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Identification of Fatty Acid Profile, Lipid Characterization and Nutritional Status of Clarias Batrachus

Rafiquel Islam
Department of Applied Chemistry and Chemical Technology, Islamic University, Kushtia, Bangladesh

Liton Kumar Mondol
Department of Applied Chemistry and Chemical Technology, Islamic University, Kushtia, Bangladesh

Liton Sheikh
Department of Applied Chemistry and Chemical Technology, Islamic University, Kushtia, Bangladesh

Sk. Shahinur Rahman
Department of Applied Nutrition and Food Technology, Islamic University, Kushtia, Bangladesh.

Mominul Islam
Department of Applied Nutrition and Food Technology, Islamic University, Kushtia, Bangladesh.

Atiqur Rahman
Department of Applied Chemistry and Chemical Technology, Islamic University, Kushtia, Bangladesh

Abstract/ Executive Summary:

The present study was carried out to characterize the lipids isolated from Clarias batrachus and to analyze the nutrient contents ?of these fishes. The specific gravity (0.87 at 30°C), refractive index (5.19 at 30°C), co-efficient of viscosity (448.26) were measured. Besides, the quantitative analysis of acid value (1.93), percent of free fatty acids (0.97%), saponification value (179.88), saponification equivalent (311.87), iodine value (53.93), peroxide value (43.63), ester value (177.95), unsaponificable matter (1.96%), acetyl value (16.09), reichert-meissel value (3.13), cholesterol (15.46%) were evaluated. Likewise, carbohydrate, protein and lipids were 6.26, 14.14 and 6.91% respectively. Moisture contents were 78% but ashes were (0.1%). Further, the mineral studied as calcium, phosphorus and iron were 210.10, 70.05 and 7.06 mg/kg respectively, whereas zinc contents were trace amount. The fatty acids profile of C. batrachus was identified as lauric acid (2.6%), palmitic acid (37.41%), oleic acid (49.1%) and stearic acid (3.6%) arachidic acid (3.04%) and behanic acid (4.21%) respectively.

Keywords: Clarias batrachus, Lipids, Fatty acids, Nutrients

Location: Kustia district.

Start Date:

End Date:

Program Area: Quality and Nutrition

Commodity: Cat fish

Objectives:

To characterize the lipids isolated from Clarias batrachus and to analyze the nutrient contents ?of these fishes.

Methodology:

Collection of fish sample: Fresh water 10 fish (Clarias batrachus) samples were collected from Municipal fish Market, Kushtia, Bangladesh at the time of Late Autumn. The average weight of each piece of fish was 300g and the age of C. batrachus was approximately 3.5 years. Extraction of lipid using organic solvents: After collection, 50 g fish fillet was cleansed by discarding their bones, liver, stomach and viscera. Then it was transferred to a volumetric flask. Then lipids were extracted by solvent extraction method using chloroform-methanol solvent. Characterization of lipid of Clarias batrachus: After extraction, the solvent was removed from the lipid at low temperature (to prevent oxidation) with the help of a rotary vacuum evaporator. The lipid was stored at 4⁰C in the presence of an inert gas until analyzed. The physico-chemical characterizations were determined using standard methods. Determination of Fatty acids profile: Preparation of the methyl ester mixture from the lipid The lipid (5 g) was taken in a round bottom flask (125 ml) and saponified with alcoholic potassium hydroxide solution (50 ml). The mixture was then refluxed for 45 minutes on a water bath until it became clear. The reaction mixture was allowed to cool and then neutralized with HCl (5 N). Alcohol was removed from the neutralized solution by evaporation over a steam bath. 25 ml water was added to this alcohol free solution and pH of the solution was adjusted by adding concentrated HCl. The acidified aqueous mixture was then extracted with 20 ml ether in a separating funnel and the extraction was repeated for three times. The combined ether extract was washed with water in order to remove any adhering HCl. Ether was then removed from the extract to give the fatty acid mixture. The fatty acid mixture was then esterified with methanolic solution of sulfuric acid (0.25 M, 5 ml/g acid). After esterification, the mixture was dissolved in ether (25 ml) in a separating funnel and washed with dilute sodium carbonate solution until the effervescence ceased. It was then washed with water, dried over anhydrous Na₂SO₄ and finally ether was removed to give methyl ester mixture. Gas liquid chromatographic examination of methyl esters: The experiment was carried out with a “PUE UNICAM” 4500 U model gas chromatograph equipped with a flame ionization detector. A glass coiled column (3 mm, I.D. 2.1 m) packed with 70-100 mesh chromosorb after impregnating it with 10% diethylene glycol succinate was used for the regular packed column GLC. The temperature programming in the oven was from 130⁰C to 230⁰C with the rate of rising 4⁰C per minute. The oven, injector and detector temperature were 190, 200 and 205⁰C respectively with a nitrogen carrier gas flow rate 30 ml/min. The speed of the chromatogram was at 0.5 cm/min. The fatty acids in the mixture were identified by comparing its relative retention volume. The area of each chromatogram peak was determined by multiplying the height of the peak by the width of the peak at one-half of the height. The percentage of fatty acid contributing to each peak was calculated and the results have been computed. Nutritional status of Clarias batrachus: Estimation of minerals by Atomic Absorption Spectrophotometer (AAS) For the estimation of the trace elements (Ca, P, Fe and Zn) of Clarias batrachus, the chemical analyses of fish were performed with flame atomic absorption spectrophotometer. The technique involved the following steps: Preparation of the standard solution: The stock standard solutions of 100 ppm were prepared from analytical grade reagents of Ca, P Fe and Zn salt with distilled demonized water. The stock standard solution were preserved in clean polythene bottles and kept in a refrigerator. Standard solutions of these metal ions were prepared by suitable dilution of the stock standard solution. Dilution was made freshly with distilled demonized water. Analysis of fish sample: The sample of fish were diluted to a known volume and analyzed by a flame atomic absorption spectrophotometer (Flame AAS). The samples were analyzed against standard solution of each element. A reagent blank was also maintained, and the absorption due to reagent was subtracted. All the glasses were Pyrex and were cleaned before use with detergent, 1:1 HNO₃ and demonized water to avoid any contamination. All the reagents and chemicals were of analytical grade, reagents were prepared with distilled deionized water. The water was prepared by passing distilled water through a mixed-bed ion-exchange resin column. Samples were subsequently analyzed for trace elements by “AAS-680”Atomic Absorption/Flame Emission spectrophotometer (Shimadzu, Japan). A single hollow cathode lamp for each element was used with an air-acetylene and nitrous-oxide-acetylene. Statistical analysis: The identification of fatty acid profile, physico-chemical characterization the lipids and nutrients of Clarias batrachus were analyzed. Each experiment was run in triplicate, and mean values were calculated.

Source of Information: Nutritional Science and Food Technology 2013. 2(1) 1-5

Article Link (Online Journal):

Funding Source:

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Achievement/Findings:

The results obtained from this study suggested that lipids of Clarias batrachus are suitable for edible purpose as it contained higher amount of unsaturated fatty acids. These unsaturated fatty acids always play an important role in the metabolism of living organism. Moreover, fishes also contain higher amount of macro and micro nutrients which may take part in the buildup of human good health.

Knowledge Management: Journal