Incomplete Hydrolysis of Cholesteryl Esters during the Enzymatic Cholesterol Determination as Evidenced by Aqueous Cholesteryl Ester Solutions: Comparison of Six Enzymatic Procedures with the Liebermann-Burchard Method

Summary: Aqueous solutions of cholesterol and some cholesteryl esters were prepared. The hydrolysis of cholesteryl esters with enzymatic methods could therefore be studied in some detail. The total cholesterol concentration of the aqueous cholesterol and cholesteryl ester solutions was determined by 6 different enzymatic procedures as well as the Liebermann-Burchard method. For some esters (acetate and arachidonate esters) the esterase reaction is not complete within the usual reaction time, whereas most other esters gave analytical results lower than the theoretical. With the Liebermann-Burchard method all esters reacted completely within the reaction time. The esterases have very different specificities for the various cholesteiyl esters. With the enzymatic method several commercial control sera as well as human sera gave lower cholesterol concentrations than the Liebermann-Burchard method. These differences can be explained mainly by this incomplete hydrolysis. Some practical recommendations are given. Unvollständige Hydrolyse von Cholesterinestern bei der enzymätischen Cholesterinbestimmung in wäßrigen Lösungen von Cholesterinestern