Rapid method for the quantitation of cholesterol in human serum lipoproteins by high performance liquid chromatography.

A simple and rapid method for the quantitation of cholesterol in human serum lipoproteins (VLDL, LDL, HDL2, and HDL3) was developed (1). The content of cholesterol in each lipoprotein fraction was determined by means of a commercial enzymatic reaction kit after separation by high performance liquid chromatography with gel permeation columns. The quantitation of cholesterol could be performed with only 10-20 mu l of serum in less than 50 min by measuring A550 after passing the mixed eluate and enzyme solution through an on-line reactor system of a high-speed chemical derivatization liquid chromatograph. The precision, reproducibility and sensitivity of the quantitation of cholesterol with this method were acceptable, and the values of HDL-cholesterol determined by this method correlated well with those found by the heparin-manganese chloride precipitation method (r=0.958, n=93).