Simple Method for Analysis of Apolipoproteins in High Density Lipoproteins by High Performance Liquid Chromatography1

A simple and rapid method has been developed for the separation of apolipoproteins in high density lipoprotein (HDL) fractions by high performance liquid chromatography (HPLC) with gel permeation columns (G3000SW TSK GEL). The HPLC pattern monitored by A280 for a mixed solution of the HDL fraction (10 microliter) and an eluent buffer (200 microliter, 0.1 M sodium phosphate buffer containing 0.1% SDS) incubated at 60 degrees C for 5 min showed two completely separated peaks which corresponded to the major components of human HDL, apolipoprotein A-I and A-II. Moreover, quantitation of apolipoprotein A-I by our method was found to correlate well with that by a single radial immunodiffusion (SRID) assay.