Kinetic enzymatic method for automated determination of HDL cholesterol in plasma.
A sensitive fixed-time kinetic enzymatic method for the measurement of high density lipoprotein cholesterol is described, as adapted for the Cobas Bio centrifugal analyser (Hoffmann LaRoche). The Boehringer Mannheim cholesterol esterase/cholesterol oxidase/peroxidase/3,4-dichlorophenol kinetic reagent was modified by the inclusion of 2,4,6-tribromo-3-hydroxybenzoic acid which reacts with hydrogen peroxide and 4-aminophenazone to produce a quinone-imine dye with a greater molar absorptivity than that produced with phenol. The method has been developed for the determination of HDL fractions isolated with polyethylene glycol 6000, for which a reagent of high sensitivity is required. The method is linear to at least 3.88 mmol/l of HDL cholesterol and the coefficients of variation for within-run and day-to-day precision were less than 3.0%. Correlation of the kinetic HDL cholesterol method with an equilibrium method was good (R = 0.9980). The assay is rapid, inexpensive and large numbers of specimens can be processed conveniently.
Moshides
J S
JS
Department of Clinical Chemistry, Prince of Wales Hospital, Sydney, Australia.
D001322:Autoanalysis; D001970:Bromobenzoates; D008076:Cholesterol, HDL / Q000097:blood*; D006801:Humans; D062385:Hydroxybenzoates; D007202:Indicators and Reagents; D007700:Kinetics
D001970:Bromobenzoates; D008076:Cholesterol, HDL; D062385:Hydroxybenzoates; D007202:Indicators and Reagents; C043713:2,4,6-tribromo-3-hydroxybenzoic acid