INTERNATIONAL JOURNAL OF FOOD AND NUTRITION SCIENCE VOL1 NO2 MARCH 2012

Extraction and Physicochemical Determination of Garlic (Allium sativum L) Oil

Gafar M. K., A. U. Itodo, A. A. Warra and L. Abdullahi
AbstractThe oil was extracted from powder of garlic cloves using soxhlet extraction method with n-hexane as the solvent. The garlic cloves give 22.5% oil yield with specific gravity density of 0.90g/cm3. The oil is light yellow in colour with a pungent smell. The chemical analyses revealed saponificacation value of 192 1.00mgKOH/g, peroxide value of 2.5 0.50mg/100g, acid value of 4.18 0.01mgKOH/g, free fatty acid value of 2.10 0.05% and Iodine value of 12.69 0.05g/100g. These results obtained shown that the oil could be of benefit for soap production. Index Terms chemical analysis, Extraction, Garlic oil, oil yield, Industrial uses. I. INTRODUCTION

llium the Latin word given to garlic[1], 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 vegetables[2]. One of the species belong to this genus is the Allium sativum L. also known as the cultivated garlic, which belong to the onion family Alliaceae and closely related to the onion, shallot, leek, chive, and rakkyo[3]. It has been used throughout recorded history for both culinary and medicinal propose[4]. Garlic plant is moderately tall (up to three 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 with glistering and transparent covering, the plant grows as a vegetable rosette close to the ground, the leaves is broad like kidney shaped. The plants also produce an erect flowering stem with numerous quitter inch. The majority of Allium species are native to the Northern hemisphere, mainly in Asia. A few species are native to Africa and South America. 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[5].

Allium sativum L. consist of sulfur containing compounds such as allicin, alliin, ajoene, dially disulfide, dithiin and Sallylcysteine.These large number of sulfur compounds contributes to the smell and taste of garlic. Dially disulfide is believed to be an important odour component in the garlic. Allicin has been found to be the compound most responsible for the spiciness of the raw garlic and being a powerful antibiotic and antifungal compound, it believe to be the agent responsible for the speed recovery from strep throat or other mild ailments when garlic is used[6]. Garlic contain oil which is an essential oil, the oil is extracted by process of steam distillation of the garlic cloves using n-hexane as solvent. Essential garlic oil contains variety of sulfide such as dially disulfide and dilly trisulfide. During the process allicin is completely eliminated from the oil[7]. 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 extract[8]. Garlic oil has numerous uses in the world today, its uses include the flavouring some cuisine like salads, and sauces[9]. The regular consumption of garlic oil can reduce blood pressure, prevent heart disease including atherosclerosis, high cholesterol and cancer[10]. 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[11]. II. MATERIALS AND METHODS

Manuscript received January 23, 2012.

Lecturers; Gafar M. K., Dr .A. U. Itodo and Research student L. Abdullahi are from the Department of Chemistry, Kebbi State University of Science and Technology, Aliero, Nigeria. Researcher A.A. Warra is a Senior Laboratory Technologist from the Department of Biochemistry, Kebbi State University of Science and Technology,Aliero Nigeria.

Sampling and sample treatment: The garlic cloves were obtained from Kurya Madaro town Kaura Namada Local government area of Zamfara State and identified by a taxonomist at Botany department in Kebbi State University of Science and Technology, Aliero as cultivated garlic Allium sativum L. The transparent covering of garlic was removed manually and sun-dried for two weeks. Mortar and pestle was used to pound the dry garlic cloves into powder, then sieved and stored in a covered plastic container for further uses. All reagents were of analytical reagent grade unless otherwise stated. Distilled water was used in the preparation of solutions and dilution unless otherwise stated. The physiochemical analyses were carried out in triplicates unless otherwise stated.

Dr. A.U. Itodo (e-mail: itodoson2002@gmail.com).

INTERNATIONAL JOURNAL OF FOOD AND NUTRITION SCIENCE VOL1 NO2 MARCH 2012 Oil extraction The extraction of garlic oil was conducted with a soxhlet extractor using n-hexane (boiling point of 40oC - 60oC) for six hours. The oils were obtained after the solvent was removed under reduced temperature and pressure and refluxing at 70 oC so as to remove any excess solvent used for the oil extracted. The extracted garlic oil was stored in refrigerator freezer at 2oC for subsequent physicochemical analyses[12]. Determination of percentage yield The oil which was recovered by complete distilling of the solvent on a heating mantle was then transferred into a measuring cylinder. The measuring cylinder is then placed over water bath for complete evaporation of solvent for about 2-3 hours in accordance with the method reported[12] and volume of the oil was recorded and expressed as oil content (%) as follow[12]. Oil content (%) = Volume of the oil x 100% Weight of sample Determination of specific gravity 10cm3 of the oil was measured in a pre weighed measuring cylinder. The weight of the cylinder and oil were measured, the weight of the oil was then obtained by subtracting the weight of the cylinder from the weight of the oil and cylinder. The specific gravity of oil was obtained using equations below[13]. Density of water = W1 W0 V0 Where W1 = weight of empty measuring cylinder + water W0 = weight of measuring cylinder V0 = volume of water used Density of oil = W1 W0 V0 Where W1 = weight of empty measuring cylinder + oil W0 = weight of measuring cylinder V0 = volume of oil used

Determination of peroxide value 2.0g of oil extracted was added to 22cm3 of a solution mixture of 12cm3 chloroform and 10cm3 acetic acid. 0.5cm3 of saturated potassium iodide was added to the flask. The flask was corked and allowed to stay with occasional shaking for 1 minute. 30cm3 of distilled water was then added to the mixture and titrated against 0.1M of Na2S2O3 until yellow colour is almost gone. 0.5cm3 of starch indicator was quickly added and titration continued until blue colour just disappeared. A blank titration was also carried out at the same condition[15]. (S B) X N x 1000 W Where Peroxide value =Meq peroxide per 100g of sample S = volume of titrant (cm3) for sample B = volume of titrant (cm3) for blank N = molarity of Na2S2O3 solution (mEq/cm3) 1000 = conversion of units (g/kg) W = Weight of oil sample Peroxide value = Determination of iodine value 0.4 g of the garlic oil sample was weighed into a conical flask and 20 cm3 of carbon tetrachloride was added to dissolve the oil. Then 25 cm3 of wijs reagent was added to the flask using a safety pipette in fume chamber. Stopper was then inserted and the content of the flask was vigorously swirled. The flask was then placed in the dark for 2 hours 30 minutes. At the end of this period, 20 cm3 of 10% aqueous potassium iodide and 125ml of water were added using a measuring cylinder. The content was titrated with 0.1M sodium-thiosulphate solutions until the yellow colour almost disappeared. Few drops of 1% starch indicator was added and the titration continued by adding thiosulphate drop wise until blue coloration disappeared after vigorous shaking. The same procedure was used for blank test and other samples[14]. The iodine value (I.V) is given by the expression Iodine value = 12.69C (V1-V2) M Where C = Concentration of sodium V1 = Volume of sodium thiosulphate used for blank V2 = Volume of sodium thiosulphate used for determination M = Weight of the sample.

Therefore specific gravity = Density of water used Density of oil used

Determination of saponification value Determination of free fatty acid About 2 g of the garlic oil was added to a flask with 30 ml 2.0g of of oil was measured into 250cm3 Erlenmeyer flask, ethanolic KOH and was then attached to a condenser for 30 minutes 100cm3 of ethanol was added and followed by 2cm3 of to ensure the sample was fully dissolved. After sample the had phenolphthalein indicator. The mixture was shake and titrated cooled, 1 ml of phenolphthalein was added and titrated with 0.5M against 0.1M NaOH with continuously shaking until the HCl until a pink endpoint has reached[14]. endpoint is reached, which is indicated by a slight pink color Saponification value was calculated from the equation[14] that persists for 30seconds, the free fatty acid is expressed as[16] Saponification value = (S-B) x M x 56.1 %FFA = V X N X 282 X100 Sample weight (g) W Where S = sample titre value Where %FFA = Percent free fatty acid (g/100g) B = blank titre value V = Volume of NaOH (cm3) M = molarity of the HCl N = Molarity of NaOH 56.1 = molecular weight of KOH 282 = Molecular weight of oleic acid W = weight of oil sample

INTERNATIONAL JOURNAL OF FOOD AND NUTRITION SCIENCE VOL1 NO2 MARCH 2012

3.58%[21], indicates that the oil can be kept for a very long period of time[21]. Determination of acid value 25cm3 of 5% ethanol was boiled on a water bath the heating is to ensure the removal of dissolved gases. 2.5g of garlic oil was addedAcid to value: The acid value 4.18 0.01mgKOH/g obtained 25cm3 of hot ethanol and the mixture was heated to boil. Then few from the oil is low when compared to Sheanut butter oil drops of 1% phenolphthalein indicator was added and titrated against 10.3mgKOH/g[22] and high when compared to 2.39 0.1M KOH with constantly shaking until a permanent pink colour 0.065mgKOH/g, 1.20 0.065mgKOH/g and 0.81 was obtained, the acid value is expressed as[17]. 0.01mgKOH/g for castor seed oil, jatropha oil and cotton seed Acid value = 56.1X M X V oil respectively[12]. Thus the higher the acid value of an oil, W the lower its storage quality and vice-versa[15], this shows that Where M = Concentration of KOH the garlic oil have an excellent storage quality when compared V = Titre value to that of Sheanut butter oil[22]. 56.1 = Molecular weight of KOH W = weight of oil sample Free fatty acid: - Free fatty acid (oleic acid), determine the suitability of the oil for edibility or industrial uses. The free III. RESULTS AND DISCUSSION fatty acid value is 2.10 0.05% which is low when compared The results of the physicochemical analyses conducted on the to the Hyptus spicigera seed oil 3.50%. This shows that it is garlic oil sample are presented in Tables 1and 2. suitable for eating[23]. TABLE 1 Physical properties of the garlic oil Parameters Colour of oil Specific gravity Smell Oil yield Results Light yellow 0.90 Pungent odour 22.5% Saponification value: -The saponification value of the garlic oil was found to be 192 1.00mgKOH/g which is high when compared to 183.1mgKOH/g of shea butter oil[22] and 123.3 3.428mgKOH/g, 122.49 2.591mgKOH/g and 199.42 0.53mgKOH/g for castor seed oil, jatropha oil and cotton seed oil respectively, which they have potential for soap production[12]. This indicates that the oil could be used in soap making since its saponification value is higher than these oils22. Iodine value: - This is a measure of the proportion of unsaturated acid or fat and oil present, but the test measures the amount of iodine absorbed per gram of sample. The determination of iodine value measures the reaction of the double bonds with halogen[24]. The iodine value 12.69 0.05g/100g which was obtained from the oil is low when compared to 16.0g/100g Tiger nut oil[24]. The oil shows quite degree of unsaturated fatty acid which indicates that the oil is suitable for consumption and can also be used as a non drying oil, which are useful in the manufacture of soap[24].

TABLE 2 Chemical properties of garlic oil Parameters

Saponification value (mgKOH/g) Peroxide value (mg/100g) Acid value (mgKOH/g) Free fatty acid (% oleic acid) Iodine value (gI2/100)

Results
192 1.00 2.50 0.50 4.18 0.01 2.10 0.05 12.69 0.05

Values expressed as: Mean SD Physical properties The percentage yield was found to be 22.5% which is low when compared to Bottle Gourd Lageneria siceraria seed oil 39.22%[18] and specific gravity is 0.90g/cm3 which is within 0.915g/cm3 and 0.923g/cm3 in Sesamum indicum L. seed oil[19] and 0.89g/cm3 in African Oil Bean seeds Pentaclethra marcrophylla[20], this indicate that the oil could be used on commercial scale. The colour of the oil is light yellow with a pungent odour. However the oil is liquid at room temperature which could indicate the presence of oleic acid and linoleic acid and other unsaturated fatty acids[18]. Chemical properties Peroxide value: This measured deterioration of oil from oxidation[16]. Therefore, the low peroxide value 2.50 0.50% obtained from oil when compared to palm kernel oil IV. CONCLUSION The results in this analysis indicated good quality for the garlic oil. chemical analyses presented saponification value , peroxide value, acid value ,free fatty acid and Iodine values that fell within the range of those acceptable as having good potential for soap production and with an excellent storage property. REFERENCES [1] U. Quattrocchi, CRC world dictionary of plant names volume I CRC press Baca Raton, New York pp 91, 2000. [2] G.Simonetti. Simon and schusters. Guide to herbs and species simon and Schuster Inc. 1ST Ed. Pp. 67, 1990.

INTERNATIONAL JOURNAL OF FOOD AND NUTRITION SCIENCE VOL1 NO2 MARCH 2012 [3] E. Block. Garlic and other Alliums: The lore and the science. Royal society of Chemistry. pp.190-9,2010. [4] H. McGee, The Onion Family: Onions, Garlic, Leeks. On food and cooking revised edition sceribnes pp. 310-313 , 2004. [5] M. L. Fernald. and S .Gray. Manual of Botany, 8th Edition , new York, American book co, Pp. 306-308,1950. [6] E. Block. The organosulfur chemistry of the genus Allium implications for organic sulfur chemistry. Angewandte Chemie International Edition vol. 140,pp.11581203,1992. [7] E. Block. The Chemistry of garlic and onions. Scientific American. Vol. 252. Pp.114-119, 1985. [8] H. Amagase .,B.L. Petesch H. Matsuura S. Kasuga and Y.Hakura. Intake of Garlic and its bioactive components. The Journal of nutrition. Vol. 131, pp. 955 962,2001. [9] K.L. Chadha. Handbook of Horticulture Indian council of agriculture research 6th Edition New Delhi ,pp.78, 2001. [10] A.E. Andreatta. G. Foco, G. Mabe, and S.B.Bottini . Extraction of garlic oil with Quasi-Critical Solvents. ENPROMA, Argentina. Pp 1-.9, 2005. [11] J.Turner . Spices The History of a Temptation of Vintage Books .1st Ed. Pp.16, 2004. [12] A.A. Warra, I.G. Wawata, L. G. Hassan, S.Y. Gunu, and K.M. Aujara. Extraction and physicochemical analysis of some selected Northern Nigerian industrial oils. Scholars Research Library: Archives of applied science research, vol.3, no4, pp. 536-541,2011. [13] K. John. Analytical Chemistry for Technicians, 3rd Edition, Lewis Publishers, Kalyani Publisher, New Delhi, India, Pp 347-349, 2003. [14] U.G. Akpan , D. Jimoh and A.D. Mohammed . Extraction Characterization and Modification of castor seed oil Leonardo journal of science.vol.8, pp.43-52 ,2006. [15] S. Nielsen. Food analysis, laboratory manual 3th Edition kluwer academic premium Publishers, London.Pp 88-89,2003. [16] S.L, Chopra, and J.S. Kanwar, Analytical Agricultural Chemistry, 4th Edition, New Delhi,Kalyami publishers, Pp 1215, 1991. [17] AOAC - Official methods of analysis of the Association of official analytical chemists,16th Edition, Gaithersburg Pp 13, 1998. [18] L.G. Hassan and N.A. Sani. Comparative Studies on the Physico-Chemical Properties of Bottle Gourd Lageneria

siceraria seeds oils Extracted by two methods. Nigeria journal of Basic and Applied. Sciences: vol.15, no.1&2, pp.50-51, 2007. [19] M.I .Mohammed and Z.U. Hamza . Physicochemical Properties of Oil Extracts from Sesamum Indicum L. Seeds Grown in Jigawa State-Nigeria. Journal Applied Environment Management. Vol.12 no.2, pp.99-100, 2008. [20] S.A. Odoemelam . Proximate Composition and selected Physicochemical Properties of the Seeds of African Oil Bean (Pentaclethra marcrophylla). Pakistan Journal of Nutrition vol.4, no.6. pp.382-383, 2005. [21] M.N. Ogbuagu. Inhibitory effect of onion and garlic extracts on the rancidity of palm and palmKernel Oils. Journal Chemistry Society of Nigeria, vol. 33 no. 1, Pp 43-44, 2008. [22] A. A. Warra, S.Y. Gunu, A. Jega and J.A. Aisha. Soap Production from Shea nut Butter. International Journal of Applied Sciences. Vol. 5, no. 4:410-412,2011. [23] Z. Ladan, E.M. Okonkwo, J.O. Amupitan, E.O. Ladan, and B. Aina. Physicochemical Properties and Fatty Acid Profile of Hyptus spicigera Seed Oil. Research Journal of Applied Sciences vol.5, no. 2, pp. 123-125, 2010. [24] C. I. O. Kamalu. And P. Oghome. Extration and Characterization of Tiger Nut oil. Journal of chemistry Society of Nigeria. Vol. 33, no1, pp. 79-87, 2008. Correspondence to Main Author: * Gafar M.K Department of Chemistry, Kebbi State University of Science and Technology, Aliero, Nigeria Email: maryamgmaryam2011@gmail.com Tel: +2348027415703