Improved method for determination of high-density-lipoprotein cholesterol II. Enzymic determination of cholesterol in high-density lipoprotein fractions with a sensitive reagent.
A reagent is described for the colorimetric enzymic determination of high-density-lipoprotein (HDL) cholesterol. The reagent can be used with HDL fractions isolated by the various methods of precipitation of low- and very-low-density lipoproteins we investigated. The considerable sensitivity obtained by use of Barham-Trinder's reaction allows the sample/reagent volume ratio to be decreased to 1:80, and major interferences thus eliminated. The response is linear from 100 to 2000 mg of HDL cholesterol per liter. The maximum CV obtained in precision tests was approximately 1% within series and approximately 3% between series. Most of the bilirubin interference is eliminated by adopting a reaction pH of 6.1. Because of its sensitivity, the reagent is particularly suitable for use with HDL fractions isolated after precipitation with polyethylene glycol 6000, which are characterized by a marked dilution. HDL cholesterol determination with the proposed reagent is accurate and precise. Values obtained are in line with those provided for by the Abell-Kendall method. The method can easily be automated.
D000632:Aminopyrine; D002733:Chlorophenols; D002784:Cholesterol / Q000097:blood* / Q000378:metabolism; D002789:Cholesterol Oxidase / Q000378:metabolism; D002863:Chromogenic Compounds; D003124:Colorimetry; D006801:Humans; D006863:Hydrogen-Ion Concentration; D008075:Lipoproteins, HDL / Q000032:analysis*; D010544:Peroxidases / Q000378:metabolism; D002787:Sterol Esterase / Q000378:metabolism
D002733:Chlorophenols; D002863:Chromogenic Compounds; D008075:Lipoproteins, HDL; C028879:sulfonated 2,4-dichlorophenol; D000632:Aminopyrine; D002784:Cholesterol; D002789:Cholesterol Oxidase; D010544:Peroxidases; D002787:Sterol Esterase