J. Lipid Res.
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Originally published In Press as doi:10.1194/jlr.D300024-JLR200 on October 16, 2003

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Journal of Lipid Research, Vol. 45, 396-401, February 2004
Copyright © 2004 by American Society for Biochemistry and Molecular Biology

Methods

A method of direct measurement for the enzymatic determination of cholesteryl esters

Toshimi Mizoguchi1, Toshiyuki Edano and Tomoyuki Koshi

Tokyo New Drug Research Laboratories II, Pharmaceutical Division, Kowa Co. Ltd., Tokyo 189-0022, Japan

1 To whom correspondence should be addressed. e-mail: t-mizogt{at}kowa.co.jp

A direct measurement method for the enzymatic determination of cholesteryl esters (CEs) without measuring total cholesterol (TC) and free cholesterol (FC) is described. In the first step, hydrogen peroxide generated by cholesterol oxidase from FC was decomposed by catalase. In the second step, CE was measured by enzymatic determination using a colorimetric method or a fluorometric method. The measurement sensitivity of the fluorometric method was more than 20 times that of the colorimetric method. Optimal conditions of the assay were determined, and examples of measured CE in human plasma, rat liver, and cultured cells are indicated.

The method of directly measuring CE was simple and has exceptional reproducibility compared with the technique of subtracting FC from TC using each measured TC and FC.

Supplementary key words cholesterol assay • cholesterol oxidase catalase • plasma • liver • tissue culture


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