MEDICINAL

CHEMISTRY
RESEARCH

Med Chem Res (2010) 19:10921105

DOI 10.1007/s00044-009-9255-z
ORIGINAL RESEARCH

Extraction of essential oils from garlic (Allium sativum)

using ligarine as solvent and its immunity activity
in gastric cancer rat
Rui Li Wei-chang Chen Wei-peng Wang
Wen-yan Tian Xue-guang Zhang

Received: 9 May 2008 / Accepted: 2 September 2009 / Published online: 24 December 2009
Birkhauser Boston 2009

Abstract The solvent extraction (SE) of garlic essential oil (Allium sativum) was
studied. A multivariate study based on a four-factor, three-level BoxBehnken
design (BBD) was used to evaluate the influence of four major variables affecting
the performance of the SE of garlic essential oil. The yield and the composition of
the essential oils from garlic obtained by SE were determined, and compared with
those obtained by the supercritical fluid extraction (SFE). Statistical treatment of the
results provided by the BBD revealed that the selected parameters, extraction time
and extraction temperature, were significant. The essential oils were analyzed by gas
chromatographymass spectrometry (GCMS). Major essential oil components
were 3-vinyl-4H-1,2-dithiin (31.89%), diallyl trisulfide (13.31%), diallyl sulfide
(2.22%), dially disulfide (6.87%), propyl allyl disulfide (13.89%), and dimethyl
disulfide (7.05%). Compared with SFE, the yield of essential oil obtained by SE was
slightly lower, but major essential oil components were quantitatively similar.
Moreover, residual solvent in garlic essential oil was very low (\50 mg/kg). In
addition, a significant increment in the ratio of CD4?/CD8? in serum of gastric
cancer rats was determined after administration of garlic essential oil. It can be

R. Li  W. Chen (&)  W. Tian

Department of Gastroenterology, The First Affiliated Hospital of Soochow University,
Suzhou, China
e-mail: chenwc_prof@yahoo.cn
R. Li  W. Chen  W. Tian  X. Zhang
Key Laboratory of Medicine and Clinical Immunology of Jiangsu Province, Suzhou 215006, China
W. Wang
College of Pharmacy, Soochow University, Suzhou 215007, China
X. Zhang
Institute of Medical Biotechnology, Soochow University, Suzhou 215007, China

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concluded, that the SE method offers obvious advantages over SFE. Moreover, the
results indicated that garlic essential oil may be useful for treatment of patients with
inflammatory disease, e.g., gastric cancer.
Keywords BoxBehnken design  Garlic  Essential oils  Extraction 
GCMS  Supercritical fluid extraction  CD4?/CD8?  Gastric cancer 
CD40

Introduction
Garlic (Allium sativum) is widely used for flavoring purposes in food, especially in
the Nordic countries Sweden and Norway, the United Kingdom, and in Asia. Garlic
(Allium sativum) has played an important dietary as well as medicinal role for
centuries (Chauhan 2006; Eidi et al., 2006; Khajeh et al., 2004). The major
components of garlic essential oil are 3-vinyl-4H-1,2-dithiin, diallyl trisulfide,
diallyl sulfide, dially disulfide, propyl allyl disulfide, and dimethyl disulfide. Bordia
et al., (1998) reported that diallyl disulfide and diallyl trisulfide, the two major
organosulfur compounds derived from garlic essential oil, could inhibit platelet
thromboxane formation, and hence platelet aggregation. Shenoy and Choughuley
(1992) further indicated that garlic (Allium sativum L.) essential oil were effective in
reducing the chemical formation of N-nitrosamines (NAs), and suggested that
diallyl sulfur compounds in garlic essential oil might also act as nitrite scavengers.
Supercritical fluid extraction (SFE) of active compounds from plant material is a
promising field for the industrial application of SFE (Reverchon 1997; Quynh et al.,
2008) since it has certain advantages over steam distillation and solvent extraction
(SE). SFE can be performed at lower temperatures, thereby preserving the original
extract composition and properties. Unfortunately, SFE method also has some
disadvantages that prohibit its industrial uses because of high equipment investment
and small extraction scale.
The essential oils of the native spices have been isolated using steam distillation
and/or SE (Xu et al., 2009; Khajeh et al., 2004; Kitzberger et al., 2007). Although
people conventionally think that low yield, loss of volatile compounds, long
extraction times, toxic solvent residue, degradation of unsaturated compounds, and
production of undesirable off-flavors (due to heat) are often involved in using such
techniques, the SE method offers obvious advantages over SFE. To date, there have
been a few successful reports using such techniques (Cho et al., 2007; Franco et al.,
2007; Chyau et al., 2007).
In this article, the SE of garlic essential oil has been studied to understand in
depth and to optimize this classical technique of extraction. A four-factor, threelevel BoxBehnken design (BBD) has been developed to specify the importance of
the four major factors affecting the SE. The yields and the composition of each
sample of essential oils resulting from the experiments of the BBD have been
analyzed and compared. This study still describes a comparative study of classical
SE and SFE of garlic essential oils. Chemical compositions of the SFE extract and
essential oil obtained by SE were analyzed by GC and GC/MS methods. We still

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investigated the role of garlic essential oil in inflammation and immune response by
measuring the mean (?SE) percentage of CD3?, CD4?, and CD8? and the ratio of
CD4?/CD8? in gastric cancer rats.

Materials and methods

Materials
Garlic was purchased from local market (Suzhou City, China). During the
experiment, all the samples were maintained in our lab at appropriate conditions
(dark, 25C). In each batch, 50 g of garlic was blended with 20-ml deionized water
for 2 min in a commercial blender. Four different solvents were used for the
distillations and the extractions of garlic samples: pure ethanol, n-pentane, ligarine,
and ethyl acetate.
Extraction of essential oil
The ground garlic samples (50 g) were weighed and quantitatively transferred into a
500-ml flask, then extracted with different volume (1, 2, 3, 4, 5) of organic solvent
(ethanol, ligarine, ethyl acetate, or n-pentane) for a given time (10, 20, 30, 40, 50,
60, and 70 min) when extraction temperatures were, respectively set at 20, 25, 30,
35, 40, 45, and 50C. After extraction, the organic layer was filtered and collected.
The water layer was extracted with another equal volume of the solvent. The
extraction procedure was repeated several times (15). The combined filtrate was
concentrated using rotary vacuum evaporator at 40C and weighed on an analytical
balance to give the extraction rate of essential oil.
The remaining solvent in the essential oil extract was determined by GC analysis.
Experimental design
A four-factor, three-level BoxBehnken design (Yin and Dang 2008; Muthukumar
et al., 2003) was applied to determine the best combination of extraction variable
parameters for the production of essential oil from garlic. The factorial design
consisted of three center points and six BBD Blocks, leading to 27 sets of
experiments. The variables Xi were coded as xi according to the following
Equation:
xi Xi  Xi=2=DXi;
where xi is the coded value of an independent variable, Xi is the real value of an
independent variable, Xi/2 is the real value of an independent variable at the central
point, and DXi is the step change value. The variables and their levels, with both
coded values and natural values investigated in this study, are represented in
Table 1. Table 2 lists the actual experimental parameters corresponding to the

Med Chem Res (2010) 19:10921105

Table 1 The level of variables
chosen for the BoxBehnken
design

1095

Variable

Coded levels
-1

?1

Extraction temperature X1 (C)

35

40

45

Extraction time X2 (min)

30

40

50

Ratio of liquid to raw material X3

Extraction number X4

Natural levels

Table 2 BoxBehnken design

with coded values for four
extraction parameters and results

Run

X1

X2

X3

X4

-1

-1

0.38

-1

0.53

-1

0.57

0.72
0.46

-1

-1

-1

0.59

-1

0.67

0.69

-1

-1

0.46

10

-1

0.51

11

-1

0.65

12

0.73

13

-1

-1

0.47

14

-1

0.68

15

-1

0.55

16

0.71

17

-1

-1

0.34

18

-1

0.63

19

-1

0.66

20

0.79

21

-1

-1

0.53

22

-1

0.59

23

-1

0.62

24

0.65

25

0.67

26

0.67

27

0.67

designed levels that were carried out for developing the model. Each experiment
was performed in duplicate and the average of extraction yield of essential oil was
taken as the response, Y.

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The responses obtained from each set of experimental design (Table 2) were
subjected to multiple nonlinear regression using the software SAS 8.0 (SAS
Institute, Inc) to obtain the coefficients of the second polynomial model. The quality
of the fit of the polynomial model equation was expressed by the coefficient of
determination R2, and its statistical significance was checked by an F-test. The
significances of the regression coefficient were tested by a t-test.
Supercritical fluid extraction of garlic essential oil
Supercritical fluid extraction of garlic essential oil was performed according to the
method described by Gouveia et al., (2007) and Wang et al., (2006).
The experiment was carried on a HP 7680 A supercritical fluid Extraction Unit
(Hewlett Packard, Les Ulis, France). The extraction column was packed with
powdered raw materials and glass beads. The liquid CO2 from a cylinder was
compressed to the working pressure at the temperature of a water bath. The pressure
was controlled by a back-pressure regulator, and the fluid, before reaching the
extractor, passed through a coil immersed in a water bath at a temperature above the
supercritical one. The supercritical CO2 with dissolved compounds was passed
through a heated micrometer valve, and subsequently expanded to ambient pressure,
and the solutes were collected in cooled glass U-tubes filled with glass wool. Gas
flow rate was monitored with a rotameter, and the total volume of gas was measured
with a wet test meter.
The experimental conditions were : (i) density of CO2: 0.6 g/ml; (ii) flow of CO2:
1 ml/min; (iii) equilibrium time: 5 min; (iv) experiment temperature: 35C; (v) trap
temperature: 0C; (vi) extraction time: 130 min; and (vii) extraction pressures:
10 MPa.
For each extraction test, the extractor was charged with about 50 g of ground
garlic. The oil weight was measured by precision balance until no oil was extracted
out from the ground garlic.
Gas chromatography, gas chromatographymass spectrometry (GC, GCMS)
analysis
The essential oil was analyzed using a Hewlett Packard 5890 II GC equipped with a
FID detector and HP-5ms capillary column (30 m 9 0.25 mm, film thickness
0.25 lm). For GC/MS detection, an electron ionization system with ionization
energy of 70 eV was used. Injector and detector temperatures were set at 220 and
290C, respectively. Column temperature was initially kept at 50C for 3 min, then
gradually increased to 240C with a rate of 3C/min. Helium was the carrier gas, at
a flow rate of 1 ml/min. Samples of 1 ll were injected manually and in the splitless
mode. Tentative identification of the compounds was based on the comparison of
their mass spectra with those of NIST 98 and Wiley 275 library data of the GC/MS
system. Quantitative data were obtained electronically from FID area percent data
without the use of correction factors.

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Animals and treatment

Male Wistar rats (245 10 g) were housed in steel cages. They were allowed free
access to drinking water and normal diet during the experiment. The rats were
acclimatized for 7 days and randomly allotted into four groups. All the animal
experiments were performed under the guidelines of the Laboratory Animal
Experiment Committee of China. Rat gastric cancer model induced by 1-methyl-3nitro-1-nitrosoguanidine was performed in all animals.
Rats were allotted into three groups: normal rats (group I) and essential oil-fed
rats (groups II and III). Rats of groups II and III orally received essential oil at a
single dose of 50 mg/kg and 100 mg/kg, respectively. Rats in group I orally
received an equal amount of vehicle. After 4 weeks, all the animals were sacrificed.
Four months after essential oil treatment, rats were euthanized and bled from the
abdominal aorta. Blood samples were collected in heparinized tubes and centrifuged
(4C) at 15009g for 15 min. Plasma were stored at -70C until use.
CD4/CD8 cell estimation
Two milliliters of blood was drawn from these individuals between 08:00 and
10:00 h into 5-ml collection tubes containing 60 ll of 5% EDTA. The Capcellia
CD4/CD8 whole blood kit from Sanofi DIAGNOSTICS Pasteur, (France) was
used to measure CD4 and CD8 counts. This is an immuno-capture ELISA-based
assay in which paramagnetic particles coated with anti-pan-T cell antibodies
capture T-lymphocytes. The unbound cells are washed using a manual washing
manifold connected to an electric vacuum pump while keeping the plate on a
magnetic frame. The T cell subsets are estimated using peroxidase labeled
monoclonal antibodies specific to CD4 and CD8 cells, respectively. Tetra methyl
benzidine (TMB) is the substrate used in this system. Four cell standards and a
blank each for CD4 and CD8 were included as mentioned in the protocol for the
validation and interpretation of the results. A few modifications were introduced
such as manual rotation for 18 min during the 20-min incubation after the
addition of the conjugate and gentle manual rotation of the plate for 15 min after
the addition of substrate. The readings were taken using a ELx800 Automated
Microplate ELISA reader (Bio-Tek, USA) at a wavelength of 450 nm with a
reference wavelength of 630 nm. The cell counts specified for each of the four
standards and the blank (considered to have 0 cells) as recommended by the
kit manufacturer were fed to the reader prior to taking reading thus enabling the
software to draw a line of best fit. Whenever a given standard was not close to
the expected value, that particular standard was omitted and reading was taken by
feeding the other three standard values and the blank. The calibrated reader
provided the numbers of cells corresponding to the OD values. Though the kit
manufacturer recommended each sample to be tested as singletons, we tested all
the samples in duplicate wells. The data presented in this report are of tests done
within 2 h of collection of blood samples.

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CD40 analysis
CD40 expression rate was measured according to our past method (Li et al., 2009).

Results and discussion

Effect of different solvents on extraction yield of essential oil
Table 3 showed that maximum extraction yield of garlic essential oil was, in turn,
0.71% (ethanol), 0.69% (ethyl acetate), 0.75% (ligarine), and 0.76% (n-pentane).
Results showed that yield of the extract was the highest when ligarine was used as a
solvent. Moreover, the quality of the essential oil extract was the best. Therefore, we
used ligarine as the solvent to extract garlic essential oil in this work.
Effect of extraction temperature on extraction yield of essential oil
Increasing the extraction temperature increased the essential oil extraction (Fig. 1a).
The average essential oil yields from garlic in 50 min were 0.34, 0.43, 0.52, 0.69,
0.74, 0.78, and 0.78% of total oil at extraction temperature of 20, 25, 30, 35, 40, 45,
and 50C, respectively. This indicated that the increase in extraction temperature,
within the range investigated, increased the mass transfer coefficient resulting in
higher extraction rates from garlic. Therefore, optimum extraction temperature was
chosen as 45C.
Effect of extraction time on extraction yield of essential oil
The effect of extraction time on extraction yield of essential oil was shown in
Fig. 1b. The extraction yield of essential oil increased with the increase of
extraction time in the range of 1060 min, in general. There existed maximum
extraction yield of essential oil when extraction time was 60 min. Extraction time of
60 min is recommended based on the experimental results.
Effects of the ratio of solvent to raw material on extraction yield of essential oil
The effect of the ratio of solvent to raw material on extraction yield of essential oil
is shown in Fig. 1c. As expected, yields increased with the increase in the ratio of
solvent to raw material at a given condition, in fact due to the increase of oil
solubility. This increase results from the decrease of the concentration of the extract
Table 3 Effect of different solvents on extraction yield of essential oil
Ethanol
Extraction yield (%)
Residual solvent (mg/kg)

0.71
53

n-Pentane
0.76
40

Ligarine
0.75
35

Ethyl acetate
0.69
57

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Fig. 1 Effect of extraction temperature on (a), extraction time (b), ratio of solvent to raw material (c)
extraction number (d) extraction yield of essential oil

solution, leading to the increase of its solvent power. As is shown in Fig. 1c, the
extraction yield of essential oil rapidly increased up to 0.78% when the ratio of
solvent to raw material varied from 1 to 4, and then no longer increased. Therefore,
the recommended ratio of solvent to raw material is between 3 and 5.
Effects of number of extractions on extraction yield of essential oil
The effects of varying extraction number over a range of 15 on the production of
essential oil were shown in Fig. 1d. The extraction yield of essential oil rapidly
increased up to 0.77% when extraction number varied from 1 to 4 and then no
longer increased. This indicates that essential oil in garlic sample has been
completely extracted when extraction number is 4.
Optimization of extraction conditions
A three-factor three-coded level BoxBehnken design was used to determine the
response value (Y) for different extraction parameters. Extraction temperature (X1),
extraction time (X2), ratio of solvent to raw material (X3), and extraction number
(X4) were independent variables studied to predict the response (Y). Independent
variables and their levels for the BoxBehnken design used in this study are shown
in Table 1. Using the relationships in Table 1, the actual levels of the variables for
each of the experiments in the design matrix were calculated, and the experimental
results obtained are as given in Table 2.
Table 2 shows the experimental conditions and the results of essential oil
production according to the factorial design. Maximum extraction yield of essential
oil (0.79%) was recorded under the experimental conditions of extraction

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temperature 45C, extraction time 40 min, ratio of solvent to raw material 4, and
extraction number 4 in the test set No. 20.
Multiple regression analysis was performed on the experimental data, and the
coefficients of the model were evaluated for significance with the Student t-test. The
four linear coefficients, and three quadratic terms (extraction temperature, extraction
time, and extraction number) except the ratio of solvent to raw material were
significant (P \ 0.05). All the cross-product coefficients except X1X3 were
eliminated in the refined equation as the P values of these coefficients are very
insignificant (P [ 0.05). The analysis of fit Statistics for Y is shown in Table 4. The
goodness of fit of the model was checked by determination coefficient (R2). In this
case, the value of the determination coefficient (R2 = 0.965) indicated that only
3.5% of the total variations were not explained by the model. The value of the
adjusted determination coefficient (Adj. R2 = 0.925) was also very high to advocate
for a high significance of the model. At the same time, a relatively lower value of
the coefficient of variation (CV = 5.05%) indicated a better precision and reliability
of the experiments carried out. Neglecting the insignificant terms, the final
predictive equation obtained is as given below:
Y 0:67 0:105833 X1 0:046667 X2 0:091667 X3 0:030833 X4
 0:054583 X1 X1  0:04 X1 X3  0:050833 X2 X2  0:032083 X4 X4
Statistical analysis revealed that the most relevant variables (P \ 0.001) for the
essential oil production were extraction temperature and ratio of solvent to raw
material. The predicted maximum extraction yield (0.782%) derived from RSM
regression was obtained when extraction temperature is 45C, extraction time
42 min, ratio of solvent to raw material 4, and extraction number 4.
Effect of variables on response value (Y)
In order to gain a better understanding of the results, the predicted models with one
variable kept at central level and the other two varying within the experimental
ranges are presented in Fig. 2af as the 3D response surface plots and Fig. 3af as
the 2D contour plots for extraction yield, respectively. The obvious trough in the
response surfaces indicates that the optimal conditions were exactly located inside
the design boundary (Fig. 2af, and Fig. 3af).
Figures 2a and 3a show the effect of extraction temperature and extraction time
on extraction yield. The figures show good results both near center of extraction
Table 4 Fit statistics for Y

Master model
Mean

0.59963

Predictive model
0.59963

R-square

96.52%

96.52%

Adj. R-square

92.47%

92.47%

RMSE

0.030311

0.030311

CV

5.054935

5.054935

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1101

time and high extraction temperature level studied. Figures 2b and 3b show the
effect of extraction temperature and ratio of solvent to raw material on extraction
yield and Figs. 2c and 3c show the effect of extraction temperature and extraction
number on extraction yield. The figures show good results both near center of
extraction number and high extraction temperature level studied. Figures 2d and 3d
show the effect of extraction time and ratio of solvent to raw material on extraction
yield. Figures 2e and 3e show the effect of extraction time and extraction number on
extraction yield. Figures 2f and 3f show the effect of extraction number and ratio of
solvent to raw material on extraction yield. The figures show good results both near
center of extraction number and high ratio of solvent to raw material level studied. It
shows the maximum extraction yield near center of extraction number and high
ratio of solvent to raw material.
In the contour plots for extraction yield, the more steep the curve was the more
significant the factors on the response value were. According to the principle,
extraction temperature and ratio of solvent to raw material were two significant
factors on extraction yield. That is, a slight change in extraction temperature or in
ratio of solvent to raw material could lead to a greater extraction yield.
Confirmation experiments
In order to confirm the validity of the statistical experimental strategies, one run of
additional confirmation experiments was conducted. The chosen conditions for
extraction temperature, extraction time, ratio of solvent to raw material, and
extraction number are all listed in Table 5, along with the predicted and measured
results. As shown in Table 5, the measured extraction yield were close to those

Fig. 2 Surface graphs of extraction yield showing the effect of variables: a X1X2, b X1X3, c X1X4,
d X2X3, e X2X4, and f X3X4

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Fig. 3 Contour plots of extraction yield showing the effect of variables: a X1X2, b X1X3, c X1X4, d
X2X3, e X2X4, and f X3X4

estimated using RSM. This also testifies that the RSM approach was appropriate for
optimizing the operational conditions of the extraction process.
Composition of garlic oil obtained by different methods
Table 6 listed the composition of garlic essential oil obtained by two different
methods according to the results of GCMS. It could be seen that 23 compounds in
the garlic essential oil had been identified, in which 3-vinyl-4H-1,2-dithiin, diallyl
trisulfide, propyl allyl disulfide, and dimethyl disulfide were the main components
of garlic essential oil. The composition of the garlic essential oil extracted by two
different methods was mostly similar, and relative concentrations of the identified
main compounds were slightly different. This result is slightly different from the
other authors findings (Qiao et al., 2001; Lin et al., 2005). We postulate that
different solvents used in the SE method is responsible for the difference.

Table 5 Optimum conditions, predicted, and experimental value of response at that condition
Optimum condition

Extraction yield (%)

Extraction
temperature (C)

Extraction time Ratio of solvent to raw Extraction

(min)
material
number

Experimental Predicted

45

42

0.776

0.782

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Table 6 Chemical composition of garlic essential oil

NO.

Compounds

3-methylthio propanal

0.3

1,2-epithiopropane

0.34

Diallyl sulfide

2.3

2.22

Methyl propyl disulfide

1.12

1.06

3-methyl-2-cyclopentene-1-thione

0.52

Vinylfuran

0.98

0.92

Methyl allyl disulfide

1.92

1.87

Dimethyl disulfide

7.44

7.05

Methyl butyl trisulfide

10

Propyl allyl disulfide

13.95

13.89

11

Dially disulfide

8.02

6.87

12

3-vinyl-4H-1,2-dithiin

32.17

31.89

13

Methyl allyl trisulfide

4.67

5.02

14

Diallyl trisulfide

12.94

13.31

15

3,5-diethyl-1,2,4-trithiolane

2.62

2.56

16

Methyl 2-propenyl disulfide

0.34

17

2-vinyl-4H-1,3-dithiin

1.78

Extraction yield (%) = (essential oil/garlic sample) 9 100

Extraction time (min)

SFE (%)

SE (%)

0.04

0.14

1.10

0.81

0.78

130 min

42 min

SFE supercritical fluid extraction, SE solvent extraction

Extraction yield, extraction time, product quality, process cost, and scale are the
important factors for the industrialization. Therefore, comprehensive comparisons
of the garlic essential oils obtained by two different methods were further done. It
could be seen from Table 6 that the extraction yield (0.78%) of garlic by employing
SE method was slightly lower than that (0.81%) obtained by SFE. In addition, the
extract time (42 min) by SE method was shorter than that (130 min) obtained by
SFE. However, extraction yield and product quality obtained by SE was basically
similar to the ones obtained by SFE. Furthermore, the process cost and scale of SE
in the extracted oil was by far lower and larger than that of SFE, respectively.
Effect of garlic oil on mean (?SE) percentage of CD3?, CD4?, and CD8?, ratio
of CD4?/CD8? and CD40 expression in gastric cancer rats
In contrast to control rats (group I), essential oil-treated rats dose-dependently and
significantly (P \ 0.01) increased the percent of CD3? and CD4? leukocytes in
groups II and III. However, the percent of CD8? leukocytes and CD40 expression in
groups II and III was significantly (P \ 0.01) lower than that in group I rats
(control). One consequence of this bidirectional effect was a significant (P \ 0.01)
increase in the CD4?/CD8? ratio in groups II and III rats after the essential oil
treatment as compared to the group I rats (Table 7).

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Table 7 Effect of garlic essential oil on mean (?SE) percentage of CD3?, CD4? and CD8?, ratio of
CD4?/CD8? and CD40 expression in gastric cancer rats
CD3?

CD4?

CD8?

CD4?/CD8?

CD40

(I) Control

40.1 2.5

22.7 1.3

45.9 1.9

0.49 0.05

1.87 0.01

(II) Low dose

48.8 3.2a

29.3 1.2a

39.1 1.8a

0.73 0.04a

1.68 0.03a

1.61 0.02a

Group

(III) High dose

a

57.8 4.4

37.1 2.4

32.5 2.5

1.16 0.09

P \ 0.01, compared with control rats

Conclusions
The objectives of this study were to evaluate the effect of four major variables
(extraction temperature, extraction time, ratio of solvent to raw material, and the
number of extraction) on the performance of the SE of garlic essential oil. Results of
optimal experiment indicate that both extraction temperature and ratio of solvent to
raw material had the most apparent effect on the yield of essential oil. According to
extraction yield, the best optimal extraction conditions were extraction temperature
45C, extraction time 42 min, ratio of solvent to raw material 4, and extraction
number 4, when the yield was 0.78%.
The compositions of garlic essential oil obtained by SE and SFE methods were
compared. Although main compositions of the essential oils obtained by SE and
SFE are basically similar, their minor compositions do differ quantitatively. In
addition, extraction yield of SE was slightly higher than that obtained by SFE.
However, comprehensively considering various factors, it can be concluded that the
SE method offers obvious advantages over SFE. Therefore, SE is considered as the
optimum process for obtaining garlic essential oil with high quality.
In addition, the result, that is, the detection of a significant increment in ratio of
CD4?/CD8? in serum of rats after administration of garlic essential oil, indicated
that garlic essential oil may be useful for treatment of patients with inflammatory
disease, e.g., gastric cancer.
Acknowledgment The work was supported by the National Natural Science Foundation of China (No.
30872943) and the social development projects of Suzhou (No. SS0711).

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