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ISSN 2229-5054

International Journal of Drug Formulation & Research Mar.-Apr. 2011, Vol. 2(2) 94-110

International Journal of Drug Formulation & Research

Vineet Singla , Radhika Bhaskar* , Rahul Bhaskar
Department of Pharmaceutical Sciences, Lovely School of Pharmaceutical
Sciences , Chaheru, Phagwara (Punjab)

Garlic and its preparations have been widely recognized as an agent for prevention and treatment of cardiovascular
diseases and other metabolic disorders, atherosclerosis, hyperlipidemia, thrombosis, hypertension and
hypoglycemia. This review discusses the possible mechanism of therapeutic actions of garlic, different extraction
procedures along with determination of its constituents, its stability and dissolution method of garlic tablet.
Keywords: Garlic, Stability, Tablet, Alliin. Allicin, Antioxidant, Antimicrobial, Anticancer, Antihyperlipidemia,
Antihypertension and Hypoglycemia.

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ISSN 2229-5054
International Journal of Drug Formulation & Research Mar.-Apr. 2011, Vol. 2(2) 94-110

Garlic consists of fresh or dried compound bulbs of Allium Sativum Linn. (Family Lilliaceae).1
It contains not less than 0.2% of alliin (the main constituent present in the garlic), calculated on
the dried basis. Its bulbs are made up of cloves and are wrapped in a white papery sheath with
pungent taste and odour. 2
1) Sanskrit

: Rasona, Yavanesta

2) English

: Garlic

3) Gujrati

: Lasan, Lassun

4) Hindi

: Lahasun

5) Punjabi

: Lasan

6) Urdu

: Lahsan, Seer

7) Spanish

: Ajo 3

Constituents present in different garlic preparations: 2) Garlic homogenate

: Allicin

3) Heat treated garlic

: Alliin

4) Garlic powder

: Alliin

5) Aged garlic extract

: S-allyl cystine

6) Steam distilled garlic oil

: Diallyl disulfide

7) Oil-macerated/ether-extracted garlic oil

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

8) Major metabolite of raw garlic on blood

: Allyl mercapton 4

Major organo-sulphur compounds present in different garlic preparations: - The varieties

of garlic and manufacturing processes are of important considerations when choosing a garlic
supplement, since products with different biologically active compounds are (fig. 1)4
Formation of organo-sulphur compounds during metabolic pathways in processed garlic
(fig. 2)4
Reported Activities: - Garlic preparations are used as multipurpose and these are
1) Antioxidant and Antiatherosclerotic effect
2) Antihyperlipidimic effect
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ISSN 2229-5054
International Journal of Drug Formulation & Research Mar.-Apr. 2011, Vol. 2(2) 94-110

3) Platelet inhibitory and fibrinolytic effect

4) Antihypertensive effect
5) Anticancer effect
6) Antimicrobial effect
7) Hypoglycemic effect
Antioxidant: - The antioxidant effects of certain substances (garlic metabolites) of the secondary
plant metabolism belong to the non-enzymatic protective mechanisms. Garlic enhances the
serum levels of two antioxidant enzymes viz catalase and glutathione peroxidase5. It also
decreases plasma malondialdehyde concentration and thus plays a role in the prevention of
diseases of cardiovascular as well as providing protection against environmental sources of free
Antiatherosclerotic effect: - Atherosclerosis results by accumulation of intra and extra cellular
lipids. Garlic powder extract inhibits biosynthesis of cholesteryl esters and triglycerides in
atherosclerotic cells and it also inhibits the activity of acyl-CoA cholesterol acyltransferase, the
enzyme involved in the formation of cholesteryl esters, the main component of the excessive fat
accumulated by cells. On the other hand, garlic extract stimulates cholesteryl ester hydrolase that
degrades cholesteryl esters in atherosclerotic cells. 7
Antihyperlipidimic effect: - The inhibition of cholesterol synthesis in the liver is exerted at the
level of 3-Hydroxy-3-Methyl-Glutaryl Coenzyme A (HMG-CoA) reductase, an enzyme involved
in cholesterol biosynthesis. A low concentration of garlic compounds has no direct inhibition of
HMG-CoA reductase. This activity is carried out by an indirect mechanism.8
Platelet inhibitory and fibrinolytic effect: -Some form of garlic which increases cyclic
adenosine monophosphate levels and thus decreases thromboxane formation. There may be a
direct inhibition of thromboxane. -thromboglobulin release is decreased, which suggests that
the effect may be more on the platelet activation phase. 9
Antihypertensive effect: - The hypotensive action of garlic may be due to a direct relaxant
effect on smooth muscles. Aqueous garlic extract and its individual components, allicin and
ajoene also open K+ channels, causing a membrane hyperpolarization and thus lead to a decrease

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ISSN 2229-5054
International Journal of Drug Formulation & Research Mar.-Apr. 2011, Vol. 2(2) 94-110

of the Ca2+ inward current into the vascular smooth muscle cell and finally induce vasodilation
resulting from intracellular Ca2+ decrease.9
Anticancer effect: - Treatment of human melanoma cells with S-allylcysteine reduces
expression of cell-surface ganglisides, the tumor associated markers of differentiation and
transformation. S180 tumor cells when exposed to a garlic extract displayed delayed progression
to S phase. 10
Antimicrobial effect: - The antimicrobial activity of allicin is due to the inhibition of thiolcontaining enzymes in the microorganisms. 11
Hypoglycemic effect: - The hypoglycemic action of garlic could possibly be due to
potentiation of the insulin effect of plasma by increasing the pancreatic secretion of insulin from
existing -cells or its release from bound insulin and enhancement of insulin sensitivity. It has
been previously suggested that garlic (allicin) can enhance serum insulin by effectively
combining with compounds like cysteine, due to spare insulin from SH group. 12-14

Purity Tests: - To ensure that all the analytical procedures performed allow an
accurate statement of the content of impurities of an analyte, i.e. related substances test, heavy
metals, residual solvents content, etc and the limits of garlic are.
1) Total ash

- Not more than 5.0%.

2) Acid-insoluble ash

- Not more than 1.0%.

3) Water-soluble extractive

- Not less than 5.0%.

4) Alcohol-soluble extractive

- Not less than 4.0%.

5) Moisture

- Not more than 7% 15

Extraction of allicin by different methods: - There are different methods to extract the allicin
from the garlic. These are:I) 50 g of the peel garlic is cut into small pieces and homogenize in 70 ml of cold, sterile 0.9%
NaCl in the presence of some crushed ice. The homogenization is carried out in a blender at high
speed using 30 second bursts for a total of 10 minutes. The homogenized mixture is filtered 3
times through cheesecloth, the filtrate is centrifuge at 2000 RCF (relative centrifugal force) for
10 minutes and the clear supernatant is dilute to 100 ml with normal saline. The concentration of
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ISSN 2229-5054
International Journal of Drug Formulation & Research Mar.-Apr. 2011, Vol. 2(2) 94-110

this garlic preparation is considered to be 500 mg/ml on the basis of the weight of the starting
material (50 g/100 ml). The aqueous extract of garlic is store in small aliquots at -20C until
used. Determination is carried out by HPLC by taking allicin as standard. 13

II) Dried and ground bulbs (about 100 g) are submitting to extraction with 300 ml ethanol (80%)
in a Soxhlet apparatus for 72 h. After extraction, the solvent is filter and then evaporated by
Rotavapor. The obtained garlic alcoholic extract is store at -20

C until being used.

Determination is done by HPLC and allicin is taken as standard. 14

III) Garlic powder (125 mg) is transfer to a 25 ml volumetric flask, diluted with water to volume,
vortex for 15 sec, sonicated at 8.0oC for 5 min, allow to stand at room temperature for 10 min
(conversion of alliin into allicin) and centrifuged for 5 min. Then 5 ml of the supernatant is
transfer to a 10 ml volumetric flask, 0.3 ml portion of internal standard 500 g/ml (Ethyl
paraben) is add (final concentration 15 g/ml) and dilute with cold methanol to volume and store
at -20 0C until being used. Determination is carried out by HPLC and ethyl paraben is used as
internal standard. 16

IV) To 0.800 g of garlic powder, add 20.0 ml of water R and homogenise the mixture in an
ultrasonic bath at 4C for 5 min. Allow to stand at room temperature for 30 min. Then centrifuge
for 30 min. Dilute 10.0 ml of the supernatant to 25.0 ml with a mixture of 40 volumes of a 1 %
V/V solution of anhydrous formic acid R and 60 volumes of methanol R (stock solution). Shake
and centrifuge for 5 min. Place 0.50 ml of the internal standard solution into a volumetric flask
and dilute to 10.0 ml with the stock solution. 17
This is the only method in which allicin is not used as an internal standard and no need to store at
20 0C.

Determination by HPLC
Column dimension : 0.25m long & 4mm internal diameter
Stationary phase

: Silanised octadecylsilyl silica gel

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ISSN 2229-5054
International Journal of Drug Formulation & Research Mar.-Apr. 2011, Vol. 2(2) 94-110

Mobile phase

: Anhydrous formic acid: Methanol (4: 6)

Flow rate

: 0.8 ml/min

Internal standard

: Butyl para hydroxybenzoate


: Spectrophotometer at 254nm

Garlic powder
Homogenization 0.8gm powder with20ml of water at 4oC for 5 minutes in
Ultrasonic bath
Homogenize powder
Incubate at room temperature for 30inutes
Allicin containing Homogenize powder
Centrifugation for 30 minutes 6000 Xg
Dilute 10ml of the supernatant to 25ml stock solution
Centrifugation for 5 minutes

0.50ml of the internal standard solution in a volumetric flask and dilute

to 10ml with the stock solution

Determination by HPLC and Butyl parahydroxy benzoate as standard

Calculate the percentage of allicin from the expression:

22.75 X S1 M2/ S2 M1
S1 = area of the peak corresponding to allicin (most prominent peak)
S2 = area of the peak corresponding to butyl para hydroxybenzoate in the chromatogram
obtained with the test solution
M1 = mass of the drug in grams
M2 = mass of butyl para hydroxybenzoate in grams in 100.0 ml of the internal standard
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ISSN 2229-5054
International Journal of Drug Formulation & Research Mar.-Apr. 2011, Vol. 2(2) 94-110

1 mg of butyl para hydroxybenzoate corresponds to 8.65 mg of allicin.18

Stability: - Stability is defined as the capacity of a drug substance or drug product to remain
within established specifications to maintain its identity, strength, quality, and purity throughout
the retest or expiration dating periods. Physical, chemical, and microbiological data are
generated as a function of time and storage conditions (e.g., temperature and relative humidity

Thermo stability of allicin: Allicin is separated from garlic by using HPLC in a C18 column of MG-II (5m, 4.6mm x
Solvent used

: 0.02M phosphate buffer (pH6.5): Acetonitrile: 1, 4 dioxane (7:1:2)

Flow rate

: 0.5 ml/min


: 220nm

Then compare the peak area produced by authentic allicin with that of the garlic extract.
Authentic allicin is a reagent-grade preparation with 99.39% purity and is kept at -70oC until
being use (fig. 3). 19
Determination of degradation rate constant and half life of allicin by biological assays
The biological half life of allicin is calculated according to the equation:
t1/2 = 0.693/k
The biological activity of allicin decreases proportionally to the incubation time as shown in
Table 119
Stability of allicin in different solvent: Ethanolic extract of allicin is more efficient than aqueous extract but the levels decreased
gradually at room temperature, and allicin disappeared within a half-month, especially in 100 %
ethanol or water. The 20 % and 50 % aqueous ethanol solutions are the most suitable solvents to
maintain allicin for a couple of weeks at room temperature. In 20 % alcohol, allicin keeps its
biological activity for longer time than in other ethanolic or aqueous solutions. In the 50 % and
70 % ethanolic solutions, the antibacterial activity against E. coli was kept longer than in water
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ISSN 2229-5054
International Journal of Drug Formulation & Research Mar.-Apr. 2011, Vol. 2(2) 94-110

or similar to that in water against S. aureus. In n-hexane and the vegetable oil, the amount of
allicin and its biological activity decreased more rapidly than those in ethanolic aqueous
solutions. Allicin is more unstable in vegetable oil than in n-hexane (Table 2). 20
Dissolution method: - Under the simulated gastrointestinal condition, the release of allicin and
other allyl thiosulfinates is to be observed from the delayed-release garlic tablets. Using USP
apparatus-II (paddle) dissolution method equilibrated at 37oC, one tablet is placed into round
bottom glass vessels containing 750ml of 0.1N HCl and paddle stirred at 100 rpm for 2 h, after
which 250ml of 0.2M Na3PO4 is added and the pH slightly adjusted if necessary, giving 1000ml
at pH 6.80 0.05. 1ml of the solution transfer to a test tube containing 0.05 ml of 0.21M
carboxymethoxylamine hemihydrochloride immediately upon removal from the dissolution
vessel in order to inhibit the alliinase enzyme and followed by HPLC analysis. The time to
achieve complete disintegration is determined by observation during the dissolution test.2,


Contraindications: Garlic is contraindicated in patients with a known allergy to the drug. During pregnancy and
lactation the use of garlic is not recommended and rare cases gastro-intestinal irritation or
allergic reactions. 15
Precautions: Patients on warfarin therapy should be warned that garlic supplements may increase bleeding
times. Blood clotting times have been reported to be double in patients taking warfarin and garlic
supplements. 15

Recommended daily doses in humans: 1) Fresh garlic

: 4g approx 1 clove (412 mg of alliin or 2

5 mg of Allicin)

2) Dehydrated garlic powder

: 6001200 mg in divided doses

3) Aged garlic

: 17.2 g/day

4) Fresh air-dried bulb

: 25 g

5) Garlic oil

: 25 mg
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ISSN 2229-5054
International Journal of Drug Formulation & Research Mar.-Apr. 2011, Vol. 2(2) 94-110

6) Dried bulb

: 24 g three times daily

7) Tincture (1:5 in 45% alcohol)

: 24 ml three times daily 9

Marketed preparation of garlic (Table 3)

Garlic has many health benefits and has been traditionally used worldwide. The wealth of
scientific literature supports the proposal that garlic consumption have significant
cardioprotective effect. Garlic is a magical medicinal herb and if consumed at regular basis, it
has got the prophylactic but also curing effect.
References: 1. Indian Pharmacopoeia (2007). Garlic. The Indian Pharmacopoeia Commission,
Ghaziabad, 2047-2048.
2. United States Pharmacopoeia (2004). Garlic. United States Pharmacopeial Consention,
Inc, Rockville. 1997-2002.
3. The Ayurvedic Pharmacopoeia of India (2001). Garlic. The Controller of Publications
Civil Lines, Delhi. 108-109.
4. Martinez, M.C., Corzo, N. & Villamiel, M., (2007). Biological properties of onions and
garlic. Trends in Food Science & Technology. 18, 609-625.
5. Al-Numair, K.S., (2009). Hypocholesteremic and Antioxidant Effects of Garlic (Allium
sativum L.) Extract in Rats Fed High Cholesterol Diet. Pakistan Journal of Nutrition. 8,
6. Mathew, B.C. & Biju, R.S., (2007). Neuroprotective effects of garlic a review. Libyan
Jounal of Medicine. 23-33.
7. Alexander, N.O. & Grunwald, J., (1996). Effects of Garlic on Atherosclerosis. Nutrition.
13, 656-663.
8. Ackermann, R.T., Mulrow, C.D., Ramirez, G., Gardner, C.D., Morbidoni, L. &
Lawrence, V.A., (2001). Garlic shows promise for improving some cardiovascular risk
factors. Archives of Internal Medicine. 161, 813-824.

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ISSN 2229-5054
International Journal of Drug Formulation & Research Mar.-Apr. 2011, Vol. 2(2) 94-110

9. Bathaei, F.S. & Akhondzadeh, S. (2008). Cardiovascular Effects of Allium Sativum

(Garlic): An Evidence-Based Review. The Journal of Tehran University Heart Center. 1,
10. Riazati, N., (1998). The Stinking Rose. Nutrition Bytes. 4, 1-8.
11. Shadkchan, Y., Shemesh, E. & Mirelman, D., (2004) Efficacy of allicin, the reactive
molecule of garlic, in inhibiting Aspergillus spp. in vitro, and in a murine model of
disseminated aspergillosis. Journal of Antimicrobial Chemotherapy. 1-5.
12. Shariatzadeh, S.M.A., Mehranjani, M.S. & Mahmoodi, M., (2008) Effect of Garlic
(Allium Sativum) on Blood Sugar and Nephropathy in Diabetic Rats. Journal of
Biological Sciences. 1-6.
13. Thomson, M., Al-Amin, Z.M., Al-Qattan, K.K., Shaban, L.H. & Ali, M., (2007) Antidiabetic and hypolipidaemic properties of garlic (Allium Sativum) in streptozotocininduced diabetic rats. International Journal of Diabetes & Metabolism. 15, 108-115.
14. Eidia, A., Eidib, M. & Esmaeilia, E., (2006). Antidiabetic effect of garlic (Allium sativum
L.) in normal and streptozotocin-induced diabetic rats. Phytomedicine. 13, 624629.
15. WHO monographs on selected medicinal plants. Bulbus Allii Sativi., vol. 1, 1999.
16. Diego, M.D., Avello, M., Mennickent, S., Fernandez, M. & Fernandez, P., (2007)
Validated liquid chromatographic method for quantitative determination of allicin in
garlic powder and tablets. Journal of Separation Science. 30, 2703-2707.
17. Baghalian, K., Ziai, S.A., Naghavi, M.R., Badi, HN. & Khalighi, A., (2005). Evaluation
of allicin and botanical traits in Iranian garlic (Allium Sativum L.) Ecotypes. Scientia
Horiculture. 10, 155-166.
18. British Pharmacopoeia (2000). Garlic. Stationery Office, London. 49-50.
19. Fujisawa, H., Suma, K., Origuchi, K., Seki, T. & Ariga, T., (2008). Thermostability of
allicin determined by chemical and biological assays. Bioscience Biotechnology and
Biochemistry. 72, 2877-2883.
20. Fujisawa, H., Suma, K., Origuchi, K., Kumagal, H., Seki, T. & Ariga, T., (2008).
Biological and chemical stability of garlic-derived allicin. Journal of Agriculture and
Food Chemistry. 56, 4229-4235.
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International Journal of Drug Formulation & Research Mar.-Apr. 2011, Vol. 2(2) 94-110

21. Blatt, Y., Cohen, D., Kimmelman, E., Friedman, O. & Rotman, A., (2007). US Patent
No. 6270803 B1.

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ISSN 2229-5054
International Journal of Drug Formulation & Research Mar.-Apr. 2011, Vol. 2(2) 94-110



Heat treated



Aged garlic

Steam distilled


Major metabolite



garlic oil

extracted garlic oil

of raw garlic In blood


S-allyl cysteine

Diallyl disulfide 2-vinyl-4H-1,3-dithiin Allyl mercaptan


Methyl-allyl thiosulfinates

1-propenyl allyl thiosulfinate

S-allyl mercaptocysteine Diallyl trisulfide

Allyl methyl trisulfide



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ISSN 2229-5054
International Journal of Drug Formulation & Research Mar.-Apr. 2011, Vol. 2(2) 94-110


Allyl methyl disulfide


Diallyl tetrasulfide (DATS)

Allyl methyl tetrasulfide

Dimethyl trisulfide

Diallyl sulfide (DAS)

Figure 1 Organo-sulphur compounds present in different garlic preparations


Hydrolysis and

S-alk-(en)yl-L-cysteine sulphoxides (CSS):

Methiin (S-methyl-L-cysteine sulphoxide)

Alliin (S-allyl-L-cysteine sulphoxide). This one is most abundant and
parental sulphur compound responsible for the majority of odoriferous

When garlic is cut, crushed, chewed or dehydrated,

the vacuolar enzyme, alliinase, rapidly lyses the cytosolic
cysteine sulphoxides (CSS), giving sulphenic acids,
highly reactive organo-sulphur intermediates.

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ISSN 2229-5054
International Journal of Drug Formulation & Research Mar.-Apr. 2011, Vol. 2(2) 94-110

S-allyl cysteine (SAC)

2-propenesulphenic acid

Self - condensation

Thiosulphinates, mainly alliicin, very

volatile and extremely unstable compounds

Instantaneous decomposition

DAS, DADS, DATS, dithiins, ajoene, etc.

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ISSN 2229-5054
International Journal of Drug Formulation & Research Mar.-Apr. 2011, Vol. 2(2) 94-110

Figure 2 Formation of organo-sulphur compounds during metabolic pathways in processed garlic

Figure 3 HPLC pattern of authentic allicin 20

Table 1 Degradation rate constant and half life of allicin by biological assays
S. aureus
Temperature (oC)





t1/2 (d)





















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ISSN 2229-5054
International Journal of Drug Formulation & Research Mar.-Apr. 2011, Vol. 2(2) 94-110















Table 2 Stability of allicin in different solvent


Biological half-life

Biological half-life

Organism: S. aureus

Organism: E. coli

Chemical half life


6.5 0.09 days

10.3 0.3 days

3.7 0.2 days

Ethanol 20%

12.0 0.14 days

17.6 0.3 days

14.7 0.8 days

Ethanol 50%

11.8 0.30 days

17.7 0.2 days

8.3 0.3 days

Ethanol 70%

6.6 0.07 days

8.7 0.9 days

7.8 0.6 days

Ethanol 100%

3.2 0.06 days

6.5 0.3 days

7.2 1.6 days


3.4 0.13 h

8.6 1.0 h

3.3 0.5 h

Vegetable oil

3.1 0.05 h

2.4 0.2 h

0.8 0.1 h

Biological and Chemical half-lives of Allicin in Garlic Extracts made with various solvents at 23oC.

Table 3 Marketed preparation of garlic

Brand Name

Dosage Form


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ISSN 2229-5054
International Journal of Drug Formulation & Research Mar.-Apr. 2011, Vol. 2(2) 94-110

Kwai Heart Fit Garlic


Lichtwer Pharma, USA

Super Garlic


Schiff Products, USA

Garlic XTRA


Sundown Herbals, USA



Chattem, USA

Kyolic Nutritional Supplement


Wakunaga of America Co. Ltd.,


One A Day Dietary Supplement


Bayer Corporation, USA

Your life Advanced Strength


Leiner Health Produts, USA



Natrol, USA

Daily Garlic


Rite Aid Corporation, USA

Enteric Coated Garlic

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