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Clinical Chemistry 42: 915-926, 1996;
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Clinical Chemistry, Vol 42, 915-926, Copyright © 1996 by American Association for Clinical Chemistry

Sequential microenzymatic assay of cholesterol, triglycerides, and phospholipids in a single aliquot

MN Nanjee and NE Miller
Department of Cardiovascular Biochemistry, The Medical College of St. Bartholomew's Hospital, Charterhouse Square, London, UK.

The assay of multiple analytes in a single aliquot can be advantageous from both measurement and economic standpoints. The objective of this study was to develop a simple and sensitive microenzymatic method for the determination of three biologically important lipids. Triglycerides (as glycerol), phospholipids (as choline), and total cholesterol (as unesterified cholesterol) were assayed, in that order, by sequential addition of sample, reagents, and microbial enzymes directly into a single microtiter plate well, accompanied by continuous monitoring of a common reporter reaction in which hydrogen peroxide is quantified either by colorimetry with 4-aminoantipyrine and 3-hydroxy-2,4,6- triiodobenzoic acid or by ultraviolet fluorometry with p- hydroxyphenylacetic acid. The detection limit of the method is in the subnanomole mass range for all three lipids. Results obtained with either fluorescence or colored endpoints were in excellent agreement with alternative individual chemical and enzymatic procedures.


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M. N. Nanjee, C. J. Cooke, W. L. Olszewski, and N. E. Miller
Lipid and apolipoprotein concentrations in prenodal leg lymph of fasted humans: associations with plasma concentrations in normal subjects, lipoprotein lipase deficiency, and LCAT deficiency
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M. N. Nanjee and E. A. Brinton
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M. N. Nanjee, J. E. Doran, P. G. Lerch, and N. E. Miller
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Copyright © 1996 by the American Association for Clinical Chemistry.