Research Article Phytochemical Contents and Antioxidant ...downloads.hindawi.com/journals/bmri/2016/8495630.pdf · T : Description and moisture contents of six sesame seeds varieties

Research ArticlePhytochemical Contents and Antioxidant and AntiproliferativeActivities of Selected Black and White Sesame Seeds

Lin Zhou12 Xiaohui Lin2 Arshad Mehmood Abbasi23 and Bisheng Zheng2

1Guangdong Province Key Laboratory for Biotechnology Drug Candidates School of Biosciences and BiopharmaceuticsGuangdong Pharmaceutical University Guangzhou 510006 China2School of Light Industry and Food Sciences South China University of Technology Guangzhou 510641 China3Department of Environmental Sciences COMSATS Institute of Information Technology Abbottabad 22060 Pakistan

Correspondence should be addressed to Lin Zhou zhoulingdpueducn

Received 15 April 2016 Revised 7 June 2016 Accepted 30 June 2016

Academic Editor Michael D Coleman

Copyright copy 2016 Lin Zhou et alThis is an open access article distributed under the Creative CommonsAttribution License whichpermits unrestricted use distribution and reproduction in any medium provided the original work is properly cited

Sesame (Sesamum indicum L) seeds are popular nutritional food but with limited knowledge about their antioxidant andantiproliferative activities of various varieties Phytochemical profiles and antioxidant and antiproliferative activities of six varietiesof sesame (Sesamum indicum L) seeds were studied Fenheizhi3 (black) cultivar exhibited the maximum contents of total phenolicsand lignans and values of total oxygen radical absorbance capacity (ORAC) and antiproliferative activity (EC

50) against HepG2

cells Bound ORAC values showed strong associations with bound phenolics contents (119903 = 0976 119901 lt 001) in bound phenolicextracts EC

50values showed strong negative associations with phenolic contents (119903 = minus0869 119901 lt 005) and ORAC values

(119903 = minus0918 119901 lt 001) Moreover the contents of free phenolics were higher than that of the bound phenolics and the threeblack sesame seeds generally depicted higher total phenolics compared to the three white varietiesThe antioxidant (ORAC values)and antiproliferation activities of six sesame seeds were both associated with contents of bound phenolics (119903 gt 08 119901 lt 005)Interestingly nonlignan components in bound phenolics contributed to the antioxidant and antiproliferative activities This studysuggested that Fenheizhi3 variety is superior to the other five varieties as antioxidant supplements

1 Introduction

Sesame ldquoSesamum indicum Lrdquo is a commonly growing plantspecies mainly in tropical and subtropical regions of theworld particularly in Burma India China and Sudan [1]Sesame seeds are preferably used along with bread biscuitscrackers and so forth and as seasoning in food around theworld [2] Sesame seeds are important sources of oil proteincarbohydrates and minerals for human nutrition [3]

Sesame seeds color varied from cream-white to charcoal-black whereas white and black are the typical skin colorBlack sesame seed is superior to white one as food forhealth in traditional beliefs of Asian countries and is includedin Pharmacopoeia of the Peoplersquos Republic of China (PPRC2015) as liver and kidney benefiting traditional Chinesemedicine (TCM) It has been reported that the seed colors ofsesame affect the phytochemical contents and their biologicalactivities [4 5] Phytochemical compounds in sesame seed

such as sesamin sesamol and anthrasesamone F have beenproved to have in vitroin vivo antioxidant and antiagingactivity [6ndash9] Moreover sesamin and sesamolin showedanti-inflammatory antihypertensive and anticarcinogeniceffects in numerous studies [10ndash12]

The common nutritional evaluations of sesame seeds arebased on the contents of proteins oils and lignans How-ever little work was conducted on the correlations betweenmulticomponents and antioxidant activity and multicompo-nents and antiproliferative activity in various sesame seedsvarieties Therefore the phytochemical profiles includingthe contents of phenolics flavonoids and lignans in vitroantioxidant activities and antiproliferative activities againstHepG2 cells of six varieties of sesame seeds in China werestudied in the present work Particularly the six sesame seeds(three of black and three of white skin color nonscaledplanting) are newly bred varieties in midwest of China forimproved nutrition values The knowledge of the antioxidant

Hindawi Publishing CorporationBioMed Research InternationalVolume 2016 Article ID 8495630 9 pageshttpdxdoiorg10115520168495630

2 BioMed Research International

and antiproliferative activities of various sesame seeds willbenefit the sesame seed planters relevant manufacturers andordinary consumers

2 Materials and Methods

21 Chemicals and Materials Methanol (MeOH) ethanol(EtOH) n-hexane ethyl acetate hydrochloric acid (HCl)and acetic acid (HAC) were purchased from GuanghuaSci-Tech Co Ltd (Guangdong China) Potassium chloride(KCl) sodium acetate (NaAC) sodium carbonate (NaCO

3)

sodiumhydroxide (NaOH) potassiumphosphatemonobasic(KH2PO4) and potassiumphosphate dibasic (K

2HPO4) were

of analytical grade and were purchased from Sangon BiotechCo Ltd (Shanghai China) 221015840-Azobis(2-amidinopropane)dihydrochloride (ABAP) 2101584071015840-dichlorofluorescin diacetate(DCFH-DA) fluorescein disodium salt catechin hydrate(HPLC ge98) Folin-Ciocalteu reagent ascorbic acid6-hydroxy-2578-tetramethylchroman-2-carboxylic acid(Trolox) formic acid (chromatographic grade) andmethanol(chromatographic grade) were purchased from Sigma-Aldrich Inc (St LouisMOUSA) Trifluoroacetic acid (TFAanalytical grade) and aluminum chloride (AlCl

3sdot6H2O

analytical grade) were purchased from Fisher Scientific(Fair Lawn NJ USA) Sodium borohydride (NaBH

4 reagent

grade) chloranil (analytical grade) vanillin (analytical grade99) and gallic acid (analytical grade 99) were purchasedfrom Aladdin Inc (Shanghai China) Sesamol sesamolinand sesamin (HPLC ge98) were purchased from ChengduPufei De Biotech Co Ltd (Sichuan China) HepG2 humanliver cancer cells were purchased from American TypeCulture Collection (Rockville MD USA) WME mediumfoetal bovine serum (FBS) insulin and other cell culturereagents were purchased from Gibco Life TechnologiesCo (Grand Island NY USA) Six sesame seed varieties(three black and three white) were kindly donated by ShanxiAcademy of Agricultural Sciences (Taiyuan city China)which were harvested in 2014 and stored in desiccator atroom temperature until analysis

22 Moisture Content The moisture content was analyzedby oven-dry method [13] Briefly 1 g of sample was driedin electric oven at 105∘C to a constant weight and moisturecontent was expressed as percent of dry weight (DW) intriplicate

23 Phenolics Extraction and Determination

231 Free Phenolics Extraction Free phenolic compoundswere extracted using the modified method reported pre-viously [14] Briefly 1 g of sesame seed was blended with5mL of hexane for 1min After centrifugation at 2700timesgfor 3min residue was defatted two more times Add 10mLof 80 chilled acetone to the residue and blend 1min Thesupernatant was collected after being centrifuged at 2700timesgfor 3min repeated twice The supernatant was pooled andevaporated to dryness at 45∘C under vacuum The solution

was reconstituted in 10mLof 70methanol and free phenolicextracts (100mg mLminus1) were stored at minus40∘C until analysis

232 Bound Phenolics Extraction Bound phenolics wereextracted using themethod described by Chen et al [14] withmodifications Briefly the residues obtained after extractionof free phenolics were flushed with nitrogen gas for 2minsealed and digested with 20mL of 4MNaOH at roomtemperature for 1 h The mixture was neutralized with 10Mconcentrated HCl Then 20mL hexane was added to extractresidual lipids in the mixture for 10min (not necessaryif no residual lipids) After centrifuging at 2700timesg for5min remaining residues were extracted five times withethyl acetate The ethyl acetate fractions were pooled andevaporated to dryness at 45∘C under vacuum The boundphenolics were reconstituted to 10mL of 70 methanol andbound phenolic extracts (100mgmLminus1) were stored at minus40∘Cuntil analysis

233 Determination of Total Phenolic Contents Total pheno-lic contents of each sample were determined using the mod-ified Folin-Ciocalteu colorimetric method [14 15] Brieflyfree phenolic and bound phenolic extracts were diluted 10ndash20 times with distilled water and were reacted with Folin-Ciocalteu reagent and then neutralized with Na

2CO3 After

90min incubation the absorbance of the resulting solutionwas recorded at 760 nm using MRX II Dynex plate reader(Dynex Technologies Inc Chantilly VA USA) Total pheno-lic contents were expressed as grams of gallic acid equivalentsper kg of sample on DW (gGAE kgminus1)

24 Determination of Total Flavonoid Contents Totalflavonoid contents were determined by sodium borohydride-chloranil (SBC) protocol [16] Briefly 1mL of free and 1mLof bound phenolic extracts were added to test tubes (15 times150mm) andwere dried under nitrogen gasThe residues andcatechin hydrate standards (01ndash100mM) were prepared in1mL of THFEtOH (1 1 vv) Extracts or standard solutionwasmixed with 05mL of 50mMNaBH

4solution and 05mL

of 746mMAlCl3solution After 30min shaking in an orbital

shaker at room temperature 01mL of 500mM NaBH4

solution was added with continued shaking for 30min atroom temperature 04mL of cold 08M acetic acid solutionwas added and the mixtures were kept in the dark for 15minafter thorough mixing 02mL of 200mM chloranil wasadded and the mixture was heated at 95∘C with shaking for60min Then the reaction solutions were cooled using tapwater and were brought to 1mL using methanol Afterward02mL of 16 (wv) vanillin was added and mixed followedby addition of 04mL of 12MHCl and the reaction solutionswere kept in the dark for 15min after thorough mixing Atlast the absorbance was recorded at 490 nm using MRXmicroplate reader with Revelation workstation (DynexTechnologies Inc) Total flavonoid contents were expressedas grams of catechin equivalents per kg of sample on DW(gCE kgminus1)

BioMed Research International 3

Table 1 Description and moisture contents of six sesame seeds varieties

Varieties Abbreviation Harvest year Color Origin Moisture content ()Zhenzhou1 B1 2014 Black Zhenzhou Henan 163 plusmn 013

Fenheizhi3 B2 2014 Black Agriculture academy of Shanxi 514 plusmn 009

05H27 B3 2014 Black Shanxi Academy of Agricultural Sciences 522 plusmn 001

Jizhi157 W1 2014 White Hebei 450 plusmn 014

Fenzhi2 W2 2014 White Shanxi 463 plusmn 013

Jizhi1 W3 2014 White Shanxi Academy of Agricultural Sciences 463 plusmn 003

Data were reported as mean plusmn SD

25 Estimation of Lignan Contents Both the free and boundphenolic extracts (100mgmLminus1) were used for determina-tion of lignan contents The chromatographic analysis wasperformed to estimate lignan contents in sesame seeds asdescribed by Reshma et al [17] with some modificationsBriefly three representative lignans (sesamol sesamin andsesamolin) were analyzed for the contents using WatersHPLC system (Waters Corp Milford MA) with a WatersC18

column (5 120583m 250mm times 46mm) The mobile phaseconsisted of methanolwater (75 25 vv) with 01 formicacid inwater at a flow rate of 1mLminminus1TheUVdetectorwasset at 290 nmwith column temperature of 30∘C and injectionvolume of 10 120583L Peak was identified by calibration standardsof sesamol sesamin and sesamolin with a retention time of34 83 and 102min respectively The concentration rangeof standards of sesamol sesamin and sesamolin was from 2to 200120583gmLminus1 with recovery at 9950 plusmn 100 9947 plusmn 102and 9962 plusmn 113 respectively The results were expressed asmilligrams per kg on DW (mg kgminus1)

26 Determination of Total Antioxidant Activity

261 Oxygen Radical Absorbance Capacity (ORAC) ORACvalues were determined using a Fluoroskan Ascent fluores-cent spectrophotometer (Molecular Devices Sunnyvale CA)[18 19] Briefly 20120583L extracts (100mgmLminus1) or Trolox wasmixed with 200120583L of fluorescein and incubated at 37∘Cfor 20min followed by adding 20120583L of freshly prepared1194mM ABAP into black walled 96-well plates Fluores-cence intensity was recorded automatically at 37∘C excitationof 485 nm and emission of 535 nm for 35 cycles every 5minFinal values ofORACwere expressed asmicromoles of Troloxequivalents (TE) per gram of sesame seeds on DW basis(120583mol TE gminus1DW)

262 Peroxyl Radical Scavenging Capacity (PSC) PSC valueswere determined by the method described by Adom andLiu [20] Briefly 100 120583L extracts (100mgmLminus1) or standardssolutions (freshly prepared ascorbic acid solutions) wereadded in each well of the 96-well plate Then 100 120583L ofDCFH dye (1326 120583M) hydrolyzing with 1mM KOH justbefore being used in the reaction was also added Afteradding 50120583L of peroxyl radicals producer (ABAP 40mM)the mixtures were kept at 37∘C for 40min Fluorescent wasdynamically recorded at excitation of 485 nm and emissionof 538 nm using a multimode microplate reader (FilterMax

F5 Molecular Devices USA) The results were calculatedas micromoles of vitamin C equivalents (VCE) per gram ofsesame seeds on DW basis (120583molVCE gminus1DW)

27 Determination of Antiproliferative Activity The antipro-liferative effects of sesame seeds extracts were assessed bymodified methylene blue assay [21] Briefly HepG2 humanliver cancer cell was grown in WME medium supplementedwith 5 FBS 10mM N-2-hydroxyethylpiperazine-N1015840-2-ethanesulfonic acid (HEPES) 2mMLminus1 glutamine 5 gmLminus1insulin 005 gmLminus1 hydrocortisone 50 unitsmLminus1 penicillin50 gmLminus1 streptomycin and 100 gmLminus1 gentamycin at 37∘Cand 5 CO

2as described by Liu et al [22] HepG2 cells

were cultured at a density of 25 times 104cellswell in a 96-well microplate with 100120583L of growth medium After 4 h ofincubation at 37∘C growth medium was displaced by 100120583Lof fresh medium containing 10ndash150mgmLminus1 of sesame seedsextractsThe wells containing fresh medium without extractswere set as the control After 72 h of incubation the number ofthe viable cells was counted by the methylene blue assay Theantiproliferative effects were assessed by the EC

50values and

expressed asmilligrams of sesame seeds extracts permilliliter

28 Statistical Analysis Statistical analysis was performedusing SPSS software version 190 (LEAD Technologies IncChicago IL) Variation of means was analyzed by ANOVAand Duncanrsquos test Significance of correlations was calcu-lated by the Pearson coefficient Dose-Effect analysis wasperformed using CalcuSyn software version 20 (BiosoftCambridge UK) Statistical significance was set at 119901 lt005 All data were expressed as the mean plusmn SD for threereplications

3 Results and Discussion

31 Moisture Content Moisture content and description ofsix varieties of sesame seeds are given in Table 1 ExceptB1 variety the average moisture content of the other fivesesame seed varieties ranged from 436 to 523 which wasconsistent with the published data [3 13]The possible reasonfor the lower moisture content in B1 variety was the moistureloss during storage before transportation


BCD

H

DEF

BCD

H

DEFCDE

H

EFG

B

GH

DEF

FG

A

B

BC

H

Free phenolicsBound phenolicsTotal phenolics

DE

B2 B3 W1 W2 W3B1Varieties

0

1

2

3

4

5

6

7

8

9Ph

enol

ic co

nten

t (gG

AE

kgminus1

DW

)

(a)

BC

EE

AB

FGHFGHFGH

FGH FG

CD

GH

AA

EFEF

HI

D

I

Free flavonoidsBound flavonoidsTotal flavonoids


0

1

2

3

4

5

6

7

8

9

Flav

onoi

d co

nten

ts (g

CEkgminus1

DW

)

(b)

Figure 1 Free bound and total phenolic and flavonoid contents in six varieties of sesame seeds (B1 B2 B3 W1 W2 andW3) Bars with noletters in common are significant difference at 119901 lt 005 (a) Phenolic contents (b) Flavonoid contents

32 Phytochemical Composition of Selected Sesame Seeds

321 Contents of Phenolics and Flavonoids Measured levelsof total phenolic and total flavonoid contents of six sesameseed varieties were presented in Figures 1(a) and 1(b) FromFigure 1(a) free phenolic contents were higher than that ofthe bound phenolic Black sesame seed variety B2 exhib-ited maximum free and bound phenolics (499 plusmn 047 and233 plusmn 036 gGAE kgminus1 resp) while minimum amount wasdetermined in free phenolics of W1 (258 plusmn 007 g GAE kgminus1)and bound phenolics of W2 (080 plusmn 011 gGAE kgminus1) In B2cultivar free phenolics were 6819 and bound phenolicswere 3181 of total phenolic contents Total phenolic con-tents in black sesame varieties (B1 B2 and B3) ranged from454 to 732 gGAE kgminus1 with maximum value in B2 whereasamong white sesame seed W3 exhibited the highest totalphenolic contents of 404 plusmn 013 gGAE kgminus1 followed by W2(400 plusmn 018 gGAE kgminus1) and W1 (356 plusmn 008 gGAE kgminus1)variety On the whole the black sesame varieties containmore phenolics compared to the white varieties Howeverno significant differences between free bound and totalphenolics were observed between black and white sesamevarieties Nadeem et al [23] reported the total phenolicsextracted from sesame cake were 172 gGAE kgminus1 Shahidiet al [4] reported that the total phenolic contents of twosesame seed cultivars (black and white) were 299 plusmn 06 and106 plusmn 16 g catechin equivalents kgminus1 crude ethanolic extractrespectively These studies were not comparable with ourdata for different samples (sesame cake) or reference control(catechin) used in the determination

From Figure 1(b) the contents of free flavonoid rangedfrom 288 (B3) to 461 (W3) gCE kgminus1 and the boundflavonoids were between 224 (W2) and 352 (B2) g CE kgminus1while total flavonoid contents ranged from 580 (B3) to 804

SesamolinSesamin

Sesamol

2 4 6 8 10 12 14 160t (min)

000

005

010

015

020

025

030

Figure 2 Elution curve of lignans (sesamol sesamin andsesamolin) of free phenolic extracts of B2 variety (mobile phasemethanolwater 75 25 flow rate 1mLminminus1 wavenumber 290 nmand Waters C

18column)

(W3) gCE kgminus1 W3 variety exhibited the highest level oftotal flavonoids (804 plusmn 026) and free flavonoids (461 plusmn006) g CE kgminus1 while the highest levels of bound flavonoidswere present in B2 (352 plusmn 008 gCE kgminus1) On the whole nosignificant differences of the flavonoid contents (free boundand total flavonoids) were observed between black and whitesesame varieties

322 Contents of Lignans Lignans here including sesamolsesamin and sesamolin in free and bound extracts of sesameseeds were analyzed by HPLC (Figure 2 Table 2) Overallfree phenolic extracts contain more levels of lignans (gt89)


Table 2 Lignans (sesamol sesamin and sesamolin) contents of free and bound phenolics extracts in six sesame seeds varieties

Varieties Sesamol (mg kgminus1) Sesamin (mg kgminus1) Sesamolin (mg kgminus1) Total (mg kgminus1)Free extracts Bound extracts Free extracts Bound extracts Free extracts Bound extracts

B1 7921 plusmn 362 714 plusmn 023 ND ND ND ND 8635B2 18725 plusmn 1056 3054 plusmn 298 3955 plusmn 038 ND 2512 plusmn 095 ND 28246B3 12148 plusmn 328 ND 495 plusmn 020 ND 1030 plusmn 020 ND 13672W1 3458 plusmn 199 ND 1425 plusmn 174 ND 494 plusmn 021 ND 5376W2 715 plusmn 075 ND 3957 plusmn 126 ND 1286 plusmn 076 ND 5957W3 2732 plusmn 346 ND ND ND 197 plusmn 022 ND 2928ND means not detectedData were reported as mean plusmn SDLimit of detection was 006mg kgminus1 for sesamol 083mg kgminus1 for sesamin and 101mg kgminus1 for sesamolin

compared to the bound extracts Sesamol and sesamolincontents were detected in the free extracts of all varietiesThe lignans in free phenolic extracts of white and blacksesame varieties ranged from 2928 to 5376 and 8283 to25191mg kgminus1 respectivelyThe average lignan content of thethree black cultivars (16734mg kgminus1) was three times higherthan that of the white cultivars (4754mg kgminus1) B2 varietyshowed the highest sesamol (18725 plusmn 1056mg kgminus1) in freephenolic extracts followed by B3 (12148 plusmn 328mg kgminus1)and B1 (7921 plusmn 362mg kgminus1) In white varieties sesamol infree phenolic extracts ranged from 715 plusmn 075 (W2) to 3458plusmn 199mg kgminus1 (W1) Likewise B2 variety had the highestsesamolin content (2512 plusmn 095mg kgminus1) in free phenolicextracts followed by B3 (1030 plusmn 020mg kgminus1) and B1 (notdetected) Sesamin was detected in the free extracts of fourvarieties with the highest value in W2 (3957 plusmn 126mg kgminus1)and B2 (3955 plusmn 038mg kgminus1) followed by W1 (1425 plusmn174mg kgminus1) andB3 (495plusmn 020mg kgminus1)However sesaminwas not detected in bound phenolic extracts

It has been reported [24ndash26] that sesamin and sesamolincontents in sesame seeds ranged from 1550 to 4200mg kgminus1and 620 to 3590mg kgminus1 respectively In the present studythe possible reason for lower sesamin and sesamolin contents(lt40mg kgminus1) was the extraction solvents Although 70sim80methanol is common solvent for the extraction of total phe-nolics and flavonoids [14 20] lipid-soluble lignans (sesaminand sesamolin) are partially soluble in 70 methanol basedon visual observation Likewise the relatively higher contentof sesamol compared to sesamin and sesamolin was observedin the free and bound extracts which is possibly relatedto the relatively higher dissolving ability of sesamol in70 methanol Mostly previous studies were designed todetermine the maximal content of lignans in sesame seedsso the extraction organic solvents commonly used wereweak polar ethanol methanol and acetonitrile Here ouraim was to evaluate the lignan levels and their contributionto in vitro antioxidant activity in extracted fractions ofsesame seeds which leads to the relatively lower contents oflignans determined Sesamin and sesamolinwere successfullyseparated in setting chromatographic parameters (Figure 2)but the baseline separation of sesamol was not attained in thepresent work

33 In Vitro Antioxidant Activity Due to the complexcomponents in sesame seeds two in vitro antioxidant activityassays (ORAC and PSC) were used in the present studyThe common principle of determination was to measureradical chain breaking ability of antioxidants by monitoringthe inhibition of peroxyl radical while different oxidant-fluorescein probes were used ABAP-fluorescein in ORACassay and ABAP-DCFH in PSC assay [18 20]

Measured values of ORAC and PSC based on DWare given in Figures 3(a) and 3(b) ORAC values rangedfrom 3485 (B1) to 9548 (W2) 120583mol TE gminus1 in freefractions (Figure 3(a)) while in bound fractions it wasbetween 1497 (W2) and 4074 (B3) 120583mol TE gminus1 Blacksesame variety B2 exhibited the highest total ORAC value(13233 120583mol TE gminus1) whereas B1 showed the lowest totalORAC value (5951 120583mol TE gminus1) White sesame variety W1had the highest total PSC value among all varieties whichwas 4 times higher than the lowest one (W2) followed by B3B1 B2 W3 andW2 (Figure 3(b)) PSC value of free fractionsranged from 255 (W2) to 1227 (B3) 120583molVCE gminus1 whilein bound fractions it ranged from 133 (W3) to 645 (W1)120583molVCE gminus1 The total PSC values ranged from 450 (W2)to 1855 (W1) 120583molVCE gminus1 Ishiyama et al [27] reported thatORAC values of two sesame seed varieties (Japan Kanto No1 and Gomazou) were 6583 and 827mgTE gminus1 respectivelyThese values correspond to 2630 and 3304 120583mol TE gminus1 ofsesame seed which was comparable to the free fractionsof B1 and B3 but was nearly one-fourth to one-third of thecorresponding values of varieties B2 W1 W2 and W3Othman et al [28] recently reported that the average ORACvalues of polar-soluble crude extracts for white and goldsesame seed were 34720 and 21700120583mol TE gminus1 respectivelyThese reported data were nearly 26- and 16-fold higher thantotal ORAC value in B2 (the highest in this study) whichverified the significant antioxidant activity of polar-solubleextracts

Correlation analysis of the in vitro antioxidant activitywith the multicomponents of selected sesame seeds wasshown in Table 3 Bound ORAC values showed strongassociations with contents of bound phenolics (119903 = 0976119901 lt 001) Moreover total ORAC values showed strong asso-ciation with contents of total phenolics (119903 = 0701 119901 lt 001)


Table 3 Correlation of multicomponents (phenolics flavonoids and lignans) with antioxidant (ORAC PSC) and antiproliferation (EC50)

activities of six sesame seed varieties

Free phenolics Bound phenolics Total phenolics Free flavonoids Bound flavonoids Total flavonoids LignansFree PSC values minus0406 NC NC minus0265 NC NC NCBound PSC values NC 0604 NC NC 0294 NC NCTotal PSC values NC NC 0020 NC NC 0130 0542Free ORAC values 0286 NC NC 0634 NC NC NCBound ORAC value NC 0976

lowastlowast NC NC 0417 NC NCTotal ORAC values NC NC 0701lowastlowast NC NC 0412lowast 0533Free EC

50minus0750 NC NC 0465 NC NC NC

Bound EC50

NC minus0869lowast NC NC minus0666 NC NC

lowastlowastmeans significant difference at 119901 lt 001lowastmeans significant difference at 119901 lt 005NC means not computed

BC

J

E

H

B

FG

CD

IJ

DE

IJ

HGH

CD

A

F

H

DE

HI

Free ORACBound ORACTotal ORAC


0

20

40

60

80

100

120

140

160

ORA

C va

lues

(120583m

olTE

gminus1

DW

)

(a)

D

I

E

G

HIH

A

EF

B

A

F

BB

FF

HI

B

C

Free PSCBound PSCTotal PSC


02468

101214161820

PSC

valu

es (120583

mol

VCEgminus

1D

W)

(b)

Figure 3 In vitro antioxidant activities of six sesame seed varieties (B1 B2 B3 W1 W2 andW3) (mean plusmn SD 119899 = 3) Bars with no letters incommon are significant difference at 119901 lt 005 (a) ORAC values (b) PSC value

Moderate associations were observed between total ORACvalues and contents of total flavonoids (119903 = 0412 119901 lt 005)However associations between total lignans and antioxidantactivities (ORAC and PSC values) were not observed FromORAC values of B2 W1 W2 and W3 PSC values of B1 B3W1 and W3 and their corresponding contents of lignans(Figure 3 Table 3) it can be inferred that the relatively higherantioxidant activities of free phenolic extracts compared tobound phenolic extracts were related to the relatively highercontents of lignans in free phenolic extracts Comparableantioxidant activity was observed in PSC values of B2 andW2 variety and ORAC values of B2 and B3 variety infree and bound phenolic extracts (Figure 3 Table 3) evennone or less lignans determined in bound phenolic extractswhich indicated that nonlignan components contribute totheir corresponding antioxidant activities There were nosignificant associations between PSC values (free bound andtotal) with corresponding contents of phenolic and flavonoidThough specific reason is not clear synergistic effects ofvarious phytochemicals may involve complex antioxidant

behavior of these constituents Many similar reports wereobserved in selected components of sesame seed or sesameseed oil such as synergistic antioxidant activity betweenlignans (sesamin sesamol sesamolin) and tocopherols (120572120573 and 120574) in various test system [29 30] Except the weakpolar lignan components thewater soluble extracts of sesameseed raised concern Moazzami et al [31] quantified lignanglucosides in 65 different sesame seed cultivars but no signif-icant difference between black and white seeds was observedOthman et al [28] reported that extracts from white sesameseed had relatively higher antioxidant capacity compared toextracts from gold sesame seeds and ascribed the possiblereason to the differences in the contents of antioxidants Ide etal [32] compared the physiological activities of sesame seedswith different concentration of lignans in rats and found thatsesame seeds rich in lignans irrespective of composition oflignans greatly affect hepatic fatty acid oxidation and serumtriacylglycerol levels However the researchers stated thatthere is a possibility that compounds other than lignans areinvolved in the physiological activities of tested sesame seeds


Table 4 Antiproliferative activities of the free and bound phenolicextracts from six sesame seeds varieties against HepG2 cell

Varieties Antiproliferative capacity to HepG2 EC50(mgmLminus1)

Free phenolic extract Bound phenolic extractB1 6608 plusmn 035

b5549 plusmn 901

b

B2 2104 plusmn 060a

2357 plusmn 088a

B3 6345 plusmn 085b

3006 plusmn 082a

W1 8729 plusmn 357c

7774 plusmn 129c

W2 8212 plusmn 310c

10953 plusmn 123d

W3 12491 plusmn 279d

6718 plusmn 192c

Data were reported as mean plusmn SDValues in the same rows with different letters differ significantly at 119901 lt 005

These reports indicate the complex antioxidant activity ofmulticomponents of sesame seeds

34 Antiproliferation in HepG2 Cancer Cell Cell culturemodels provide efficient approach that imitates the uptakedistribution and metabolism of antioxidant compounds invivo [33] and human liver cancer HepG2 cell model is widelyused in the antioxidant antiproliferative activity of variouscompounds [14 34 35] The antiproliferative effect of freeand bound phenolic extracts against HepG2 cells was presentin Table 4 The EC

50of free phenolic extracts ranged from

2104 plusmn 060 to 12491 plusmn 279mgmLminus1 for HepG2 cells whilecorresponding values for bound phenolic extracts rangedfrom 2357 plusmn 088 to 10953 plusmn 123mgmLminus1 Lower values ofEC50indicate a higher antiproliferative activity Both free and

bound phenolic extracts of B2 variety showed antiprolifera-tive activityThis result is consistent with the relatively highercontents of phenolics flavonoids and lignans and ORACvalues of B2 variety Interestingly the black sesame seedvarieties showed higher antiproliferative activity comparedto the white varieties in both free phenolic and bound phe-nolic extracts Moreover in bound phenolic extracts highlynegative significant correlations were observed between EC

50

and bound phenolics (119903 = minus0869 119901 lt 005) and EC50

andbound ORAC values (119903 = minus0918 119901 lt 001) in freephenolic extracts highly negative significant correlationswere observed between EC

50and sesamol contents (119903 =

minus0858 119901 lt 005) (data not shown)Sesamin and sesamol have been reported to have anti-

cancer activities in many cancer cell lines including HepG2Sesamin of 25ndash125120583M could induce HepG2 cell deathinvolving apoptosis [36] Content of sesamin in free phe-nolic extract of B2 variety was nearly 3955mg kgminus1DWwhich is roughly equivalent to 11 120583M sesamin As forsesamol at dosage of 50120583M apoptosis against HepG2 cellcould be observed by Liu et al [37] while the content ofsesamol in free phenolic extracts of B2 variety was nearly18725mg kgminus1DW which is roughly equivalent to 135 120583M ofsesamol These results were consistent with the significantcorrelation observed between antiproliferation activity andcontent of sesamol in free phenolic extracts Additionallyin bound phenolic extracts of B1 B2 W1 and W2 variety

contents of lignans (sesamol sesamin and sesamolin) werefar less compared to their free phenolic extracts (Table 2)However the corresponding antiproliferative activities in freephenolic extracts and boundphenolic extractswere compara-ble (Table 4)Thus it can be inferred that the antiproliferativeactivities of sesame seed were not only related to the contentsof phenolics and lignans (sesamol) of free phenolic extractsbut also related to nonlignan components in bound phenolicextracts which need further investigation

4 Conclusions

Sesame seed variety of Fenheizhi3 (black B2) could bemore valuable than the others (Zhenzhou1 05H27 Jizhi157Fenzhi2 and Jizhi1) based on the ex vivo antioxidant andantiproliferative activities In selected sesame seeds free phe-nolic contents were higher than the bound phenolic contentsblack sesame seeds generally depicted higher contents oftotal phenolic compared to the white varieties Moreover theantioxidant activities (total ORAC values) were associatedwith the contents of total phenolics and flavonoids (119903 gt 04119901 lt 005) the antiproliferation activities (bound EC

50) were

associated with contents of bound phenolics and sesamol(119903 gt 08 119901 lt 005) Additionally nonlignan componentsin bound phenolic extracts contributed to the correspondingantioxidant and antiproliferative activities

Competing Interests

All authors declared that they have no conflict of interests

Authorsrsquo Contributions

Lin Zhou designed and performed the experiments Xiao-hui Lin participated in the experiments Arshad MehmoodAbbasi and Bisheng Zhen analyzed data and providedreagents materials and analytical tools Lin Zhou providedfinancial support

Acknowledgments

The authors are grateful for the financial support fromthe Fundamental Research Funds for the Central Univer-sities (2015 D2155140) and Natural Science Foundation ofGuangdong Province (44555019) Professor Rui hai Liu fromDepartment of Food Science Cornel University providedtechnical support on the antioxidant determination

References

[1] H A Abou-Gharbia A A Y Shehata and F Shahidi ldquoEffect ofprocessing on oxidative stability and lipid classes of sesame oilrdquoFood Research International vol 33 no 5 pp 331ndash340 2000

[2] M Namiki ldquoNutraceutical functions of sesame a reviewrdquoCritical Reviews in Food Science and Nutrition vol 47 no 7 pp651ndash673 2007

[3] M Elleuch S Besbes O Roiseux C Blecker and H AttialdquoQuality characteristics of sesame seeds and by-productsrdquo FoodChemistry vol 103 no 2 pp 641ndash650 2007


[4] F Shahidi C M Liyana-Pathirana and D SWall ldquoAntioxidantactivity of white and black sesame seeds and their hull frac-tionsrdquo Food Chemistry vol 99 no 3 pp 478ndash483 2006

[5] H Zhang H Miao L Wei C Li R Zhao and C WangldquoGenetic analysis andQTLmapping of seed coat color in sesame(Sesamum indicum L)rdquo PLoS ONE vol 8 no 5 Article IDe63898 2013

[6] Y Zuo C Peng Y Liang et al ldquoSesamin extends the meanlifespan of fruit fliesrdquo Biogerontology vol 14 no 2 pp 107ndash1192013

[7] M Tada Y Ono M Nakai et al ldquoEvaluation of antioxidativeeffects of sesamin on the in vivo hepatic reducing abilities by aradiofrequency ESR methodrdquo Analytical Sciences vol 29 no 1pp 89ndash94 2013

[8] M Srisayam N Weerapreeyakul S Barusrux and KKanokmedhakul ldquoAntioxidant antimelanogenic and skin-protective effect of sesamolrdquo Journal of Cosmetic Science vol65 no 2 pp 69ndash79 2014

[9] T Furumoto and K Nishimoto ldquoIdentification of a charac-teristic antioxidant anthrasesamone F in black sesame seedsand its accumulation at different seed developmental stagesrdquoBioscience Biotechnology and Biochemistry vol 80 no 2 pp350ndash355 2016

[10] D-Z Hsu C-T Liu P-Y Chu Y-H Li S Periasamy and M-Y Liu ldquoSesame oil attenuates ovalbumin-induced pulmonaryedema and bronchial neutrophilic inflammation in micerdquoBioMed Research International vol 2013 Article ID 905670 7pages 2013

[11] P Xu F Cai X Liu and L Guo ldquoSesamin inhibits lipopo-lysaccharide-induced proliferation and invasion through thep38-MAPK and NF-120581B signaling pathways in prostate cancercellsrdquo Oncology Reports vol 33 no 6 pp 3117ndash3123 2015

[12] A A Dar and N Arumugam ldquoLignans of sesame purifica-tion methods biological activities and biosynthesismdasha reviewrdquoBioorganic Chemistry vol 50 pp 1ndash10 2013

[13] P J Kanu ldquoBiochemical analysis of black and white sesameseeds from Chinardquo American Journal of Biochemistry andMolecular Biology vol 1 no 2 pp 145ndash157 2011

[14] Y Chen G Chen X Fu and R-H Liu ldquoPhytochemical profilesand antioxidant activity of different varieties of Adinandra tea(Adinandra Jack)rdquo Journal of Agricultural and Food Chemistryvol 63 no 1 pp 169ndash176 2015

[15] V L Singleton R Orthofer and R M Lamuela-RaventosldquoAnalysis of total phenols and other oxidation substrates andantioxidants by means of Folin-Ciocalteu reagentrdquo Methods inEnzymology vol 299 pp 152ndash178 1998

[16] X He D Liu and R H Liu ldquoSodium borohydridechloranil-based assay for quantifying total flavonoidsrdquo Journal of Agricul-tural and Food Chemistry vol 56 no 20 pp 9337ndash9344 2008

[17] M V Reshma C Balachandran C Arumughan et al ldquoExtrac-tion separation and characterisation of sesame oil lignan fornutraceutical applicationsrdquo Food Chemistry vol 120 no 4 pp1041ndash1046 2010

[18] D Huang B Ou M Hampsch-Woodill J A Flanagan and RL Prior ldquoHigh-throughput assay of oxygen radical absorbancecapacity (ORAC) using a multichannel liquid handling systemcoupled with a microplate fluorescence reader in 96-wellformatrdquo Journal of Agricultural and Food Chemistry vol 50 no16 pp 4437ndash4444 2002

[19] MW Zhang R F Zhang F X Zhang and R H Liu ldquoPhenolicprofiles and antioxidant activity of black rice bran of different

commercially available varietiesrdquo Journal of Agricultural andFood Chemistry vol 58 no 13 pp 7580ndash7587 2010

[20] K K Adom and R H Liu ldquoRapid peroxyl radical scav-enging capacity (PSC) assay for assessing both hydrophilicand lipophilic antioxidantsrdquo Journal of Agricultural and FoodChemistry vol 53 no 17 pp 6572ndash6580 2005

[21] D L Felice J Sun and R H Liu ldquoA modified methylene blueassay for accurate cell countingrdquo Journal of Functional Foodsvol 1 no 1 pp 109ndash118 2009

[22] R H Liu J R Jacob J H Hotchkiss P J Cote J L Gerinand B C Tennant ldquoWoodchuck hepatitis virus surface antigeninduces nitric oxide synthesis in hepatocytes possible role inhepatocarcinogenesisrdquo Carcinogenesis vol 15 no 12 pp 2875ndash2877 1994

[23] M Nadeem C Situ A Mahmud et al ldquoAntioxidant activity ofsesame (Sesamum indicum L) cake extract for the stabilizationof olein based butterrdquo Journal of the American Oil ChemistsrsquoSociety vol 91 no 6 pp 967ndash977 2014

[24] J H Kim W D Seo S K Lee et al ldquoComparative assessmentof compositional components antioxidant effects and lignanextractions from Korean white and black sesame (Sesamumindicum L) seeds for different crop yearsrdquo Journal of FunctionalFoods vol 7 no 1 pp 495ndash505 2014

[25] S K Kwan S H Park and M G Choung ldquoNondestructivedetermination of lignans and lignan glycosides in sesameseeds by near infrared reflectance spectroscopyrdquo Journal ofAgricultural and Food Chemistry vol 54 no 13 pp 4544ndash45502006

[26] N Rangkadilok N Pholphana C Mahidol et al ldquoVariationof sesamin sesamolin and tocopherols in sesame (Sesamumindicum L) seeds and oil products in Thailandrdquo Food Chem-istry vol 122 no 3 pp 724ndash730 2010

[27] K Ishiyama M Nagashima T Yasumoto and Y FukudaldquoChanges in amounts of lignan compounds and radical scav-enging activities in improved high-lignan breed of sesamelsquoGomazoursquo during germinationrdquo Journal of the Japanese Societyfor Food Science and Technology vol 53 no 1 pp 8ndash16 2006

[28] S B Othman N Katsuno Y Kanamaru and T Yabe ldquoWater-soluble extracts from defatted sesame seed flour show antiox-idant activity in vitrordquo Food Chemistry vol 175 pp 306ndash3142015

[29] M Dachtler F H M Van De Put F V Stijn C M Beindorffand J Fritsche ldquoOn-line LC-NMR-MS characterization ofsesame oil extracts and assessment of their antioxidant activityrdquoEuropean Journal of Lipid Science and Technology vol 105 no9 pp 488ndash496 2003

[30] Ghafoorunissa SHemalatha andMVV Rao ldquoSesame lignansenhance antioxidant activity of vitamin E in lipid peroxidationsystemsrdquoMolecular and Cellular Biochemistry vol 262 no 1-2pp 195ndash202 2004

[31] A A Moazzami S L Haese and A Kamal-Eldin ldquoLignancontents in sesame seeds and productsrdquo European Journal ofLipid Science and Technology vol 109 no 10 pp 1022ndash10272007

[32] T Ide A Azechi S Kitade et al ldquoComparative effects of sesameseeds differing in lignan contents and composition on fatty acidoxidation in rat liverrdquo Journal of Oleo Science vol 64 no 2 pp211ndash222 2015

[33] K L Wolfe and H L Rui ldquoCellular antioxidant activity(CAA) assay for assessing antioxidants foods and dietarysupplementsrdquo Journal of Agricultural and Food Chemistry vol55 no 22 pp 8896ndash8907 2007


[34] K J Meyers C B Watkins M P Pritts and R H LiuldquoAntioxidant and antiproliferative activities of strawberriesrdquoJournal of Agricultural and Food Chemistry vol 51 no 23 pp6887ndash6892 2003

[35] L Z Zhang andRH Liu ldquoPhenolic and carotenoid profiles andantiproliferative activity of foxtail milletrdquo Food Chemistry vol174 pp 495ndash501 2015

[36] P Deng C Wang L Chen et al ldquoSesamin induces cell cyclearrest and apoptosis through the inhibition of signal transducerand activator of transcription 3 signalling in human hepatocel-lular carcinoma cell line HepG2rdquo Biological and PharmaceuticalBulletin vol 36 no 10 pp 1540ndash1548 2013

[37] Z Liu Q Xiang L Du G Song Y Wang and X Liu ldquoTheinteraction of sesamol with DNA and cytotoxicity apoptosisand localization in HepG2 cellsrdquo Food Chemistry vol 141 no1 pp 289ndash296 2013

Submit your manuscripts athttpwwwhindawicom

Hindawi Publishing Corporationhttpwwwhindawicom Volume 2014

Anatomy Research International

PeptidesInternational Journal of


Hindawi Publishing Corporation httpwwwhindawicom

International Journal of

Volume 2014

Zoology


Molecular Biology International

GenomicsInternational Journal of


The Scientific World JournalHindawi Publishing Corporation httpwwwhindawicom Volume 2014


BioinformaticsAdvances in

Marine BiologyJournal of



Signal TransductionJournal of


BioMed Research International

Evolutionary BiologyInternational Journal of



Biochemistry Research International

ArchaeaHindawi Publishing Corporationhttpwwwhindawicom Volume 2014


Genetics Research International


Advances in

Virolog y

Hindawi Publishing Corporationhttpwwwhindawicom

Nucleic AcidsJournal of

Volume 2014

Stem CellsInternational



Enzyme Research



Microbiology


and antiproliferative activities of various sesame seeds willbenefit the sesame seed planters relevant manufacturers andordinary consumers

2 Materials and Methods

21 Chemicals and Materials Methanol (MeOH) ethanol(EtOH) n-hexane ethyl acetate hydrochloric acid (HCl)and acetic acid (HAC) were purchased from GuanghuaSci-Tech Co Ltd (Guangdong China) Potassium chloride(KCl) sodium acetate (NaAC) sodium carbonate (NaCO

3)

sodiumhydroxide (NaOH) potassiumphosphatemonobasic(KH2PO4) and potassiumphosphate dibasic (K

2HPO4) were

of analytical grade and were purchased from Sangon BiotechCo Ltd (Shanghai China) 221015840-Azobis(2-amidinopropane)dihydrochloride (ABAP) 2101584071015840-dichlorofluorescin diacetate(DCFH-DA) fluorescein disodium salt catechin hydrate(HPLC ge98) Folin-Ciocalteu reagent ascorbic acid6-hydroxy-2578-tetramethylchroman-2-carboxylic acid(Trolox) formic acid (chromatographic grade) andmethanol(chromatographic grade) were purchased from Sigma-Aldrich Inc (St LouisMOUSA) Trifluoroacetic acid (TFAanalytical grade) and aluminum chloride (AlCl

3sdot6H2O

analytical grade) were purchased from Fisher Scientific(Fair Lawn NJ USA) Sodium borohydride (NaBH

4 reagent

grade) chloranil (analytical grade) vanillin (analytical grade99) and gallic acid (analytical grade 99) were purchasedfrom Aladdin Inc (Shanghai China) Sesamol sesamolinand sesamin (HPLC ge98) were purchased from ChengduPufei De Biotech Co Ltd (Sichuan China) HepG2 humanliver cancer cells were purchased from American TypeCulture Collection (Rockville MD USA) WME mediumfoetal bovine serum (FBS) insulin and other cell culturereagents were purchased from Gibco Life TechnologiesCo (Grand Island NY USA) Six sesame seed varieties(three black and three white) were kindly donated by ShanxiAcademy of Agricultural Sciences (Taiyuan city China)which were harvested in 2014 and stored in desiccator atroom temperature until analysis

22 Moisture Content The moisture content was analyzedby oven-dry method [13] Briefly 1 g of sample was driedin electric oven at 105∘C to a constant weight and moisturecontent was expressed as percent of dry weight (DW) intriplicate

23 Phenolics Extraction and Determination

231 Free Phenolics Extraction Free phenolic compoundswere extracted using the modified method reported pre-viously [14] Briefly 1 g of sesame seed was blended with5mL of hexane for 1min After centrifugation at 2700timesgfor 3min residue was defatted two more times Add 10mLof 80 chilled acetone to the residue and blend 1min Thesupernatant was collected after being centrifuged at 2700timesgfor 3min repeated twice The supernatant was pooled andevaporated to dryness at 45∘C under vacuum The solution

was reconstituted in 10mLof 70methanol and free phenolicextracts (100mg mLminus1) were stored at minus40∘C until analysis

232 Bound Phenolics Extraction Bound phenolics wereextracted using themethod described by Chen et al [14] withmodifications Briefly the residues obtained after extractionof free phenolics were flushed with nitrogen gas for 2minsealed and digested with 20mL of 4MNaOH at roomtemperature for 1 h The mixture was neutralized with 10Mconcentrated HCl Then 20mL hexane was added to extractresidual lipids in the mixture for 10min (not necessaryif no residual lipids) After centrifuging at 2700timesg for5min remaining residues were extracted five times withethyl acetate The ethyl acetate fractions were pooled andevaporated to dryness at 45∘C under vacuum The boundphenolics were reconstituted to 10mL of 70 methanol andbound phenolic extracts (100mgmLminus1) were stored at minus40∘Cuntil analysis

233 Determination of Total Phenolic Contents Total pheno-lic contents of each sample were determined using the mod-ified Folin-Ciocalteu colorimetric method [14 15] Brieflyfree phenolic and bound phenolic extracts were diluted 10ndash20 times with distilled water and were reacted with Folin-Ciocalteu reagent and then neutralized with Na

2CO3 After

90min incubation the absorbance of the resulting solutionwas recorded at 760 nm using MRX II Dynex plate reader(Dynex Technologies Inc Chantilly VA USA) Total pheno-lic contents were expressed as grams of gallic acid equivalentsper kg of sample on DW (gGAE kgminus1)

24 Determination of Total Flavonoid Contents Totalflavonoid contents were determined by sodium borohydride-chloranil (SBC) protocol [16] Briefly 1mL of free and 1mLof bound phenolic extracts were added to test tubes (15 times150mm) andwere dried under nitrogen gasThe residues andcatechin hydrate standards (01ndash100mM) were prepared in1mL of THFEtOH (1 1 vv) Extracts or standard solutionwasmixed with 05mL of 50mMNaBH

4solution and 05mL

of 746mMAlCl3solution After 30min shaking in an orbital

shaker at room temperature 01mL of 500mM NaBH4

solution was added with continued shaking for 30min atroom temperature 04mL of cold 08M acetic acid solutionwas added and the mixtures were kept in the dark for 15minafter thorough mixing 02mL of 200mM chloranil wasadded and the mixture was heated at 95∘C with shaking for60min Then the reaction solutions were cooled using tapwater and were brought to 1mL using methanol Afterward02mL of 16 (wv) vanillin was added and mixed followedby addition of 04mL of 12MHCl and the reaction solutionswere kept in the dark for 15min after thorough mixing Atlast the absorbance was recorded at 490 nm using MRXmicroplate reader with Revelation workstation (DynexTechnologies Inc) Total flavonoid contents were expressedas grams of catechin equivalents per kg of sample on DW(gCE kgminus1)



















50values and






BCD

H

DEF

BCD

H

DEFCDE

H

EFG

B

GH

DEF

FG

A

B

BC

H


DE


0

1

2

3

4

5

6

7

8

9Ph

enol

ic co

nten

t (gG

AE

kgminus1

DW

)

(a)

BC

EE

AB

FGHFGHFGH

FGH FG

CD

GH

AA

EFEF

HI

D

I



0

1

2

3

4

5

6

7

8

9

Flav

onoi

d co

nten

ts (g

CEkgminus1

DW

)

(b)





SesamolinSesamin

Sesamol

2 4 6 8 10 12 14 160t (min)

000

005

010

015

020

025

030


18column)


















Bound EC50



BC

J

E

H

B

FG

CD

IJ

DE

IJ

HGH

CD

A

F

H

DE

HI



0

20

40

60

80

100

120

140

160

ORA

C va

lues

(120583m

olTE

gminus1

DW

)

(a)

D

I

E

G

HIH

A

EF

B

A

F

BB

FF

HI

B

C



02468

101214161820

PSC

valu

es (120583

mol

VCEgminus

1D

W)

(b)








b5549 plusmn 901

b

B2 2104 plusmn 060a

2357 plusmn 088a

B3 6345 plusmn 085b

3006 plusmn 082a

W1 8729 plusmn 357c

7774 plusmn 129c

W2 8212 plusmn 310c

10953 plusmn 123d


6718 plusmn 192c







50







4 Conclusions


50) were


Competing Interests




Acknowledgments


References
















































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology



















50values and






BCD

H

DEF

BCD

H

DEFCDE

H

EFG

B

GH

DEF

FG

A

B

BC

H


DE


0

1

2

3

4

5

6

7

8

9Ph

enol

ic co

nten

t (gG

AE

kgminus1

DW

)

(a)

BC

EE

AB

FGHFGHFGH

FGH FG

CD

GH

AA

EFEF

HI

D

I



0

1

2

3

4

5

6

7

8

9

Flav

onoi

d co

nten

ts (g

CEkgminus1

DW

)

(b)





SesamolinSesamin

Sesamol

2 4 6 8 10 12 14 160t (min)

000

005

010

015

020

025

030


18column)


















Bound EC50



BC

J

E

H

B

FG

CD

IJ

DE

IJ

HGH

CD

A

F

H

DE

HI



0

20

40

60

80

100

120

140

160

ORA

C va

lues

(120583m

olTE

gminus1

DW

)

(a)

D

I

E

G

HIH

A

EF

B

A

F

BB

FF

HI

B

C



02468

101214161820

PSC

valu

es (120583

mol

VCEgminus

1D

W)

(b)








b5549 plusmn 901

b

B2 2104 plusmn 060a

2357 plusmn 088a

B3 6345 plusmn 085b

3006 plusmn 082a

W1 8729 plusmn 357c

7774 plusmn 129c

W2 8212 plusmn 310c

10953 plusmn 123d


6718 plusmn 192c







50







4 Conclusions


50) were


Competing Interests




Acknowledgments


References
















































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology


BCD

H

DEF

BCD

H

DEFCDE

H

EFG

B

GH

DEF

FG

A

B

BC

H


DE


0

1

2

3

4

5

6

7

8

9Ph

enol

ic co

nten

t (gG

AE

kgminus1

DW

)

(a)

BC

EE

AB

FGHFGHFGH

FGH FG

CD

GH

AA

EFEF

HI

D

I



0

1

2

3

4

5

6

7

8

9

Flav

onoi

d co

nten

ts (g

CEkgminus1

DW

)

(b)





SesamolinSesamin

Sesamol

2 4 6 8 10 12 14 160t (min)

000

005

010

015

020

025

030


18column)


















Bound EC50



BC

J

E

H

B

FG

CD

IJ

DE

IJ

HGH

CD

A

F

H

DE

HI



0

20

40

60

80

100

120

140

160

ORA

C va

lues

(120583m

olTE

gminus1

DW

)

(a)

D

I

E

G

HIH

A

EF

B

A

F

BB

FF

HI

B

C



02468

101214161820

PSC

valu

es (120583

mol

VCEgminus

1D

W)

(b)








b5549 plusmn 901

b

B2 2104 plusmn 060a

2357 plusmn 088a

B3 6345 plusmn 085b

3006 plusmn 082a

W1 8729 plusmn 357c

7774 plusmn 129c

W2 8212 plusmn 310c

10953 plusmn 123d


6718 plusmn 192c







50







4 Conclusions


50) were


Competing Interests




Acknowledgments


References
















































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology
















Bound EC50



BC

J

E

H

B

FG

CD

IJ

DE

IJ

HGH

CD

A

F

H

DE

HI



0

20

40

60

80

100

120

140

160

ORA

C va

lues

(120583m

olTE

gminus1

DW

)

(a)

D

I

E

G

HIH

A

EF

B

A

F

BB

FF

HI

B

C



02468

101214161820

PSC

valu

es (120583

mol

VCEgminus

1D

W)

(b)








b5549 plusmn 901

b

B2 2104 plusmn 060a

2357 plusmn 088a

B3 6345 plusmn 085b

3006 plusmn 082a

W1 8729 plusmn 357c

7774 plusmn 129c

W2 8212 plusmn 310c

10953 plusmn 123d


6718 plusmn 192c







50







4 Conclusions


50) were


Competing Interests




Acknowledgments


References
















































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology







Bound EC50



BC

J

E

H

B

FG

CD

IJ

DE

IJ

HGH

CD

A

F

H

DE

HI



0

20

40

60

80

100

120

140

160

ORA

C va

lues

(120583m

olTE

gminus1

DW

)

(a)

D

I

E

G

HIH

A

EF

B

A

F

BB

FF

HI

B

C



02468

101214161820

PSC

valu

es (120583

mol

VCEgminus

1D

W)

(b)








b5549 plusmn 901

b

B2 2104 plusmn 060a

2357 plusmn 088a

B3 6345 plusmn 085b

3006 plusmn 082a

W1 8729 plusmn 357c

7774 plusmn 129c

W2 8212 plusmn 310c

10953 plusmn 123d


6718 plusmn 192c







50







4 Conclusions


50) were


Competing Interests




Acknowledgments


References
















































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology





b5549 plusmn 901

b

B2 2104 plusmn 060a

2357 plusmn 088a

B3 6345 plusmn 085b

3006 plusmn 082a

W1 8729 plusmn 357c

7774 plusmn 129c

W2 8212 plusmn 310c

10953 plusmn 123d


6718 plusmn 192c







50







4 Conclusions


50) were


Competing Interests




Acknowledgments


References
















































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology













































Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology













Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology








Volume 2014

Zoology






















Advances in

Virolog y



Volume 2014




Enzyme Research



Microbiology

Documents

Research Article Phytochemical Contents and Antioxidant ...downloads.hindawi.com/journals/bmri/2016/8495630.pdf · T : Description and moisture contents of six sesame seeds varieties