Enzymatic assay of total cholesterol in serum or plasma by amperometric measurement of rate of oxygen depletion following saponification

Clinica Chimica Acta; International Journal of Clinical Chemistry
A Kumar, G D Christian

Abstract

A method for serum or plasma cholesterol assay involving amperometric measurement of the rate of oxygen depletion in the cholesterol oxidase-catalyzed oxidation of cholesterol is described. The hydrolysis of the serum cholesterol esters is accomplished by saponification of 50 mul of sample with 0.2 ml of ethanolic KOH (1.0 mol/1) containing 0.5% Triton X-100 for 5 min at 75 degrees C. The rate of oxygen consumption in a 25-mul aliquot of this is measured with a Clark electrode in a Beckman Glucose Analyzer and the assay takes about one minute after incubation; results are read digitally on the instrument. The analyzer cell contains 1 ml of 1 M phosphate buffer, pH 7.4, with 100 mg sodium cholate/100 ml and 0.1-0.2 U cholesterol oxidase.

References

Dec 1, 1972·Scandinavian Journal of Clinical and Laboratory Investigation·A Kallner

Citations

Dec 1, 1990·Baillière's Clinical Endocrinology and Metabolism·M F Laker, F L Game
Jan 1, 1981·Annals of the New York Academy of Sciences·G G Guilbault
Jan 1, 1982·Applied Biochemistry and Biotechnology·G G Guilbault

Related Concepts

Ampholytes
Epicholesterol
Cholic Acids
Electrochemistry
Hydrogen-Ion Concentration
Hydrolysis
Hydroxysteroid Dehydrogenases
Study, Methodological
Oxygen Consumption
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