Influence of incomplete cholesteryl ester hydrolysis on enzymic measurements of cholesterol

Esterolytic activity of three enzymic cholesterol reagents was evaluated with use of human serum-based reference materials. Primary cholesterol standards were used to calibrate the enzymic methods, and incomplete cholesteryl ester hydrolysis was measured as the bias between the enzymic cholesterol values and corresponding reference values. The average biases observed with eight reference specimens were 0.15, -1.16, and -7.24%, respectively, for a commercial enzymic reagent with microbial cholesterol esterase, a commercial reagent with an animal source of esterase, and a Centers for Disease Control (CDC) reagent preparation with a microbial cholesterol esterase. By liquid chromatography, we examined the respective specificities of the three esterase reagent preparations and found that the preparations differ in the rate and specificity of catalysis of cholesteryl ester hydrolysis. The CDC reagent was only partly active towards cholesteryl arachidonate, which probably accounts for most of the negative bias of this reagent. Esterolytic activity of the various enzymic reagents is related to the source and matrix of the cholesterol esterase and must be considered with respect to proper selection of calibration materials for serum cholesterol measurement.