Advance Journal of Food Science and Technology 9(4): 269-277, 2015

DOI: 10.19026/ajfst.9.2007
ISSN: 2042-4868; e-ISSN: 2042-4876
© 2015 Maxwell Scientific Publication Corp.
Submitted: December 14, 2014 Accepted: January 27, 2015 Published: August 15, 2015

Research Article
Quantitation of Allicin in Garlic-based Products: Comparisons among Spectrophotometry,
GC and HPLC
1
Chunli Zhou, 1Xueyan Hu, 2Chen Chao, 2Hui Li, 1Shuiyin Zhang, 1Xueming Yan,
1
Fengmei Yang and 2Quanhong Li
1
School of Life Science, Jiangxi Science and Technology Normal University, Nanchang 330013, China
2
College of Food Science and Nutritional Engineering, China Agriculture University,
Beijing 100083, China

Abstract: Spectrophotometric, GC and HPLC methods were used to determinate allicin concentration in garlic-
based products (garlic powder, garlic oil and garlic tablets). Allicin was extracted using a mixture of water and
ethanol and analyzed by the three methods. The results revealed that the GC method was unsuitable for allicin
quantitation because of high coefficient of variations and high temperatures. The spectrophotometric method was the
simplest and most effective method for solid garlic-based products. The HPLC method was more accurate for allicin
quantitation in all garlic-based products, especially liquids. Furthermore, the HPLC method allowed the
simultaneous quantitation of allicin and allicin degradation products such as diallyl sulfide, dimethyl trisulfide,
diallyl disulfide and diallyl trisulfide. Based on the spectrophotometric and HPLC method, garlic oil contained the
highest allicin concentration, followed by the garlic tablets and garlic powder.

Keywords: Allicin, garlic products, gas chromatography, high pressure liquid chromatography, spectrophotomety

INTRODUCTION

Garlic (Allium sativum linn.), a common food

condiment, is considered a dietary supplement due to its
cholesterol-lowering effects. Garlic-based products have
become increasingly popular over the past several years.
For individuals who cannot tolerate the consumption of
raw garlic, garlic-based products constitute a means of
consuming allicin, the main bioactive compound in
garlic. Allicin (S- (2-propenyl) 2-propene-1-
sulfinothioate), which is also known as diallyl
thiosulfinate, is a flavor compound. As shown in Fig. 1,
the structure of allicin has a Sulfur Oxide (SO) group
(Ali et al., 2000). Allicin is not present in whole garlic. Fig. 1: Structure of allicin (S- (2-propenyl) 2-propene-1-
Allicin is instantaneously synthesized when garlic sulfinothioate or diallyl thiosulfinate)
cloves are crushed or damaged and the enzyme alliinase
and the compound alliin, which appear to be stored in Heat induces the degradation of allicin into several
separate compartments in whole garlic, interact diallyl sulfide compounds (Lawson and Gardner, 2005):
(Bachrach et al., 2011). The synthesis of allicin takes Diallyl Sulfide (DAS), Dimethyl Trisulfide (DAT),
place through a series of reactions detailed in Scheme 1 Diallyl Disulfide (DADS) and Diallyl Trisulfide
(Miron et al., 2000). Extracted allicin, however, loses its (DATS). These compounds are stable and used in the
beneficial properties within hours through its conversion treatment of cancer and bacterial and fungal infections
into other sulfur-containing compounds (Sigounas et al., (Song and John, 2001; Tripathi, 2009). Among the
1997; Hirsch et al., 2000). As shown in Fig. 2, allicin diallyl sulfide compounds, DADS is the most potent. A
(diallyl thiosulfinate) is formed by attaching an allyl study reported that garlic extract, allyl alcohol and
group and an oxygen atom to the sulfur atom of cysteine DADS increase oxidative stress and reduce glutathione
(Khar et al., 2011). concentrations in Candida albicans (Lemar et al., 2007).

Corresponding Author: Quanhong Li, College of Food Science and Nutritional Engineering, China Agriculture University,
Beijing 100083, China, Tel.: +86-0791-88537850
This work is licensed under a Creative Commons Attribution 4.0 International License (URL: http://creativecommons.org/licenses/by/4.0/).
269
 Adv. J. Food Sci. Technol., 9(4): 269-277, 2015

Fig. 2: Structure of degradation products of allicin (Lawson and Gardner, 2005)

Furthermore, allicin and its degradation products are garlic-based products/pharmaceutical preparations
important due to their antimicrobial, antithrombotic, should be standardized. The purpose of this study was to
hypolipidemic, antiarthritic, hypoglycemic and compare spectrophotometry, GC and HPLC in the
antitumor activities (Oommen et al., 2004). Allicin quantitation of allicin in garlic oil, garlic powder and
belongs to the family of phytochemicals, which are garlic tablets:
useful in the treatment and cure of several cancers.
According to Baneriee and Maulik (2002), allicin
prevents the risk of cardiovascular diseases and cancers
and has antimicrobial and antioxidant effects.
Additionally, allicin protects against dental caries and
periodontitis (Bachrach et al., 2011).
The quantitative determination of allicin and its
degradation products is important due to their
pharmacologic activities. Currently, quantitative
determination methods rely on spectrophotometry, High
Pressure Liquid Chromatography (HPLC) and Gas
Chromatography (GC) (Han et al., 1995; Miron et al.,
2002; Baghalian et al., 2005; His and Wu, 1989). Other Scheme 1
methods including GC-Mass Spectrometry (GC-MS)
(Mondy et al., 2001), Supercritical Fluid MATERIALS AND METHODS
Chromatography Mass Spectrometry (SFC-MS) (Calvey
et al., 1994), Ultra-Performance Liquid Materials: Eight garlic-based products were purchased
Chromatography with photo Diode Array Detection from different areas of Shandong province and Jiangsu
(UPLC-DAD) (Khar et al., 2011) have been used. GC- province in China. This sampling was performed
MS, SFC-MS and UPLC-DAD methods can be used for according to information obtained from local
scientific research; however, they are very expensive. agricultural extension offices. The garlic products
Chromatographic methods are reliable, selective and included garlic tablets, garlic powder and garlic oil. The
accurate because possible interferences from other geographical origins of the garlic-based products are
sulfur-containing compounds can be easily eliminated. shown in Table 1. DAS, DAT, DADS and DATS were
However, the accuracy of allicin determination from purchased from Sigma Chemical Inc. (USA). L-
these chromatographic methods ultimately depends on Cysteine and 5, 5-dithiobis- (2-nitrobenzoic acid)
the extraction procedures and purification steps. (DTNB) were obtained from Beijing Land Bridge
Pharmaceutical preparations of garlic, which is Technology, Co., Ltd. (Beijing, China).
considered one of the safest herbal plant medicines, are
currently available in the market (Velisek et al., 1997). Allicin extraction: The garlic tablets were ground.
Therefore, the quantitative determination of allicin in Approximately, 0.5 g of garlic-based product was mixed
270
 Adv. J. Food Sci. Technol., 9(4): 269-277, 2015

Table 1: Geographical origins of the garlic products

Sample number Region originated Product Climatea
Gp1 Jinxiang county Garlic oil Warm temperate-semi humid
Gp2 Shouguang county Garlic powder Warm temperate-semi humid
Gp3 Chengwu county Garlic powder Warm temperate-semi humid
Gp4 Yutai county Garlic powder Warm temperate-semi humid
Gp5 Guangrao county Garlic piece Warm temperate-semi humid
Gp6 Shanghe county Garlic piece Warm temperate-semi humid
Gp7 Sheyang county Garlic piece Subtropics transition temperate-humid
Gp8 Cangshan county Garlic piece Warm temperate-semi humid

with 5 mL of double distilled water in a 100-mL conical GC method: GC was performed using an Agilent GC-
glass bottle. Enzymatic hydrolysis was allowed to take 6890 system equipped with a flame ionization detector.
place in a 50°C water bath for 60 min. A volume of 15 Air and H2 flow rates were 350 and 50 mL/min,
M ethanol was added and the mixture was placed in a respectively. The flow rate of the carrier gas (N2) was
45°C oscillator water bath at 150 rpm for 90 min. Every set to 1.0 mL/min. The temperature of the injection port
5 min, the bottle containing the mixture was open to and detector were set to 160 and 180°C, respectively.
release air. The mixture was filtered using filter paper
The oven temperature was set to 50°C for 3 min,
and the filtrate (allicin) was collected.
increased to 160°C at a rate of 10°C/min and held at this
Spectrophotometric method: Reactions were temperature for 3 min and finally increased to 230°C at
performed at 30°C. Five milliliters of 10 mM L-cysteine a rate of 30°C/min and held at this temperature for 3
in 50 mM Tris-HCL buffer (pH 7.5) was either mixed min. The injection volume was 1 µL (splitless mode). A
with 1 mL of double distilled water (blank) or 1 mL of capillary column (SPD-1 fused-silica capillary; 30
extracted allicin; 1 mL of the resulting solution was m×0.53 mm I.D., 0.01 µm; Supelco, Bellefonte, PA,
diluted to 100 mL. The diluted solution (4.5 mL) was USA) was used. Allicin was quantified using the
mixed with 1.5 mM DTNB (0.5 mL) in 50 mM Tris- following equations:
HCL buffer (pH 7.5). Absorbance was measured at 412
nm after 15 min. Allicin was quantified using the ρ × A i × V × V2 × 100%
following equations: Allicin (%, w/w) =
mA 0 V1 × (10000μg/mg)
△A412 = A0-A
Callicin (mmol/mL) = (△A412×β) / (2×14, 150) where,
ρ = The allicin concentration in the standard (µg/mL)
where, Ai = The allicin peak area of the standard
β = The dilution of L-Cysteine
V = The final sample volume (mL)
14,150 = The molar Extinction coefficient (E) of
allicin in water V1 = The extration volume of the sample (mL)
A0 = The absorbance reading of water V2 = The allicin volume in the standard (mL)
A = The absorbance reading of allicin m = The sample weight (g)

HPLC method: HPLC was performed using a Waters Statistical analyses: Data analyses were performed
616 pump and UV-1575/VIS absorbance detector. using SPSS software. Simple statistics (i.e., mean,
Compounds were separated on a 250×4 mm i.d.×5 µm maximum and minimum values, standard deviations and
LiChrospher 100 RP18 (C18) column (Hewlett- coefficient of variation) were used to compare the three
Packard). The mobile phase consisted of methanol-water different methods. Analysis of variance (ANOVA) and
(70:30, v/v). DAS, DAT, DADS and DATS were used Duncan's multiple range tests were performed.
as standards. HPLC was performed at ambient Statistical significance was set at pU0.05.
temperature at a flow rate of 0.8 mL/min. Allicin in the
eluent was detected at 202 and 210 nm. Allicin was
RESULTS AND DISCUSSION
quantified using the following equation:
Allicin is not available in the market because of its
C × FV × D × 100% instability. DADS, DATS, DAS, DAT and allyl methyl
Allicin (%, w/w) =
W (1000 μ g/mg) disulfide are the main degradation products of allicin
and allyl methyl thiosulfinates (Vernin et al., 1986;
where, Lawson et al., 1991). In this study, we used DADS,
C = The allicin concentration (µg/mL) obtained from DATS, DAS and DAT as standards. Unlike the HPLC
the standard curve and GC methods, the spectrophotometry method did not
FV = The final sample volume (mL) require the use of standards. In the GC method (Fig. 3),
D = The dilution of the sample the standards were separated and identified by three
W = The sample weight (mg) discrete peaks: DAS with a retention time of 3.549 min,
271
 Adv. J. Food Sci. Technol., 9(4): 269-277, 2015

Fig. 3: GC chromatogram of standards

DAS: Retention time of 3.549 min; DADS: Retention time of 6.077 min; DATS: Retention time of 8.932 min

(a)

(b)

Fig. 4: HPLC chromatogram of standards. DAS (peak 1; retention time of 6.941 min) and DAT (peak 2; retention time of 8.706
min); (a): were detected at 202 nm. DADS (peak 3; retention time of 12.634 min) and DATS (peak 4; retention time of
17.794 min); (b): were detected at 210 nm

DADS with a retention time of 6.077 min and DATS the GC method; the lowest detection limit of HPLC was
with a retention time of 8.932 min. However, a DAT lower than that of GC. The average Coefficient of
peak was not observed. On the other hand, in the HPLC Variation (CV) of the spectrophotometric, GC and
method, all four standards were separated and identified HPLC method was 3.44, 8.10 and 4.68, respectively
by four distinct peaks (Fig. 4a and b). DAS (retention (Table 2). When 5%<CV<10%, the precision of the
time of 6.941 min) and DAT (retention time of 8.706 instrument is adequate (Osman and Chin, 2006). Based
min) had maximum absorbance readings at 202 nm; on the results, the spectrophotometric and HPLC
DADS (retention time of 12.634 min) and DATS methods were more precise than the GC method.
(retention time of 17.794 min) had maximum The allicin concentration of the garlic-based
absorbance readings at 210 nm (Fig. 5). The results products was determined by the three methods. In the
revealed that the HPLC method was more sensitive than spectrophotometric method, allicin concentration in the
272
 Adv. J. Food Sci. Technol., 9(4): 269-277, 2015

Fig. 5: Ultraviolet spectra of standards. DAS and DAT were detected at 202 nm, DADS and DATS were detected at 210 nm

Table 2: Allicin concentration in the eight garlic products

Allicin (%) Sp CV GC CV HPLC CV
Gp1 61.150 4.82 59.40 8.99 76.39 5.24
Gp2 0.410 3.97 0.36 7.67 0.39 3.80
Gp3 0.480 4.76 0.46 7.94 0.47 6.48
Gp4 0.430 2.46 0.41 6.84 0.43 7.24
Gp5 0.619 4.28 0.56 10.55 0.61 4.16
Gp6 0.570 4.79 0.54 6.23 0.55 2.08
Gp7 0.810 1.06 0.67 12.34 0.68 3.17
Gp8 0.610 1.41 0.58 4.25 0.60 5.25
CV: Coefficient of variation; Sp: Spectrophotometric

products was 0.41-61.15%. Based on the

spectrophotometric method, garlic oil contained the
highest allicin concentration (61.15%) followed by the
garlic tablets (0.65%) and garlic powder (0.44%).
According to the GC method, garlic oil had the highest
allicin concentration (59.41%); garlic tablets and garlic
powder contained 0.59 and 0.41% allicin, respectively.
According to the HPLC method, garlic oil contained the
highest allicin concentration (76.39%) followed by
garlic tablets (0.61%) and garlic powder (0.43%).
Regardless of the quantitative method, allicin
concentration was significantly different between garlic
oil and garlic powder and between garlic oil and garlic Fig. 6: Allicin concentration in the garlic products based the
tablets. However, there were no significant differences spectrophotometric, CG and HPLC methods. In Gp1,
in allicin concentration between garlic powder and allicin concentration was multiplied by 100
garlic tablets (p>0.05). Garlic oil had an allicin
concentration of 76.39% based on the HPLC method from the HPLC method. Unlike the GC and HPLC
and of 59.41% based on the GC method. The allicin methods, the spectrophotometric method did not require
concentrations in the garlic tablets and powder were allicin standards for the quantitation of allicin because
higher by the spectrophotometric method than by the this method is based on the quantitation of cysteine
GC method (Fig. 6). The spectrophotometric method is using the established DTNB/NTB system (Li and Xu,
simple and provided similar results to those obtained 2007). Compared to the GC and HPLC methods, the
273
 Adv. J. Food Sci. Technol., 9(4): 269-277, 2015

spectrophotometric method was simpler, easier and The results revealed that, while allicin can be
more efficient. The spectrophotometric method is based detected by GC, most of the compound was degraded
on the formation of disulfide compounds from the during chromatography. Therefore, the HPLC method is
reaction between sulfhydryl-containing compounds and more sensitive and accurate for the analysis of sulfur-
activated disulfide bonds of thiosulfinates. Therefore, containing compounds in garlic-based products. The
the spectrophotometric method can only provide the HPLC chromatogram was characterized by four peaks
concentration of total allicin and not of specific-sulfur with the characteristic UV maxima of DAS, DAT,
containing compounds (Han et al., 1995; Miron et al., DADS and DATS. The HPLC chromatograms of garlic
2002). However, the GC and HPLC methods can oil (Gp1), garlic powder (Gp3) and garlic tablets (Gp7)
provide the concentration of allicin and of specific- are shown in Fig. 7a to f. According to the HPLC
sulfur containing compounds (Table 3 and 4). The results, garlic oil consisted of 43.12% DADS, 17.58%
results revealed that the GC method had the lowest DATS, 14.22% DAT and 1.47% DAS (Table 3). A
efficiency for the quantitation of allicin in garlic-based typical commercial preparation of garlic oil contains
products. A comparison between the HPLC and GC 26% DADS, 19% DATS, 15% allyl methyl trisulfide
methods revealed that garlic oil might have been and 13% allyl methyl disulfide (Lawson, 1998). Garlic
distilled and that the compounds were degraded at high oil from Jinxiang County contained a higher
temperature. The allicin concentration in garlic oil by concentration of allicin than commercial garlic oil.
the HPLC method was higher than that by the CG According to the British Pharmacopoeia, the minimum
method. Regardless of the method (GC or HPLC), allicin concentration required to ensure its
DADS and DATS were the major allicin pharmaceutical and economical viability is 0.45% in
compounds in the garlic-based products especially in garlic powder (Baghalian et al., 2005). The total allicin
the garlic tablets and garlic powder. On the other hand, concentration in the garlic- based products was higher
the concentrations of DAS and DAT were insignificant than required for pharmaceutical purposes, with the
(Table 4). exception of Gp2 and Gp4 garlic powder.

Table 3: DAS, DAT, DADS and DATS concentration by the GC method

Sample DAS DAT DADS DAT TAC (%)
Gp1 5.82 0 11.41 42.20 59.41±0.39
Gp2 0 0 0.16 0.20 0.36±1.20
Gp3 0 0 0.18 0.28 0.46±0.06
Gp4 0 0 0.15 0.26 0.41±0.48
Gp5 0 0 0.27 0.29 0.56±0.97
Gp6 0 0 0.30 0.24 0.54±0.30
Gp7 0 0 0.35 0.32 0.67±0.27
Gp8 0 0 0.31 0.27 0.58±0.48
TAC: Total allicin concentration

Table 4: DAS, DAT, DADS and DATS concentration by HPLC method

Sample DAS DAT DADS DATS TAC (%)
Gp1 1.470 14.220 43.120 17.580 76.39±0.12
Gp2 0.037 0.017 0.270 0.066 0.39±1.01
Gp3 0.047 0.038 0.290 0.095 0.47±0.28
Gp4 0.049 0.032 0.265 0.084 0.43±0.23
Gp5 0.053 0.029 0.445 0.083 0.61±0.52
Gp6 0.069 0.030 0.368 0.083 0.55±2.98
Gp7 0.076 0.030 0.477 0.097 0.68±0.34
Gp8 0.036 0.021 0.436 0.107 0.60±0.16
TAC: Total allicin concentration

mAU
202nm4nm (1.00)

3000

2000
2#

1000
1#

0
0.0 5.0 10.0 15.0 20.0 25.0 30.0 min

(a)
274
 Adv. J. Food Sci. Technol., 9(4): 269-277, 2015

mAU

3#
4000 210nm4nm (1.00)

3000

2000

1000

4#
0

0.0 5.0 10.0 15.0 20.0 25.0 30.0 min

(b)

mAU
202nm4nm (1.00)

300

200
2#

100
1#

0.0 5.0 10.0 15.0 20.0 25.0 30.0 min

(c)
mAU
3#

210nm4nm (1.00)
300

200
4#

100

0.0 5.0 10.0 15.0 20.0 25.0 30.0 min

(d)
mAU
202nm4nm (1.00)

500

250
1#

2#

0.0 5.0 10.0 15.0 20.0 25.0 30.0 min

(e)

275
 Adv. J. Food Sci. Technol., 9(4): 269-277, 2015

mAU
400 210nm4nm (1.00)

300

200

3#
100

4#
0

0.0 5.0 10.0 15.0 20.0 25.0 30.0 min

(f)

Fig. 7: HPLC chromatogram of samples; (a): HPLC chromatogram of DAS (peak 1) and DAT (peak 2); These compounds were
detected at 202 nm in garlic oil (Gp1); (b): HPLC chromatogram of DADS (peak 3) and DATS (peak 4); These compounds
were detected at 210 nm in garlic oil (Gp1); Dilution of garlic oil: 10,000; (c): HPLC chromatogram of DAS (peak 1) and
DAT (peak 2); These compounds were detected at 202 nm in garlic powder (Gp3); (d): HPLC chromatogram of DADS
(peak 3) and DATS (peak 4); These compounds were detected at 210 nm in garlic powder (Gp3); (e): HPLC chromatogram
of DAS (peak 1) and DAT (peak 2); These compounds were detected at 202 nm in garlic tablets (Gp7); (f): HPLC
chromatogram of DADS (peak 3) and DATS (peak 4); These compounds were detected at 210 nm in garlic oil (Gp7)

The spectrophotometric method was efficient to DAS : Diallyl sulfide

measure total allicin concentration. The low costs DAT : Dimethyl trisulfide
associated with this quantitation method make it useful DADS : Diallyl disulfide
to assess the pharmaceutical properties of garlic-based DATS : Diallyl trisulfide
products. For component analysis and international
trade, the HPLC method is more adequate. Therefore, ACKNOWLEDGMENT
the spectrophotometric method can be used to measure
total allicin concentration, which is an important index This work was supported by Doctoral Research
of garlic quality. However, the HPLC method is more Start-up Fund Project of Jiangxi Science and
applicable for the analysis of allicin. Consequently, Technology Normal University (2015), the Natural
laboratories that do not have HPLC instruments can Science Foundation of Jiangxi Province, China (Grant
assess allicin concentration in garlic and garlic-based
No. 20151BAB204039), Quality engineering projects of
products by the spectrophotometric method.
Jiangxi Science and Technology Normal University
(Biological chemistry curriculums, 2010) and the Public
CONCLUSION
Sector (Agriculture) Project Special Funds for Scientific
Research (201303112).
The results of this study revealed that the GC
method was not suitable for the quantitation of allicin,
whereas the spectrophotometric method was simpler and REFERENCES
suitable for solid garlic products. The HPLC method
was more accurate for the quantitation of allicin in all Ali, M., M. Thomson and M. Afza, 2000. Garlic and
garlic-based products, especially liquid garlic-based onions: Their effect on eicosanoid metabolism and
products. Regardless of the quantitative method, it could its clinical relevance. Prostag. Leukotr. Ess.,
be noticed that allicin content in garlic oil was higher 62(2): 55-73.
than that of garlic tablets and garlic powder. Based on Bachrach, G., A. Jamil, R. Naor, G. Tai, Z. Ludmer and
the spectrophotometric and HPLC method, garlic oil D. Steinberg, 2011. Garlic allicin as a potential
contained the highest allicin concentration (61.15 and agent for controlling oral pathogens. J. Med. Food,
76.39%) followed by the garlic tablets (0.65 and 0.61%) 14: 1338-1343.
and garlic powder (0.44 and 0.43%), respectively. Baghalian, K., S.A. Ziai, M.R. Naghavi, H.N. Badi and
A. Khalighi, 2005. Evaluation of allicin content
ABBREVIATIONS and botanical traits in Iranian garlic (Allium
sativum L.) ecotypes. Sci. Hortic., 103: 155-166.
GC : Gas chromatography Baneriee, S.K. and S.K. Maulik, 2002. Effect of garlic
HPLC : High pressure liquid chromatography on cardiovascular disorders: A review. Nutr. J., 1:
CV : Coefficient of variation 1-14.
276
 Adv. J. Food Sci. Technol., 9(4): 269-277, 2015

Calvey, E.M., J.A. Roach and E. Block, 1994. Miron, T., A. Rabinkov, D. Mirelman, M. Wilchek and
Supercritical fluid chromatography of garlic L .Weiner, 2000. The mode of acton of allicin: Its
(Allium sativum) extracts with mass spectrometric ready permeability through phospholipid
identification of Allicin. J. Chromatogr. Sci., 32: membranes may contribute to its biological
93-96. activity. Biochim. Biophys. Acta, 1463: 20-30.
Han, J., L. Lawson, G. Han and P. Han, 1995. A Miron, T., I. Shin, G. Feigenblat, L. Weiner,
spectrophotometric method for quantitative D. Mirelman, M. Wilchek and A. Rabinkov, 2002.
determination of allicin and total garlic A spectrophotometric assay for allicin, alliin and
thiosulfinates. Anal. Biochem., 225: 157-160. alliinase (alliin lyase) with a chromogenic thiol:
Reaction of 4-mercaptopyridine with thiosufinates.
Hirsch, K., M. Danilenko, J. Giat, T.A. Miron,
Anal. Biochem., 307: 76-83.
Rabinkov, M. Wilchek, D. Mirelman, J. Levy and Mondy, N., A. Naudin, J.P. Christides, N. Mandon and
Y. Sharoni, 2000. Effect of purified allicin, the J. Auger, 2001. Comparison of GC-MS and HPLC
major ingredient of freshly crushed garlic, on for the analysis of Allium volatiles.
cancer cell proliferation. Nutr. Cancer, 38: Chromatographia, 53: 356-360.
245-254. Oommen, S., R.J. Anto, G. Srinivas and
His, Y. and C. Wu, 1989. Stability of Allicin in garlic D. Karunagaran, 2004. Allicin (from garlic)
juice. Food Sci., 54: 977-981. induces caspase-mediated apoptosis in cancer cells.
Khar, A., K. Banerjee, R.J. Manjusha and Eur. J. Pharmacol., 485: 97-103.
K.E. Lawande, 2011. Evaluation of garlic ecotypes Osman, H. and Y.K. Chin, 2006. Comparative
for allicin and other allyl thiosulphinates. Food sensitivities of cholesterol analysis using GC,
Chem., 128: 988-996. HPLC and spectrophotometric methods. Malays.
Lawson, L.D., 1998. Garlic: A review of its medicinal J. Anal. Sci., 10: 205-210.
effects and indicated active compounds. In: Sigounas, G., J. Hooker, A. Anagnostou and M. Steiner,
Lawson, L.D. and R. Bauer (Ed.), Phytomedicines 1997. S-allylmercaptocysteine inhibits cell
of Europe: Chemistry and Biological Activity. proliferation and reduces the viability of
ACS Symposium Series 691, American Chemical erythroleukemia, breast and prostate cancer cell
Society, Washington, DC, pp: 176-209. lines. Nutr. Cancer, 27: 186-191.
Lawson, L.D. and C.D. Gardner, 2005. Composition, Song, K. and M.A. John, 2001. The influence of
stability and bioavailability of garlic products heating on the garlic anticancer properties. Am.
being used in a clinical trial. J. Agr. Food Chem., Soc. Nutr. Sci., 31: 1054S-1057S.
53: 6254-6261. Tripathi, K., 2009. A review-garlic, the spice of life-
Lawson, L.D., Z.J. Wang and B.G. Hughes, 1991. (Part-I). Asian J. Res. Chem., 2: 8-13.
Identification and HPLC quantitation of the Velisek, J., R. Kubec and J. Davidk, 1997. Chemical
sulfides and dialk (en) yl thiosulfinates in composition and classification of culinary and
commercial garlic products. Planta Med., 57: pharmaceutical garlic-based products. Z. Lebensm.
363-370 (PubMed: 1775579). Unters. F. A, 204: 161-164.
Lemar, K.M., M.A. Aon, S. Cortassa, B.O. Rourke, Vernin, G., J. Metzger, D. Fraisse and C. Scharff, 1986.
C.T. Muller and D. Lioyd, 2007. Diallyl disulphide GC-MS (EI, PCI, NCI) computer analysis of
depletes glutathione in Candida albicans: Oxidative volatile sulfur compounds in garlic essential oils
stress-mediated cell death studied by two-photon Application of the mass fragmentometry SIM
microscopy. Yeast, 24: 695-706. technique. Planta Med., 52: 96-101 (PubMed:
Li, Y. and S.Y. Xu, 2007. Preparation of garlic powder 17345207).
with high allicin content. Agric. Sci. China, 6:
890-898.

277