ISOLATION OF DOCOSAHEXAENOID ACID FROM FISH OIL AND ITS VALIDATION BY GAS CHROMATOGRAPHY

Abstract

Docosahexaenoic acid (DHA) was isolated from fish oil and identified by gas chromatography. The isolation of DHA
was carried out by the column chromatography. The preparation of column using silica gel 60-120 mesh and elution of
column with different solvent in increasing order of polarity was done. Isolated fractions were subjected to identification test for DHA by thin layer chromatography using solvent benzene. Determination of isolated DHA was carried out by gas chromatography using reagent hexane as a solvent. The chromatogram of isolated DHA and standard DHA were compared. It was showed that in thin layer chromatography the fraction 51-58 and 72-85 showed single spot in benzene mobile phase having Rf value 0.56 when compared with standard DHA which matched with Rf Value of standard DHA. The retention time of isolated DHA and standard DHA by gas chromatography were 5.73 and 5.01 respectively. Isolated DHA showed 95.21 percentage purity. It was observed that fish oil was used as precursor of DHA and the developed method of column chromatographic isolation of DHA and its validation by GC is simple, accurate and precise.