Estimation of polyunsaturated fatty acid content in lipids of aquatic organisms using thin-layer chromatography on a plain silica gel plate.

Abstract

We have designed a rapid method for the separation of polyunsaturated fatty acids (PUFA, > or = trienes) from non-PUFA, and for estimation of total amounts of PUFA in lipids of aquatic organisms. Lipids from thirty-one species, including marine and fresh water fishes, shell fishes, marine algae, and other aquatic animals, and from terrestrial organisms, were transesterified with sodium methoxide in methanol. The resulting fatty acid methyl esters were separated by thin-layer chromatography on commercially available plain silica gel plates with a developing solvent of n-hexane/ethyl ether/acetic acid (95:5:1, by vol). All of the methyl esters from aquatic organisms tested separated into two spots, whereas those from terrestrial sources, except for linseed oil, showed a single unresolved spot. The upper and lower spots were scraped separately from the plate, and their fatty acid compositions were determined by gas-liquid chromatography. The lower spot was composed of PUFA having more than two double bonds, whereas components of the upper spot were saturated, monoenoic, and the greater part of the dienoic fatty acids. When the spots on the silica gel plate were stained with Coomassie brilliant blue, the amounts of PUFA in aquatic organisms could be estimated satisfactorily using a scanning densitometer.