Analysis of cholesterol and cholesteryl esters in human serum using capillary supercritical fluid chromatography

Capillary supercritical fluid chromatography (SFC) using carbon dioxide as a mobile phase was applied for the determination of free cholesterol and cholesteryl esters in human serum. Serum samples were extracted with methanol-chloroform (2:1, v/v), and the extracts were analyzed by pressure programmed capillary SFC-flame-ionization detection (FID) without thermal degradation and derivatization. The total cholesterol concentrations obtained from SFC analysis were compared with those from GC or enzymatic analysis. The capillary SFC-FID method having high resolution gave an acceptable average relative standard deviation of 2.6%, and a detection limit of 4-6 pg. The quantitative results were acceptable for the simultaneous analysis of cholesterol and its esters in biological fluids. The concentration profiles of each compound in various samples, normal Korean human serum, Western human serum, and from high-cholesterol patient plasma, have been compared with this method.