QUANTITATIVE DETECTION METHOD OF TRIGLYCERIDES IN SERUM-LIPOPROTEINS AND SERUM-FREE GLYCEROL BY HIGH-PERFORMANCE LIQUID-CHROMATOGRAPHY

We have developed a simple and reliable method for quantitative detect ion of triglycerides (TG) in serum lipoproteins and serum-free glycero l (FG) by high-performance liquid chromatography (HPLC). After separat ion of serum constituents using a new gel-permeation column (TSK gel L ipopropak XL, Tosoh) and a new eluent (TSK eluent LP-2, Tosoh), TG and FG were detected by on-line reaction using a modified reagent which c ontained glycerol kinase, glycerol-3-phosphate oxidase and lipoprotein lipase. HPLC patterns showed five peaks corresponding to chylomicrons , very-low-density, low-density, high-density lipoproteins and FG. Abs olute concentrations of TG in each lipoprotein fraction and serum FG w ere calculated from the corresponding peak areas using standard FG as a calibrator. Due to its very high sensitivity of peak detection, this method has become desirable for the analyses of lipoproteins of very low concentrations such as in cell culture systems. This technique wil l contribute to a better understanding of lipoprotein TG and serum FG distribution in human and nonhuman subjects. (C) 1998 Elsevier Science B.V. All rights reserved.