Qualitative assessment of fish body oil extracted from Sardinella fimbriata from Muttom coastal waters, Kanyakumari District, Southwest coast of India

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1 Int. J. Int. J. Cur. Tr. Res (2014) 3 (2):34-38 ISSN: Qualitative assessment of fish body oil extracted from Sardinella fimbriata from Muttom coastal waters, Kanyakumari District, Southwest coast of India Xavier Eugien, L..E. Anand Ganesh and S.M.Raffi* Centre of Advanced Study in Marine Biology, Faculty of Marine Sciences, Annamalai University Parangipettai , Tamil Nadu, India. Department of Biochemistry, NKP, Salve Institute of Medical Sciences, Digdoh Hills, Hingna. Nagpur , Maharashtra, India. Received: 5 January 2014 Accepted: 22 February 2014/ Published online 28 February INJCTR 2014 ABSTRACT The present study dealt with the extraction and purification of fish body oil from a low value fish Sardinella fimbriata collected from Muttom coastal waters of Kanyakumari district, Tamilnadu, India. Direct steaming method was employed for the production of fish oil which produced an average yield of 130 ± 4.5 ml for every 1000g fish tissues. The quantity of fish oil produced after each steps of purification, namely degumming, neutralization, bleaching and deodourization were worked out separately. Purified fish oil showed notable improvement in free fatty acid, iodine value, peroxide value and saponification values that exhibited lower values than that of crude ones. In the case of fresh and stocked samples, these values were slightly increased in 30 days old (Stock) samples than that of fresh ones. Keywords: Fish body oil, Sardinella fimbriata, qualitative assessment. Introduction Fish oil is an excellent dietary source, rich in essential fatty acids, especially with Polyunsaturated Fatty Acid (PUFA) in like ω-3 Fatty Acid, Eicosapentaenoic acid (EPA) and Docosahexaenoic acid (DHA) ( Conner, 2000; Zhao et al., 2010). Fish oil has fetched more importance in the recent years because of its wider application prospects. Production of fish oil from low value fishes has gained increased momentum in recent past. Various processing methods have been adopted for the extraction of fish oil from the liver and whole body, such as Soxhlet method, Bligh and Dyer method, direct steaming method, solar extraction method and Mc Gill and Moffat method (Immanuel et al., 2002). The conventional method of extraction (direc t steaming method) is considered the finest extraction process due to its winsome qualities such as higher yield, economic viability, less laborious and less time consumption etc. ( Bimbo, 1990; Raffi, 2012). The production of the fish oil deals with the separation of lipids from other constituents of the fish. Generally, the production involves a series of steps for the purification of the product. According to Firestone (1989) of American Oil Chemist s Society, (AOCS), the major steps in fish oil refining are degumming, neutralizing, bleaching and deodourizing. Both soluble and insoluble impurities are removed through degumming process, followed by neutralization of crude oil with Sodium hydroxide that removes Free Fatty Acids (FFAs). Bleaching aids in the removal of soap, trace metals, sulfurous compounds and part of the more stable pigments and pigment-breakdown Corresponding author* E.mail: raffi_cas@yahoo.co.in

2 35 Xavier et al. Int. J. Cur. Tr. Res (2014) 3 (2):34-38 products, aldehydes and ketones. The final step is deodourization that removes residual FFA, aldehydes and ketones, which are responsible for an unacceptable oil odour and flavour (Young, 1978; Gavin, 1977). Many species of marine fish have been studied for fish oil production, but little attention has been paid to the production of fish oil from low value fishes. Understanding the nutraceuticals properties of fish oil and its wide application, an attempt has been made in the present study in which fish body oil was extracted from low value fish, namely, Sardinella fimbriata (lesser sardine) by direct steaming method and its yield and quality assessment were determined for the crude and purified oil. Materials and Methods Sardinella fimbriata (lesser sardines) were collected from fish landings of Muttom, Kanyakumari district, Tamilnadu, southwest coast of India (lat N; long E). The fishes were washed thoroughly in running water for the removal of sand and external debris; scales, head, fins, spines, digestive system and excretory system were removed and the tissues alone were taken for extraction of oil. The tissues were subjected for extraction of oil by Direct steaming method ( Immanuel et al., 2002). The homogenized fish tissues was taken in a muslin bag and kept in Steam Boiler at C for 30minutes. The boiled fish tissues were then pressed using Fish Oil Extractor, so as to remove the liquid content from the tissues (containing oil and water). The oil was separated from the water by centrifuging at 2000rpm for 15minutes and by using separating funnel. The filtered oil was stored separately in opaque dark bottle and placed in deep freezer at -20 C. Purification of oil Purification/Refining is a series of process that includes 4 steps, namely degumming, neutralization, bleaching and deodourization. Degumming was carried out following the method of Dijkstra and Opstal (1989) with modifications. A sample of crude oil (100g ) placed in a 600ml beaker and heated to 70 C in oven for 1min. 3ml of 3% aqueous citric acid solution was added to the oil, and the mixture was thoroughly mixed at 70 C for 1minute. The oil was then cooled to room temperature and centrifuged at 2,560 g for 10minutes to remove impurities. The degummed oil was neutralized following to AOCS Official Method (Firestone, 1989). Sodium hydroxide (12.6g of 9.5% NaOH solution) was added to 100g of degummed oil and the mixture was heated at 65 C for 30minutes with constant stirring using a magnetic stirrer bar. The sample was then cooled to room temperature and kept undisturbed for six hours. After centrifugation at 2,560 g for 10minutes, the oil was decanted from the precipitated soap. 50ml of demineralized water was added to the centrifuged sample to wash out any remaining soap. This operation was repeated three times. The remaining water and impurities were removed by centrifugation at 2,560 g for 10minutes. Table 1 Quantity of fish body oil obtained from S. fimbriata after different steps of purification S.No Purification process Oil used (ml) Oil obtained (ml) 1 Degumming ± Neutralization ± Bleaching ± Deodourization ± 0.5 Table 2 Quality Analysis of crude and purified fish oil S.No Analytical Para meters Moisture Content (%%%) Free fatty acid (mg KOH/g) Iodine Value (I2/100g) Peroxide Value (meq/kg) Saponificati on value (mg/ KOH/g) 6 Colour Fresh (0 days) 0.7 ± ± ± ± ± 1.6 Slight Brow nish Crude oil Stock (30days) 0.4 ± ± ± ± ± 2.1 Slight Browni sh Purified oil Fresh (0 days) 0.3 ± ± ± ± ± 2.05 Bright Stock (30 days) 0.32 ± ± ± ± ± 3.2 Bright

3 Int. J. Cur. Tr. Res (2014) 3 (2): Xavier et al. 36 Fig. 1 Percentage wise yield of fish oil obtained during different steps of purification Crude oil Degummed oil Neutralised oil Bleached oil Deodourised oil ml of fish oil was subjected to each stage of purification and the recovered yield was worked out (Table 1). 100ml of crude fish oil subjected to degumming produced 83 ± 0.5 ml, 100ml of degummed oil subjected to neutralization produced 87 ± 0.5 ml, 100ml of neutralized oil subjected to bleaching produced 95 ± 1.0 ml oil and from 100ml bleached oil subjected to deodourization yielded 82 ± 0.5 ml of purified oil. The percentage wise loss of oil removed as impurities in each step of purification was calculated (Fig.1). The total recovery percentage of oil at the final stage of purification process was 55% i.e., 55 ml of refined oil was obtained from 100ml of crude oil. Analytical Properties of oil The neutralized oil was bleached following the method of Makhoukhi et al. (2009 ). The 100ml of neutralized oil was bleached with 1g of acid activated bentonite at 100 C for 20minutes with constant stirring with a magnetic stirrer bar. The activated bentonite with the absorbed impurities was removed immediately by filtration to avoid colour changes to the oil. The bleached oil was deodourized following to the method of Firestone (1989) with modifications. The bleached oil was deodourized using a laboratory distillation unit, consisting of a 500ml roundbottomed boiling flask with three outlets. One outlet was connected to a vacuum pump and outlet was connected to a glass distillation column, and the third outlet was sealed with a thermometer inserted. The flask was placed on a heating system. The bleached oil (100m l) was added to the flask and heated at 100 C for 30minutes under vacuum (5mm Hg). The temperature was controlled manually. Volatile products were condensed in a cooling system installed on the vacuum line and are removed. Quality Assessment of Fish oil Quality assessment of crude and purified oil was worked out separately for fresh and 30 days old samples were kept in refrigerator. Free fatty acids (FFA) were assessed following the methodology of Cox and Pearson (1962), iodine value (IV) by Horowitx (1975), peroxide value (PV) by Cox and Pearson (1962), saponification value (SV) by Horowitx (1975) and Observation of colour (Immanuel et al., 2002). The fish oil extracted from the tissues of Sardinella fimbriata employing Direct steaming method produced an average of 132 ± 4.5 ml for every 1000g of Sardinella fimbriata fish tissues. The qualitative estimation of crude and refined fish oil were evaluated separately for freshly prepared samples and for 30 days old samples stored in refrigerator at 0 C (Table 2). These analytical values fall well within the acceptable standard values. The results showed that purified fish oil showed notable improvement in the analytical properties in the case of free fatty acid, Iodine value, Peroxide value and saponification values that exhibited lower values than that of crude ones. In the case of fresh and stocked samples, these values were slightly increased in 30 days old (Stock) samples than that of fresh ones. Discussion During last few years, fish oil is being considered as cheap and chief nutraceutical which warrants an ever growing demand for it. Direct steaming is considered as a good old traditional and economic technique for extraction of oil. The present experiment supports the suggestions of Sunarya et al. (1991) that oil extraction by direct steaming is easier, cheaper, and quicker and also affordable to laymen and rural communities. The oil extracted from Sardinella fimbriata in the present study (13%), was almost double the quantity of oil extracted from Sardinella lemuru (6%) by Khoddami et al. (2009) in Malaysian waters. The results of purification were more or less similar to the work carried out by Sathivel et al. (2003) and the possible reason for slight difference in deodourization process might be due to the difference in bleaching steps adopted, where activated bentonite was used in the present investigation instead of activated earth. The standard values for fish oil have been emphasised by FAO (2005).The results obtained in the present study were

4 37 Xavier et al. Int. J. Cur. Tr. Res (2014) 3 (2):34-38 similar to the standard values; nevertheless some values were pretty close to upper limit of standard range.results of the present study confirmed the findings of Summers et a1. (1991) where purified oil produced showed very low percentage of free fatty acids and peroxides. The appreciably lower values of free fatty acids in freshly purified oil could be attributed to the fact that the lesser the period of exposure of oil to atmosphere, the lesser the rate of the oxidation of the oil and consequently the lesser values of FFA ( Cmolik and Pokorny, 2000). The results of Free Fatty acids, Iodine value, Peroxide value and Saponification value were increased from fresh to 30 days old samples; which are in line with the findings of Boran et al. (2006). Due to its high content of unsaturated fatty acids, fish oil is highly susceptible to oxidative spoilage (Huss, 1988). The reduction in free fatty acids during the deodourization process has been due to the vaporizability of free fatty acids (Gavin, 1978 and Norris, 1982). The appearance of the crude fish oil was brownish, which might be due to the prolonged heating period during, that often oxidizes the oil and imparts a brownish colour (Hall, 1992). After refining, the oil turned bright colour, which indicates the significant removal of pigments. Adeniyi and Bawa (2006) suggested that bleaching contributed to the physical improvement of the refined oil. The impurities were completely removed during the purification process, which resulted in bright colouration to refined oil. The present study recommends that being a low value fish, Sardinella fimbriata should be considered as a candidate species for the commercial production of fish body oil as it ensures excellent qualities both quantitatively and qualitatively Acknowledgements The authors are thankful to the authorities of Annamalai University for the facilities provided and Department of Biotechnology, Government of India for the financial support. References Adeniyi, O. D and A. A. Bawa (2006). Mackerel (Scomber Scrombrus) Oil Extraction and Evaluation as Raw Materials for Industrial Utilization. Leonardo J Sci. 8: Bimbo, A. P. (1990). Production of fish oil. In: Fish oil and nutrition. Ed. M. E. Stansby, Reinhold Publishing Co. Ltd., New York, Boran, G., H. Karac and M. Boran (2006). Changes in the quality of fish oils due to storage temperature and time. Food Chem. 98: Cmolik, J and J. Pokorny (2000). Physical refining of edible oils. European J Lipid Science and Tech. 102: Colin, F. M., S. M. Alister, H. Roy and S. A. Robert (1993). The Production of Fish Oil Enriched in Polyunsaturated Fatty Acid Containing Triglycerides. J Am. Oil Chem. Soc., 70(2): Conner, W.E. ( 2000). Importance of omega-3 fatty acids in health and disease. American J. Clin. Nutrit., 71: Cox. H. E and D. Pearson (1962). The chemical analysis of foods. Chemical publishing Co. Inc, New York, 41 pp Dijkstra, A and M. V. Opstal (1989). The Total Degumming Process. Chemical publishing Co. Inc, New York, 66 pp FAO (2005). United Nations Food & Agriculture Organization, Nutritional elements of fish. FAO, Rome.pp 68 Firestone, D. (1989). AOCS, Official Methods and Recommended Practices of the American Oil Chemists Society. American Oil Chemists Society, pp Gavin, A. M.(1977). Edible Oil Deodorizing Systems, J. Am. Oil Chem. Soc., 54: Hall, G. M. (1992). Fishery by products in Fish Processing Technology. Blackie Academic & Professional Publishers, pp:179. Horowitx, W. (1975). Official methods of analysis of AOAC. Association of official analytical chemists Washington (12 th Ed), pp: Huss, H. H. (1988). Fresh fish quality and quality changes. Food and Agricultural Organisation U.N., Rome, special publication, pp:48. Immanuel, G., V. Menenthira, A. Palavesam and M. Peter Marian (2002). Physio chemical properties and fatty acid profile of Odonus niger liver oil. Ind. J. Fish., 49(2): ISI. (1974). Indian Standard Methods of Sampling and tests for oil and fats. Part II. Purity tests (Second Revision), Indian Standard Institute (ISI), New Delhi, pp Khoddami, A., A. A. Ariffin, J. Bakar and H. M. Ghazali (2009). Fatty Acid Profile of the Oil Extracted from Sardinella lemuru Fish Waste (Head, Intestine and Liver). World App Sciences J., 7 (1): Makhoukhi, B., M. A. Didi, D. Villeminb and A. Azzouzc (2009). Acid activation of Bentonite for use as a vegetable oil bleaching agent. Grasas Y Aceites, 60 (4): Moffat, C. F., A. S. McGill, R. Hardy and R. S. Anderson (1993). The production of fish oil enriched in polyunsaturated fatty acid containing triglycerides. JAOCS, 70: Raffi, S.M. (2012). By catch Resources. In: UNU- UNESCO, Training manual, Annamalai University, India, pp: 468. Sathivel, S., W. Prinyawiwatkul, J. M. King, C. C. Grimm and S. Lloyd (2003). Oil Production from Catfish Viscera. JAOCS, 80 (4):

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