Enzymatic Determination of Triglycerides in Conjunction with High Performance Liquid Chromatography

Abstract

The glycerol kinase (GK) catalyzed reaction involving the conversion of glycerol and adenosine triphosphate (ATP) to glycerol-3-phosphate and adenosine diphosphate (ADP) has been used in conjunction with HPLC for the determination of triglycerides. After alkaline hydrolysis of the triglycerides to glycerol, the enzyme reaction was carried out. The ADP formed and the remaining ATP were then separated by HPLC and the ADP peak area correlated to the concentration of triglycerides originally present in the sample. Linearity of the method was established from 28–180 mg/dl with a reproducibility of 6.5% RSD. A comparison between the HPLC method and the standard coupled enzyme system for triglycerides in real serum indicated a correlation coefficient of 0.977.