Quantitation of plasma free cholesterol and cholesteryl esters by high performance liquid chromatography. Study of a...

We describe a convenient method for the separation and quantitation of plasma free cholesterol and cholesteryl esters by high performance liquid chromatography (HPLC). After extraction of 100 microliters plasma with isopropanol the plasma cholesteryl esters were resolved on a Zorbax ODS reversed-phase column by isocratic elution with acetonitrile/isopropanol (50:50, v/v). Baseline separation of the plasma cholesteryl esters including the internal standard was obtained within a 25-min run. The intra- and interassay CV was less than 4%. The results obtained by HPLC show good agreement with enzymatic and gas-liquid chromatographic methods. High performance liquid chromatography provides a simple method for the quantitation of individual cholesteryl esters avoiding tedious chromatographic and derivatisation steps inherent to GLC. Our HPLC method was applied to the monitoring of plasma cholesteryl esters in a normal population and can also be used for the study of cholesteryl esters from lipid extracts of biological samples.