶藶子] extract a good peak

shape of the gluconapin was achieved as shown in Figure 22b And its corresponding area

502612 was within the range of the calibration curve

44


Figure 22b Chromatogram of twenty-fold dilution of Seed of Lepidium apetalum Willd [葶藶子] extract

min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion

The developed reversed-phase HPLC method using C18 column provides sufficient retention

and baseline separation for analyzing twelve intact glucosinolates (glucoiberin glucocheirolin


glucotropaeolin glucoerucin gluconasturtiin) in three vegetable and twelve Traditional

Chinese Medicine (TCM) samples Combined the method with ESI-QTOF-MS in negative

mode and MSMS analysis the glucosinolates in the sample extract can be detected and

identified Glucosinolate concentrations in the sample extract can be successfully determined

by an external calibration method

At least two glucosinoates were found in the three vegetable and cruciferous TCM samples

None of the detectable glucosinolates were found in the non-cruciferous TCM samples

Concentrations of the total glucosinolates were found to be very high in seed TCM samples

more than 9000microgg

Seed of Lepidium apetalum Willd [葶藶子] contained the highest concentration of total

glucosinolates 1319982microgg Root of Baphicacanthus cusia (Nees) Bremek

[南板藍根] Patrinia scabiosaefolia Fisch ex Trev [敗醬草] and Leaf of Baphicacanthus

cusia (Nees) Bremek [廣東大青葉] contained the least concentration of total glucosinolates

000microgg The range of the total glucosinolates in the sample extracts was found to be

000microgg to 1319982microgg respectively

46

6 Future plan

Fresh vegetable samples were analyzed in this project The water content was different in

different vegetable samples Therefore a drying freezer can be used to dry the vegetable

samples before analysis

Glucosinolate concentrations in the reproductive tissues (florets flowers and seeds) are often

as much as 10-40 times higher than those in vegetative tissues [2] And the seed TCM

samples were found with relatively high glucosinolate levels Therefore reproductive tissues

of the cruciferous plants can be analyzed and compared with the vegetative tissues

Different forms of 板藍根 products in the market can be analyzed for example 板藍根

extract to investigate whether they are made of Root of Isatis indigotica Fort [北板藍根] or

Root of Baphicacanthus cusia (Nees) Bremek [南板藍根]

47

5 References

1 Tsiafoulis CG Prodromidis MI Karayannis MI Anal Chem 2003 75 927-934

2 Bennett RN Mellon FA Kroon PA J Agric Food Chem 2004 52 428-438

3 Arguello LG Sensharma DK Qiu F Nurtaeva A Rassi ZE J of AOAC int

1999 82(5) 1115-1127

4 Warton B Matthiessen JN Shackleton MA J Agric Food Chem 2001 49

5244-5250

5 Szmigielska AM Schoenau JJ J Agric Food Chem 2000 48 5190-5194

6 Nastruzzi C Cortesi R Esposito E Menegatti E Leoni O Iori R Palmieri S

J Agric Food Chem 1996 44 1014-1021

7 Leoni O Iori R Palmieri S J Agric Food Chem 1991 39 2322-2326

8 Karcher A Melouk HA Rassi ZE J Agric Food Chem 1999 47 4267-4274

9 Karcher A Melouk HA Rassi ZE Analytical Biochemistry 1999 267 92-99

10 Mellon FA Bennett RN Holst B Williamson G Analytical Biochemistry 2002

306 83-91

11 Nastruzzi C Cortesi R Esposito E Menegatti E Leoni O Iori R Palmieri J

Agric Food Chem 2000 48 3572-3575

12 Tolra RP Alonso R Poschenrieder C Barcelo D Barcelo J J of

Chromatograph A 2000 889 75-81

13 Cai ZW Cheung CY Ma WT Au WM Zhang XY Lee A Anal Bioanal

Chem 2004 378 827-833

14 Kiddle G Bennett RN Botting NP Davidson NE Robertson AAB

Wallsgrove RM Phytochemical Analysis 2001 12 226-242

15 Botting CH Davidson NE Griffiths DW Bennett RN Botting NP J Agric

48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX

Figure A1 MSMS spectrum of glucoiberin standard Figure A2 MSMS spectrum of glucocheirolin standard

-TOF Product (4220) 30 MCA scans from Sample 6 (glucoriderin-MS2) of chung2wiffa=356258132228446760e-004 t0=570181404275608660e+001

Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O

Glucoiberin standard MW = 4230327

[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

-TOF Product (4380) 30 MCA scans from Sample 32 (GlucocheirolinMS2) of Chungwiffa=356036418804506270e-004 t0=568918465398965050e+001

Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O

Glucocheirolin standardMW = 4390277 [M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50

Figure A3 MSMS spectrum of progoitrin standard

Figure A4 MSMS spectrum of sinigrin standard

-TOF Product (3880) 30 MCA scans from Sample 8 (progoitrin-MS2) of chung2wiffa=356258132228446760e-004 t0=570181404275608660e+001

Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH

Progoitrin standard MW = 3890450

[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

-TOF Product (3580) 30 MCA scans from Sample 4 (sinigrinMS2) of Chungwiffa=356036418804506270e-004 t0=568918465398965050e+001

Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2

Sinigrin standard MW = 3590345

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51

Figure A5 MSMS spectrum of epiprogoitrin standard

Figure A6 MSMS spectrum of glucoraphenin standard

-TOF Product (3880) 30 MCA scans from Sample 1 (epiprogoitrin-MS2) of chung2wiffa=356258132228446760e-004 t0=570181404275608660e+001

Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H

Epiprogoitrin standard MW = 3890450

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

-TOF Product (4340) 30 MCA scans from Sample 18 (GlucorapheninMS2) of Chungwiffa=356036418804506270e-004 t0=568918465398965050e+001

Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3

Glucoraphenin standard MW = 4350327

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52

Figure A7 MSMS spectrum of sinalbin standard

Figure A8 MSMS spectrum of gluconapin standard

-TOF Product (4240) 30 MCA scans from Sample 57 (SinalbinMS2) of Chungwiffa=356036418804506270e-004 t0=568918465398965050e+001

Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH

Sinalbin standard MW = 4250450

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53

Figure A9 MSMS spectrum of glucosibarin standard Figure A10 MSMS spectrum of glucotropaeolin standard

-TOF Product (4080) 30 MCA scans from Sample 38 (GlucotropaeolinMS2) of Chungwiffa=356036418804506270e-004 t0=568918465398965050e+001

Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C

Glucotropaeolin standard MW = 4090501

-TOF Product (4380) 30 MCA scans from Sample 12 (glucosibarinMS2) of Chungwiffa=356036418804506270e-004 t0=568918465398965050e+001

Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH

Glucosibarin standard MW = 4390607

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54

Figure A11 MSMS spectrum of glucoerucin standard

Figure A12 MSMS spectrum of gluconasturtiin standard

-TOF Product (4220) 30 MCA scans from Sample 27 (GluconasturtiinMS2) of Chungwiffa=356036418804506270e-004 t0=568918465398965050e+001

Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

Gluconasturtiin standardMW = 4230658

[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

-TOF Product (4200) 30 MCA scans from Sample 22 (GlucoerucinMS2) of Chungwiffa=356036418804506270e-004 t0=568918465398965050e+001

Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3

Glucoerucin standard MW = 4210535

[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55

Figure A13a ESI-QTOF-MS spectrum of Lycopersicon esculentum [番茄(Tomato)]

extract at 8729min Figure A13b MSMS spectrum of Lycopersicon esculentum [番茄(Tomato)] extract at

8729min

-TOF Product (4340) 30 MCA scans from Sample 2 (t-rt-694-MS2) of chung1wiffa=356264499558618230e-004 t0=571842229067915470e+001

Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3

Glucoraphenin in Lcopersicon esculentum [番茄(Tomato)] extract at 8729min MW = 4350327

-TOF MS 0050 min from Sample 1 (t-rt-694) of chung1wiffa=356264499558618230e-004 t0=571842229067915470e+001

Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3

Glucoraphenin in Lycopersicon esculentum [番茄(Tomato)] extract at 8729min MW = 4350327

[M-H]-

56

Figure A14a ESI-QTOF-MS spectrum of Root of Isatis indigotica Fort [北板藍根]

extract at 5834min Figure A14b MSMS spectrum of Root of Isatis indigotica Fort [北板藍根] extract at

5834min

-TOF MS 40 MCA scans from Sample 21 (Rt5435) of Chung291104wiffa=355978894933761710e-004 t0=566641529975095180e+001

Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH

Progoitrin in Root of Isatis indigotica Fort [北板藍根] extract at 5834min MW = 3890450

[M-H]-

-TOF Product (3880) 60 MCA scans from Sample 22 (Rt5435(MSMS)-38801-1) of Chung291104wia=355978894933761710e-004 t0=566641529975095180e+001

Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57

Figure A15 ESI-QTOF-MS spectrum of Root of Isatis indigotica Fort [北板藍根]

extract at 6276min Figure A16a ESI-QTOF-MS spectrum of Root of Isatis indigotica Fort [北板藍根]

extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2

Sinigrin in Root of Isatis indigotica Fort [北板藍根] extract at 6276min

[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H

Epiprogoitrin in Root of Isatis indigotica Fort [北板藍根] extract at 6971min MW = 3890450

[M-H]-

58

Figure A16b MSMS spectrum of Root of Isatis indigotica Fort [北板藍根] extract at

6971min Figure A17a ESI-QTOF-MS spectrum of Root of Isatis indigotica Fort [北板藍根]

extract at 11290min

-TOF Product (3880) 100 MCA scans from Sample 47 (Rt6369(MSMS)-388) of Chung291104wiffa=355978894933761710e-004 t0=566641529975095180e+001

Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H

Epiprogoitrin in Root of Isatis indigotica Fort [北板藍根] extract at 6971min MW = 3890450 [M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH

Sinalbin in Root of Isatis indigotica Fort [北板藍根] extract at 11290min MW = 4250450

[M-H]-

59


11290min

Figure A18a ESI-QTOF-MS spectrum of Thlaspi arvense L [菥蓂] extract at 6005min

-TOF MS 0400 min from Sample 3 (z-rt-615) of chung1wiffa=356264499558618230e-004 t0=571842229067915470e+001

Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492

-TOF Product (4240) 60 MCA scans from Sample 34 (Rt11258(MSMS)-424-1) of Chung291104wiffa=355978894933761710e-004 t0=566641529975095180e+001

Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH

Sinalbin in Root of Isatis indigotica Fort [北板藍根] extract at 11290min MW = 4250450 [M-H]-

HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2

Sinigrin in Thlaspi arvensis L [菥蓂] extract at 6005min MW = 3590345

[M-H]-

60

Figure A18b MSMS spectrum of Thlaspi arvense L [菥蓂] extract at 6005min


-TOF MS 30 MCA scans from Sample 11 (z85) of chung2wiffa=356258132228446760e-004 t0=570181404275608660e+001

Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410

-TOF Product (3580) 30 MCA scans from Sample 4 (z-rt-615-MS2) of chung1wiffa=356264499558618230e-004 t0=571842229067915470e+001

Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3

Glucoraphenin in Thlaspi arvensis L [菥蓂] extract at 8347min MW = 4350327 [M-H]-

61

Figure A20a ESI-QTOF-MS spectrum of Seed of Lepidium apetalum Willd [葶藶子]

extract at 11875min

Figure A20b MSMS spectrum of Seed of Lepidium apetalum Willd [葶藶子] extract at

11875min

-TOF Product (3720) 22 MCA scans from Sample 5 (t-rt-118-ms2) of chung1wiffa=356264499558618230e-004 t0=571842229067915470e+001

Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2

Gluconapin in Seed of Lepidium apetalum Willd [葶藶子] extract at 11875min MW = 3730501

[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

II

ACKNOWLEDGEMENT

This is an ongoing project and has been done by graduated students Miss CY Cheung Miss

WM Au and Mr CK Kwong The negative electrospray ionization-quadrupole time-of- flight

mass spectrometry and MSMS analysis of glucosinolate standards vegetable and Traditional

Chinese Medicine (TCM) samples were done by Dr Zongwei Cairsquos M Phil students Mr WT

Ma and Mr W Chan All other experiments described in this thesis were my own original work

and were carried out by myself under the supervision of Dr Zongwei Cai Thank you for his

valuable advice and guidance for me in this project

Thank you for Prof Albert WM Lee as my observer and provide me Rorippa indica (Linn)

Hiern sample [蔊菜]

Thank you for Dr Zhong-zhen Zhao provide me Leaf of Isatis indigotica Fort sample [大青葉]

Thank you for Dr Zongwei Cairsquos M Phil students Mr WT Ma and Mr W Chan as well as his

PhD student Miss Q Luo and laboratory technician Mr John Ng who have helped me a lot in

this project

Signature of Student

Student Name

Department of Chemistry Hong Kong Baptist University

Date

III


By



ABSTRACT

















n=3) respectively

IV

Contents






















V




4 Discussion

41 Extraction P40





5 Conclusion P45



1

1 Introduction







ΟΟΗ

OH

OH

CH2OH

S C

R

NOSO3













2


R C

N

S

OSO3

β⎯D⎯glucose

R C

S

N

ΟΟ Η

O H

O H

C H 2 O H

O H















+ +

gt pH7 pH3 Fe2+

Myrosinase H2O

3






























4









isothiocyanates



















5











6







4230327



4390277



3890450


Prop-2-enyl-GS

3590345



3890450



4350327


p-Hydroxybenzyl-GS

4250450





4390607


Benzyl-GS

4090501



4210535


Phenethyl-GS

4230658



H2C

H2C

H2CS

O

H3C

H2C

H2C

H2CS

O

H3C

O

H 2CC

O H

H

CH

H 2C

(2R)

H2CC

HH2C

(2S)

H2CC

H

H2CC

HS

O

H3C

H2CHO

H2C

H2CC

HH2C

H2CC

OH

H

H2C

H2C

H2C

H2C

H2CSH3C

H2C

H2C

H 2CC

H

O H

CH

H 2 C

7




















8






















9
























10




















11









27 HPLC analysis













12

Gradient program

0

5

10

15

20

25

30

35

0 5 10 15 20 25 30

Time(min)

B

1
















13















min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800

1000

1200


47

72

51

50

58

37 6

244

70

42

84

92

11

234

11

899

15

084

15

811

16

160

19

101

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

14























15















gluconasturtiin



respectively

16








min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


48

05

52

12

58

34

62

76

69

71

85

48

11

306

11

883

15

799

16

026

19

255

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

Glucoerucin

17



















respectively

18



extract at 11883min


Max 39820 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3000

3200

3400

3600

3800

3982 3720233

2641464 3740192

315054126721773830960 44000993410834 4559759 484019639399712972460


Max 44870 counts

200 210 220 230 240 250 260 270 280 290 300 310 320 330 340 350 360 370 380 390 400mz amu

0

500

1000

1500

2000

2500

3000

3500

4000

4487 3720536

3740497

25523932201537 2272081 2832709 388065535803532931867 32518753392078

[M-H]- Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


19


















20




Max 1240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120124 3720156

969548

749854

1950168

25899591300262

274971617897167995151289233

850283 29205781450374 2270103 2409984 3009815564612


Max 1180 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

118 749866

969584

37206001300304799501 25901761950337 27499751789801850244 1289378 2920796590074 17599271630819

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S


Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S[M-H]-

[M-HSO4-H]-

[M-SO3-H]-

21











standards spiked

Progoitrin G

lucocheirolin

Glucoiberin

Glucoerucin

Glucosibarin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


47

60

51

78

58

40

62

42

70

83

86

16

11

271

11

861

15

077

15

834

16

103

19

110

Glucoiberin

Glucocheirolin


Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Progoitrin

Glucoraphenin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


46

75

51

48

61

86

87

09

15

122

16

142

22





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

11

49

71

56

31

60

64

68

38

82

40

11

051

11

718

14

935

15

656

15

991 1

890

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


82

67

14

950

18

873

Glucoraphenin

Glucosibarin

Gluconasturtiin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


58

53

87

29

16

168

19

153

Progoitrin

Glucoraphenin

Glucoerucin

Gluconasturtiin

23




standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

78

51

58

58

18

62

57

70

91

86

68

11

291

11

881

15

111

15

847

16

159

19

138

Glucoiberin

Glucocheirolin

Progoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Epiprogoitrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


52

12

58

34

62

76

69

71

85

48

11

306

118

83

15

799

19

255

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

24




min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

61

50

60

56

98

61

03

69

18

84

01

11

169

11

741

14

944 1

569

2 1

601

1

18

963

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


25




min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


49

78

55

68

60

05

14

842

835

0

Glucosibarin

Glucoraphenin

Sinigrin

Progoitrin G

lucocheirolin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


46

57

50

54 56

93

60

99

69

01

83

67

11

161

11

743 14

941

15

688

16

006

18

908

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

-50 0

50 100 150 200 250 300 350


46

43

50

21

56

67

60

04

68

75

83

47

11

137

11

724

14

931

15

684

16

005

18

962

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

26




extract

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


47

63

51

69 5

823

6

238

70

65

86

68

11

293

11

873

15

081 1

583

2 1

614

1

19

126

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150

200 250 300 350


55

88

84

78

14

918

Progoitrin

Glucoraphenin

Glucosibarin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


27





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

59

50

50

56

94 6

094

6

901

83

64

11

141

11

736 1

494

0 1

570

8 1

601

0 18

967

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


62

09

19

122

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

04

49

88

56

02

59

96

67

77

82

20

11

064

11

629

14

850 1

561

1 1

594

1

18

920

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin Sinigrin

Gluconasturtiin

28




min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

69

51

46 5

819

6

006

70

49

85

26

11

128

11

722

15

166

15

899

16

228 1

919

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

72

58

21

59

99

70

50

11

138

11

723

15

177

16

204 1

919

2


Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

53

58

18

61

76

70

27

11

212

11

833

15

119

16

145

19

093


Sinigrin

Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

29





standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150

175


47

47

51

30

58

12

617

4

702

3

84

67

11

208

11

834

15

077

158

18 1

614

0

19

084

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


57

50 60

92

69

29

79

53

11

809

Progoitrin

Epiprogoitrin

Glucoraphenin

Gluconapin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

33

50

10

56

77 60

70

68

57

79

24

11

128

11

732

14

924 1

567

7 1

599

2

18

929

Glucoiberin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Glucocheirolin

30




min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

III


By



ABSTRACT

















n=3) respectively

IV

Contents






















V




4 Discussion

41 Extraction P40





5 Conclusion P45



1

1 Introduction







ΟΟΗ

OH

OH

CH2OH

S C

R

NOSO3













2


R C

N

S

OSO3

β⎯D⎯glucose

R C

S

N

ΟΟ Η

O H

O H

C H 2 O H

O H















+ +

gt pH7 pH3 Fe2+

Myrosinase H2O

3






























4









isothiocyanates



















5











6







4230327



4390277



3890450


Prop-2-enyl-GS

3590345



3890450



4350327


p-Hydroxybenzyl-GS

4250450





4390607


Benzyl-GS

4090501



4210535


Phenethyl-GS

4230658



H2C

H2C

H2CS

O

H3C

H2C

H2C

H2CS

O

H3C

O

H 2CC

O H

H

CH

H 2C

(2R)

H2CC

HH2C

(2S)

H2CC

H

H2CC

HS

O

H3C

H2CHO

H2C

H2CC

HH2C

H2CC

OH

H

H2C

H2C

H2C

H2C

H2CSH3C

H2C

H2C

H 2CC

H

O H

CH

H 2 C

7




















8






















9
























10




















11









27 HPLC analysis













12

Gradient program

0

5

10

15

20

25

30

35

0 5 10 15 20 25 30

Time(min)

B

1
















13















min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800

1000

1200


47

72

51

50

58

37 6

244

70

42

84

92

11

234

11

899

15

084

15

811

16

160

19

101

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

14























15















gluconasturtiin



respectively

16








min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


48

05

52

12

58

34

62

76

69

71

85

48

11

306

11

883

15

799

16

026

19

255

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

Glucoerucin

17



















respectively

18



extract at 11883min


Max 39820 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3000

3200

3400

3600

3800

3982 3720233

2641464 3740192

315054126721773830960 44000993410834 4559759 484019639399712972460


Max 44870 counts

200 210 220 230 240 250 260 270 280 290 300 310 320 330 340 350 360 370 380 390 400mz amu

0

500

1000

1500

2000

2500

3000

3500

4000

4487 3720536

3740497

25523932201537 2272081 2832709 388065535803532931867 32518753392078

[M-H]- Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


19


















20




Max 1240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120124 3720156

969548

749854

1950168

25899591300262

274971617897167995151289233

850283 29205781450374 2270103 2409984 3009815564612


Max 1180 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

118 749866

969584

37206001300304799501 25901761950337 27499751789801850244 1289378 2920796590074 17599271630819

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S


Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S[M-H]-

[M-HSO4-H]-

[M-SO3-H]-

21











standards spiked

Progoitrin G

lucocheirolin

Glucoiberin

Glucoerucin

Glucosibarin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


47

60

51

78

58

40

62

42

70

83

86

16

11

271

11

861

15

077

15

834

16

103

19

110

Glucoiberin

Glucocheirolin


Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Progoitrin

Glucoraphenin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


46

75

51

48

61

86

87

09

15

122

16

142

22





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

11

49

71

56

31

60

64

68

38

82

40

11

051

11

718

14

935

15

656

15

991 1

890

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


82

67

14

950

18

873

Glucoraphenin

Glucosibarin

Gluconasturtiin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


58

53

87

29

16

168

19

153

Progoitrin

Glucoraphenin

Glucoerucin

Gluconasturtiin

23




standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

78

51

58

58

18

62

57

70

91

86

68

11

291

11

881

15

111

15

847

16

159

19

138

Glucoiberin

Glucocheirolin

Progoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Epiprogoitrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


52

12

58

34

62

76

69

71

85

48

11

306

118

83

15

799

19

255

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

24




min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

61

50

60

56

98

61

03

69

18

84

01

11

169

11

741

14

944 1

569

2 1

601

1

18

963

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


25




min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


49

78

55

68

60

05

14

842

835

0

Glucosibarin

Glucoraphenin

Sinigrin

Progoitrin G

lucocheirolin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


46

57

50

54 56

93

60

99

69

01

83

67

11

161

11

743 14

941

15

688

16

006

18

908

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

-50 0

50 100 150 200 250 300 350


46

43

50

21

56

67

60

04

68

75

83

47

11

137

11

724

14

931

15

684

16

005

18

962

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

26




extract

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


47

63

51

69 5

823

6

238

70

65

86

68

11

293

11

873

15

081 1

583

2 1

614

1

19

126

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150

200 250 300 350


55

88

84

78

14

918

Progoitrin

Glucoraphenin

Glucosibarin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


27





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

59

50

50

56

94 6

094

6

901

83

64

11

141

11

736 1

494

0 1

570

8 1

601

0 18

967

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


62

09

19

122

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

04

49

88

56

02

59

96

67

77

82

20

11

064

11

629

14

850 1

561

1 1

594

1

18

920

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin Sinigrin

Gluconasturtiin

28




min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

69

51

46 5

819

6

006

70

49

85

26

11

128

11

722

15

166

15

899

16

228 1

919

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

72

58

21

59

99

70

50

11

138

11

723

15

177

16

204 1

919

2


Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

53

58

18

61

76

70

27

11

212

11

833

15

119

16

145

19

093


Sinigrin

Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

29





standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150

175


47

47

51

30

58

12

617

4

702

3

84

67

11

208

11

834

15

077

158

18 1

614

0

19

084

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


57

50 60

92

69

29

79

53

11

809

Progoitrin

Epiprogoitrin

Glucoraphenin

Gluconapin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

33

50

10

56

77 60

70

68

57

79

24

11

128

11

732

14

924 1

567

7 1

599

2

18

929

Glucoiberin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Glucocheirolin

30




min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

IV

Contents






















V




4 Discussion

41 Extraction P40





5 Conclusion P45



1

1 Introduction







ΟΟΗ

OH

OH

CH2OH

S C

R

NOSO3













2


R C

N

S

OSO3

β⎯D⎯glucose

R C

S

N

ΟΟ Η

O H

O H

C H 2 O H

O H















+ +

gt pH7 pH3 Fe2+

Myrosinase H2O

3






























4









isothiocyanates



















5











6







4230327



4390277



3890450


Prop-2-enyl-GS

3590345



3890450



4350327


p-Hydroxybenzyl-GS

4250450





4390607


Benzyl-GS

4090501



4210535


Phenethyl-GS

4230658



H2C

H2C

H2CS

O

H3C

H2C

H2C

H2CS

O

H3C

O

H 2CC

O H

H

CH

H 2C

(2R)

H2CC

HH2C

(2S)

H2CC

H

H2CC

HS

O

H3C

H2CHO

H2C

H2CC

HH2C

H2CC

OH

H

H2C

H2C

H2C

H2C

H2CSH3C

H2C

H2C

H 2CC

H

O H

CH

H 2 C

7




















8






















9
























10




















11









27 HPLC analysis













12

Gradient program

0

5

10

15

20

25

30

35

0 5 10 15 20 25 30

Time(min)

B

1
















13















min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800

1000

1200


47

72

51

50

58

37 6

244

70

42

84

92

11

234

11

899

15

084

15

811

16

160

19

101

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

14























15















gluconasturtiin



respectively

16








min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


48

05

52

12

58

34

62

76

69

71

85

48

11

306

11

883

15

799

16

026

19

255

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

Glucoerucin

17



















respectively

18



extract at 11883min


Max 39820 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3000

3200

3400

3600

3800

3982 3720233

2641464 3740192

315054126721773830960 44000993410834 4559759 484019639399712972460


Max 44870 counts

200 210 220 230 240 250 260 270 280 290 300 310 320 330 340 350 360 370 380 390 400mz amu

0

500

1000

1500

2000

2500

3000

3500

4000

4487 3720536

3740497

25523932201537 2272081 2832709 388065535803532931867 32518753392078

[M-H]- Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


19


















20




Max 1240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120124 3720156

969548

749854

1950168

25899591300262

274971617897167995151289233

850283 29205781450374 2270103 2409984 3009815564612


Max 1180 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

118 749866

969584

37206001300304799501 25901761950337 27499751789801850244 1289378 2920796590074 17599271630819

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S


Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S[M-H]-

[M-HSO4-H]-

[M-SO3-H]-

21











standards spiked

Progoitrin G

lucocheirolin

Glucoiberin

Glucoerucin

Glucosibarin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


47

60

51

78

58

40

62

42

70

83

86

16

11

271

11

861

15

077

15

834

16

103

19

110

Glucoiberin

Glucocheirolin


Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Progoitrin

Glucoraphenin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


46

75

51

48

61

86

87

09

15

122

16

142

22





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

11

49

71

56

31

60

64

68

38

82

40

11

051

11

718

14

935

15

656

15

991 1

890

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


82

67

14

950

18

873

Glucoraphenin

Glucosibarin

Gluconasturtiin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


58

53

87

29

16

168

19

153

Progoitrin

Glucoraphenin

Glucoerucin

Gluconasturtiin

23




standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

78

51

58

58

18

62

57

70

91

86

68

11

291

11

881

15

111

15

847

16

159

19

138

Glucoiberin

Glucocheirolin

Progoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Epiprogoitrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


52

12

58

34

62

76

69

71

85

48

11

306

118

83

15

799

19

255

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

24




min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

61

50

60

56

98

61

03

69

18

84

01

11

169

11

741

14

944 1

569

2 1

601

1

18

963

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


25




min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


49

78

55

68

60

05

14

842

835

0

Glucosibarin

Glucoraphenin

Sinigrin

Progoitrin G

lucocheirolin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


46

57

50

54 56

93

60

99

69

01

83

67

11

161

11

743 14

941

15

688

16

006

18

908

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

-50 0

50 100 150 200 250 300 350


46

43

50

21

56

67

60

04

68

75

83

47

11

137

11

724

14

931

15

684

16

005

18

962

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

26




extract

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


47

63

51

69 5

823

6

238

70

65

86

68

11

293

11

873

15

081 1

583

2 1

614

1

19

126

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150

200 250 300 350


55

88

84

78

14

918

Progoitrin

Glucoraphenin

Glucosibarin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


27





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

59

50

50

56

94 6

094

6

901

83

64

11

141

11

736 1

494

0 1

570

8 1

601

0 18

967

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


62

09

19

122

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

04

49

88

56

02

59

96

67

77

82

20

11

064

11

629

14

850 1

561

1 1

594

1

18

920

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin Sinigrin

Gluconasturtiin

28




min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

69

51

46 5

819

6

006

70

49

85

26

11

128

11

722

15

166

15

899

16

228 1

919

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

72

58

21

59

99

70

50

11

138

11

723

15

177

16

204 1

919

2


Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

53

58

18

61

76

70

27

11

212

11

833

15

119

16

145

19

093


Sinigrin

Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

29





standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150

175


47

47

51

30

58

12

617

4

702

3

84

67

11

208

11

834

15

077

158

18 1

614

0

19

084

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


57

50 60

92

69

29

79

53

11

809

Progoitrin

Epiprogoitrin

Glucoraphenin

Gluconapin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

33

50

10

56

77 60

70

68

57

79

24

11

128

11

732

14

924 1

567

7 1

599

2

18

929

Glucoiberin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Glucocheirolin

30




min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

V




4 Discussion

41 Extraction P40





5 Conclusion P45



1

1 Introduction







ΟΟΗ

OH

OH

CH2OH

S C

R

NOSO3













2


R C

N

S

OSO3

β⎯D⎯glucose

R C

S

N

ΟΟ Η

O H

O H

C H 2 O H

O H















+ +

gt pH7 pH3 Fe2+

Myrosinase H2O

3






























4









isothiocyanates



















5











6







4230327



4390277



3890450


Prop-2-enyl-GS

3590345



3890450



4350327


p-Hydroxybenzyl-GS

4250450





4390607


Benzyl-GS

4090501



4210535


Phenethyl-GS

4230658



H2C

H2C

H2CS

O

H3C

H2C

H2C

H2CS

O

H3C

O

H 2CC

O H

H

CH

H 2C

(2R)

H2CC

HH2C

(2S)

H2CC

H

H2CC

HS

O

H3C

H2CHO

H2C

H2CC

HH2C

H2CC

OH

H

H2C

H2C

H2C

H2C

H2CSH3C

H2C

H2C

H 2CC

H

O H

CH

H 2 C

7




















8






















9
























10




















11









27 HPLC analysis













12

Gradient program

0

5

10

15

20

25

30

35

0 5 10 15 20 25 30

Time(min)

B

1
















13















min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800

1000

1200


47

72

51

50

58

37 6

244

70

42

84

92

11

234

11

899

15

084

15

811

16

160

19

101

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

14























15















gluconasturtiin



respectively

16








min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


48

05

52

12

58

34

62

76

69

71

85

48

11

306

11

883

15

799

16

026

19

255

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

Glucoerucin

17



















respectively

18



extract at 11883min


Max 39820 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3000

3200

3400

3600

3800

3982 3720233

2641464 3740192

315054126721773830960 44000993410834 4559759 484019639399712972460


Max 44870 counts

200 210 220 230 240 250 260 270 280 290 300 310 320 330 340 350 360 370 380 390 400mz amu

0

500

1000

1500

2000

2500

3000

3500

4000

4487 3720536

3740497

25523932201537 2272081 2832709 388065535803532931867 32518753392078

[M-H]- Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


19


















20




Max 1240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120124 3720156

969548

749854

1950168

25899591300262

274971617897167995151289233

850283 29205781450374 2270103 2409984 3009815564612


Max 1180 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

118 749866

969584

37206001300304799501 25901761950337 27499751789801850244 1289378 2920796590074 17599271630819

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S


Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S[M-H]-

[M-HSO4-H]-

[M-SO3-H]-

21











standards spiked

Progoitrin G

lucocheirolin

Glucoiberin

Glucoerucin

Glucosibarin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


47

60

51

78

58

40

62

42

70

83

86

16

11

271

11

861

15

077

15

834

16

103

19

110

Glucoiberin

Glucocheirolin


Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Progoitrin

Glucoraphenin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


46

75

51

48

61

86

87

09

15

122

16

142

22





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

11

49

71

56

31

60

64

68

38

82

40

11

051

11

718

14

935

15

656

15

991 1

890

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


82

67

14

950

18

873

Glucoraphenin

Glucosibarin

Gluconasturtiin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


58

53

87

29

16

168

19

153

Progoitrin

Glucoraphenin

Glucoerucin

Gluconasturtiin

23




standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

78

51

58

58

18

62

57

70

91

86

68

11

291

11

881

15

111

15

847

16

159

19

138

Glucoiberin

Glucocheirolin

Progoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Epiprogoitrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


52

12

58

34

62

76

69

71

85

48

11

306

118

83

15

799

19

255

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

24




min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

61

50

60

56

98

61

03

69

18

84

01

11

169

11

741

14

944 1

569

2 1

601

1

18

963

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


25




min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


49

78

55

68

60

05

14

842

835

0

Glucosibarin

Glucoraphenin

Sinigrin

Progoitrin G

lucocheirolin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


46

57

50

54 56

93

60

99

69

01

83

67

11

161

11

743 14

941

15

688

16

006

18

908

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

-50 0

50 100 150 200 250 300 350


46

43

50

21

56

67

60

04

68

75

83

47

11

137

11

724

14

931

15

684

16

005

18

962

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

26




extract

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


47

63

51

69 5

823

6

238

70

65

86

68

11

293

11

873

15

081 1

583

2 1

614

1

19

126

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150

200 250 300 350


55

88

84

78

14

918

Progoitrin

Glucoraphenin

Glucosibarin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


27





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

59

50

50

56

94 6

094

6

901

83

64

11

141

11

736 1

494

0 1

570

8 1

601

0 18

967

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


62

09

19

122

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

04

49

88

56

02

59

96

67

77

82

20

11

064

11

629

14

850 1

561

1 1

594

1

18

920

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin Sinigrin

Gluconasturtiin

28




min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

69

51

46 5

819

6

006

70

49

85

26

11

128

11

722

15

166

15

899

16

228 1

919

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

72

58

21

59

99

70

50

11

138

11

723

15

177

16

204 1

919

2


Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

53

58

18

61

76

70

27

11

212

11

833

15

119

16

145

19

093


Sinigrin

Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

29





standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150

175


47

47

51

30

58

12

617

4

702

3

84

67

11

208

11

834

15

077

158

18 1

614

0

19

084

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


57

50 60

92

69

29

79

53

11

809

Progoitrin

Epiprogoitrin

Glucoraphenin

Gluconapin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

33

50

10

56

77 60

70

68

57

79

24

11

128

11

732

14

924 1

567

7 1

599

2

18

929

Glucoiberin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Glucocheirolin

30




min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

1

1 Introduction







ΟΟΗ

OH

OH

CH2OH

S C

R

NOSO3













2


R C

N

S

OSO3

β⎯D⎯glucose

R C

S

N

ΟΟ Η

O H

O H

C H 2 O H

O H















+ +

gt pH7 pH3 Fe2+

Myrosinase H2O

3






























4









isothiocyanates



















5











6







4230327



4390277



3890450


Prop-2-enyl-GS

3590345



3890450



4350327


p-Hydroxybenzyl-GS

4250450





4390607


Benzyl-GS

4090501



4210535


Phenethyl-GS

4230658



H2C

H2C

H2CS

O

H3C

H2C

H2C

H2CS

O

H3C

O

H 2CC

O H

H

CH

H 2C

(2R)

H2CC

HH2C

(2S)

H2CC

H

H2CC

HS

O

H3C

H2CHO

H2C

H2CC

HH2C

H2CC

OH

H

H2C

H2C

H2C

H2C

H2CSH3C

H2C

H2C

H 2CC

H

O H

CH

H 2 C

7




















8






















9
























10




















11









27 HPLC analysis













12

Gradient program

0

5

10

15

20

25

30

35

0 5 10 15 20 25 30

Time(min)

B

1
















13















min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800

1000

1200


47

72

51

50

58

37 6

244

70

42

84

92

11

234

11

899

15

084

15

811

16

160

19

101

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

14























15















gluconasturtiin



respectively

16








min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


48

05

52

12

58

34

62

76

69

71

85

48

11

306

11

883

15

799

16

026

19

255

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

Glucoerucin

17



















respectively

18



extract at 11883min


Max 39820 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3000

3200

3400

3600

3800

3982 3720233

2641464 3740192

315054126721773830960 44000993410834 4559759 484019639399712972460


Max 44870 counts

200 210 220 230 240 250 260 270 280 290 300 310 320 330 340 350 360 370 380 390 400mz amu

0

500

1000

1500

2000

2500

3000

3500

4000

4487 3720536

3740497

25523932201537 2272081 2832709 388065535803532931867 32518753392078

[M-H]- Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


19


















20




Max 1240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120124 3720156

969548

749854

1950168

25899591300262

274971617897167995151289233

850283 29205781450374 2270103 2409984 3009815564612


Max 1180 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

118 749866

969584

37206001300304799501 25901761950337 27499751789801850244 1289378 2920796590074 17599271630819

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S


Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S[M-H]-

[M-HSO4-H]-

[M-SO3-H]-

21











standards spiked

Progoitrin G

lucocheirolin

Glucoiberin

Glucoerucin

Glucosibarin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


47

60

51

78

58

40

62

42

70

83

86

16

11

271

11

861

15

077

15

834

16

103

19

110

Glucoiberin

Glucocheirolin


Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Progoitrin

Glucoraphenin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


46

75

51

48

61

86

87

09

15

122

16

142

22





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

11

49

71

56

31

60

64

68

38

82

40

11

051

11

718

14

935

15

656

15

991 1

890

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


82

67

14

950

18

873

Glucoraphenin

Glucosibarin

Gluconasturtiin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


58

53

87

29

16

168

19

153

Progoitrin

Glucoraphenin

Glucoerucin

Gluconasturtiin

23




standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

78

51

58

58

18

62

57

70

91

86

68

11

291

11

881

15

111

15

847

16

159

19

138

Glucoiberin

Glucocheirolin

Progoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Epiprogoitrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


52

12

58

34

62

76

69

71

85

48

11

306

118

83

15

799

19

255

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

24




min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

61

50

60

56

98

61

03

69

18

84

01

11

169

11

741

14

944 1

569

2 1

601

1

18

963

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


25




min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


49

78

55

68

60

05

14

842

835

0

Glucosibarin

Glucoraphenin

Sinigrin

Progoitrin G

lucocheirolin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


46

57

50

54 56

93

60

99

69

01

83

67

11

161

11

743 14

941

15

688

16

006

18

908

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

-50 0

50 100 150 200 250 300 350


46

43

50

21

56

67

60

04

68

75

83

47

11

137

11

724

14

931

15

684

16

005

18

962

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

26




extract

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


47

63

51

69 5

823

6

238

70

65

86

68

11

293

11

873

15

081 1

583

2 1

614

1

19

126

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150

200 250 300 350


55

88

84

78

14

918

Progoitrin

Glucoraphenin

Glucosibarin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


27





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

59

50

50

56

94 6

094

6

901

83

64

11

141

11

736 1

494

0 1

570

8 1

601

0 18

967

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


62

09

19

122

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

04

49

88

56

02

59

96

67

77

82

20

11

064

11

629

14

850 1

561

1 1

594

1

18

920

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin Sinigrin

Gluconasturtiin

28




min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

69

51

46 5

819

6

006

70

49

85

26

11

128

11

722

15

166

15

899

16

228 1

919

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

72

58

21

59

99

70

50

11

138

11

723

15

177

16

204 1

919

2


Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

53

58

18

61

76

70

27

11

212

11

833

15

119

16

145

19

093


Sinigrin

Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

29





standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150

175


47

47

51

30

58

12

617

4

702

3

84

67

11

208

11

834

15

077

158

18 1

614

0

19

084

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


57

50 60

92

69

29

79

53

11

809

Progoitrin

Epiprogoitrin

Glucoraphenin

Gluconapin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

33

50

10

56

77 60

70

68

57

79

24

11

128

11

732

14

924 1

567

7 1

599

2

18

929

Glucoiberin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Glucocheirolin

30




min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

2


R C

N

S

OSO3

β⎯D⎯glucose

R C

S

N

ΟΟ Η

O H

O H

C H 2 O H

O H















+ +

gt pH7 pH3 Fe2+

Myrosinase H2O

3






























4









isothiocyanates



















5











6







4230327



4390277



3890450


Prop-2-enyl-GS

3590345



3890450



4350327


p-Hydroxybenzyl-GS

4250450





4390607


Benzyl-GS

4090501



4210535


Phenethyl-GS

4230658



H2C

H2C

H2CS

O

H3C

H2C

H2C

H2CS

O

H3C

O

H 2CC

O H

H

CH

H 2C

(2R)

H2CC

HH2C

(2S)

H2CC

H

H2CC

HS

O

H3C

H2CHO

H2C

H2CC

HH2C

H2CC

OH

H

H2C

H2C

H2C

H2C

H2CSH3C

H2C

H2C

H 2CC

H

O H

CH

H 2 C

7




















8






















9
























10




















11









27 HPLC analysis













12

Gradient program

0

5

10

15

20

25

30

35

0 5 10 15 20 25 30

Time(min)

B

1
















13















min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800

1000

1200


47

72

51

50

58

37 6

244

70

42

84

92

11

234

11

899

15

084

15

811

16

160

19

101

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

14























15















gluconasturtiin



respectively

16








min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


48

05

52

12

58

34

62

76

69

71

85

48

11

306

11

883

15

799

16

026

19

255

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

Glucoerucin

17



















respectively

18



extract at 11883min


Max 39820 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3000

3200

3400

3600

3800

3982 3720233

2641464 3740192

315054126721773830960 44000993410834 4559759 484019639399712972460


Max 44870 counts

200 210 220 230 240 250 260 270 280 290 300 310 320 330 340 350 360 370 380 390 400mz amu

0

500

1000

1500

2000

2500

3000

3500

4000

4487 3720536

3740497

25523932201537 2272081 2832709 388065535803532931867 32518753392078

[M-H]- Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


19


















20




Max 1240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120124 3720156

969548

749854

1950168

25899591300262

274971617897167995151289233

850283 29205781450374 2270103 2409984 3009815564612


Max 1180 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

118 749866

969584

37206001300304799501 25901761950337 27499751789801850244 1289378 2920796590074 17599271630819

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S


Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S[M-H]-

[M-HSO4-H]-

[M-SO3-H]-

21











standards spiked

Progoitrin G

lucocheirolin

Glucoiberin

Glucoerucin

Glucosibarin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


47

60

51

78

58

40

62

42

70

83

86

16

11

271

11

861

15

077

15

834

16

103

19

110

Glucoiberin

Glucocheirolin


Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Progoitrin

Glucoraphenin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


46

75

51

48

61

86

87

09

15

122

16

142

22





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

11

49

71

56

31

60

64

68

38

82

40

11

051

11

718

14

935

15

656

15

991 1

890

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


82

67

14

950

18

873

Glucoraphenin

Glucosibarin

Gluconasturtiin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


58

53

87

29

16

168

19

153

Progoitrin

Glucoraphenin

Glucoerucin

Gluconasturtiin

23




standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

78

51

58

58

18

62

57

70

91

86

68

11

291

11

881

15

111

15

847

16

159

19

138

Glucoiberin

Glucocheirolin

Progoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Epiprogoitrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


52

12

58

34

62

76

69

71

85

48

11

306

118

83

15

799

19

255

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

24




min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

61

50

60

56

98

61

03

69

18

84

01

11

169

11

741

14

944 1

569

2 1

601

1

18

963

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


25




min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


49

78

55

68

60

05

14

842

835

0

Glucosibarin

Glucoraphenin

Sinigrin

Progoitrin G

lucocheirolin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


46

57

50

54 56

93

60

99

69

01

83

67

11

161

11

743 14

941

15

688

16

006

18

908

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

-50 0

50 100 150 200 250 300 350


46

43

50

21

56

67

60

04

68

75

83

47

11

137

11

724

14

931

15

684

16

005

18

962

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

26




extract

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


47

63

51

69 5

823

6

238

70

65

86

68

11

293

11

873

15

081 1

583

2 1

614

1

19

126

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150

200 250 300 350


55

88

84

78

14

918

Progoitrin

Glucoraphenin

Glucosibarin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


27





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

59

50

50

56

94 6

094

6

901

83

64

11

141

11

736 1

494

0 1

570

8 1

601

0 18

967

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


62

09

19

122

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

04

49

88

56

02

59

96

67

77

82

20

11

064

11

629

14

850 1

561

1 1

594

1

18

920

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin Sinigrin

Gluconasturtiin

28




min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

69

51

46 5

819

6

006

70

49

85

26

11

128

11

722

15

166

15

899

16

228 1

919

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

72

58

21

59

99

70

50

11

138

11

723

15

177

16

204 1

919

2


Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

53

58

18

61

76

70

27

11

212

11

833

15

119

16

145

19

093


Sinigrin

Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

29





standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150

175


47

47

51

30

58

12

617

4

702

3

84

67

11

208

11

834

15

077

158

18 1

614

0

19

084

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


57

50 60

92

69

29

79

53

11

809

Progoitrin

Epiprogoitrin

Glucoraphenin

Gluconapin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

33

50

10

56

77 60

70

68

57

79

24

11

128

11

732

14

924 1

567

7 1

599

2

18

929

Glucoiberin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Glucocheirolin

30




min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

3






























4









isothiocyanates



















5











6







4230327



4390277



3890450


Prop-2-enyl-GS

3590345



3890450



4350327


p-Hydroxybenzyl-GS

4250450





4390607


Benzyl-GS

4090501



4210535


Phenethyl-GS

4230658



H2C

H2C

H2CS

O

H3C

H2C

H2C

H2CS

O

H3C

O

H 2CC

O H

H

CH

H 2C

(2R)

H2CC

HH2C

(2S)

H2CC

H

H2CC

HS

O

H3C

H2CHO

H2C

H2CC

HH2C

H2CC

OH

H

H2C

H2C

H2C

H2C

H2CSH3C

H2C

H2C

H 2CC

H

O H

CH

H 2 C

7




















8






















9
























10




















11









27 HPLC analysis













12

Gradient program

0

5

10

15

20

25

30

35

0 5 10 15 20 25 30

Time(min)

B

1
















13















min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800

1000

1200


47

72

51

50

58

37 6

244

70

42

84

92

11

234

11

899

15

084

15

811

16

160

19

101

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

14























15















gluconasturtiin



respectively

16








min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


48

05

52

12

58

34

62

76

69

71

85

48

11

306

11

883

15

799

16

026

19

255

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

Glucoerucin

17



















respectively

18



extract at 11883min


Max 39820 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3000

3200

3400

3600

3800

3982 3720233

2641464 3740192

315054126721773830960 44000993410834 4559759 484019639399712972460


Max 44870 counts

200 210 220 230 240 250 260 270 280 290 300 310 320 330 340 350 360 370 380 390 400mz amu

0

500

1000

1500

2000

2500

3000

3500

4000

4487 3720536

3740497

25523932201537 2272081 2832709 388065535803532931867 32518753392078

[M-H]- Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


19


















20




Max 1240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120124 3720156

969548

749854

1950168

25899591300262

274971617897167995151289233

850283 29205781450374 2270103 2409984 3009815564612


Max 1180 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

118 749866

969584

37206001300304799501 25901761950337 27499751789801850244 1289378 2920796590074 17599271630819

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S


Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S[M-H]-

[M-HSO4-H]-

[M-SO3-H]-

21











standards spiked

Progoitrin G

lucocheirolin

Glucoiberin

Glucoerucin

Glucosibarin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


47

60

51

78

58

40

62

42

70

83

86

16

11

271

11

861

15

077

15

834

16

103

19

110

Glucoiberin

Glucocheirolin


Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Progoitrin

Glucoraphenin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


46

75

51

48

61

86

87

09

15

122

16

142

22





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

11

49

71

56

31

60

64

68

38

82

40

11

051

11

718

14

935

15

656

15

991 1

890

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


82

67

14

950

18

873

Glucoraphenin

Glucosibarin

Gluconasturtiin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


58

53

87

29

16

168

19

153

Progoitrin

Glucoraphenin

Glucoerucin

Gluconasturtiin

23




standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

78

51

58

58

18

62

57

70

91

86

68

11

291

11

881

15

111

15

847

16

159

19

138

Glucoiberin

Glucocheirolin

Progoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Epiprogoitrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


52

12

58

34

62

76

69

71

85

48

11

306

118

83

15

799

19

255

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

24




min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

61

50

60

56

98

61

03

69

18

84

01

11

169

11

741

14

944 1

569

2 1

601

1

18

963

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


25




min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


49

78

55

68

60

05

14

842

835

0

Glucosibarin

Glucoraphenin

Sinigrin

Progoitrin G

lucocheirolin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


46

57

50

54 56

93

60

99

69

01

83

67

11

161

11

743 14

941

15

688

16

006

18

908

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

-50 0

50 100 150 200 250 300 350


46

43

50

21

56

67

60

04

68

75

83

47

11

137

11

724

14

931

15

684

16

005

18

962

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

26




extract

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


47

63

51

69 5

823

6

238

70

65

86

68

11

293

11

873

15

081 1

583

2 1

614

1

19

126

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150

200 250 300 350


55

88

84

78

14

918

Progoitrin

Glucoraphenin

Glucosibarin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


27





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

59

50

50

56

94 6

094

6

901

83

64

11

141

11

736 1

494

0 1

570

8 1

601

0 18

967

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


62

09

19

122

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

04

49

88

56

02

59

96

67

77

82

20

11

064

11

629

14

850 1

561

1 1

594

1

18

920

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin Sinigrin

Gluconasturtiin

28




min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

69

51

46 5

819

6

006

70

49

85

26

11

128

11

722

15

166

15

899

16

228 1

919

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

72

58

21

59

99

70

50

11

138

11

723

15

177

16

204 1

919

2


Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

53

58

18

61

76

70

27

11

212

11

833

15

119

16

145

19

093


Sinigrin

Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

29





standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150

175


47

47

51

30

58

12

617

4

702

3

84

67

11

208

11

834

15

077

158

18 1

614

0

19

084

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


57

50 60

92

69

29

79

53

11

809

Progoitrin

Epiprogoitrin

Glucoraphenin

Gluconapin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

33

50

10

56

77 60

70

68

57

79

24

11

128

11

732

14

924 1

567

7 1

599

2

18

929

Glucoiberin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Glucocheirolin

30




min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

4









isothiocyanates



















5











6







4230327



4390277



3890450


Prop-2-enyl-GS

3590345



3890450



4350327


p-Hydroxybenzyl-GS

4250450





4390607


Benzyl-GS

4090501



4210535


Phenethyl-GS

4230658



H2C

H2C

H2CS

O

H3C

H2C

H2C

H2CS

O

H3C

O

H 2CC

O H

H

CH

H 2C

(2R)

H2CC

HH2C

(2S)

H2CC

H

H2CC

HS

O

H3C

H2CHO

H2C

H2CC

HH2C

H2CC

OH

H

H2C

H2C

H2C

H2C

H2CSH3C

H2C

H2C

H 2CC

H

O H

CH

H 2 C

7




















8






















9
























10




















11









27 HPLC analysis













12

Gradient program

0

5

10

15

20

25

30

35

0 5 10 15 20 25 30

Time(min)

B

1
















13















min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800

1000

1200


47

72

51

50

58

37 6

244

70

42

84

92

11

234

11

899

15

084

15

811

16

160

19

101

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

14























15















gluconasturtiin



respectively

16








min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


48

05

52

12

58

34

62

76

69

71

85

48

11

306

11

883

15

799

16

026

19

255

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

Glucoerucin

17



















respectively

18



extract at 11883min


Max 39820 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3000

3200

3400

3600

3800

3982 3720233

2641464 3740192

315054126721773830960 44000993410834 4559759 484019639399712972460


Max 44870 counts

200 210 220 230 240 250 260 270 280 290 300 310 320 330 340 350 360 370 380 390 400mz amu

0

500

1000

1500

2000

2500

3000

3500

4000

4487 3720536

3740497

25523932201537 2272081 2832709 388065535803532931867 32518753392078

[M-H]- Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


19


















20




Max 1240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120124 3720156

969548

749854

1950168

25899591300262

274971617897167995151289233

850283 29205781450374 2270103 2409984 3009815564612


Max 1180 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

118 749866

969584

37206001300304799501 25901761950337 27499751789801850244 1289378 2920796590074 17599271630819

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S


Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S[M-H]-

[M-HSO4-H]-

[M-SO3-H]-

21











standards spiked

Progoitrin G

lucocheirolin

Glucoiberin

Glucoerucin

Glucosibarin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


47

60

51

78

58

40

62

42

70

83

86

16

11

271

11

861

15

077

15

834

16

103

19

110

Glucoiberin

Glucocheirolin


Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Progoitrin

Glucoraphenin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


46

75

51

48

61

86

87

09

15

122

16

142

22





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

11

49

71

56

31

60

64

68

38

82

40

11

051

11

718

14

935

15

656

15

991 1

890

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


82

67

14

950

18

873

Glucoraphenin

Glucosibarin

Gluconasturtiin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


58

53

87

29

16

168

19

153

Progoitrin

Glucoraphenin

Glucoerucin

Gluconasturtiin

23




standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

78

51

58

58

18

62

57

70

91

86

68

11

291

11

881

15

111

15

847

16

159

19

138

Glucoiberin

Glucocheirolin

Progoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Epiprogoitrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


52

12

58

34

62

76

69

71

85

48

11

306

118

83

15

799

19

255

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

24




min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

61

50

60

56

98

61

03

69

18

84

01

11

169

11

741

14

944 1

569

2 1

601

1

18

963

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


25




min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


49

78

55

68

60

05

14

842

835

0

Glucosibarin

Glucoraphenin

Sinigrin

Progoitrin G

lucocheirolin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


46

57

50

54 56

93

60

99

69

01

83

67

11

161

11

743 14

941

15

688

16

006

18

908

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

-50 0

50 100 150 200 250 300 350


46

43

50

21

56

67

60

04

68

75

83

47

11

137

11

724

14

931

15

684

16

005

18

962

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

26




extract

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


47

63

51

69 5

823

6

238

70

65

86

68

11

293

11

873

15

081 1

583

2 1

614

1

19

126

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150

200 250 300 350


55

88

84

78

14

918

Progoitrin

Glucoraphenin

Glucosibarin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


27





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

59

50

50

56

94 6

094

6

901

83

64

11

141

11

736 1

494

0 1

570

8 1

601

0 18

967

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


62

09

19

122

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

04

49

88

56

02

59

96

67

77

82

20

11

064

11

629

14

850 1

561

1 1

594

1

18

920

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin Sinigrin

Gluconasturtiin

28




min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

69

51

46 5

819

6

006

70

49

85

26

11

128

11

722

15

166

15

899

16

228 1

919

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

72

58

21

59

99

70

50

11

138

11

723

15

177

16

204 1

919

2


Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

53

58

18

61

76

70

27

11

212

11

833

15

119

16

145

19

093


Sinigrin

Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

29





standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150

175


47

47

51

30

58

12

617

4

702

3

84

67

11

208

11

834

15

077

158

18 1

614

0

19

084

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


57

50 60

92

69

29

79

53

11

809

Progoitrin

Epiprogoitrin

Glucoraphenin

Gluconapin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

33

50

10

56

77 60

70

68

57

79

24

11

128

11

732

14

924 1

567

7 1

599

2

18

929

Glucoiberin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Glucocheirolin

30




min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

5











6







4230327



4390277



3890450


Prop-2-enyl-GS

3590345



3890450



4350327


p-Hydroxybenzyl-GS

4250450





4390607


Benzyl-GS

4090501



4210535


Phenethyl-GS

4230658



H2C

H2C

H2CS

O

H3C

H2C

H2C

H2CS

O

H3C

O

H 2CC

O H

H

CH

H 2C

(2R)

H2CC

HH2C

(2S)

H2CC

H

H2CC

HS

O

H3C

H2CHO

H2C

H2CC

HH2C

H2CC

OH

H

H2C

H2C

H2C

H2C

H2CSH3C

H2C

H2C

H 2CC

H

O H

CH

H 2 C

7




















8






















9
























10




















11









27 HPLC analysis













12

Gradient program

0

5

10

15

20

25

30

35

0 5 10 15 20 25 30

Time(min)

B

1
















13















min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800

1000

1200


47

72

51

50

58

37 6

244

70

42

84

92

11

234

11

899

15

084

15

811

16

160

19

101

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

14























15















gluconasturtiin



respectively

16








min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


48

05

52

12

58

34

62

76

69

71

85

48

11

306

11

883

15

799

16

026

19

255

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

Glucoerucin

17



















respectively

18



extract at 11883min


Max 39820 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3000

3200

3400

3600

3800

3982 3720233

2641464 3740192

315054126721773830960 44000993410834 4559759 484019639399712972460


Max 44870 counts

200 210 220 230 240 250 260 270 280 290 300 310 320 330 340 350 360 370 380 390 400mz amu

0

500

1000

1500

2000

2500

3000

3500

4000

4487 3720536

3740497

25523932201537 2272081 2832709 388065535803532931867 32518753392078

[M-H]- Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


19


















20




Max 1240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120124 3720156

969548

749854

1950168

25899591300262

274971617897167995151289233

850283 29205781450374 2270103 2409984 3009815564612


Max 1180 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

118 749866

969584

37206001300304799501 25901761950337 27499751789801850244 1289378 2920796590074 17599271630819

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S


Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S[M-H]-

[M-HSO4-H]-

[M-SO3-H]-

21











standards spiked

Progoitrin G

lucocheirolin

Glucoiberin

Glucoerucin

Glucosibarin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


47

60

51

78

58

40

62

42

70

83

86

16

11

271

11

861

15

077

15

834

16

103

19

110

Glucoiberin

Glucocheirolin


Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Progoitrin

Glucoraphenin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


46

75

51

48

61

86

87

09

15

122

16

142

22





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

11

49

71

56

31

60

64

68

38

82

40

11

051

11

718

14

935

15

656

15

991 1

890

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


82

67

14

950

18

873

Glucoraphenin

Glucosibarin

Gluconasturtiin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


58

53

87

29

16

168

19

153

Progoitrin

Glucoraphenin

Glucoerucin

Gluconasturtiin

23




standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

78

51

58

58

18

62

57

70

91

86

68

11

291

11

881

15

111

15

847

16

159

19

138

Glucoiberin

Glucocheirolin

Progoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Epiprogoitrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


52

12

58

34

62

76

69

71

85

48

11

306

118

83

15

799

19

255

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

24




min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

61

50

60

56

98

61

03

69

18

84

01

11

169

11

741

14

944 1

569

2 1

601

1

18

963

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


25




min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


49

78

55

68

60

05

14

842

835

0

Glucosibarin

Glucoraphenin

Sinigrin

Progoitrin G

lucocheirolin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


46

57

50

54 56

93

60

99

69

01

83

67

11

161

11

743 14

941

15

688

16

006

18

908

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

-50 0

50 100 150 200 250 300 350


46

43

50

21

56

67

60

04

68

75

83

47

11

137

11

724

14

931

15

684

16

005

18

962

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

26




extract

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


47

63

51

69 5

823

6

238

70

65

86

68

11

293

11

873

15

081 1

583

2 1

614

1

19

126

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150

200 250 300 350


55

88

84

78

14

918

Progoitrin

Glucoraphenin

Glucosibarin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


27





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

59

50

50

56

94 6

094

6

901

83

64

11

141

11

736 1

494

0 1

570

8 1

601

0 18

967

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


62

09

19

122

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

04

49

88

56

02

59

96

67

77

82

20

11

064

11

629

14

850 1

561

1 1

594

1

18

920

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin Sinigrin

Gluconasturtiin

28




min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

69

51

46 5

819

6

006

70

49

85

26

11

128

11

722

15

166

15

899

16

228 1

919

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

72

58

21

59

99

70

50

11

138

11

723

15

177

16

204 1

919

2


Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

53

58

18

61

76

70

27

11

212

11

833

15

119

16

145

19

093


Sinigrin

Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

29





standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150

175


47

47

51

30

58

12

617

4

702

3

84

67

11

208

11

834

15

077

158

18 1

614

0

19

084

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


57

50 60

92

69

29

79

53

11

809

Progoitrin

Epiprogoitrin

Glucoraphenin

Gluconapin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

33

50

10

56

77 60

70

68

57

79

24

11

128

11

732

14

924 1

567

7 1

599

2

18

929

Glucoiberin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Glucocheirolin

30




min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

6







4230327



4390277



3890450


Prop-2-enyl-GS

3590345



3890450



4350327


p-Hydroxybenzyl-GS

4250450





4390607


Benzyl-GS

4090501



4210535


Phenethyl-GS

4230658



H2C

H2C

H2CS

O

H3C

H2C

H2C

H2CS

O

H3C

O

H 2CC

O H

H

CH

H 2C

(2R)

H2CC

HH2C

(2S)

H2CC

H

H2CC

HS

O

H3C

H2CHO

H2C

H2CC

HH2C

H2CC

OH

H

H2C

H2C

H2C

H2C

H2CSH3C

H2C

H2C

H 2CC

H

O H

CH

H 2 C

7




















8






















9
























10




















11









27 HPLC analysis













12

Gradient program

0

5

10

15

20

25

30

35

0 5 10 15 20 25 30

Time(min)

B

1
















13















min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800

1000

1200


47

72

51

50

58

37 6

244

70

42

84

92

11

234

11

899

15

084

15

811

16

160

19

101

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

14























15















gluconasturtiin



respectively

16








min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


48

05

52

12

58

34

62

76

69

71

85

48

11

306

11

883

15

799

16

026

19

255

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

Glucoerucin

17



















respectively

18



extract at 11883min


Max 39820 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3000

3200

3400

3600

3800

3982 3720233

2641464 3740192

315054126721773830960 44000993410834 4559759 484019639399712972460


Max 44870 counts

200 210 220 230 240 250 260 270 280 290 300 310 320 330 340 350 360 370 380 390 400mz amu

0

500

1000

1500

2000

2500

3000

3500

4000

4487 3720536

3740497

25523932201537 2272081 2832709 388065535803532931867 32518753392078

[M-H]- Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


19


















20




Max 1240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120124 3720156

969548

749854

1950168

25899591300262

274971617897167995151289233

850283 29205781450374 2270103 2409984 3009815564612


Max 1180 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

118 749866

969584

37206001300304799501 25901761950337 27499751789801850244 1289378 2920796590074 17599271630819

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S


Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S[M-H]-

[M-HSO4-H]-

[M-SO3-H]-

21











standards spiked

Progoitrin G

lucocheirolin

Glucoiberin

Glucoerucin

Glucosibarin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


47

60

51

78

58

40

62

42

70

83

86

16

11

271

11

861

15

077

15

834

16

103

19

110

Glucoiberin

Glucocheirolin


Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Progoitrin

Glucoraphenin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


46

75

51

48

61

86

87

09

15

122

16

142

22





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

11

49

71

56

31

60

64

68

38

82

40

11

051

11

718

14

935

15

656

15

991 1

890

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


82

67

14

950

18

873

Glucoraphenin

Glucosibarin

Gluconasturtiin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


58

53

87

29

16

168

19

153

Progoitrin

Glucoraphenin

Glucoerucin

Gluconasturtiin

23




standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

78

51

58

58

18

62

57

70

91

86

68

11

291

11

881

15

111

15

847

16

159

19

138

Glucoiberin

Glucocheirolin

Progoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Epiprogoitrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


52

12

58

34

62

76

69

71

85

48

11

306

118

83

15

799

19

255

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

24




min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

61

50

60

56

98

61

03

69

18

84

01

11

169

11

741

14

944 1

569

2 1

601

1

18

963

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


25




min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


49

78

55

68

60

05

14

842

835

0

Glucosibarin

Glucoraphenin

Sinigrin

Progoitrin G

lucocheirolin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


46

57

50

54 56

93

60

99

69

01

83

67

11

161

11

743 14

941

15

688

16

006

18

908

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

-50 0

50 100 150 200 250 300 350


46

43

50

21

56

67

60

04

68

75

83

47

11

137

11

724

14

931

15

684

16

005

18

962

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

26




extract

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


47

63

51

69 5

823

6

238

70

65

86

68

11

293

11

873

15

081 1

583

2 1

614

1

19

126

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150

200 250 300 350


55

88

84

78

14

918

Progoitrin

Glucoraphenin

Glucosibarin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


27





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

59

50

50

56

94 6

094

6

901

83

64

11

141

11

736 1

494

0 1

570

8 1

601

0 18

967

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


62

09

19

122

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

04

49

88

56

02

59

96

67

77

82

20

11

064

11

629

14

850 1

561

1 1

594

1

18

920

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin Sinigrin

Gluconasturtiin

28




min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

69

51

46 5

819

6

006

70

49

85

26

11

128

11

722

15

166

15

899

16

228 1

919

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

72

58

21

59

99

70

50

11

138

11

723

15

177

16

204 1

919

2


Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

53

58

18

61

76

70

27

11

212

11

833

15

119

16

145

19

093


Sinigrin

Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

29





standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150

175


47

47

51

30

58

12

617

4

702

3

84

67

11

208

11

834

15

077

158

18 1

614

0

19

084

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


57

50 60

92

69

29

79

53

11

809

Progoitrin

Epiprogoitrin

Glucoraphenin

Gluconapin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

33

50

10

56

77 60

70

68

57

79

24

11

128

11

732

14

924 1

567

7 1

599

2

18

929

Glucoiberin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Glucocheirolin

30




min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

7




















8






















9
























10




















11









27 HPLC analysis













12

Gradient program

0

5

10

15

20

25

30

35

0 5 10 15 20 25 30

Time(min)

B

1
















13















min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800

1000

1200


47

72

51

50

58

37 6

244

70

42

84

92

11

234

11

899

15

084

15

811

16

160

19

101

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

14























15















gluconasturtiin



respectively

16








min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


48

05

52

12

58

34

62

76

69

71

85

48

11

306

11

883

15

799

16

026

19

255

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

Glucoerucin

17



















respectively

18



extract at 11883min


Max 39820 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3000

3200

3400

3600

3800

3982 3720233

2641464 3740192

315054126721773830960 44000993410834 4559759 484019639399712972460


Max 44870 counts

200 210 220 230 240 250 260 270 280 290 300 310 320 330 340 350 360 370 380 390 400mz amu

0

500

1000

1500

2000

2500

3000

3500

4000

4487 3720536

3740497

25523932201537 2272081 2832709 388065535803532931867 32518753392078

[M-H]- Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


19


















20




Max 1240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120124 3720156

969548

749854

1950168

25899591300262

274971617897167995151289233

850283 29205781450374 2270103 2409984 3009815564612


Max 1180 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

118 749866

969584

37206001300304799501 25901761950337 27499751789801850244 1289378 2920796590074 17599271630819

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S


Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S[M-H]-

[M-HSO4-H]-

[M-SO3-H]-

21











standards spiked

Progoitrin G

lucocheirolin

Glucoiberin

Glucoerucin

Glucosibarin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


47

60

51

78

58

40

62

42

70

83

86

16

11

271

11

861

15

077

15

834

16

103

19

110

Glucoiberin

Glucocheirolin


Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Progoitrin

Glucoraphenin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


46

75

51

48

61

86

87

09

15

122

16

142

22





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

11

49

71

56

31

60

64

68

38

82

40

11

051

11

718

14

935

15

656

15

991 1

890

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


82

67

14

950

18

873

Glucoraphenin

Glucosibarin

Gluconasturtiin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


58

53

87

29

16

168

19

153

Progoitrin

Glucoraphenin

Glucoerucin

Gluconasturtiin

23




standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

78

51

58

58

18

62

57

70

91

86

68

11

291

11

881

15

111

15

847

16

159

19

138

Glucoiberin

Glucocheirolin

Progoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Epiprogoitrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


52

12

58

34

62

76

69

71

85

48

11

306

118

83

15

799

19

255

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

24




min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

61

50

60

56

98

61

03

69

18

84

01

11

169

11

741

14

944 1

569

2 1

601

1

18

963

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


25




min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


49

78

55

68

60

05

14

842

835

0

Glucosibarin

Glucoraphenin

Sinigrin

Progoitrin G

lucocheirolin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


46

57

50

54 56

93

60

99

69

01

83

67

11

161

11

743 14

941

15

688

16

006

18

908

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

-50 0

50 100 150 200 250 300 350


46

43

50

21

56

67

60

04

68

75

83

47

11

137

11

724

14

931

15

684

16

005

18

962

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

26




extract

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


47

63

51

69 5

823

6

238

70

65

86

68

11

293

11

873

15

081 1

583

2 1

614

1

19

126

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150

200 250 300 350


55

88

84

78

14

918

Progoitrin

Glucoraphenin

Glucosibarin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


27





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

59

50

50

56

94 6

094

6

901

83

64

11

141

11

736 1

494

0 1

570

8 1

601

0 18

967

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


62

09

19

122

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

04

49

88

56

02

59

96

67

77

82

20

11

064

11

629

14

850 1

561

1 1

594

1

18

920

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin Sinigrin

Gluconasturtiin

28




min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

69

51

46 5

819

6

006

70

49

85

26

11

128

11

722

15

166

15

899

16

228 1

919

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

72

58

21

59

99

70

50

11

138

11

723

15

177

16

204 1

919

2


Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

53

58

18

61

76

70

27

11

212

11

833

15

119

16

145

19

093


Sinigrin

Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

29





standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150

175


47

47

51

30

58

12

617

4

702

3

84

67

11

208

11

834

15

077

158

18 1

614

0

19

084

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


57

50 60

92

69

29

79

53

11

809

Progoitrin

Epiprogoitrin

Glucoraphenin

Gluconapin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

33

50

10

56

77 60

70

68

57

79

24

11

128

11

732

14

924 1

567

7 1

599

2

18

929

Glucoiberin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Glucocheirolin

30




min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

8






















9
























10




















11









27 HPLC analysis













12

Gradient program

0

5

10

15

20

25

30

35

0 5 10 15 20 25 30

Time(min)

B

1
















13















min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800

1000

1200


47

72

51

50

58

37 6

244

70

42

84

92

11

234

11

899

15

084

15

811

16

160

19

101

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

14























15















gluconasturtiin



respectively

16








min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


48

05

52

12

58

34

62

76

69

71

85

48

11

306

11

883

15

799

16

026

19

255

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

Glucoerucin

17



















respectively

18



extract at 11883min


Max 39820 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3000

3200

3400

3600

3800

3982 3720233

2641464 3740192

315054126721773830960 44000993410834 4559759 484019639399712972460


Max 44870 counts

200 210 220 230 240 250 260 270 280 290 300 310 320 330 340 350 360 370 380 390 400mz amu

0

500

1000

1500

2000

2500

3000

3500

4000

4487 3720536

3740497

25523932201537 2272081 2832709 388065535803532931867 32518753392078

[M-H]- Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


19


















20




Max 1240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120124 3720156

969548

749854

1950168

25899591300262

274971617897167995151289233

850283 29205781450374 2270103 2409984 3009815564612


Max 1180 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

118 749866

969584

37206001300304799501 25901761950337 27499751789801850244 1289378 2920796590074 17599271630819

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S


Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S[M-H]-

[M-HSO4-H]-

[M-SO3-H]-

21











standards spiked

Progoitrin G

lucocheirolin

Glucoiberin

Glucoerucin

Glucosibarin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


47

60

51

78

58

40

62

42

70

83

86

16

11

271

11

861

15

077

15

834

16

103

19

110

Glucoiberin

Glucocheirolin


Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Progoitrin

Glucoraphenin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


46

75

51

48

61

86

87

09

15

122

16

142

22





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

11

49

71

56

31

60

64

68

38

82

40

11

051

11

718

14

935

15

656

15

991 1

890

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


82

67

14

950

18

873

Glucoraphenin

Glucosibarin

Gluconasturtiin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


58

53

87

29

16

168

19

153

Progoitrin

Glucoraphenin

Glucoerucin

Gluconasturtiin

23




standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

78

51

58

58

18

62

57

70

91

86

68

11

291

11

881

15

111

15

847

16

159

19

138

Glucoiberin

Glucocheirolin

Progoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Epiprogoitrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


52

12

58

34

62

76

69

71

85

48

11

306

118

83

15

799

19

255

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

24




min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

61

50

60

56

98

61

03

69

18

84

01

11

169

11

741

14

944 1

569

2 1

601

1

18

963

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


25




min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


49

78

55

68

60

05

14

842

835

0

Glucosibarin

Glucoraphenin

Sinigrin

Progoitrin G

lucocheirolin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


46

57

50

54 56

93

60

99

69

01

83

67

11

161

11

743 14

941

15

688

16

006

18

908

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

-50 0

50 100 150 200 250 300 350


46

43

50

21

56

67

60

04

68

75

83

47

11

137

11

724

14

931

15

684

16

005

18

962

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

26




extract

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


47

63

51

69 5

823

6

238

70

65

86

68

11

293

11

873

15

081 1

583

2 1

614

1

19

126

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150

200 250 300 350


55

88

84

78

14

918

Progoitrin

Glucoraphenin

Glucosibarin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


27





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

59

50

50

56

94 6

094

6

901

83

64

11

141

11

736 1

494

0 1

570

8 1

601

0 18

967

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


62

09

19

122

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

04

49

88

56

02

59

96

67

77

82

20

11

064

11

629

14

850 1

561

1 1

594

1

18

920

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin Sinigrin

Gluconasturtiin

28




min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

69

51

46 5

819

6

006

70

49

85

26

11

128

11

722

15

166

15

899

16

228 1

919

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

72

58

21

59

99

70

50

11

138

11

723

15

177

16

204 1

919

2


Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

53

58

18

61

76

70

27

11

212

11

833

15

119

16

145

19

093


Sinigrin

Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

29





standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150

175


47

47

51

30

58

12

617

4

702

3

84

67

11

208

11

834

15

077

158

18 1

614

0

19

084

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


57

50 60

92

69

29

79

53

11

809

Progoitrin

Epiprogoitrin

Glucoraphenin

Gluconapin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

33

50

10

56

77 60

70

68

57

79

24

11

128

11

732

14

924 1

567

7 1

599

2

18

929

Glucoiberin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Glucocheirolin

30




min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

9
























10




















11









27 HPLC analysis













12

Gradient program

0

5

10

15

20

25

30

35

0 5 10 15 20 25 30

Time(min)

B

1
















13















min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800

1000

1200


47

72

51

50

58

37 6

244

70

42

84

92

11

234

11

899

15

084

15

811

16

160

19

101

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

14























15















gluconasturtiin



respectively

16








min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


48

05

52

12

58

34

62

76

69

71

85

48

11

306

11

883

15

799

16

026

19

255

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

Glucoerucin

17



















respectively

18



extract at 11883min


Max 39820 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3000

3200

3400

3600

3800

3982 3720233

2641464 3740192

315054126721773830960 44000993410834 4559759 484019639399712972460


Max 44870 counts

200 210 220 230 240 250 260 270 280 290 300 310 320 330 340 350 360 370 380 390 400mz amu

0

500

1000

1500

2000

2500

3000

3500

4000

4487 3720536

3740497

25523932201537 2272081 2832709 388065535803532931867 32518753392078

[M-H]- Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


19


















20




Max 1240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120124 3720156

969548

749854

1950168

25899591300262

274971617897167995151289233

850283 29205781450374 2270103 2409984 3009815564612


Max 1180 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

118 749866

969584

37206001300304799501 25901761950337 27499751789801850244 1289378 2920796590074 17599271630819

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S


Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S[M-H]-

[M-HSO4-H]-

[M-SO3-H]-

21











standards spiked

Progoitrin G

lucocheirolin

Glucoiberin

Glucoerucin

Glucosibarin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


47

60

51

78

58

40

62

42

70

83

86

16

11

271

11

861

15

077

15

834

16

103

19

110

Glucoiberin

Glucocheirolin


Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Progoitrin

Glucoraphenin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


46

75

51

48

61

86

87

09

15

122

16

142

22





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

11

49

71

56

31

60

64

68

38

82

40

11

051

11

718

14

935

15

656

15

991 1

890

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


82

67

14

950

18

873

Glucoraphenin

Glucosibarin

Gluconasturtiin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


58

53

87

29

16

168

19

153

Progoitrin

Glucoraphenin

Glucoerucin

Gluconasturtiin

23




standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

78

51

58

58

18

62

57

70

91

86

68

11

291

11

881

15

111

15

847

16

159

19

138

Glucoiberin

Glucocheirolin

Progoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Epiprogoitrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


52

12

58

34

62

76

69

71

85

48

11

306

118

83

15

799

19

255

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

24




min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

61

50

60

56

98

61

03

69

18

84

01

11

169

11

741

14

944 1

569

2 1

601

1

18

963

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


25




min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


49

78

55

68

60

05

14

842

835

0

Glucosibarin

Glucoraphenin

Sinigrin

Progoitrin G

lucocheirolin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


46

57

50

54 56

93

60

99

69

01

83

67

11

161

11

743 14

941

15

688

16

006

18

908

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

-50 0

50 100 150 200 250 300 350


46

43

50

21

56

67

60

04

68

75

83

47

11

137

11

724

14

931

15

684

16

005

18

962

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

26




extract

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


47

63

51

69 5

823

6

238

70

65

86

68

11

293

11

873

15

081 1

583

2 1

614

1

19

126

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150

200 250 300 350


55

88

84

78

14

918

Progoitrin

Glucoraphenin

Glucosibarin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


27





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

59

50

50

56

94 6

094

6

901

83

64

11

141

11

736 1

494

0 1

570

8 1

601

0 18

967

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


62

09

19

122

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

04

49

88

56

02

59

96

67

77

82

20

11

064

11

629

14

850 1

561

1 1

594

1

18

920

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin Sinigrin

Gluconasturtiin

28




min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

69

51

46 5

819

6

006

70

49

85

26

11

128

11

722

15

166

15

899

16

228 1

919

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

72

58

21

59

99

70

50

11

138

11

723

15

177

16

204 1

919

2


Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

53

58

18

61

76

70

27

11

212

11

833

15

119

16

145

19

093


Sinigrin

Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

29





standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150

175


47

47

51

30

58

12

617

4

702

3

84

67

11

208

11

834

15

077

158

18 1

614

0

19

084

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


57

50 60

92

69

29

79

53

11

809

Progoitrin

Epiprogoitrin

Glucoraphenin

Gluconapin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

33

50

10

56

77 60

70

68

57

79

24

11

128

11

732

14

924 1

567

7 1

599

2

18

929

Glucoiberin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Glucocheirolin

30




min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

10




















11









27 HPLC analysis













12

Gradient program

0

5

10

15

20

25

30

35

0 5 10 15 20 25 30

Time(min)

B

1
















13















min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800

1000

1200


47

72

51

50

58

37 6

244

70

42

84

92

11

234

11

899

15

084

15

811

16

160

19

101

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

14























15















gluconasturtiin



respectively

16








min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


48

05

52

12

58

34

62

76

69

71

85

48

11

306

11

883

15

799

16

026

19

255

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

Glucoerucin

17



















respectively

18



extract at 11883min


Max 39820 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3000

3200

3400

3600

3800

3982 3720233

2641464 3740192

315054126721773830960 44000993410834 4559759 484019639399712972460


Max 44870 counts

200 210 220 230 240 250 260 270 280 290 300 310 320 330 340 350 360 370 380 390 400mz amu

0

500

1000

1500

2000

2500

3000

3500

4000

4487 3720536

3740497

25523932201537 2272081 2832709 388065535803532931867 32518753392078

[M-H]- Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


19


















20




Max 1240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120124 3720156

969548

749854

1950168

25899591300262

274971617897167995151289233

850283 29205781450374 2270103 2409984 3009815564612


Max 1180 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

118 749866

969584

37206001300304799501 25901761950337 27499751789801850244 1289378 2920796590074 17599271630819

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S


Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S[M-H]-

[M-HSO4-H]-

[M-SO3-H]-

21











standards spiked

Progoitrin G

lucocheirolin

Glucoiberin

Glucoerucin

Glucosibarin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


47

60

51

78

58

40

62

42

70

83

86

16

11

271

11

861

15

077

15

834

16

103

19

110

Glucoiberin

Glucocheirolin


Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Progoitrin

Glucoraphenin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


46

75

51

48

61

86

87

09

15

122

16

142

22





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

11

49

71

56

31

60

64

68

38

82

40

11

051

11

718

14

935

15

656

15

991 1

890

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


82

67

14

950

18

873

Glucoraphenin

Glucosibarin

Gluconasturtiin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


58

53

87

29

16

168

19

153

Progoitrin

Glucoraphenin

Glucoerucin

Gluconasturtiin

23




standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

78

51

58

58

18

62

57

70

91

86

68

11

291

11

881

15

111

15

847

16

159

19

138

Glucoiberin

Glucocheirolin

Progoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Epiprogoitrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


52

12

58

34

62

76

69

71

85

48

11

306

118

83

15

799

19

255

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

24




min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

61

50

60

56

98

61

03

69

18

84

01

11

169

11

741

14

944 1

569

2 1

601

1

18

963

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


25




min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


49

78

55

68

60

05

14

842

835

0

Glucosibarin

Glucoraphenin

Sinigrin

Progoitrin G

lucocheirolin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


46

57

50

54 56

93

60

99

69

01

83

67

11

161

11

743 14

941

15

688

16

006

18

908

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

-50 0

50 100 150 200 250 300 350


46

43

50

21

56

67

60

04

68

75

83

47

11

137

11

724

14

931

15

684

16

005

18

962

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

26




extract

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


47

63

51

69 5

823

6

238

70

65

86

68

11

293

11

873

15

081 1

583

2 1

614

1

19

126

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150

200 250 300 350


55

88

84

78

14

918

Progoitrin

Glucoraphenin

Glucosibarin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


27





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

59

50

50

56

94 6

094

6

901

83

64

11

141

11

736 1

494

0 1

570

8 1

601

0 18

967

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


62

09

19

122

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

04

49

88

56

02

59

96

67

77

82

20

11

064

11

629

14

850 1

561

1 1

594

1

18

920

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin Sinigrin

Gluconasturtiin

28




min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

69

51

46 5

819

6

006

70

49

85

26

11

128

11

722

15

166

15

899

16

228 1

919

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

72

58

21

59

99

70

50

11

138

11

723

15

177

16

204 1

919

2


Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

53

58

18

61

76

70

27

11

212

11

833

15

119

16

145

19

093


Sinigrin

Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

29





standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150

175


47

47

51

30

58

12

617

4

702

3

84

67

11

208

11

834

15

077

158

18 1

614

0

19

084

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


57

50 60

92

69

29

79

53

11

809

Progoitrin

Epiprogoitrin

Glucoraphenin

Gluconapin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

33

50

10

56

77 60

70

68

57

79

24

11

128

11

732

14

924 1

567

7 1

599

2

18

929

Glucoiberin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Glucocheirolin

30




min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

11









27 HPLC analysis













12

Gradient program

0

5

10

15

20

25

30

35

0 5 10 15 20 25 30

Time(min)

B

1
















13















min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800

1000

1200


47

72

51

50

58

37 6

244

70

42

84

92

11

234

11

899

15

084

15

811

16

160

19

101

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

14























15















gluconasturtiin



respectively

16








min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


48

05

52

12

58

34

62

76

69

71

85

48

11

306

11

883

15

799

16

026

19

255

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

Glucoerucin

17



















respectively

18



extract at 11883min


Max 39820 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3000

3200

3400

3600

3800

3982 3720233

2641464 3740192

315054126721773830960 44000993410834 4559759 484019639399712972460


Max 44870 counts

200 210 220 230 240 250 260 270 280 290 300 310 320 330 340 350 360 370 380 390 400mz amu

0

500

1000

1500

2000

2500

3000

3500

4000

4487 3720536

3740497

25523932201537 2272081 2832709 388065535803532931867 32518753392078

[M-H]- Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


19


















20




Max 1240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120124 3720156

969548

749854

1950168

25899591300262

274971617897167995151289233

850283 29205781450374 2270103 2409984 3009815564612


Max 1180 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

118 749866

969584

37206001300304799501 25901761950337 27499751789801850244 1289378 2920796590074 17599271630819

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S


Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S[M-H]-

[M-HSO4-H]-

[M-SO3-H]-

21











standards spiked

Progoitrin G

lucocheirolin

Glucoiberin

Glucoerucin

Glucosibarin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


47

60

51

78

58

40

62

42

70

83

86

16

11

271

11

861

15

077

15

834

16

103

19

110

Glucoiberin

Glucocheirolin


Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Progoitrin

Glucoraphenin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


46

75

51

48

61

86

87

09

15

122

16

142

22





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

11

49

71

56

31

60

64

68

38

82

40

11

051

11

718

14

935

15

656

15

991 1

890

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


82

67

14

950

18

873

Glucoraphenin

Glucosibarin

Gluconasturtiin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


58

53

87

29

16

168

19

153

Progoitrin

Glucoraphenin

Glucoerucin

Gluconasturtiin

23




standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

78

51

58

58

18

62

57

70

91

86

68

11

291

11

881

15

111

15

847

16

159

19

138

Glucoiberin

Glucocheirolin

Progoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Epiprogoitrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


52

12

58

34

62

76

69

71

85

48

11

306

118

83

15

799

19

255

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

24




min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

61

50

60

56

98

61

03

69

18

84

01

11

169

11

741

14

944 1

569

2 1

601

1

18

963

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


25




min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


49

78

55

68

60

05

14

842

835

0

Glucosibarin

Glucoraphenin

Sinigrin

Progoitrin G

lucocheirolin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


46

57

50

54 56

93

60

99

69

01

83

67

11

161

11

743 14

941

15

688

16

006

18

908

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

-50 0

50 100 150 200 250 300 350


46

43

50

21

56

67

60

04

68

75

83

47

11

137

11

724

14

931

15

684

16

005

18

962

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

26




extract

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


47

63

51

69 5

823

6

238

70

65

86

68

11

293

11

873

15

081 1

583

2 1

614

1

19

126

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150

200 250 300 350


55

88

84

78

14

918

Progoitrin

Glucoraphenin

Glucosibarin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


27





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

59

50

50

56

94 6

094

6

901

83

64

11

141

11

736 1

494

0 1

570

8 1

601

0 18

967

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


62

09

19

122

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

04

49

88

56

02

59

96

67

77

82

20

11

064

11

629

14

850 1

561

1 1

594

1

18

920

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin Sinigrin

Gluconasturtiin

28




min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

69

51

46 5

819

6

006

70

49

85

26

11

128

11

722

15

166

15

899

16

228 1

919

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

72

58

21

59

99

70

50

11

138

11

723

15

177

16

204 1

919

2


Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

53

58

18

61

76

70

27

11

212

11

833

15

119

16

145

19

093


Sinigrin

Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

29





standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150

175


47

47

51

30

58

12

617

4

702

3

84

67

11

208

11

834

15

077

158

18 1

614

0

19

084

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


57

50 60

92

69

29

79

53

11

809

Progoitrin

Epiprogoitrin

Glucoraphenin

Gluconapin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

33

50

10

56

77 60

70

68

57

79

24

11

128

11

732

14

924 1

567

7 1

599

2

18

929

Glucoiberin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Glucocheirolin

30




min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

12

Gradient program

0

5

10

15

20

25

30

35

0 5 10 15 20 25 30

Time(min)

B

1
















13















min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800

1000

1200


47

72

51

50

58

37 6

244

70

42

84

92

11

234

11

899

15

084

15

811

16

160

19

101

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

14























15















gluconasturtiin



respectively

16








min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


48

05

52

12

58

34

62

76

69

71

85

48

11

306

11

883

15

799

16

026

19

255

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

Glucoerucin

17



















respectively

18



extract at 11883min


Max 39820 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3000

3200

3400

3600

3800

3982 3720233

2641464 3740192

315054126721773830960 44000993410834 4559759 484019639399712972460


Max 44870 counts

200 210 220 230 240 250 260 270 280 290 300 310 320 330 340 350 360 370 380 390 400mz amu

0

500

1000

1500

2000

2500

3000

3500

4000

4487 3720536

3740497

25523932201537 2272081 2832709 388065535803532931867 32518753392078

[M-H]- Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


19


















20




Max 1240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120124 3720156

969548

749854

1950168

25899591300262

274971617897167995151289233

850283 29205781450374 2270103 2409984 3009815564612


Max 1180 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

118 749866

969584

37206001300304799501 25901761950337 27499751789801850244 1289378 2920796590074 17599271630819

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S


Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S[M-H]-

[M-HSO4-H]-

[M-SO3-H]-

21











standards spiked

Progoitrin G

lucocheirolin

Glucoiberin

Glucoerucin

Glucosibarin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


47

60

51

78

58

40

62

42

70

83

86

16

11

271

11

861

15

077

15

834

16

103

19

110

Glucoiberin

Glucocheirolin


Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Progoitrin

Glucoraphenin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


46

75

51

48

61

86

87

09

15

122

16

142

22





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

11

49

71

56

31

60

64

68

38

82

40

11

051

11

718

14

935

15

656

15

991 1

890

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


82

67

14

950

18

873

Glucoraphenin

Glucosibarin

Gluconasturtiin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


58

53

87

29

16

168

19

153

Progoitrin

Glucoraphenin

Glucoerucin

Gluconasturtiin

23




standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

78

51

58

58

18

62

57

70

91

86

68

11

291

11

881

15

111

15

847

16

159

19

138

Glucoiberin

Glucocheirolin

Progoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Epiprogoitrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


52

12

58

34

62

76

69

71

85

48

11

306

118

83

15

799

19

255

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

24




min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

61

50

60

56

98

61

03

69

18

84

01

11

169

11

741

14

944 1

569

2 1

601

1

18

963

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


25




min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


49

78

55

68

60

05

14

842

835

0

Glucosibarin

Glucoraphenin

Sinigrin

Progoitrin G

lucocheirolin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


46

57

50

54 56

93

60

99

69

01

83

67

11

161

11

743 14

941

15

688

16

006

18

908

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

-50 0

50 100 150 200 250 300 350


46

43

50

21

56

67

60

04

68

75

83

47

11

137

11

724

14

931

15

684

16

005

18

962

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

26




extract

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


47

63

51

69 5

823

6

238

70

65

86

68

11

293

11

873

15

081 1

583

2 1

614

1

19

126

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150

200 250 300 350


55

88

84

78

14

918

Progoitrin

Glucoraphenin

Glucosibarin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


27





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

59

50

50

56

94 6

094

6

901

83

64

11

141

11

736 1

494

0 1

570

8 1

601

0 18

967

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


62

09

19

122

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

04

49

88

56

02

59

96

67

77

82

20

11

064

11

629

14

850 1

561

1 1

594

1

18

920

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin Sinigrin

Gluconasturtiin

28




min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

69

51

46 5

819

6

006

70

49

85

26

11

128

11

722

15

166

15

899

16

228 1

919

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

72

58

21

59

99

70

50

11

138

11

723

15

177

16

204 1

919

2


Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

53

58

18

61

76

70

27

11

212

11

833

15

119

16

145

19

093


Sinigrin

Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

29





standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150

175


47

47

51

30

58

12

617

4

702

3

84

67

11

208

11

834

15

077

158

18 1

614

0

19

084

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


57

50 60

92

69

29

79

53

11

809

Progoitrin

Epiprogoitrin

Glucoraphenin

Gluconapin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

33

50

10

56

77 60

70

68

57

79

24

11

128

11

732

14

924 1

567

7 1

599

2

18

929

Glucoiberin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Glucocheirolin

30




min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

13















min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800

1000

1200


47

72

51

50

58

37 6

244

70

42

84

92

11

234

11

899

15

084

15

811

16

160

19

101

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

14























15















gluconasturtiin



respectively

16








min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


48

05

52

12

58

34

62

76

69

71

85

48

11

306

11

883

15

799

16

026

19

255

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

Glucoerucin

17



















respectively

18



extract at 11883min


Max 39820 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3000

3200

3400

3600

3800

3982 3720233

2641464 3740192

315054126721773830960 44000993410834 4559759 484019639399712972460


Max 44870 counts

200 210 220 230 240 250 260 270 280 290 300 310 320 330 340 350 360 370 380 390 400mz amu

0

500

1000

1500

2000

2500

3000

3500

4000

4487 3720536

3740497

25523932201537 2272081 2832709 388065535803532931867 32518753392078

[M-H]- Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


19


















20




Max 1240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120124 3720156

969548

749854

1950168

25899591300262

274971617897167995151289233

850283 29205781450374 2270103 2409984 3009815564612


Max 1180 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

118 749866

969584

37206001300304799501 25901761950337 27499751789801850244 1289378 2920796590074 17599271630819

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S


Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S[M-H]-

[M-HSO4-H]-

[M-SO3-H]-

21











standards spiked

Progoitrin G

lucocheirolin

Glucoiberin

Glucoerucin

Glucosibarin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


47

60

51

78

58

40

62

42

70

83

86

16

11

271

11

861

15

077

15

834

16

103

19

110

Glucoiberin

Glucocheirolin


Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Progoitrin

Glucoraphenin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


46

75

51

48

61

86

87

09

15

122

16

142

22





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

11

49

71

56

31

60

64

68

38

82

40

11

051

11

718

14

935

15

656

15

991 1

890

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


82

67

14

950

18

873

Glucoraphenin

Glucosibarin

Gluconasturtiin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


58

53

87

29

16

168

19

153

Progoitrin

Glucoraphenin

Glucoerucin

Gluconasturtiin

23




standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

78

51

58

58

18

62

57

70

91

86

68

11

291

11

881

15

111

15

847

16

159

19

138

Glucoiberin

Glucocheirolin

Progoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Epiprogoitrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


52

12

58

34

62

76

69

71

85

48

11

306

118

83

15

799

19

255

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

24




min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

61

50

60

56

98

61

03

69

18

84

01

11

169

11

741

14

944 1

569

2 1

601

1

18

963

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


25




min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


49

78

55

68

60

05

14

842

835

0

Glucosibarin

Glucoraphenin

Sinigrin

Progoitrin G

lucocheirolin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


46

57

50

54 56

93

60

99

69

01

83

67

11

161

11

743 14

941

15

688

16

006

18

908

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

-50 0

50 100 150 200 250 300 350


46

43

50

21

56

67

60

04

68

75

83

47

11

137

11

724

14

931

15

684

16

005

18

962

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

26




extract

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


47

63

51

69 5

823

6

238

70

65

86

68

11

293

11

873

15

081 1

583

2 1

614

1

19

126

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150

200 250 300 350


55

88

84

78

14

918

Progoitrin

Glucoraphenin

Glucosibarin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


27





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

59

50

50

56

94 6

094

6

901

83

64

11

141

11

736 1

494

0 1

570

8 1

601

0 18

967

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


62

09

19

122

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

04

49

88

56

02

59

96

67

77

82

20

11

064

11

629

14

850 1

561

1 1

594

1

18

920

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin Sinigrin

Gluconasturtiin

28




min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

69

51

46 5

819

6

006

70

49

85

26

11

128

11

722

15

166

15

899

16

228 1

919

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

72

58

21

59

99

70

50

11

138

11

723

15

177

16

204 1

919

2


Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

53

58

18

61

76

70

27

11

212

11

833

15

119

16

145

19

093


Sinigrin

Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

29





standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150

175


47

47

51

30

58

12

617

4

702

3

84

67

11

208

11

834

15

077

158

18 1

614

0

19

084

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


57

50 60

92

69

29

79

53

11

809

Progoitrin

Epiprogoitrin

Glucoraphenin

Gluconapin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

33

50

10

56

77 60

70

68

57

79

24

11

128

11

732

14

924 1

567

7 1

599

2

18

929

Glucoiberin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Glucocheirolin

30




min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

14























15















gluconasturtiin



respectively

16








min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


48

05

52

12

58

34

62

76

69

71

85

48

11

306

11

883

15

799

16

026

19

255

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

Glucoerucin

17



















respectively

18



extract at 11883min


Max 39820 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3000

3200

3400

3600

3800

3982 3720233

2641464 3740192

315054126721773830960 44000993410834 4559759 484019639399712972460


Max 44870 counts

200 210 220 230 240 250 260 270 280 290 300 310 320 330 340 350 360 370 380 390 400mz amu

0

500

1000

1500

2000

2500

3000

3500

4000

4487 3720536

3740497

25523932201537 2272081 2832709 388065535803532931867 32518753392078

[M-H]- Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


19


















20




Max 1240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120124 3720156

969548

749854

1950168

25899591300262

274971617897167995151289233

850283 29205781450374 2270103 2409984 3009815564612


Max 1180 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

118 749866

969584

37206001300304799501 25901761950337 27499751789801850244 1289378 2920796590074 17599271630819

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S


Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S[M-H]-

[M-HSO4-H]-

[M-SO3-H]-

21











standards spiked

Progoitrin G

lucocheirolin

Glucoiberin

Glucoerucin

Glucosibarin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


47

60

51

78

58

40

62

42

70

83

86

16

11

271

11

861

15

077

15

834

16

103

19

110

Glucoiberin

Glucocheirolin


Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Progoitrin

Glucoraphenin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


46

75

51

48

61

86

87

09

15

122

16

142

22





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

11

49

71

56

31

60

64

68

38

82

40

11

051

11

718

14

935

15

656

15

991 1

890

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


82

67

14

950

18

873

Glucoraphenin

Glucosibarin

Gluconasturtiin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


58

53

87

29

16

168

19

153

Progoitrin

Glucoraphenin

Glucoerucin

Gluconasturtiin

23




standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

78

51

58

58

18

62

57

70

91

86

68

11

291

11

881

15

111

15

847

16

159

19

138

Glucoiberin

Glucocheirolin

Progoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Epiprogoitrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


52

12

58

34

62

76

69

71

85

48

11

306

118

83

15

799

19

255

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

24




min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

61

50

60

56

98

61

03

69

18

84

01

11

169

11

741

14

944 1

569

2 1

601

1

18

963

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


25




min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


49

78

55

68

60

05

14

842

835

0

Glucosibarin

Glucoraphenin

Sinigrin

Progoitrin G

lucocheirolin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


46

57

50

54 56

93

60

99

69

01

83

67

11

161

11

743 14

941

15

688

16

006

18

908

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

-50 0

50 100 150 200 250 300 350


46

43

50

21

56

67

60

04

68

75

83

47

11

137

11

724

14

931

15

684

16

005

18

962

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

26




extract

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


47

63

51

69 5

823

6

238

70

65

86

68

11

293

11

873

15

081 1

583

2 1

614

1

19

126

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150

200 250 300 350


55

88

84

78

14

918

Progoitrin

Glucoraphenin

Glucosibarin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


27





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

59

50

50

56

94 6

094

6

901

83

64

11

141

11

736 1

494

0 1

570

8 1

601

0 18

967

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


62

09

19

122

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

04

49

88

56

02

59

96

67

77

82

20

11

064

11

629

14

850 1

561

1 1

594

1

18

920

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin Sinigrin

Gluconasturtiin

28




min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

69

51

46 5

819

6

006

70

49

85

26

11

128

11

722

15

166

15

899

16

228 1

919

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

72

58

21

59

99

70

50

11

138

11

723

15

177

16

204 1

919

2


Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

53

58

18

61

76

70

27

11

212

11

833

15

119

16

145

19

093


Sinigrin

Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

29





standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150

175


47

47

51

30

58

12

617

4

702

3

84

67

11

208

11

834

15

077

158

18 1

614

0

19

084

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


57

50 60

92

69

29

79

53

11

809

Progoitrin

Epiprogoitrin

Glucoraphenin

Gluconapin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

33

50

10

56

77 60

70

68

57

79

24

11

128

11

732

14

924 1

567

7 1

599

2

18

929

Glucoiberin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Glucocheirolin

30




min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

15















gluconasturtiin



respectively

16








min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


48

05

52

12

58

34

62

76

69

71

85

48

11

306

11

883

15

799

16

026

19

255

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

Glucoerucin

17



















respectively

18



extract at 11883min


Max 39820 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3000

3200

3400

3600

3800

3982 3720233

2641464 3740192

315054126721773830960 44000993410834 4559759 484019639399712972460


Max 44870 counts

200 210 220 230 240 250 260 270 280 290 300 310 320 330 340 350 360 370 380 390 400mz amu

0

500

1000

1500

2000

2500

3000

3500

4000

4487 3720536

3740497

25523932201537 2272081 2832709 388065535803532931867 32518753392078

[M-H]- Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


19


















20




Max 1240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120124 3720156

969548

749854

1950168

25899591300262

274971617897167995151289233

850283 29205781450374 2270103 2409984 3009815564612


Max 1180 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

118 749866

969584

37206001300304799501 25901761950337 27499751789801850244 1289378 2920796590074 17599271630819

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S


Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S[M-H]-

[M-HSO4-H]-

[M-SO3-H]-

21











standards spiked

Progoitrin G

lucocheirolin

Glucoiberin

Glucoerucin

Glucosibarin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


47

60

51

78

58

40

62

42

70

83

86

16

11

271

11

861

15

077

15

834

16

103

19

110

Glucoiberin

Glucocheirolin


Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Progoitrin

Glucoraphenin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


46

75

51

48

61

86

87

09

15

122

16

142

22





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

11

49

71

56

31

60

64

68

38

82

40

11

051

11

718

14

935

15

656

15

991 1

890

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


82

67

14

950

18

873

Glucoraphenin

Glucosibarin

Gluconasturtiin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


58

53

87

29

16

168

19

153

Progoitrin

Glucoraphenin

Glucoerucin

Gluconasturtiin

23




standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

78

51

58

58

18

62

57

70

91

86

68

11

291

11

881

15

111

15

847

16

159

19

138

Glucoiberin

Glucocheirolin

Progoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Epiprogoitrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


52

12

58

34

62

76

69

71

85

48

11

306

118

83

15

799

19

255

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

24




min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

61

50

60

56

98

61

03

69

18

84

01

11

169

11

741

14

944 1

569

2 1

601

1

18

963

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


25




min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


49

78

55

68

60

05

14

842

835

0

Glucosibarin

Glucoraphenin

Sinigrin

Progoitrin G

lucocheirolin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


46

57

50

54 56

93

60

99

69

01

83

67

11

161

11

743 14

941

15

688

16

006

18

908

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

-50 0

50 100 150 200 250 300 350


46

43

50

21

56

67

60

04

68

75

83

47

11

137

11

724

14

931

15

684

16

005

18

962

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

26




extract

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


47

63

51

69 5

823

6

238

70

65

86

68

11

293

11

873

15

081 1

583

2 1

614

1

19

126

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150

200 250 300 350


55

88

84

78

14

918

Progoitrin

Glucoraphenin

Glucosibarin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


27





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

59

50

50

56

94 6

094

6

901

83

64

11

141

11

736 1

494

0 1

570

8 1

601

0 18

967

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


62

09

19

122

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

04

49

88

56

02

59

96

67

77

82

20

11

064

11

629

14

850 1

561

1 1

594

1

18

920

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin Sinigrin

Gluconasturtiin

28




min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

69

51

46 5

819

6

006

70

49

85

26

11

128

11

722

15

166

15

899

16

228 1

919

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

72

58

21

59

99

70

50

11

138

11

723

15

177

16

204 1

919

2


Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

53

58

18

61

76

70

27

11

212

11

833

15

119

16

145

19

093


Sinigrin

Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

29





standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150

175


47

47

51

30

58

12

617

4

702

3

84

67

11

208

11

834

15

077

158

18 1

614

0

19

084

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


57

50 60

92

69

29

79

53

11

809

Progoitrin

Epiprogoitrin

Glucoraphenin

Gluconapin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

33

50

10

56

77 60

70

68

57

79

24

11

128

11

732

14

924 1

567

7 1

599

2

18

929

Glucoiberin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Glucocheirolin

30




min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

16








min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


48

05

52

12

58

34

62

76

69

71

85

48

11

306

11

883

15

799

16

026

19

255

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

Glucoerucin

17



















respectively

18



extract at 11883min


Max 39820 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3000

3200

3400

3600

3800

3982 3720233

2641464 3740192

315054126721773830960 44000993410834 4559759 484019639399712972460


Max 44870 counts

200 210 220 230 240 250 260 270 280 290 300 310 320 330 340 350 360 370 380 390 400mz amu

0

500

1000

1500

2000

2500

3000

3500

4000

4487 3720536

3740497

25523932201537 2272081 2832709 388065535803532931867 32518753392078

[M-H]- Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


19


















20




Max 1240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120124 3720156

969548

749854

1950168

25899591300262

274971617897167995151289233

850283 29205781450374 2270103 2409984 3009815564612


Max 1180 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

118 749866

969584

37206001300304799501 25901761950337 27499751789801850244 1289378 2920796590074 17599271630819

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S


Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S[M-H]-

[M-HSO4-H]-

[M-SO3-H]-

21











standards spiked

Progoitrin G

lucocheirolin

Glucoiberin

Glucoerucin

Glucosibarin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


47

60

51

78

58

40

62

42

70

83

86

16

11

271

11

861

15

077

15

834

16

103

19

110

Glucoiberin

Glucocheirolin


Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Progoitrin

Glucoraphenin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


46

75

51

48

61

86

87

09

15

122

16

142

22





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

11

49

71

56

31

60

64

68

38

82

40

11

051

11

718

14

935

15

656

15

991 1

890

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


82

67

14

950

18

873

Glucoraphenin

Glucosibarin

Gluconasturtiin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


58

53

87

29

16

168

19

153

Progoitrin

Glucoraphenin

Glucoerucin

Gluconasturtiin

23




standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

78

51

58

58

18

62

57

70

91

86

68

11

291

11

881

15

111

15

847

16

159

19

138

Glucoiberin

Glucocheirolin

Progoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Epiprogoitrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


52

12

58

34

62

76

69

71

85

48

11

306

118

83

15

799

19

255

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

24




min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

61

50

60

56

98

61

03

69

18

84

01

11

169

11

741

14

944 1

569

2 1

601

1

18

963

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


25




min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


49

78

55

68

60

05

14

842

835

0

Glucosibarin

Glucoraphenin

Sinigrin

Progoitrin G

lucocheirolin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


46

57

50

54 56

93

60

99

69

01

83

67

11

161

11

743 14

941

15

688

16

006

18

908

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

-50 0

50 100 150 200 250 300 350


46

43

50

21

56

67

60

04

68

75

83

47

11

137

11

724

14

931

15

684

16

005

18

962

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

26




extract

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


47

63

51

69 5

823

6

238

70

65

86

68

11

293

11

873

15

081 1

583

2 1

614

1

19

126

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150

200 250 300 350


55

88

84

78

14

918

Progoitrin

Glucoraphenin

Glucosibarin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


27





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

59

50

50

56

94 6

094

6

901

83

64

11

141

11

736 1

494

0 1

570

8 1

601

0 18

967

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


62

09

19

122

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

04

49

88

56

02

59

96

67

77

82

20

11

064

11

629

14

850 1

561

1 1

594

1

18

920

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin Sinigrin

Gluconasturtiin

28




min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

69

51

46 5

819

6

006

70

49

85

26

11

128

11

722

15

166

15

899

16

228 1

919

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

72

58

21

59

99

70

50

11

138

11

723

15

177

16

204 1

919

2


Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

53

58

18

61

76

70

27

11

212

11

833

15

119

16

145

19

093


Sinigrin

Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

29





standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150

175


47

47

51

30

58

12

617

4

702

3

84

67

11

208

11

834

15

077

158

18 1

614

0

19

084

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


57

50 60

92

69

29

79

53

11

809

Progoitrin

Epiprogoitrin

Glucoraphenin

Gluconapin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

33

50

10

56

77 60

70

68

57

79

24

11

128

11

732

14

924 1

567

7 1

599

2

18

929

Glucoiberin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Glucocheirolin

30




min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

17



















respectively

18



extract at 11883min


Max 39820 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3000

3200

3400

3600

3800

3982 3720233

2641464 3740192

315054126721773830960 44000993410834 4559759 484019639399712972460


Max 44870 counts

200 210 220 230 240 250 260 270 280 290 300 310 320 330 340 350 360 370 380 390 400mz amu

0

500

1000

1500

2000

2500

3000

3500

4000

4487 3720536

3740497

25523932201537 2272081 2832709 388065535803532931867 32518753392078

[M-H]- Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


19


















20




Max 1240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120124 3720156

969548

749854

1950168

25899591300262

274971617897167995151289233

850283 29205781450374 2270103 2409984 3009815564612


Max 1180 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

118 749866

969584

37206001300304799501 25901761950337 27499751789801850244 1289378 2920796590074 17599271630819

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S


Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S[M-H]-

[M-HSO4-H]-

[M-SO3-H]-

21











standards spiked

Progoitrin G

lucocheirolin

Glucoiberin

Glucoerucin

Glucosibarin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


47

60

51

78

58

40

62

42

70

83

86

16

11

271

11

861

15

077

15

834

16

103

19

110

Glucoiberin

Glucocheirolin


Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Progoitrin

Glucoraphenin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


46

75

51

48

61

86

87

09

15

122

16

142

22





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

11

49

71

56

31

60

64

68

38

82

40

11

051

11

718

14

935

15

656

15

991 1

890

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


82

67

14

950

18

873

Glucoraphenin

Glucosibarin

Gluconasturtiin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


58

53

87

29

16

168

19

153

Progoitrin

Glucoraphenin

Glucoerucin

Gluconasturtiin

23




standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

78

51

58

58

18

62

57

70

91

86

68

11

291

11

881

15

111

15

847

16

159

19

138

Glucoiberin

Glucocheirolin

Progoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Epiprogoitrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


52

12

58

34

62

76

69

71

85

48

11

306

118

83

15

799

19

255

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

24




min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

61

50

60

56

98

61

03

69

18

84

01

11

169

11

741

14

944 1

569

2 1

601

1

18

963

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


25




min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


49

78

55

68

60

05

14

842

835

0

Glucosibarin

Glucoraphenin

Sinigrin

Progoitrin G

lucocheirolin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


46

57

50

54 56

93

60

99

69

01

83

67

11

161

11

743 14

941

15

688

16

006

18

908

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

-50 0

50 100 150 200 250 300 350


46

43

50

21

56

67

60

04

68

75

83

47

11

137

11

724

14

931

15

684

16

005

18

962

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

26




extract

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


47

63

51

69 5

823

6

238

70

65

86

68

11

293

11

873

15

081 1

583

2 1

614

1

19

126

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150

200 250 300 350


55

88

84

78

14

918

Progoitrin

Glucoraphenin

Glucosibarin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


27





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

59

50

50

56

94 6

094

6

901

83

64

11

141

11

736 1

494

0 1

570

8 1

601

0 18

967

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


62

09

19

122

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

04

49

88

56

02

59

96

67

77

82

20

11

064

11

629

14

850 1

561

1 1

594

1

18

920

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin Sinigrin

Gluconasturtiin

28




min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

69

51

46 5

819

6

006

70

49

85

26

11

128

11

722

15

166

15

899

16

228 1

919

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

72

58

21

59

99

70

50

11

138

11

723

15

177

16

204 1

919

2


Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

53

58

18

61

76

70

27

11

212

11

833

15

119

16

145

19

093


Sinigrin

Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

29





standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150

175


47

47

51

30

58

12

617

4

702

3

84

67

11

208

11

834

15

077

158

18 1

614

0

19

084

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


57

50 60

92

69

29

79

53

11

809

Progoitrin

Epiprogoitrin

Glucoraphenin

Gluconapin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

33

50

10

56

77 60

70

68

57

79

24

11

128

11

732

14

924 1

567

7 1

599

2

18

929

Glucoiberin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Glucocheirolin

30




min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

18



extract at 11883min


Max 39820 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3000

3200

3400

3600

3800

3982 3720233

2641464 3740192

315054126721773830960 44000993410834 4559759 484019639399712972460


Max 44870 counts

200 210 220 230 240 250 260 270 280 290 300 310 320 330 340 350 360 370 380 390 400mz amu

0

500

1000

1500

2000

2500

3000

3500

4000

4487 3720536

3740497

25523932201537 2272081 2832709 388065535803532931867 32518753392078

[M-H]- Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


19


















20




Max 1240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120124 3720156

969548

749854

1950168

25899591300262

274971617897167995151289233

850283 29205781450374 2270103 2409984 3009815564612


Max 1180 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

118 749866

969584

37206001300304799501 25901761950337 27499751789801850244 1289378 2920796590074 17599271630819

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S


Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S[M-H]-

[M-HSO4-H]-

[M-SO3-H]-

21











standards spiked

Progoitrin G

lucocheirolin

Glucoiberin

Glucoerucin

Glucosibarin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


47

60

51

78

58

40

62

42

70

83

86

16

11

271

11

861

15

077

15

834

16

103

19

110

Glucoiberin

Glucocheirolin


Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Progoitrin

Glucoraphenin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


46

75

51

48

61

86

87

09

15

122

16

142

22





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

11

49

71

56

31

60

64

68

38

82

40

11

051

11

718

14

935

15

656

15

991 1

890

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


82

67

14

950

18

873

Glucoraphenin

Glucosibarin

Gluconasturtiin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


58

53

87

29

16

168

19

153

Progoitrin

Glucoraphenin

Glucoerucin

Gluconasturtiin

23




standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

78

51

58

58

18

62

57

70

91

86

68

11

291

11

881

15

111

15

847

16

159

19

138

Glucoiberin

Glucocheirolin

Progoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Epiprogoitrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


52

12

58

34

62

76

69

71

85

48

11

306

118

83

15

799

19

255

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

24




min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

61

50

60

56

98

61

03

69

18

84

01

11

169

11

741

14

944 1

569

2 1

601

1

18

963

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


25




min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


49

78

55

68

60

05

14

842

835

0

Glucosibarin

Glucoraphenin

Sinigrin

Progoitrin G

lucocheirolin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


46

57

50

54 56

93

60

99

69

01

83

67

11

161

11

743 14

941

15

688

16

006

18

908

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

-50 0

50 100 150 200 250 300 350


46

43

50

21

56

67

60

04

68

75

83

47

11

137

11

724

14

931

15

684

16

005

18

962

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

26




extract

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


47

63

51

69 5

823

6

238

70

65

86

68

11

293

11

873

15

081 1

583

2 1

614

1

19

126

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150

200 250 300 350


55

88

84

78

14

918

Progoitrin

Glucoraphenin

Glucosibarin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


27





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

59

50

50

56

94 6

094

6

901

83

64

11

141

11

736 1

494

0 1

570

8 1

601

0 18

967

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


62

09

19

122

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

04

49

88

56

02

59

96

67

77

82

20

11

064

11

629

14

850 1

561

1 1

594

1

18

920

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin Sinigrin

Gluconasturtiin

28




min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

69

51

46 5

819

6

006

70

49

85

26

11

128

11

722

15

166

15

899

16

228 1

919

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

72

58

21

59

99

70

50

11

138

11

723

15

177

16

204 1

919

2


Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

53

58

18

61

76

70

27

11

212

11

833

15

119

16

145

19

093


Sinigrin

Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

29





standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150

175


47

47

51

30

58

12

617

4

702

3

84

67

11

208

11

834

15

077

158

18 1

614

0

19

084

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


57

50 60

92

69

29

79

53

11

809

Progoitrin

Epiprogoitrin

Glucoraphenin

Gluconapin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

33

50

10

56

77 60

70

68

57

79

24

11

128

11

732

14

924 1

567

7 1

599

2

18

929

Glucoiberin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Glucocheirolin

30




min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

19


















20




Max 1240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120124 3720156

969548

749854

1950168

25899591300262

274971617897167995151289233

850283 29205781450374 2270103 2409984 3009815564612


Max 1180 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

118 749866

969584

37206001300304799501 25901761950337 27499751789801850244 1289378 2920796590074 17599271630819

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S


Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S[M-H]-

[M-HSO4-H]-

[M-SO3-H]-

21











standards spiked

Progoitrin G

lucocheirolin

Glucoiberin

Glucoerucin

Glucosibarin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


47

60

51

78

58

40

62

42

70

83

86

16

11

271

11

861

15

077

15

834

16

103

19

110

Glucoiberin

Glucocheirolin


Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Progoitrin

Glucoraphenin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


46

75

51

48

61

86

87

09

15

122

16

142

22





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

11

49

71

56

31

60

64

68

38

82

40

11

051

11

718

14

935

15

656

15

991 1

890

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


82

67

14

950

18

873

Glucoraphenin

Glucosibarin

Gluconasturtiin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


58

53

87

29

16

168

19

153

Progoitrin

Glucoraphenin

Glucoerucin

Gluconasturtiin

23




standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

78

51

58

58

18

62

57

70

91

86

68

11

291

11

881

15

111

15

847

16

159

19

138

Glucoiberin

Glucocheirolin

Progoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Epiprogoitrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


52

12

58

34

62

76

69

71

85

48

11

306

118

83

15

799

19

255

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

24




min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

61

50

60

56

98

61

03

69

18

84

01

11

169

11

741

14

944 1

569

2 1

601

1

18

963

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


25




min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


49

78

55

68

60

05

14

842

835

0

Glucosibarin

Glucoraphenin

Sinigrin

Progoitrin G

lucocheirolin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


46

57

50

54 56

93

60

99

69

01

83

67

11

161

11

743 14

941

15

688

16

006

18

908

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

-50 0

50 100 150 200 250 300 350


46

43

50

21

56

67

60

04

68

75

83

47

11

137

11

724

14

931

15

684

16

005

18

962

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

26




extract

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


47

63

51

69 5

823

6

238

70

65

86

68

11

293

11

873

15

081 1

583

2 1

614

1

19

126

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150

200 250 300 350


55

88

84

78

14

918

Progoitrin

Glucoraphenin

Glucosibarin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


27





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

59

50

50

56

94 6

094

6

901

83

64

11

141

11

736 1

494

0 1

570

8 1

601

0 18

967

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


62

09

19

122

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

04

49

88

56

02

59

96

67

77

82

20

11

064

11

629

14

850 1

561

1 1

594

1

18

920

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin Sinigrin

Gluconasturtiin

28




min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

69

51

46 5

819

6

006

70

49

85

26

11

128

11

722

15

166

15

899

16

228 1

919

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

72

58

21

59

99

70

50

11

138

11

723

15

177

16

204 1

919

2


Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

53

58

18

61

76

70

27

11

212

11

833

15

119

16

145

19

093


Sinigrin

Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

29





standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150

175


47

47

51

30

58

12

617

4

702

3

84

67

11

208

11

834

15

077

158

18 1

614

0

19

084

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


57

50 60

92

69

29

79

53

11

809

Progoitrin

Epiprogoitrin

Glucoraphenin

Gluconapin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

33

50

10

56

77 60

70

68

57

79

24

11

128

11

732

14

924 1

567

7 1

599

2

18

929

Glucoiberin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Glucocheirolin

30




min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

20




Max 1240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120124 3720156

969548

749854

1950168

25899591300262

274971617897167995151289233

850283 29205781450374 2270103 2409984 3009815564612


Max 1180 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

118 749866

969584

37206001300304799501 25901761950337 27499751789801850244 1289378 2920796590074 17599271630819

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S


Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


HSO4-

SO3-

ΟΟΗ

OH

OH

CH2OH

S[M-H]-

[M-HSO4-H]-

[M-SO3-H]-

21











standards spiked

Progoitrin G

lucocheirolin

Glucoiberin

Glucoerucin

Glucosibarin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


47

60

51

78

58

40

62

42

70

83

86

16

11

271

11

861

15

077

15

834

16

103

19

110

Glucoiberin

Glucocheirolin


Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Progoitrin

Glucoraphenin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


46

75

51

48

61

86

87

09

15

122

16

142

22





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

11

49

71

56

31

60

64

68

38

82

40

11

051

11

718

14

935

15

656

15

991 1

890

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


82

67

14

950

18

873

Glucoraphenin

Glucosibarin

Gluconasturtiin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


58

53

87

29

16

168

19

153

Progoitrin

Glucoraphenin

Glucoerucin

Gluconasturtiin

23




standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

78

51

58

58

18

62

57

70

91

86

68

11

291

11

881

15

111

15

847

16

159

19

138

Glucoiberin

Glucocheirolin

Progoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Epiprogoitrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


52

12

58

34

62

76

69

71

85

48

11

306

118

83

15

799

19

255

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

24




min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

61

50

60

56

98

61

03

69

18

84

01

11

169

11

741

14

944 1

569

2 1

601

1

18

963

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


25




min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


49

78

55

68

60

05

14

842

835

0

Glucosibarin

Glucoraphenin

Sinigrin

Progoitrin G

lucocheirolin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


46

57

50

54 56

93

60

99

69

01

83

67

11

161

11

743 14

941

15

688

16

006

18

908

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

-50 0

50 100 150 200 250 300 350


46

43

50

21

56

67

60

04

68

75

83

47

11

137

11

724

14

931

15

684

16

005

18

962

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

26




extract

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


47

63

51

69 5

823

6

238

70

65

86

68

11

293

11

873

15

081 1

583

2 1

614

1

19

126

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150

200 250 300 350


55

88

84

78

14

918

Progoitrin

Glucoraphenin

Glucosibarin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


27





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

59

50

50

56

94 6

094

6

901

83

64

11

141

11

736 1

494

0 1

570

8 1

601

0 18

967

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


62

09

19

122

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

04

49

88

56

02

59

96

67

77

82

20

11

064

11

629

14

850 1

561

1 1

594

1

18

920

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin Sinigrin

Gluconasturtiin

28




min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

69

51

46 5

819

6

006

70

49

85

26

11

128

11

722

15

166

15

899

16

228 1

919

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

72

58

21

59

99

70

50

11

138

11

723

15

177

16

204 1

919

2


Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

53

58

18

61

76

70

27

11

212

11

833

15

119

16

145

19

093


Sinigrin

Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

29





standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150

175


47

47

51

30

58

12

617

4

702

3

84

67

11

208

11

834

15

077

158

18 1

614

0

19

084

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


57

50 60

92

69

29

79

53

11

809

Progoitrin

Epiprogoitrin

Glucoraphenin

Gluconapin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

33

50

10

56

77 60

70

68

57

79

24

11

128

11

732

14

924 1

567

7 1

599

2

18

929

Glucoiberin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Glucocheirolin

30




min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

21











standards spiked

Progoitrin G

lucocheirolin

Glucoiberin

Glucoerucin

Glucosibarin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


47

60

51

78

58

40

62

42

70

83

86

16

11

271

11

861

15

077

15

834

16

103

19

110

Glucoiberin

Glucocheirolin


Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Progoitrin

Glucoraphenin

min0 2 4 6 8 10 12 14 16 18

mAU

0

50

100

150

200

250

300


46

75

51

48

61

86

87

09

15

122

16

142

22





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

11

49

71

56

31

60

64

68

38

82

40

11

051

11

718

14

935

15

656

15

991 1

890

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


82

67

14

950

18

873

Glucoraphenin

Glucosibarin

Gluconasturtiin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


58

53

87

29

16

168

19

153

Progoitrin

Glucoraphenin

Glucoerucin

Gluconasturtiin

23




standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

78

51

58

58

18

62

57

70

91

86

68

11

291

11

881

15

111

15

847

16

159

19

138

Glucoiberin

Glucocheirolin

Progoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Epiprogoitrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


52

12

58

34

62

76

69

71

85

48

11

306

118

83

15

799

19

255

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

24




min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

61

50

60

56

98

61

03

69

18

84

01

11

169

11

741

14

944 1

569

2 1

601

1

18

963

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


25




min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


49

78

55

68

60

05

14

842

835

0

Glucosibarin

Glucoraphenin

Sinigrin

Progoitrin G

lucocheirolin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


46

57

50

54 56

93

60

99

69

01

83

67

11

161

11

743 14

941

15

688

16

006

18

908

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

-50 0

50 100 150 200 250 300 350


46

43

50

21

56

67

60

04

68

75

83

47

11

137

11

724

14

931

15

684

16

005

18

962

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

26




extract

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


47

63

51

69 5

823

6

238

70

65

86

68

11

293

11

873

15

081 1

583

2 1

614

1

19

126

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150

200 250 300 350


55

88

84

78

14

918

Progoitrin

Glucoraphenin

Glucosibarin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


27





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

59

50

50

56

94 6

094

6

901

83

64

11

141

11

736 1

494

0 1

570

8 1

601

0 18

967

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


62

09

19

122

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

04

49

88

56

02

59

96

67

77

82

20

11

064

11

629

14

850 1

561

1 1

594

1

18

920

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin Sinigrin

Gluconasturtiin

28




min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

69

51

46 5

819

6

006

70

49

85

26

11

128

11

722

15

166

15

899

16

228 1

919

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

72

58

21

59

99

70

50

11

138

11

723

15

177

16

204 1

919

2


Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

53

58

18

61

76

70

27

11

212

11

833

15

119

16

145

19

093


Sinigrin

Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

29





standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150

175


47

47

51

30

58

12

617

4

702

3

84

67

11

208

11

834

15

077

158

18 1

614

0

19

084

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


57

50 60

92

69

29

79

53

11

809

Progoitrin

Epiprogoitrin

Glucoraphenin

Gluconapin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

33

50

10

56

77 60

70

68

57

79

24

11

128

11

732

14

924 1

567

7 1

599

2

18

929

Glucoiberin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Glucocheirolin

30




min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

22





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

11

49

71

56

31

60

64

68

38

82

40

11

051

11

718

14

935

15

656

15

991 1

890

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


82

67

14

950

18

873

Glucoraphenin

Glucosibarin

Gluconasturtiin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


58

53

87

29

16

168

19

153

Progoitrin

Glucoraphenin

Glucoerucin

Gluconasturtiin

23




standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

78

51

58

58

18

62

57

70

91

86

68

11

291

11

881

15

111

15

847

16

159

19

138

Glucoiberin

Glucocheirolin

Progoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Epiprogoitrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


52

12

58

34

62

76

69

71

85

48

11

306

118

83

15

799

19

255

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

24




min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

61

50

60

56

98

61

03

69

18

84

01

11

169

11

741

14

944 1

569

2 1

601

1

18

963

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


25




min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


49

78

55

68

60

05

14

842

835

0

Glucosibarin

Glucoraphenin

Sinigrin

Progoitrin G

lucocheirolin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


46

57

50

54 56

93

60

99

69

01

83

67

11

161

11

743 14

941

15

688

16

006

18

908

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

-50 0

50 100 150 200 250 300 350


46

43

50

21

56

67

60

04

68

75

83

47

11

137

11

724

14

931

15

684

16

005

18

962

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

26




extract

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


47

63

51

69 5

823

6

238

70

65

86

68

11

293

11

873

15

081 1

583

2 1

614

1

19

126

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150

200 250 300 350


55

88

84

78

14

918

Progoitrin

Glucoraphenin

Glucosibarin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


27





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

59

50

50

56

94 6

094

6

901

83

64

11

141

11

736 1

494

0 1

570

8 1

601

0 18

967

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


62

09

19

122

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

04

49

88

56

02

59

96

67

77

82

20

11

064

11

629

14

850 1

561

1 1

594

1

18

920

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin Sinigrin

Gluconasturtiin

28




min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

69

51

46 5

819

6

006

70

49

85

26

11

128

11

722

15

166

15

899

16

228 1

919

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

72

58

21

59

99

70

50

11

138

11

723

15

177

16

204 1

919

2


Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

53

58

18

61

76

70

27

11

212

11

833

15

119

16

145

19

093


Sinigrin

Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

29





standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150

175


47

47

51

30

58

12

617

4

702

3

84

67

11

208

11

834

15

077

158

18 1

614

0

19

084

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


57

50 60

92

69

29

79

53

11

809

Progoitrin

Epiprogoitrin

Glucoraphenin

Gluconapin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

33

50

10

56

77 60

70

68

57

79

24

11

128

11

732

14

924 1

567

7 1

599

2

18

929

Glucoiberin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Glucocheirolin

30




min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

23




standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

96

51

68

58

30

62

72

69

72

85

57

11

290 1

188

3

15

137

15

881

16

210

19

182

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

78

51

58

58

18

62

57

70

91

86

68

11

291

11

881

15

111

15

847

16

159

19

138

Glucoiberin

Glucocheirolin

Progoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Epiprogoitrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


52

12

58

34

62

76

69

71

85

48

11

306

118

83

15

799

19

255

Glucocheirolin

Progoitrin

Epiprogoitrin

Sinigrin

Glucoraphenin

Gluconapin

Glucotropaeolin

Gluconasturtiin

Sinalbin

24




min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

61

50

60

56

98

61

03

69

18

84

01

11

169

11

741

14

944 1

569

2 1

601

1

18

963

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


25




min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


49

78

55

68

60

05

14

842

835

0

Glucosibarin

Glucoraphenin

Sinigrin

Progoitrin G

lucocheirolin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


46

57

50

54 56

93

60

99

69

01

83

67

11

161

11

743 14

941

15

688

16

006

18

908

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

-50 0

50 100 150 200 250 300 350


46

43

50

21

56

67

60

04

68

75

83

47

11

137

11

724

14

931

15

684

16

005

18

962

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

26




extract

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


47

63

51

69 5

823

6

238

70

65

86

68

11

293

11

873

15

081 1

583

2 1

614

1

19

126

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150

200 250 300 350


55

88

84

78

14

918

Progoitrin

Glucoraphenin

Glucosibarin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


27





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

59

50

50

56

94 6

094

6

901

83

64

11

141

11

736 1

494

0 1

570

8 1

601

0 18

967

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


62

09

19

122

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

04

49

88

56

02

59

96

67

77

82

20

11

064

11

629

14

850 1

561

1 1

594

1

18

920

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin Sinigrin

Gluconasturtiin

28




min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

69

51

46 5

819

6

006

70

49

85

26

11

128

11

722

15

166

15

899

16

228 1

919

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

72

58

21

59

99

70

50

11

138

11

723

15

177

16

204 1

919

2


Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

53

58

18

61

76

70

27

11

212

11

833

15

119

16

145

19

093


Sinigrin

Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

29





standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150

175


47

47

51

30

58

12

617

4

702

3

84

67

11

208

11

834

15

077

158

18 1

614

0

19

084

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


57

50 60

92

69

29

79

53

11

809

Progoitrin

Epiprogoitrin

Glucoraphenin

Gluconapin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

33

50

10

56

77 60

70

68

57

79

24

11

128

11

732

14

924 1

567

7 1

599

2

18

929

Glucoiberin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Glucocheirolin

30




min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

24




min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

61

50

60

56

98

61

03

69

18

84

01

11

169

11

741

14

944 1

569

2 1

601

1

18

963

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


25




min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


49

78

55

68

60

05

14

842

835

0

Glucosibarin

Glucoraphenin

Sinigrin

Progoitrin G

lucocheirolin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


46

57

50

54 56

93

60

99

69

01

83

67

11

161

11

743 14

941

15

688

16

006

18

908

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

-50 0

50 100 150 200 250 300 350


46

43

50

21

56

67

60

04

68

75

83

47

11

137

11

724

14

931

15

684

16

005

18

962

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

26




extract

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


47

63

51

69 5

823

6

238

70

65

86

68

11

293

11

873

15

081 1

583

2 1

614

1

19

126

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150

200 250 300 350


55

88

84

78

14

918

Progoitrin

Glucoraphenin

Glucosibarin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


27





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

59

50

50

56

94 6

094

6

901

83

64

11

141

11

736 1

494

0 1

570

8 1

601

0 18

967

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


62

09

19

122

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

04

49

88

56

02

59

96

67

77

82

20

11

064

11

629

14

850 1

561

1 1

594

1

18

920

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin Sinigrin

Gluconasturtiin

28




min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

69

51

46 5

819

6

006

70

49

85

26

11

128

11

722

15

166

15

899

16

228 1

919

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

72

58

21

59

99

70

50

11

138

11

723

15

177

16

204 1

919

2


Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

53

58

18

61

76

70

27

11

212

11

833

15

119

16

145

19

093


Sinigrin

Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

29





standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150

175


47

47

51

30

58

12

617

4

702

3

84

67

11

208

11

834

15

077

158

18 1

614

0

19

084

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


57

50 60

92

69

29

79

53

11

809

Progoitrin

Epiprogoitrin

Glucoraphenin

Gluconapin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

33

50

10

56

77 60

70

68

57

79

24

11

128

11

732

14

924 1

567

7 1

599

2

18

929

Glucoiberin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Glucocheirolin

30




min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

25




min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


49

78

55

68

60

05

14

842

835

0

Glucosibarin

Glucoraphenin

Sinigrin

Progoitrin G

lucocheirolin

min0 25 5 75 10 125 15 175 20

mAU

0

100

200

300

400


46

57

50

54 56

93

60

99

69

01

83

67

11

161

11

743 14

941

15

688

16

006

18

908

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 2 4 6 8 10 12 14 16 18

mAU

-50 0

50 100 150 200 250 300 350


46

43

50

21

56

67

60

04

68

75

83

47

11

137

11

724

14

931

15

684

16

005

18

962

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

26




extract

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


47

63

51

69 5

823

6

238

70

65

86

68

11

293

11

873

15

081 1

583

2 1

614

1

19

126

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150

200 250 300 350


55

88

84

78

14

918

Progoitrin

Glucoraphenin

Glucosibarin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


27





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

59

50

50

56

94 6

094

6

901

83

64

11

141

11

736 1

494

0 1

570

8 1

601

0 18

967

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


62

09

19

122

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

04

49

88

56

02

59

96

67

77

82

20

11

064

11

629

14

850 1

561

1 1

594

1

18

920

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin Sinigrin

Gluconasturtiin

28




min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

69

51

46 5

819

6

006

70

49

85

26

11

128

11

722

15

166

15

899

16

228 1

919

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

72

58

21

59

99

70

50

11

138

11

723

15

177

16

204 1

919

2


Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

53

58

18

61

76

70

27

11

212

11

833

15

119

16

145

19

093


Sinigrin

Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

29





standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150

175


47

47

51

30

58

12

617

4

702

3

84

67

11

208

11

834

15

077

158

18 1

614

0

19

084

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


57

50 60

92

69

29

79

53

11

809

Progoitrin

Epiprogoitrin

Glucoraphenin

Gluconapin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

33

50

10

56

77 60

70

68

57

79

24

11

128

11

732

14

924 1

567

7 1

599

2

18

929

Glucoiberin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Glucocheirolin

30




min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

26




extract

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


47

63

51

69 5

823

6

238

70

65

86

68

11

293

11

873

15

081 1

583

2 1

614

1

19

126

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150

200 250 300 350


55

88

84

78

14

918

Progoitrin

Glucoraphenin

Glucosibarin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


27





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

59

50

50

56

94 6

094

6

901

83

64

11

141

11

736 1

494

0 1

570

8 1

601

0 18

967

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


62

09

19

122

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

04

49

88

56

02

59

96

67

77

82

20

11

064

11

629

14

850 1

561

1 1

594

1

18

920

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin Sinigrin

Gluconasturtiin

28




min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

69

51

46 5

819

6

006

70

49

85

26

11

128

11

722

15

166

15

899

16

228 1

919

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

72

58

21

59

99

70

50

11

138

11

723

15

177

16

204 1

919

2


Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

53

58

18

61

76

70

27

11

212

11

833

15

119

16

145

19

093


Sinigrin

Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

29





standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150

175


47

47

51

30

58

12

617

4

702

3

84

67

11

208

11

834

15

077

158

18 1

614

0

19

084

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


57

50 60

92

69

29

79

53

11

809

Progoitrin

Epiprogoitrin

Glucoraphenin

Gluconapin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

33

50

10

56

77 60

70

68

57

79

24

11

128

11

732

14

924 1

567

7 1

599

2

18

929

Glucoiberin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Glucocheirolin

30




min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

27





min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

59

50

50

56

94 6

094

6

901

83

64

11

141

11

736 1

494

0 1

570

8 1

601

0 18

967

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


62

09

19

122

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


46

04

49

88

56

02

59

96

67

77

82

20

11

064

11

629

14

850 1

561

1 1

594

1

18

920

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin Sinigrin

Gluconasturtiin

28




min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

69

51

46 5

819

6

006

70

49

85

26

11

128

11

722

15

166

15

899

16

228 1

919

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

72

58

21

59

99

70

50

11

138

11

723

15

177

16

204 1

919

2


Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

53

58

18

61

76

70

27

11

212

11

833

15

119

16

145

19

093


Sinigrin

Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

29





standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150

175


47

47

51

30

58

12

617

4

702

3

84

67

11

208

11

834

15

077

158

18 1

614

0

19

084

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


57

50 60

92

69

29

79

53

11

809

Progoitrin

Epiprogoitrin

Glucoraphenin

Gluconapin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

33

50

10

56

77 60

70

68

57

79

24

11

128

11

732

14

924 1

567

7 1

599

2

18

929

Glucoiberin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Glucocheirolin

30




min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

28




min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

69

51

46 5

819

6

006

70

49

85

26

11

128

11

722

15

166

15

899

16

228 1

919

4

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50 100 150 200 250 300 350


47

72

58

21

59

99

70

50

11

138

11

723

15

177

16

204 1

919

2


Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

53

58

18

61

76

70

27

11

212

11

833

15

119

16

145

19

093


Sinigrin

Epiprogoitrin

Sinalbin

Gluconapin

Glucosibarin

Glucoerucin

Gluconasturtiin

29





standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150

175


47

47

51

30

58

12

617

4

702

3

84

67

11

208

11

834

15

077

158

18 1

614

0

19

084

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


57

50 60

92

69

29

79

53

11

809

Progoitrin

Epiprogoitrin

Glucoraphenin

Gluconapin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

33

50

10

56

77 60

70

68

57

79

24

11

128

11

732

14

924 1

567

7 1

599

2

18

929

Glucoiberin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Glucocheirolin

30




min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

29





standards spiked

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150

175


47

47

51

30

58

12

617

4

702

3

84

67

11

208

11

834

15

077

158

18 1

614

0

19

084

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


57

50 60

92

69

29

79

53

11

809

Progoitrin

Epiprogoitrin

Glucoraphenin

Gluconapin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50 0

50

100 150 200 250 300 350


46

33

50

10

56

77 60

70

68

57

79

24

11

128

11

732

14

924 1

567

7 1

599

2

18

929

Glucoiberin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

Glucocheirolin

30




min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

30




min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


59

24

63

00

71

80

85

92

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-25

0

25

50

75

100

125

150


46

36

50

16

56

79 6

088

6

871

82

16

11

115

11

735

14

924

15

672

15

990

18

921

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


51

57

620

2

11

110

11

413

16

027

Glucocheirolin

Sinalbin

Gluconapin

Sinigrin

Glucoerucin

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

31




min0 25 5 75 10 125 15 175 20

mAU

-25 0

25 50 75

100 125 150 175


47

67

51

47

58

03

62

09

69

91

84

16

11

234

11

777

15

093

15

854

16

186

19

203

Glucoiberin


Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200


52

45 6

336

11

201

11

875

16

112

Glucocheirolin

Sinalbin

Gluconapin

Glucoerucin

Sinigrin

min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


46

16 4

997

56

57 6

021

68

37

82

50

10

946

11

256

15

177

15

666

15

990

18

917

Glucoiberin

Glucocheirolin

Progoitrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin

Sinigrin

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

32




Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin



















33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

33











500ppm








34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

34


y = 11916x + 28152R2 = 09997

0

1000

2000

3000

4000

5000

6000

7000

0 100 200 300 400 500 600


Are

a



Glucoiberin y = 11758x + 27812 R2 = 09996


Progoitrin y = 13687x + 38843 R2 = 09996

Sinigrin y = 20569x + 63207 R2 = 09996



Sinalbin y = 28268 x+ 39627 R2 = 09997

Gluconapin y = 12954x + 2635 R2 = 09996



Glucoerucin y = 11927x + 57822 R2 = 09994



35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

35




CL=kSBb









Glucoiberin 11758 4933 1259 2517 252

Glucocheirolin 10588 1375 390 779 078

Progoitrin 13687 410 090 180 018

Sinigrin 20569 1952 285 569 057

Epiprogoitrin 11916 1642 413 827 083

Glucoraphenin 9290 1002 539 1079 108

Sinalbin 28268 11380 1208 2415 242

Gluconapin 12954 4272 989 1979 198

Glucosibarin 13585 742 164 328 033


Glucoerucin 11927 862 217 434 043




36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

36




















37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

37









Recoveries RSD

Glucoiberin 51398 53496 53640 59528 8877 1024

Glucocheirolin 49638 50196 51235 53342 9440 661

Progoitrin 70511 71287 71792 69325 10270 527

Sinigrin 102582 101919 103370 104688 9803 593

Epiprogoitrin 54048 56265 55110 60221 9156 905

Glucoraphenin 55686 56322 59052 46658 12221 1460

Sinalbin 126954 125547 128299 136218 9318 1124

Gluconapin 77646 78820 81159 64987 12188 1460

Glucosibarin 57780 60260 60420 69012 8610 1209


Glucoerucin 60414 58620 58489 61448 9630 878



Average 9980 928

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

38







found




= 4650145ppm x 10 x 10-3L 9803

= 47436mg



= 47436mg 5g

=09487mgg

=94870microgg


94870microgg

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

39






Glucoiberin

Glucocheirolin

Progoitrin

Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucosibarin

Glucotropaeolin

Glucoerucin

Gluconasturtiin


















[葶藶子] X 3992 X 43070 X X 6655 1264894 X X 1371 X 1319982



40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

40

4 Discussion

41 Extraction









42 Clean-up process












41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

41







respectively










min0 25 5 75 10 125 15 175 20

mAU

0

50

100

150

200

250

300


Area 557971 49

06

Area 62819

55

60

Area 295705 59

19

Area 134655

66

54

Area 156009 82

66

Area 237494 10

990

Area 558152

11

645 Area 148905

15

480

Area 410646 18

953

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

glucotropaeolin w

ith interferences

Gluconasturtiin

Glucocheirolin

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

42













44 Gradient program






min0 25 5 75 10 125 15 175 20

mAU

-50

0

50

100

150

200

250

300

350


Area 523553

56

24

Area 885345 60

40

Area 107929

66

99

Area 848149 82

32

Area 162052 11

013

Area 459709

11

660 Area 105774

15

483

Area 573498 18

929

Progoitrin Sinigrin

Epiprogoitrin

Glucoraphenin

Sinalbin

Gluconapin

Glucotropaeolin

with

interferences

Gluconasturtiin

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

43




















44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

44



min0 25 5 75 10 125 15 175 20

mAU

0

200

400

600

800


524

5

633

6

11

201

11

875

16

119

min0 25 5 75 10 125 15 175 20

mAU

0

500

1000

1500

2000

2500


51

57

62

02

11

110

11

413

16

027

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

Glucocheirolin

Sinigrin

Sinalbin

Gluconapin

Glucoerucin

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

45

5 Conclusion



















46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

46

6 Future plan











47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

47

5 References




1999 82(5) 1115-1127


5244-5250








306 83-91


Agric Food Chem 2000 48 3572-3575




Chem 2004 378 827-833




48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

48

Food Chem 2002 50 983-988


5244-5250

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

49

APPENDIX



Max 330 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

323580605

969719

4220670

1959942

7499812590409

1800400

275023716200424070218

1389866

799645 228995834211371359759

1190577

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2

S CH3

O


[M-H]-ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2110 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

211 4380303

969576

749871

2590150

1960107

27499521289252 2449655

1389692799536 35809002419766

1190313 29098981450517 2149696590091

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH2 S

O

CH3

O


ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

50




Max 220 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

20

40

60

80

100

120

140

160

180

200

220 969659

749987

1359813

38808351950521

25903502750274

799665 1540002 1920159

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 620 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

45

50

55

6062 749844

969581

799540 35801751160077 195032316198182590135

16497001010191 27501751289360

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

51




Max 170 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

00

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170 749995

969709

3880763

1460472 1950836

1359845

27502412590273

1920093

3010257

3320249799678

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 450 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45 969527

4340301

41900802590061

749846

195046727499211290238

1920068 2409591799536 1389877 2559360

33099173700284

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH

S

O

CH3


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

52




Max 4930 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

493 4240434

2590156

18202882749930

19503142309794

22799602460136 344088524100542009738 29100051689768

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S


Max 1100 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110 749853

969586

37205051950279799513

25901851300285 27500451190339590082 178995213896072920821

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

53



Max 5240 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

524 969588

749859

4080478

1660322

2590134

1950338799525 2749888

2149826

13896831190357850259 2300145 3280864241001916306231689781

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C



Max 800 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80 969551

1359682

4380619

749869

19503172590157

27500033320046

799553 15397111389689 244990930100421450472 35810761289375850281

1989805 21501181699414

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

54




Max 6860 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

686 969574

749855

4220648

25901411800497

1950340799522 274989022899781389682

1190323850259 1650357 2409993 34210331310277 3010072

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-


Max 2270 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

0

20

40

60

80

100

120

140

160

180

200

220

969567

749853

4200475

2590160

799525 17803521950335

27498912269948

13896921190323 1630610 2240099850231 2420073 34009401310417

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C CH2 C

H2CH2

S CH3


[M-H]- ΟΟΗ

OH

OH

CH2OH

S

SO3-

HSO4-

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

55



8729min


Max 820 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

75

804340272

969605

25901834190099

749894 1950322

192016324097151689535 27499141290232

255946414504461359696935864 33949552032529 488225939376272975118 427792033174432342286186401315340121134158

[M-H]-

ΟΟΗ

OH

OH

CH2OH

SHSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3



Max 560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

554340255

265145418301241529173 41899282931837 3251753

2552330 2708373 311166821690011369300

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


[M-H]-

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

56



5834min


Max 20870 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

100

200

300

400

500

600

700

800

900

1000

1100

1200

1300

1400

1500

1600

1700

1800

1900

20002087 3880120

2641433

2812313

2832469 34108695031292

3890183 54913442792185

32509693689610 53909684559921 593155543110853061862 37705922971118 3393029 4719644

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-


Max 470 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

5

10

15

20

25

30

35

40

4547 749863

969559

1359633

19502701460211

38800602589906

1389644799524

11903092749692

15397261289249 1919868 3009982

574090 1610347

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(R)CH

CH2

H

OH


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

57



extract at 6971min


Max 28900 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600 620 640 660 680 700mz amu

0

200

400

600

800

1000

1200

1400

1600

1800

2000

2200

2400

2600

2800

3880270

2641544 3900253

267223445601043411002 54915843689640 50315134459772 5631551


Max 9180 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

550

600

650

700

750

800

850

9002641433

2812339

2832468

2672170

50312703580053

2792168

34108842702344

2842501 368954338801062989842 35901272931611 3673363 54913744129449 4390693 50513023813464 59315404650872

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


[M-H]-

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

58



extract at 11290min


Max 2070 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150

160

170

180

190

200207 3880269

749891

969602

1359680

1950321

2590038

1460223

2749864

13896331190328799575 1919884 2100027 332005730807661289266 2409916850345 1649880 2259676590129 1989876

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

(S)CH

CH2

OH

H


ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-


Max 5340 counts

260 280 300 320 340 360 380 400 420 440 460 480 500 520 540 560 580 600mz amu

0

50

100

150

200

250

300

350

400

450

500

534 2641495

2812373

2832526

2672217

42401502792205

27024002842546 3689629

34109872771950 3061949 3622543 4129511 4260113 55010184514223 4873240 5640580

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


[M-H]-

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

59


11290min



Max 760 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65

70

752651545

3580384

293185518301223091856

3532094

32519393972451

33721322812549

2552342 44127003812446321227924121681840176 2971492


Max 90 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440mz amu

00

05

10

15

20

25

30

35

40

45

50

55

60

65

70

75

80

85

90 969522

4240178

1820260749881

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C OH


HSO4-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

60




Max 390 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

2

4

6

8

10

12

14

16

18

20

22

24

26

28

30

32

34

36

382051455

2501725

22016774340624

2331742

2230486

2651727

14807002552594

1780723

3252181

31119371830295

1269162 20614802812803

2931983194956418694122412383 2590410


Max 650 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

5

10

15

20

25

30

35

40

45

50

55

60

65 749926

969621

959562

35803941950349

275000416199161160176799581259027413898041349781638038 2629085 3768085 447526431499301774611 43240782111616 4601431

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

HCH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH

S

O

CH3


61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

61


extract at 11875min


11875min


Max 1560 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

10

20

30

40

50

60

70

80

90

100

110

120

130

140

150156 749918

969639

37205981300357 2590226

241012819504102750018799621


Max 4710 counts

60 80 100 120 140 160 180 200 220 240 260 280 300 320 340 360 380 400 420 440 460 480 500mz amu

0

50

100

150

200

250

300

350

400

450471 3720464

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2CH

CH2


[M-H]-

Ο

(S)(S)

(R) (S)

ΟΗ

OH

OH

CH2OH

S C

NOSO3H

H2C C

H2

CH

CH2


[M-H]-

ΟΟΗ

OH

OH

CH2OH

S

HSO4-

SO3-

62

62

Documents

Develop a Sample Preparation Procedure for HPLC Analysis ...libproject.hkbu.edu.hk/trsimage/hp/02010364.pdf · Develop a Sample Preparation Procedure for HPLC Analysis of Glucosinolates