Simple thin-layer chromatographic purification procedure for the determination of cholesterol ester fatty acid compositions

Abstract

A mild procedure for the purification of methyl esters of the fatty acid components of cholesterol esters, from interfering free cholesterol and other contaminating residues, is described. Methyl esters and free cholesterol are formed during the methylation of cholesterol esters. When co-extracted, cholesterol and other contaminating residues interfere with the methyl esters because minute proportions of these residues tend to elute at the same retention times as palmitoleic and stearic acids, to yield unreliable but significantly higher values for palmitoleic (p less than 0.001) and stearic acids (p less than 0.0001), and correspondingly lower values for oleic acid (p less than 0.0001). Purification of methyl esters by thin-layer chromatography eliminates this problem and yields reliable analysis of cholesterol ester fatty acids, without measurable oxidation of unsaturated fatty acids.