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Shamsun Naher et al / Int. J. Pharm. Ph ytopharmacol. Res. 2014; 4 (3): 173-175
173
*Corresponding Author:
Dr. Shamsun Naher,
Associate Professor, Department Of Chem istry,
Jagannath University, Dhaka1100, Bang ladesh.
Email: shamsunnaher2002@yahoo.com
Tel: 088-01720250388
Research Article
Comparative Studies on Physicochemical Properties and GC-MS Analysis of
Essential Oil of Two Varieties of Allium sativum Linn (Garlic)
Shamsun Naher1*, Md. Mostak Ahmad1, Shahin Aziz2, S. M. Mahmudul Hassan2, Mala Khan2, Mirola Afroze2
1Department of Chemistry, Jagannath Unive rsity, Dhaka 1100, Bangladesh
2BCSIR Laboratories Dhaka, Bangladesh Council of Scientific and Industrial Research, Dr. Qudrat-E-Khuda Road, Dhaka-1205, Banglades h
1. INTRODUCTION
Garlic (Allium sativum Linn.), is one of the edible plants which has
generated a lot of interest throughout human history as a medicinal
panacea. It is a perennial plant belonging to the genus Allium.
Allium the Latin word giv en to garlic3, a flowering plant with
hundreds of distinct species; which many have been harvested
through human history, but only about a dozen are still
economically important today as crops or garden vegetables4. One
of the species belong to this genus is the Allium sativum Linn. also
known as the cultivated garlic, which belongs to the family Alliaceae
(Formerly classified in the lily (Liliaceae) family )5 and closely
related to the onion, shallot, leek, chive, and rakkyo6. It has been
used throughout recorded history for both culinary and medicinal
propose 7. Garlic plant is moderately tall (up to 3 feet) it is an erect
herb normally grown as an annual that is a plant that only last for a
year, it has adventitious roots and condensed, flattened stem and
narrow flat leaves. The bulb consists of 6 to 35 bulblets called
cloves various shapes and sizes8-10 with glistering and transparent
covering. They grow in various conditions from dry, well-drained
mineral-based soil to moist organic soil, most grow in sunny
locations but a number also grow in forests, or even in swap water
areas 11. Garlic is propagated by planting cloves or top bulblets. A
classic ingredient in many national cuisines, garlic has a powerful
onion like aroma and pungent taste 12. Though Original abode of
garlic in Central Asia 13, garlic has been cultivated in the Middle
East for over 6000 years, making one of mankind’s first cultivated
plants. The medicinal benefits and claims for garlic have awarded it
the name “wonder drug among all herbs”. Garlic contains 0.1 to
0.36% essential oil, the principal components of which are diallyl
disulfide, diallyl trisulfide, allyl propyl disulfide. European standards
specify that garlic supplements contain not less than 0.45%
Allicin14. Allicin is the active principal and a yellow liquid responsible
for the odor of freshly crushed garlic15. W hen the clove is crushed,
the enzyme alliinase is activated and breaks down alliin into allicin
and other allyl thiosulfinates16. At least 35 different compounds
have been identified in garlic17. These sulfur-containing com-
pounds have been the focus of the many quantitative and
qualitative studies on garlic and related species 18.
Commercially available garlic oil capsules generally contain
vegetable oil and a small amount of garlic essential oil because of
the pungent odors. Other garlic supplements fall into one of these
categories; dehydrated garlic powder, garlic oil macerate and aged
garlic extract19. The regular consumption of garlic oil can reduce
blood pressure; prevent heart disease including atherosclerosis,
high cholesterol and cancer 20. Garlic oil is an effective antibiotic,
anti-viral, anti-fungal agent, which could be used to prevent nausea,
diarrhea, ease coughs, even treatment in conditions such as
malaria and cholera probably an immune system enhancement,
some studies have found lower rates of certain types of cancer in
people 21. In 2007, the BBC reported Allium sativum may have
other beneficial properties, such as preventing and fighting the
common cold 22.
However many researchers have been carried out on garlic (Allium
sativum Linn.), but no systematic research on comparative studies
has been reported on the essential oil of garlic in Bangladesh and
China. Some disagreement about the presence of its constituents
was observed. Therefore, present work was undertaken to carry out
a complete investigation of the essential oil of Allium sativum Linn.
of two varieties from Bangladesh and China including its physical
properties & chemical constituents along with GC-MS analysis.
Abstract
Allium sativum Linn. is locally known as Garlic in Bangladesh. It is used as spices all over the world. In this
study essential oil of the two varieties of g arlic from Bangladesh an d China were inv estigated by GC-MS. The
regular consumption of garlic oil can reduce blood pressure; prevent heart disease including atherosclerosis,
high cholesterol and canc er1. Garlic oil is an e ffective antibiotic, anti-viral, an ti-fungal agent, which could be
used to prevent nausea, diarrhea, ease coughs, e ven treatment in conditions such as malaria and cholera
probably an immune system enhancement, some studies have found lower rates of certain types of c ancer in
people2. Garlic is one of the most popular spices in the world. Total 11 chemical constituents were found by gas
chromatography and mass spectrometry (GC-MS) analysis from both the essential oil o f Bangladeshi and
China garlic. Among them Diallyl disulfide (28.99%); Trisulfide, methyl 2-propenyl (23 .22%) and Trisulfide, di-2-
propenyl (15.36%) are the principle components of Bangladeshi ga rlic. In China garlic, Trisulfide, methyl 2-
propenyl (29 .12%); Trisulfide, di-2-propeny l (21.98%) and Diallyl disulfide (17.24 %) were the major
components. Quantification of active principles throu gh analytical tools is essential for es tablishing th e
authenticity and credibility. Steam dis tillation extraction combined with GC-MS has been shown to be a valuable
tool for the analysis of garlic co nstituents and can provide a useful guide to component variation. The main
objective of the present study was focused on identification and quantification of chemical constituents present
in the essential oil of garlic using GC-MS m ethod.
ISSN (Online) 2249-6084 (Print) 2250-1029
International Journal of Pharmaceutical and
Phytopharmacological Research (eIJPPR)
[Impact Factor – 0.852]
Journal Homepage: www.eijppr.com
Article info
Article History:
Received 4 August 2014
Accepted 18 September 2014
Keywords:
Gas chromatography and mass
spectrometry (GC-MS), Allium
sativum Linn. Quantification,
active principle, Analytical tool,
Essential oil, Chemical
constituents.
Shamsun Naher et al / Int. J. Pharm. Ph ytopharmacol. Res. 2014; 4 (3): 173-175
174
2. MATERIALS AND METHODS
The fresh Bangladeshi and China garlic are available in the local
markets of Dhaka. The collected samples were washed clearly by
water to remove dust materials. Then they were dried. Finally the
dried garlic was ground by Fritsch mortar grinder, Germany for one
hour. Then the powder was sieved prior to the extraction process.
The mean particle diameters obtained were 0.25 and 0.50mm.
2.1 Extraction of essential oil
There are a number of methods employed for the extraction of
essential oil or volatile oil from the plant. In the present study steam
distillation method was used. This extraction procedure was simple
and it could also provide a valuable means of producing flavor
extracts of two varieties from Bangladeshi and China garlic under
mild conditions which preserve the natural characteristics of the
fresh product. In the process, definite amount of sample (dirt free
powdered garlic) were taken in a distillation flask (Clevenger’s
apparatus). Then distilled water was added two third of its volume
to the flask. Then the flask was heated by electric heating mental
for 4 hours. Volatile substances of aniseed and generated steam in
the flask were condensed by water condenser. The essential oil
was lighter than water and so could be separated out. The steam
distilled essential oil layer which was collected over water, was
extracted and washed with analytical grade ether or chloroform.
The ether extract of the oil was dried over anhydrous Na2SO4 and
then filtered. It was collected in vial. The ether or chloroform was
removed in vacuum condition. Thus the essential oil of fresh garlic
was collected.
2.2 GC-MS Analysis
The essential oil of Allium sativum Linn. (Garlic) of two varieties
were analyzed by Electron Impact Ionization (EI) method on GC-
17A gas chromatograph, coupled to a GC-MS 2010 plus mass
spectrometer; fused silica capillary column temperature of 40˚ C (
was held 2 min) was maintained with carrier gas helium at a
constant pressure of 90kPa. Samples were injected by splitting with
the split ratio 10. Essential oil sample was dissolved in chloroform.
The operating condition were as follows: name of column- RTS-
5MS, diameter 30 cm, length 0.25mm, temperature of the column-
initial temperature 40˚C ( was held 2 min) , injector temperature-
220 ˚C, holding time 5 min, column packing- column packing was
done with 10% diethylene glycol succinate on 100-120 mesh
diatomic CAW, splitting- samples were injected by splitting with the
spilt ratio 10, carrier gas- helium gas at constant pressure 90 kPa,
sample dissolved- in chloroform, range of linear temperature
increase- 10˚C per min.
2.3 Preparation of essential oil samples for GC-MS analysis
Essential oil was diluted to 7% by chloroform. An inert gas (i.e.
nitrogen) was introduced, from a large gas cylinder through the
injection part, the column and the detector. The flow rate of the
carrier gas was adjusted to ensure reproducible retention time and
to minimize detector dirt. The sample was then injected by a micro
syringe through a heated injection part when it was vaporized and
carried into the column. The long tube of the column was tightly
packed with solid particles. The solid support was uniformly
covered with a thin film of a high boiling liquid (the stationary
phase). The mobile and stationary phases were then partitioned by
the samples and it was separated into the individual components.
The carrier gas and sample component was then emerging from
the column and passed through a detector. The amount of each
component as concentration by the device and generates a signal
which was registered electrically. The signal passed to a detector.
2.4 Identification of the components
The physical properties of the essential oil of Allium sativum Linn.
(Garlic) of two varieties of Bangladesh and China are presented in
Table-1. Interpretation of mass spectroscopy (GC-MS) was
conducted using data base of National Institute Standard and
Technology (NIST) having more than 62000 patterns. The
spectrum of the unknown component was compared with the
spectrum of the known component stored in the NIST library. The
retention time, molecular weight, molecular formula and
composition percentage of the sample material was recorded and
presented in Table -2 and Table-3.
3. RESULTS AND DISCUSSION
The physical characteristics such as color, appearance, specific
gravity, optical rotation, solubility, refractive index of the essential
oil were determined by conventional method. The result of the
physical properties of Allium sativum Linn. (Garlic) of two varieties
from Bangladesh and China are presented in Table-1.The
percentage yield was found to be Bangladeshi garlic contained
essential oil 0.1249 % and China garlic contained 0.1364 %. The
colour of the essential oil was light yellow with a pungent odour.
Refractive Index of essential oil was 1.56678 and 1.56470 of
Bangladeshi and China garlic. The slight variation of this oil content
and the composition of the essential oil depend on several factors
such genotype, stage of maturity, cultivation peculiarities, soil
composition and climate differences in various geographical
locations. Fluctuation of the oil composition can impart change in
the organoleptic properties of the plant belonging to the botanical
spices and variety. So far we aware till now no systemic
investigation on the Allium sativum L.(Garlic) have not been
investigated in Bangladesh by using modern analytical techniques.
GC-MS analyzed results which include the active principles with
their retention time, molecular formula, molecular weight and
composition of the essential oil of Allium sativum L.(Garlic) of two
varieties from two different countries of Bangladesh and China are
presented in Table-2 and Table-3. 11 major components of
essential oils were found from both the samples by using GC-MS.
Among them Bangladeshi garlic bulb (clove) contained 1-Propene,
3, 3’ –thiobis (0.89%); Disulfide, methyl 2-propenyl (3.27%); β-
Phenylethyl butyrate (1.85%); Diallyl disulfide (28.99%); Trisulfide,
methyl 2-propenyl (23.22%); Hydroxylamine, methyl-(1-phenylethyl)
(1.45%); Naphthalene, decahydro (2.91%); 3-Vinyl-1,2-
dithiacyclohex-4-ene (5.47%); Trisulfide, di-2-propenyl (15.36%);
3H-1,2,4-Triazole-3-thione,2,4-dihydro-4-methyl (3.57%); 2-Chloro-
4,4-dimethyl-1,3,2-oxathiaphospholane (9.11%). China garlic bulb
(clove) contained 1-Propene, 3, 3’ –thiobis (0.89%); Disulfide,
methyl 2-propenyl (4.94%); Diallyl disulfide (17.24%); Trisulfide,
methyl 2-propenyl (29.12%); Benzene, 1-methyl-3-(methylthio)
(4.01%); 2,4-Dithiahex-5-ene 2,2-dioxide (1.01%); 3-Vinyl-1,2-
dithiacyclohex-4-ene (5.65%); Trisulfide, di-2-propenyl (21.98%);
3H-1,2,4-Triazole-3-thione,2,4-dihydro-4-methyl (1.64%); Trimethyl
silyldiazomethane (4.48%); 1,2,4,6-Tetrathiepane (9.05%).
Results show that essential oil from both of the two varieties from
two different places of Bangladesh oils are a complex mixture of
numerous compounds, many of which are found in trace amount. It
is worth monitoring that there is a great variation in the chemical
composition of these two regions oil of Allium sativum Linn. (Garlic).
This confirms that the reported variation in oil is due to geographic
divergence and ecological conditions.
Table 1: Comparative studies on physical properties of essential oil
of Bangladeshi and China garlic (Allium sativum L.)
Parameters Results
Bangladeshi garlic China garlic
Colour of oil Light yellow Light yellow
Taste Pungent Pungent
Smell Spicy Spicy
Refractive Index 1.56678 1.56470
Oil yield 0.1249% 0.1364%
Moisture 62.824% 65.016%
Total ash 1.21% 1.33%
Crude fiber 2.08% 1.91%
Protein 3.49% 3.84%
Carbohydrates 26.70% 24.30%
Food energy (cal/gm) 144.474 139.896
Shamsun Naher et al / Int. J. Pharm. Ph ytopharmacol. Res. 2014; 4 (3): 173-175
175
Table 2: Chemical constituents of the essential oil of garlic from
Bangladesh
Peak
No:
Retention
Time
Area
(%)
Name of the Compound M.W
Formula
1 3.83 0.89 1-Propene, 3,3’ –thiobis 114 C
6
H
10
S
2 5.080 3.27
Disulfide, methyl 2-
propenyl 120 C4H8S2
3
6.240
1.85
β-Phenylethyl butyrate
192
C12H16O2
4 11.130 28.99
Diallyl disulfide 146 C
6
H
10
S
2
5 13.596 23.22
Trisulfide, methyl 2-
propenyl 152 C4H8S3
6 13.905 1.45 Hydroxylamine, methyl-(1-
phenylethyl)- 151 C9H13NO
7 14.091 2.91 Naphthalene, decahydro- 138 C
10
H
18
8 15.205 5.47 3-Vinyl-1, 2-
dithiacyclohex-4-ene 144 C6H8S2
9 20.381 15.36
Trisulfide, di-2-propenyl 178 C
6
H
10
S
3
10 20.899 3.57
3H-1,2,4-Triazole-3-
thione,2,4-dihydro-4-
methyl
115 C3H5N3S
11 22.933 9.11 2-Chloro-4,4-dimethyl-
1,3,2-oxathiaphospholane 170 C4H8ClOPS
Table 3: Chemical constituents of the essential oil of garlic from
China
Peak
No:
Retention
Time
Area
(%) Name of the Compound M.W
Formula
1 3.845 0.89 1-Propene, 3,3’ –thiobis 114 C
6
H
10
S
2 5.133 4.94 Disulfide, methyl 2-
propenyl 1 20 C4H8S2
3 10.839 17.24 Diallyl disulfide 146 C
6
H
10
S
2
4 13.870 29.12 Trisulfide, methyl 2-
propenyl 1 52 C4H8S3
5 14.319 4.01 Benzene, 1-methyl-3-
(methylthio)- 138 C8H10S
6 14.406 1.01 2,4-Dithiahex-5-ene 2 ,2-
dioxide 152 C4H8O2S2
7 15.356 5.65 3-Vinyl-1, 2-dithiacyclohex-
4-ene 144 C6H8S2
8 20.637 21.98 Trisulfide, di-2-propen yl 178 C
6
H
10
S
3
9 20.975 1.64 3H-1,2,4-Triazole-3-
thione,2,4-dihydro-4-methyl
115 C3H5N3S
10 21.383 4.48
Trimethyl
silyldiazomethane 114 C4H10N2Si
11
23.079
9.05
1,2,4,6-Tetrathiepane
170
C3H6S4
4. ACKNOWLEDGEMENT
Authors are thankful to Abu Anis Jahangir, Director, Dhaka
laboratories, BCSIR, Dhaka for his support during this research
work.
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