CHOLESTEROL (LIQUID) REAGENT
INTENDED USE
Cholesterol (Liquid) Reagent can be used for the quantitative determination of total cholesterol in serum.
INTRODUCTION
Cholesterol is a fatty substance found in blood, bile and brain tissue. It serves as a precursor to bile acids, steroids and vitamin D. The determination of serum cholesterol is a major aid in the diagnosis and classification of lipemias. Other conditions such as hepatic thyroid diseases influence cholesterol levels.
Enzymatic methods have replaced older methodologies involving cholesterol esterase, oxidase, and trinders color system. Allain et al. developed a total enzymatic technique in which hydrogen peroxide during the oxidation of cholesterol is used in conjunction with peroxidase, 4-aminoantipyrine and phenol to form a quinoneimine dye. This reagent employs p-hydroxy benzene sulfonic acid (p-HBS), in place of phenol to produce a quinoneimine dye with greater absorbance at 520 nm and a surfactant to facilitate the completion of reaction.
The results obtained from this replacement are equal to the results obtained based on the Abell-Kendall cholesterol reference method which is recommended by the CDC.
PRINCIPLE
Cholesterol (Liquid) Reagent
Cholesterol esters are hydrolyzed to produce cholesterol. Hydrogen peroxide is then produced from the oxidation of cholesterol by cholesterol oxidase. In a coupled reaction catalyzed by peroxidase, quinoneimine dye colored red is formed from 4-aminoantipyrine, p HBS and hydrogen peroxide. The absorption at 520 nm of the solution of this dye is proportional to the concentration of cholesterol in the sample.
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