ANION-EXCHANGE HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHIC ASSAY OF PLASMA-LIPOPROTEINS

A fast, accurate, and precise high-performance liquid chromatographic (HPLC) assay method for human plasma lipoproteins has been developed. The method includes complete separation of high-, low-, and very-low-d ensity lipoproteins within 20 min using a DEAE-glucomannan gel with st epwise elution, enzymatic oxidation of cholesterol (following ester hy drolysis) followed by a two-step reaction to form a fluorogenic produc t, and fluorometric detection as the total cholesterol (TC) level of e ach lipoprotein. The calibration range is 5-1000 mu g/ml for the TC le vel of 20-mu l plasma samples and the relative standard deviation (n = 5) of the precision at 5 mu g/ml is less than 10%. Each lipoprotein l evel determined by this optimized HPLC method agrees well with levels determined by ultracentrifugation or precipitation methods. This new m ethod was successfully applied to assays of plasma lipoproteins in var ious forms of hyperlipidemia. (C) 1995 Academic Press, Inc.