Serum fatty acids analysis by high performance liquid chromatography, after enzymatic hydrolysis and isolation by...

A simple method of high performance liquid chromatography for the determination of fatty acid composition of free fatty acid, triglyceride, cholesterol ester and phospholipid in 0.4 ml of human serum is described. The procedure includes the enzymatic hydrolysis of serum lipoproteins, the isolation of fatty acids using Sep-Pak C18 minicolumn, the p-bromo-phenacylester formation and the high performance liquid chromatography. Thirteen fatty acids including the internal standard separated into 10 peaks within a 30-min run. The recovery rates of Sep-Pak treatment were satisfactory. The coefficients of variation were 2-27%. The present method showed a limit of detection of 0.02-4 nEq fatty acid.