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

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Abstract
Aqueous solutions of cholesterol and some cholesteryl esters were prepared. The hydrolysis of cholesteryl esters with enzymatic methods could 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 incomplete within the usual reaction time; 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 cholesteryl 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.

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