Mar 1, 1976

Polarographic method for rapid microdetermination of cholesterol with cholesterol esterase and cholesterol oxidase

Clinical Chemistry
A Noma, K Nakayama

Abstract

Cholesterol concentrations in serum are enzymatically determined rapidly by use of a polarographic oxygen analyzer with a circuit modified to record simultaneously the amount and rate of oxygen consumption. The final assay system, assessed from the oxygen consumption value that we found to be optimum, consists of 1 ml of sodium phosphate buffer (0.6 mol/liter, pH 7.0) containing NaN3 (10 mg/liter), Triton X-100 surfactant (10 ml/liter), 0.4 U of cholesterol ester hydrolase, and 0.6 U of cholesterol oxidase. Oxygen consumption and cholesterol concentration are linearly related to 8.0 g/liter, and only 10 mul of serum is required. Replicate analyses of pooled serum by the present method demonstrated the following inter-run precision: mean = 1731 mg/liter, SD = 22.3 mg/liter, CV = 1.3%. Bilirubin and ascorbic acid were without effect on the present method, unlike the enzymatic colorimetric methods.

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Mentioned in this Paper

Epicholesterol
Oxygen Consumption
Orthophosphate
Lysosomal Acid Cholesterol Esterase
Hydrogen-Ion Concentration
Carboxylic Ester Hydrolases
Acid Cholesteryl Ester Hydrolase Deficiency, Type 2
Osmolality
Microchemistry

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