14 | T. Res. J. 02(01): 14-19; July, 2011 Muhammad et al., 2011 CHOLESTEROL, OMEGA-3 AND OLEIC ACID CONTENTS OF VARIOUS OILS, BUTTER AND VEGETABLE GHEE USED AS DIETARY FATS *1Muhammad Shoaib Akhtar, 2Darakhshan Jabbin, 1Hafiz Muhammad Irfan and 2 Naheed Abbas 1 Department of Pharmacy, University of Sargodha, Sargodha and 2Department of Rural Home Economics, University of Agriculture, Faisalabad-Pakistan. ABSTRACT Cholesterol, omega-3 and oleic acid contents were determined chemically in butter, vegetable (vanaspati) ghee and different oils routinely used as dietary fats in this country. The data obtained have suggested that cholesterol contents in the cooking oils were relatively much less than butter and ghee while omega-3 and oleic acid contents were higher in the oils tested; especially in the olive oil as compared to the ghee. Thus the use of the oils is safe and their use should be strongly recommended as they will reduce the risk of arteriosclerosis by lowering plasma triglyceride levels. Keywords: Cholesterol, Omega-3, Linoleic acid, Oleic acid, Arteriosclerosis, Dietary fats. Corresponding Address: Prof Dr Muhammad Shoaib Akhtar, Department of Pharmacy, University of Sargodha, Sargodha-Pakistan. Email: drmsakhtar@gmail.com INTRODUCTION Cholesterol belongs to sterol group of lipids which is abundantly present in food of animal origin like eggs, dairy products, fatty meals and animal meats (Khan,1988). Cholesterol has been well established to perform a number of vital functions in the body, such as providing an essential component of membranes and serving as a precursor of bile acids, steroid hormones and vitamin D (Harvey and Champe, 1994). Blood cholesterol level depends on several factors: sex, age, nutrition and genetic predisposition, etc. Older people have higher cholesterol in blood which promotes the development of various diseases including formation of gall bladder stones as well as arteriosclerosis, leading to hypertension and angina pectoris (Colmenero, 1996). Cholesterol balance studies in humans have indicated a wide variation in efficiency of intestinal cholesterol absorption, ranging from 20-80%, with most individuals absorbing between 40 to 60% of ingested cholesterol (Ros, 2000). 47-Tanzeem Manzil, Bahawalsher Raod, Mazung, Lahore, Pakistan. Cell: 923335272042,Tel.: 924238431969, Fax: 924237117012 15 | T. Res. J. 02(01): 14-19; July, 2011 Muhammad et al., 2011 Dietary fats and oils have been reported to play an important role in human nutrition as they supply more than twice the net energy than carbohydrates or proteins. They supply many essential fatty acids: linolenic acid (omega-3), linoleic acid (omega-6), oleic acid (omega-9, Botanical name: Olea europaea) and arachidonic acid, etc. in the diet. Omega3 essential fatty acids are very important because they have polyunsaturated carbon bonds that are easily broken and do not harm the body. In addition to providing energy, essential fatty acids are part of the structure of every cell in human body and are needed to achieve and maintain a healthy heart but they are also essential for a healthy function of other organs, eyes, skin, joints, hair and immune system (Schmidt et al, 1994). It has been observed that omega-3 essential fatty acids also reduce the risk of atherosclerosis by their lowering effect on plasma triglyceride levels (Goodnight et al, 1982). Omega-3 essential fatty acids are of different types depending upon the number of carbon atom and number of unsaturations in which eicosapentaenoic acid (EPA) containing 20-carbon atoms and decosahexaenoic acid (DHA) contains 22-carbon atoms. The monounsaturated fatty acids i.e. oleic acid is found in animal and vegetable oils which is used in food industry to make synthetic butter and cheeses and to flavor the baked goods. Cooking oil manufacture involves cleaning the seeds, grinding them, pressing, and extrading the oil from them. In extracting, a volatile hydrocarbon such as hexane is used as a solvent. After extracting, the oil is refined, mixed with an alkaline substance, and washed in a centrifuge. Further washing and refining follows, and then the oil is filtered and/or distilled. It is then ready for packaging.(Hoffman, G 2008). Therefore, present study was undertaken to determine levels of total cholesterol, omega-3 and oleic acid in the common dietary fats used by local population. MATERIALS AND METHODS An investigation was carried out to estimate the level of cholesterol in dietary fats and oils. For this purpose, seven samples of cooking oils, five samples of banaspati ghee, three samples of desi ghee and three samples of butter were purchased from local market. The samples included homemade and commercially available oils, ghee and butters which are most commonly used here. The samples of cooking oils included Kisan sunflower, Kisan canola oil (Pakistan Kisan Manufacturer), Planta (Dalda Foods Pvt. Limited), Habib (Habib Cooking Oil Pakistan), Kashmir(United industries limited), Rafhan corn oil and Olive oil. The samples of banaspati ghee included Dalda, Tullo (Wazir Ali Industries), Kisan, Kashmir and Handi banaspati ghee. Desi ghee samples were of Nirala, Habib and homemade desi ghee and butter samples were blue band, homemade and desi butter purchased from a local dairy shop. For cholesterol determination, samples of fats/oils were taken and weighed in 1g quantity. Chloroform was used as solvents to make the volume 10 ml. Samples were stirred to dissolve completely and diluted to 10 times. The 3ml of diluted sample solutions were taken and absorbance was determined with spectrophotometer (Spectronic 20-D, Milton Roy Company) after adding Liberman-Burchard reagent. The reagent reacts with the sterol to produce a characteristic green color whose absorbance is determined on spectrophotometer at 640 nm (Sabir et al, 2003). 47-Tanzeem Manzil, Bahawalsher Raod, Mazung, Lahore, Pakistan. Cell: 923335272042,Tel.: 924238431969, Fax: 924237117012 16 | T. Res. J. 02(01): 14-19; July, 2011 Muhammad et al., 2011 Fatty acids composition was determined by using gas chromatography. Methyl esters were prepared. For this purpose, approximately 200 mg (0.2g) of oil was taken in volumetric flask. Oil sample was heated to make it moisture free. In 200 mg of oil, 25 ml of methanol and 1-2 pellets of potassium hydroxide was added. Then contents of the flask were refluxed for 20-25 minutes on water bath by attaching with condenser and then let it cool for some time. Then these were extracted with hexane and two washings with water were given, the below water was discarded and residue was taken in beaker. Sodium sulphate (anhydrous) was added in it, shacked and was filtered off. After filtration, some was allowed to evaporate at room temperature and the rest was saved. Fatty acid methyl esters were prepared according to standard IUPAC Shimadzu gas chromatograph model (7-A) fitted with a methyl lignose rate coated (film thickness=0.02µm) polar capillary column SP™ -2330 (30m, 0.30mm) and a flame ionization detector. Oxygen free nitrogen was used as a carrier gas at a flow rate of 5ml/min, column length is 50mm, initial oven temperature-220◦C, injector temperature-230◦C, detector temperature-250◦C, temperature hold (2 minutes before the run and 10 minutes after the run), volume of sample-0.2µl, pressure-400Psi and the samples were injected with 1 µl syringe (SGE, Australia). Fatty acid methyl esters were identified by comparing their relative and absolute retention time to those of authentic standards of fatty acid methyl ester. A chromatographic station (CSW32) data-handling programme did all of the evaluation and quantification. The fatty acid composition was reported as a relative percentage of the total peak area. The results were analyzed statistically by using standard deviation and ANOVA (Steel and Torrie, 1984). RESULTS AND DISCUSSION The cholesterol level varied among different samples of cooking oils, vanaspati ghee, desi ghee and butter. Among cooking oils, minimum concentration of cholesterol was found in olive oil and maximum was in Habib cooking oil as shown in Table-1. The cholesterol concentration in ghee samples ranged from 12 to 37 mg/g. Results showed that Dalda and Tullo banaspati ghee contained lower amounts of cholesterol i.e. 12 and 13mg/g , respectively as shown in Table-1. The homemade and commercial samples of desi ghee and butter were also subjected to cholesterol estimation. The cholesterol contents in desi ghee were low as compared to butter as shown in Table-2. Sabir et al, (2003) have reported that Dalda, Tullo and Habib vanaspati contained relatively less amount of cholesterol (10, 14 and 15mg/g , respectively) and they also observed that industrial desi ghee had more cholesterol contents. Zeljka et al, (2000) have determined the milk fat quantity and cholesterol in butter samples originating from village household and industry and showed that homemade butter was better than the industrial as it contained low level of cholesterol. The present study findings have suggested that homemade desi ghee has less amount of cholesterol as compared to commercial products (Table-2). 47-Tanzeem Manzil, Bahawalsher Raod, Mazung, Lahore, Pakistan. Cell: 923335272042,Tel.: 924238431969, Fax: 924237117012 17 | T. Res. J. 02(01): 14-19; July, 2011 Muhammad et al., 2011 Table-1 Concentration of Cholesterol in cooking oils and vanaspati ghees Sr. No. 1 2 3 4 5 6 7 Cooking Oils Cholesterol mg/g Mean±S.D Banaspati ghees 3.5 4 5 4.1± 0.53 4.6 ±0.45 5.1± 0.46 5.2 Olive oil Rafhan corn oil Kisan canola oil Kisan sunflower Planta cooking oil Kashmir cooking oil Habib cooking oil Mean±S.D Dalda Tullo Kisan Cholesterol Mg/g 12 13 25 5.7± 0.37 Kashmir 29 29.6± 0.45 6 6.5±0.49 Handi 37 37.6± 0.53 10 9.7 ±0.54 - - - 12 12.3± 0.32 - - - 12.2± 0.45 13.5± 0.54 25.4± 0.36 Table-2 Concentration of Cholesterol in Desi ghee and Butter samples Sr.No. Desi ghee samples 1 2 Homemade desi ghee Nirala desi ghee 3 Habib desi ghee Cholesterol mg/g 4.3 6 Mean±S.D Butter samples 4.3 ±0.42 Homemade butter Blue band butter Desi Butter 6.5 ±0.43 7.5 Cholesterol mg/g 5 Mean±S.D 5.5± 0.54 8 8.4± 0.32 13 13.6± 0.51 Table-3 Percentage composition of fatty acids in different cooking oils Fatty acids Habib Cooking oil (Soybean&canola) Planta Cooking Oil (Soybean) Olive oil Kisan Sunflower Cooking oil C16:0 6.24± 0.48 6.85 ± 0.49 - 6.03 ±0.46 Kashmir Cooking oil (Soybean &Canola) 15.25± 0.54 C18:0 3.20 ±0.37 3.67 ± 0.39 3.77 ±0.39 18.09± 0.57 C18:1 42.65± 0.77 47.40 ± 0.81 54.32 ±0.96 17.19± 0.53 C18:2 24.50 ±0.65 27.04± 0.67 21.52± 0.62 13.03± 0.51 C20:0 - 7.87 ± 0.50 1.84 ±0.26 9.65 ±0.52 C18:3 9.55 ±0.50 7.12 ± 0.49 12.47 ±0.53 27.58 ±0.69 C20:1 C22:0 2.65± 0.35 10.40± 0.52 - 4.84 ±0.47 53.94 ±0.94 25.14 ±0.64 5.76 ±0.49 10.29 ±0.52 - - - Rafhan Corn oil 11.81 ±0.51 15.74 ±0.52 11.84 ±0.51 22.49 ±0.64 29.99 ±0.69 15.77 ±0.53 - Fatty acid analysis of cooking oils confirmed the presence of palmitic, stearic, oleic, linoleic, linolenic, eicosenoic, lignoceric and erucic acids. The retention time of different peaks of selected samples were then compared to the retention times of standard peaks which indicated the presence of fatty acids. The results showed that their percentages as follows: C16:0 in Rafhan Corn oil, C18:0 in Kisan sunflower cooking oil and C18:3 in Planta cooking oil. These values correlated with Meydani et al. (1991) who have reported 47-Tanzeem Manzil, Bahawalsher Raod, Mazung, Lahore, Pakistan. Cell: 923335272042,Tel.: 924238431969, Fax: 924237117012 18 | T. Res. J. 02(01): 14-19; July, 2011 Muhammad et al., 2011 fatty acid contents of soybean, sunflower, corn and canola oils as C16:0 10.9%, 6.02%, 11.4%, 4.1%; C18:0 4.0%, 4.7%, 1.9%, 2.1%; C18:1 (n-9) 24.2%, 20.5%, 25.4%, 56.7%; C18 (n-6) 54.1%, 69.0%, 60.9%, 26.8%; C18:3 (N-3) 7.2%, 0.1%, 0.7% and 10.3%, respectively. These findings have indicated that only Habib, Kisan, Kashmir, olive oil and Rafhan cooking oils contained Omega-3 fatty acids as shown in Table-3. The percentage of oleic acid C18:0 was 4.48% in olive oil which matched with the value 5.0% as reported by Giuliana et al. (1994). Vanaspati ghee is produced after hydrogenation of oils like cotton seed, canola, soybean or palm oils in the presence of a catalyst. The contents of C18:1 was found to be 30.3% that was in accordance with the value of 29.3% as reported by Wilson (1991) and 29.8% by Meydani (1991). Dalda banaspati had significantly higher amount of C16:0 than Kisan banaspati and the percentage of C18:0 was higher than the other studied banaspati ghee as shown in Table-4. Table-4 Percentage composition of fatty acids in ghee samples Fatty acids Dalda Tullo Kisan Handi vanaspati vanaspati vanaspati vanaspati ghee ghee ghee ghee C14:0 1.04± 0.25 1.74 ±0.29 C16:0 20.77 ±0.62 37.28± 0.74 17.18 ±0.57 29.48 ±0.69 C18:0 16.37± 0.44 8.71 ± 0.40 - 10.68 ±0.53 C18:1 cis 25.82 ±0.67 37.28± 0.74 46.33 ±0.80 - C18:1trans C18:2cis C18:2trans C20:0 C18:3 4.48± 0.39 14.08 ±0.42 3.91± 0.38 4.63± 0.32 16.95± 0.45 12.74 ± 0.54 1.10± 0.28 1.97± 0.30 36.48 ±0.73 - 34.74 ±0.70 14.34 ±0.43 4.13 ±0.30 4.85 ±0.33 Nirala desi ghee 2.36± 0.37 20.48± 0.60 10.40± 0.52 25.21± 0.67 5.51 ±0.41 1.02 ±0.22 It is conceivable; therefore, from the above data that the cooking oils tested contain much less cholesterol as compared to ghees. In addition, they have been observed to possess significant amounts of omega-3 and oleic acids. Thus their consumption by the human beings would be safe and even lower the risk of arteriosclerosis. REFERENCES Colmenero, F. J. (1996). 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The Chemistry & Technology of Edible Oils & Fats & Their High Fat Products. aAcademic Press Inc., 1989. www.madehow.com/Volume-1/CookingOil.html 47-Tanzeem Manzil, Bahawalsher Raod, Mazung, Lahore, Pakistan. Cell: 923335272042,Tel.: 924238431969, Fax: 924237117012