Bergonzi 2012 Food-Chemistry

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Analytical Methods

Identification and quantification of constituents of Gardenia jasminoides Ellis

(Zhizi) by HPLC-DAD–ESI–MS

M.C. Bergonzi ⇑, C. Righeschi, B. Isacchi, A.R. Bilia

University of Florence, Dept. of Pharmaceutical Sciences, Via U. Schiff 6, 50019 Sesto Fiorentino, Florence, Italy

a r t i c l e i n f o

Article history:

Received 21 July 2010

Received in revised form 25 October 2011

Accepted 26 February 2012

Available online 6 March 2012

Keywords:

Gardenia jasminoides Ellis (Zizhi)

HPLC-DAD–MS

Qualitative and quantitative analysis

Iridoids

Crocins

Caffeoyl quinic acid derivatives

Method validation

a b s t r a c t

A simple, rapid and specific HPLC method was carried out for the analysis of characteristic constituents in

Gardenia jasminoides Ellis (Zhizi), namely iridoids, caffeoyl quinic acid derivatives and crocins. The sepa-

ration was successfully obtained using a C18 column by gradient elution with mixtures of methanol and

water as mobile phases; detection wavelength was set at 240 nm for iridoid glycosides, 315 nm for quinic

acid derivatives and 438 nm for crocins.

The analytical method was validated and the quantification of active compounds, namely iridoids, was

performed. Linearity, precision, repeatability, stability, accuracy, limit of detection (LOD) and limit of

quantification (LOQ) were also reported. This assay was successfully applied for qualitative and quanti-

tative analysis of five commercial samples of G. jasminoides Ellis.

2012 Elsevier Ltd. All rights reserved.

1. Introduction

The fruits of Gardenia jasminoides Ellis (Rubiaceae) (Gardeniae

Fructus, Chinese name Zhizi) are widely used in the Traditional

Chinese Medicine. Numerous properties are reported for this her-

bal drug and its preparation, namely in the treatment of irritability

in febrile diseases, jaundice, acute conjunctivitis, epitasis, haemat-

uria, pyogenic infections and ulcers of the skin, and also, externally,

sprains and painful swellings due to blood stasis (Chang & But,

1987; Tang & Eisenbrand, 1992; Ukita, Yamasaki, Ino, Kawamoto,

& Saito, 1994; Wang, Tseng, Huang, & Tsai, 2004). Recent studies

have also attributed antiangiogenic properties to Zhizi (Park, Joo,

Kim, & Lim, 2003).

In addition to Gardeniae Fructus, Gardeniae Fructus Preparatus

is also present on the market, obtained by processing Fructus

Gardeniae which is stir-baked or broken into pieces in a hot pot

with middle heat until it becomes burnt-brown or burnt-black

externally and the inner surface and seed coats yellowish-brown

or dark brown (Pharmacopeia of People’s Republic of China,

2005).

The major constituents of Gardenia fruits are iridoid glycosides

including geniposide, gardenoside, genipin-1-O-b-gentiobioside,

geniposidic acid, acetylgeniposide, scandoside methyl ester,

shanzhiside and gardoside. Many authors have assumed that these

compounds are the molecules responsible for the biological

activities of this herbal drug and its preparations (Li et al., 2000;

Miura, Nishiyama, Ichimaru, Moriyasu, & Kato, 1996; Park et al.,

2003; Ukita et al., 1994; Yamauchi, Fujimoto, Kuwano, Inoue, &

Inoue, 1976).

Gardenia fruit is also widely used in Asian countries as a natural

colourant because of the presence of crocins, a series of mono- and

di-glucosyl esters of crocetin, crocin-1, -2, -3 (Pfister, Meyer, Steck,

& Pfander, 1996; van Calsteren et al., 1997).

Numerous studies have reported that even crocins possess a

variety of biological effects, principally the antioxidant properties

(Ahmad et al., 2005; Escribano, Alonso, Coca-Prados, & Fernández,

1996; He et al., 2005; Ochiai et al., 2004; Shen & Qian, 2006; Tseng,

Chu, Huang, Shiow, & Wang, 1995; Xiang et al., 2006).

Ubiquitous quinic acid derivatives have also been identified in

the fruit of G. jasminoides (Nishizawa & Fujimoto, 1986; Nishizawa,

Izuhara, Kaneko, & Fujimoto, 1987; Wenhao et al., 2010).

The aim of this study was to establish a reliable HPLC method

for simultaneous determination of these three classes of constitu-

ents in Gardenia jasminoides Ellis, namely iridoids, crocins and caf-

feoyl quinic acid derivatives.

Some chromatographic methods have been previously reported

(He, Cheng, Chen, & Zhou, 2006; Xu, Cao, Wang, & Luo, 2003) but

none of them can be applied to investigate quality control of the

commercial products.

0308-8146/$ - see front matter 2012 Elsevier Ltd. All rights reserved.doi:10.1016/j.foodchem.2012.02.157

⇑ Corresponding author. Tel.: +39 055 4573678; fax: +39 055 4573680.

E-mail address: [email protected] (M.C. Bergonzi).

Food Chemistry 134 (2012) 1199–1204

Contents lists available at SciVerse ScienceDirect

Food Chemistry

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http://dx.doi.org/10.1016/j.foodchem.2012.02.157mailto:[email protected]://dx.doi.org/10.1016/j.foodchem.2012.02.157http://www.sciencedirect.com/science/journal/03088146http://www.elsevier.com/locate/foodchemhttp://www.elsevier.com/locate/foodchemhttp://www.sciencedirect.com/science/journal/03088146http://dx.doi.org/10.1016/j.foodchem.2012.02.157mailto:[email protected]://dx.doi.org/10.1016/j.foodchem.2012.02.157


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In the present work, we have developed and validated a

HPLC-DAD–MS method suitable for identifying 12 compounds. A

comprehensive validation study was also carried out and covered

linearity, precision, repeatability, stability, accuracy, limits of

detection and quantification of the iridoids which represent the

main molecules responsible for the activity of Zhizi. The phenolic

compounds were also quantified using chlorogenic acid as external

standard. Only qualitative analysis of crocins is reported because of thelack of suitable standards on themarket for their quantification.

The developed HPLC method would be helpful for the quality

control of G. jasmonoides Ellis, its preparata and related Chinese

remedies.

2. Experimental procedures

2.1. Apparatus for HPLC-DAD–ESI–MS analysis

The HPLC system consisted of a HP 1100L instrument equipped

with a HP 1040 diode-array detector (DAD), an automatic injector,

an auto sampler and a column oven and managed by a HP 9000

workstation (Agilent Technologies, Palo Alto, CA, USA). The HPLC

system was interfaced with a HP 1100 MSD API-electrospray(Agilent Technologies, Palo Alto, CA, USA).

Thereverse-phase columnwas a Luna C18 (250 4.6 mm, 5 lm,Phenomenex) maintained at 27 C. The mobile phase was a two-

step linear gradient CH3OH (A)/H2O (pH 3.2, HCOOH) (B) at a flow

rate of 1 ml/min: 0.10–4.0 min 10% A and 90% of B; 4.0–40 min

90% B to 10% B. Before the HPLC analysis, each sample was filtered

through a cartridge-type sample filtration unit with a polytetrafluo-

roethylene (PTFE) membrane (d = 13 mm, porosity 0.45 lm, Lidamanufacturing Corp.) and immediately injected. The injected vol-

ume of sample was a 20-ll solution. UV–Vis spectra were recordedin the range 210–500 nm, and chromatogramswere acquiredat 240

(iridoid glycosides), 315 (quinic acid derivatives), 438 nm (crocins).

The HPLC system was interfaced with a HP 1100 MSD API-

electrospray (Agilent Technology, Palo Alto, CA, USA). The interfacegeometry, withan orthogonal position of the nebulizer with respect

to the capillary inlet, allowed the use of analytical conditions similar

to those of theHPLC-DADanalysis. The same column, mobile phase,

time period and flow rate were used. Mass spectrometry operating

conditions were optimised in order to achieve maximum sensitivity

values; gas temperature 350 C at a flow rate of 10 l/min, nebulizer

pressure 30 p.s.i., quadrupole temperature 30 C, and capillary volt-

age 3500 V. Full scan spectra from m/ z 100 to 800 in the positive ion

mode were obtained (scan time 1 s). Mass spectra were performed

in negative and positive ion mode, setting the fragmentation energy

between 80 and 120 V and applyingthe samechromatographiccon-

ditions described previously.

2.2. Standards and reagents

Geniposide CRS provided by China’s National Institute for the

Control of Pharmaceutical and Biological Products (NICPBP, Beijing,

China), purity 95.63% by elemental analysis. Chlorogenic acid

(P99%) was from Extrasynthese, Genay Cedex, France.

MeOH (HPLC grade) was purchased from Merck (Darmstadt,

Germany); 85% formic acid was provided by BDH AnalaR. Water

was purified by a Milli-Q plus system from Millipore (Milford, MA,

USA).

2.3. Samples

Five commercial samples of Gardenia Fructus were supplied by

PLANTASIA Heinrich Handel-Mazzettiplaz 1, A-5110 Oberndorf,Austria.

Name Code

Zhizi 25845

Gardeniae (Fructus) (Zhizi) 32496

Gardeniae (Fructus) (Zhizi) (Hebei) 33031

Gardeniae (Fructus) Preparata (Chaozhizi) (Hebei) 33024

Gardeniae (Fructus) Preparata (Jiaozhizi) (Hebai) 32990

The sample 32496 was used for the validation study.

2.4. Preparation of sample solutions

Ten millilitres of 50% methanol solution was added to 1 g of the

powdered herbal drug; this hydroalcoholic extract was ultrasoni-

cated for 40 min, and filtered (Wagner, 2004). The extraction was

repeated three times. The filtrate was evaporated until dryness

(Drug-extract-ratio, D.E.R. 3.0-3.6:1) and the residue, precisely

weighed, was dissolved in methanol to prepare the solutions for

method validation and analyses.

3. Results and discussion

3.1. Optimisation of chromatographic conditions

This study concerns the HPLC-DAD–ESI–MS characterisation of

extracts of Gardeniae Fructus (Zhizi). The wavelengths of 240, 315

and 438 nm were selected for acquiring chromatograms of iridoid

glycosides, quinic acid derivatives and pigments, respectively. In

order to achieve better chromatographic separation, various linear

gradients of acetonitrile–water and methanol–water were investi-

gated at a flow-rate of 1.0 mL min1. Finally, the gradient pro-

gramme described in the experimental part was chosen because

all the peaks in the chromatogram were clearly separated. The

HPLC-DAD profiles of the methanolic extract at the wavelengths

of 240, 315 and 438 nm are reported in Figs. 1–3, respectively. The

compounds identified are reported in Table 1. The method allowed

the separation of the iridoids, of quinic acid derivatives and fourcrocins.

The identification of compounds was carried out by comparing

the characteristic UV absorption spectra, retention time and MS

data of standard compounds to those already present in literature.

The peaks indicated in Fig. 2 by ⁄ are caffeoylquinic acid deriva-

tives, identified by their UV absorption spectra and the presence

of different diagnostic fragments in their MS spectrum, such as

[quinic acidH] m/ z 191, [quinic acidHH2O] m/ z 173,

[caffeoylHCOO] m/ z 135.

This HPLC method was also applied in the determination of the

Fructus Gardeniae constituents in five commercial samples, as re-

ported in Section 3.3. All the analyses were repeated in triplicate.

3.2. Method validation

The method was validated according to ICH guidelines (ICH,

1994, 1996). The method was found to be specific and suitable for

the routine analysis because of its simplicity, sensitivity, accuracy

and reproducibility. It can be conveniently used for the quantifica-

tion of iridoids in G. jasminoides Ellis commercial samples.

3.2.1. Linearity

Linearity range of response was determined for geniposide

(the main iridoid) reference standard CRS, chlorogenic acid andG. jasminoides Ellis extract. Geniposide was used to optimise the

extraction of the herbal drug, because it is the main iridoid of the

extract.

Linearity was determined on five levels of concentration withthree injections for each level. Geniposide and chlorogenic acid

1200 M.C. Bergonzi et al. / Food Chemistry 134 (2012) 1199–1204


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showed a linear response from 1.0 to 1000 lg/ml and all the curveshad coefficients of linear correlation P0.998.

A linearity of response in the range from 0.07 to 1.86 mg/ml of

G. jasminoides Ellis extract (concentrations used: 0.07, 0.14, 0.28,

0.57, 1.86 mg/ml in methanol) was observed for the main constit-

uent geniposide. Additionally, the value of R2 was from 0.9994 to

0.9990, with the slope R.S.D. values lower than 1.5%, which indi-

cated a high accuracy of the method.

3.2.2. Limit of detection (LoD) and limit of quantitation (LoQ) of

geniposide

The detection limit (LoD) and the quantitation limit (LoQ) of geniposide was determined by calculation of the signal-to-noise

ratio. A signal-to-noise ratio 3:1 is generally considered acceptable

for estimating the detection limit. The sample that produces a sig-

nal-to-noise ratio of approximately 10:1corresponds to the concen-

tration at which the analyte can be reliably quantified. Geniposide

at a concentration of 0.192 lg/ml (injected 5 ll, 0.76 ng) producesa signal-to-noise ratio of approximately 3.2; while geniposide at a

concentration of 0.532 lg/ml (injected 10 ll, 5.32 ng) produces asignal-to-noise ratio of approximately 12.1.

3.2.3. 48-Hour extract stability

Reference compounds and the G. jasminoides Ellis methanolicextract (1.80 mg/ml) were solubilised in methanol before the

min5 10 15 20 25

mAU

0

500

1000

1500

2000

2500

1

2

3

4

5

Fig. 1. HPLC profile at 240 nm of Gardenia jasminoides Ellis extract; iridoids (1–5) are listed on Table 1.

min0 5 10 15 20 25 30 35

mAU

0

50

100

150

200

250

300

350

400

450

*

*

*

7

8

*

*

*

6

Fig. 2. HPLC profile at 315 nm of Gardenia jasminoides Ellis extract; caffeoyl quinic derivatives (6–8) are listed on Table 1.

M.C. Bergonzi et al. / Food Chemistry 134 (2012) 1199–1204 1201


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analyses and their stability at room temperature was evaluated

every 4 h, up to 48 h. The standards and the extract were found

to be stable in methanol solution at room temperature for at least

48 h. In Table 2 the %D/ Z (D = determined concentration and

Z = determined concentration on hour-zero) values over the courseof the stability time period was reported.

3.2.4. Reproducibility of the injection integration

The reproducibility of the injection integration procedure was

determined for geniposide (0.96 mg/ml) and the G. jasminoides Ellis

extract. The solutions were injected ten times and the relative

standard deviation (R.S.D.) values were calculated.

R.S.D. of extract at the concentration of 1.14 mg/ml were: genip-

oside 1.07%; scandoside methylesthere 1.72%, gardenoside 1.87%,

genipin gentiobioside 1.23%; acetylgeniposide 0.64%, 3,4-dicaf-

feoyl-5-(3-hydroxy-methylglutaroyl) quinic acid 1.41% andcaffeoyl sinapoylquinic acid 0.98%.

3.2.5. Repeatability of the method

In order to evaluate the repeatability of the method, three

solutions at different concentrations (0.57, 1.14 and 1.86 mg/ml of

G. jasminoides Ellis dry extract in methanol) were prepared. Each

solution was injected three times. The contents of iridoids and

quinic acid derivatives were calculated in order to estimate the

R.S.D. Geniposide (R.S.D. 0.72%, 0.27% and 1.02% respectively),

scandoside methylesthere (R.S.D. 0.40%, 1.3%, 1.17%), gardenoside

(R.S.D. 1.71%, 1.51%, 1.41%), genipin gentiobioside (R.S.D. 1.39%,

1.76%, 1.35%), acetylgeniposide (R.S.D. 1.0%,0.98%, 0.40%) (3,4-dicaf-

feoyl-5-(3-hydroxy-methylglutaroyl) quinic acid (R.S.D. 1.6%,

0.17%, 0.60%) and caffeoyl sinapoylquinic acid (R.S.D. 1.57%, 0.48%,0.94%).

min0 5 10 15 20 25 30 35

0

200

400

600

800

1000

1200

9

10 11

12

1400

mAU

Fig. 3. HPLC profile at 438 nm of Gardenia jasminoides Ellis extract; crocins (9–12) are listed on Table 1.

Table 1

Compounds identified in the Gardenia Fructus extract.

Peak t R MS UVmax nm Compound

1 7.51 [M+Na]+ m/ z 427 240 Scandoside methylestere

2 9.03 [M+Na]+ m/ z 427 240 Gardenoside

3 13.68 [M+Na]+ m/ z 573 [M + H]+ m/ z 551 240 Genipin gentobioside

4 16.17 [M+Na]+ m/ z 411 240 Geniposide

5 24.45 [M+Na]+ m/ z 453 240 Acetylgeniposide

6 25.85 [M+Na]+ m/ z 609 250, 355 Quercetin-3-rutinoside

7 24.38 [MH] m/ z 659 315 3,4-Dicaffeoyl-5-(3-hydroxy-3-methylglutaroyl) quinic acid

8 24.76 [MH] m/ z 559 315 Caffeoyl sinapoylquinic acid9 27.15 [MH] m/ z 975 438 Crocin 1

10 29.26 [MH] m/ z 813 438 Crocin 2

11 35.06 [MH] m/ z 489 438 Crocin 3

12 35.48 [MH] m/ z 327 438 Crocetin

Table 2

%D/ Z (D = determined concentration and Z = determined concentration on hour-zero)

values over 48 h stability of standards and geniposide in the extract.

Time

(h)

Standard

geniposide

Standard clorogenic

acid

Geniposide in the

extract

0 100 100 100

4 98.58 99.93 99.30

8 98.87 99.49 99.60

12 98.89 99.05 99.74

16 99.62 98.97 99.32

20 98.81 98.43 99.15

24 99.17 98.41 98.99

28 99.05 98.21 98.83

32 99.01 97.94 98.03

36 98.97 97.67 98.04

40 98.59 97.29 97.98

44 98.71 96.98 97.87

48 98.99 96.82 97.87



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3.2.6. Intermediate precision

The same samples described in the Section 3.2.5. were injected

six times on three different days for the purpose of evaluating

intermediate precision relative to geniposide, used as compound

representative of the extract.

Precision intraday, n = 6 R.S.D. 0.93%, 2.50%, 0.59%; precision

interday n = 18 R.S.D. 1.86%, 0.76%, 1.32%.

3.2.7. Precision of the sample preparation

To evaluate the precision of the sample preparation, three solu-

tions at concentration about 0.57 mg/ml (0.57, 0.58, 0.60 mg/ml) of

dry extract in methanol were prepared. Each solution was injected

three times. The contents of iridoids and quinic acid derivatives

were calculated in order to estimate the R.S.D. All R.S.D. values of

the main constituents resulted between 0.55% and 1.2%.

3.2.8. Accuracy

In order to calculate thebiases forthe linearity data, theaccuracy

of themethod wasdeterminedby analysing thepercentagerecovery

of geniposide into the three preparations of G. jasminoides Ellis

extract. Three independent solutions of extract were prepared

(0.57, 1.14 and 1.86 mg/ml) and each was injected three times.

The average percentage recovery was calculated for each level of

concentration. The accuracy was determined by spiking 50 lg/mlof geniposide separately to the three batches of the extract. The

percentages recovery of geniposide standard spiked into three

preparations were 93.63 ± 2.15%; 96.21 ± 1.04% and 97.27 ± 2.55%,

respectively.

3.2.9. Specificity

The peak purity was investigated by inspecting the UV-spectra

and MS spectra at the beginning, at the apex and at the end of the

peaks of each constituent of the extract. No deviations were seen.

As an example, Fig. 4 showed the overlay of 3 UV-spectra at the

beginning, at the apex and at the end of the peak geniposide

(240 nm).

3.3. Sample analysis

The variability of the constituents in the different samples is

stressed, with special emphasis on iridoids, crocins and quinic acid

derivatives. The quantitative determination of iridoids was

performed at 240 nm, using a geniposide CRS as external standard.

Table 3 reports the content of iridoids, expressed as geniposide,

found in each Gardeniae Fructus sample.

No qualitative differences among the diverse samples were

found but quantitative differences in the constituents were found

and were attributable to different sources or different methods of

processing of the herbal drug. Sample 25845 consisted of thewholeherbal drug (Zhizi) and was the richest sample in iridoids content.

Consideringthe two samples 32496 and 33031, thedifferent profile

could be attributed to their different sites of collection. The two

samples of Gardeniae Fructus Preparata (33024 Chaozhizi; 32990

Jiaozhizi) have qualitative and quantitative profiles of iridoids

which are similar to those of Gardenia Fructus indicating that the

process obtained by heating the herbal drug does not influence

particularly the composition of iridoids in the drug, according to

the results previously reported in the literature (Sheu & Hsin, 1998).

The quantitative analysis of quinic acid derivatives was per-

formed at 315 nm, using chlorogenic acid as external standard.

The results were summarised in Table 4. Their content ranged from

23.16 to 44.39 mg/g of extract. Sample 33031 had the poorest con-

tent of these compounds, as well as for the iridoids.

From our studies it seems that the variability of the constitu-

ent’s content could be related to the geographic area of origin

rather than the processing of the plant material.

Thechromatographicprofiles of thecrocins were the same forall

five samples: the derivatives identified were crocetin, crocin-1,

Table 3

Content (mg/g and %) of iridoids found in Gardeniae Fructus extracts.

Sample Scandoside methylesthere mg/g of

extract, (%)

Gardenoside mg/g of

extract, (%)

Genipin gentiobioside mg/g of

extract, (%)

Geniposide mg/g of

extract, (%)

Acetylgeniposide mg/g of

extract, (%)

25845 8.76 ± 1.09 (0.87%) 20.72 ± 0.98 (0.21%) 20.07 ± 1.98 (2.00%) 213.31 ± 2.37 (21.33%) 5.14 ± 0.35 (0.51%)

32496 8.60 ± 0.48 (0.86%) 10.29 ± 0.79 (1.03%) 10.57 ± 0.28 (1.06%) 155.30 ± 0.94 (15.53%) 7.43 ± 0.06 (0.74%)

33031 1.77 ± 0.24 (0.17%) 1.70 ± 0.56 (0.17%) 9.23 ± 0.07 (0.92%) 125.49 ± 1.60 (12.55%) 3.97 ± 0.76 (0.39%)

33024 4.64 ± 1.25 (0.46%) 16.90 ± 0.55 (0.17%) 16.90 ± 0.55 (1.69%) 152.36 ± 1.98 (15.23%) 6.95 ± 0.19 (0.69%)

32990 4.92 ± 0.46 (0.49%) 11.25 ± 2.92 (1.12%) 7.73 ± 2.16 (0.77%) 157.94 ± 13.85 (15.79%) 6.12 ± 0.83 (0.61%)

Fig. 4. The overlay of 3 UV-spectra (240 nm) at the beginning, at the apex and at the end of the peak geniposide at 16.17 min.

M.C. Bergonzi et al. / Food Chemistry 134 (2012) 1199–1204 1203


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crocin-2, crocin3 (Fig.3). Thecomparison of the AUCs of singlepeaks

in the different investigated commercial samples showed once

again that the lowest content was evidenced in sample 33031, col-

lected from the district of Hebei, while the highest quantity was

present in the entire Zhizi (sample 25845).

4. Conclusions

The separation of the constituents of Gardenia fructus was suc-

cessfully obtained with a C18 column using a gradient elution with

methanol and water as mobile phases. Detection wavelengthswere set at 240 nm for iridoid glycosides, 315 nm for quinic acid

derivatives and 438 nm for crocins. The method was validated

according to ICH guidelines, taking into account that iridoids rep-

resent the characteristic molecules responsible for the activity of

Zhizi. This HPLC assay was successfully used for the determination

of constituents for the qualitative and quantitative characterisation

of five commercial samples of Gardeniae Fructus. The method was

found to be specific and suitable for the routine analysis because of

its simplicity, sensitivity, accuracy and reproducibility; it can be

conveniently used for iridoids in G. jasminoides Ellis commercial

samples. This HPLC method can be proposed for the quality control

of Gardenia jasmonoides Ellis and its related Chinese remedies. The

analytical method is also specific for other components of the ex-

tract, such as crocins and quinic acid derivatives.

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Table 4

Content (mg/g and %) of caffeoyl quinic acid derivatives found in Gardeniae Fructus extracts.

Sample 3,4-Dicaffeoyl-5-(3-hydroxy-

3-methylglutaroyl) quinic acid

Caffeoyl sinapoylquinic acid Other caffeoylquinic

derivatives (evidenced by ⁄ in Fig. 2)

25845 4.89 ± 0.22 (0.49%) 20.43 ± 0.18 (2.04%) 19.08 ± 0.28 (1.9%)

32496 5.73 ± 0.21 (0.57%) 8.64 ± 0.85 (0.86%) 17.83 ± 0.38 (1.8%)

33031 3.76 ± 1.11(0.38%) 9.25 ± 0.038 (0.92%) 10.15 ± 0.32 (1.0%)

33024 5.97 ± 0.58 (0.60%) 12.65 ± 0.41 (1.26%) 14.37 ± 0.73 (1.4%)

32990 4.17 ± 1.40 (0.42%) 6.96 ± 0.81 (0.70%) 15.59 ± 1.64 (1.6%)


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