CHOLESTEROL REAGENT SET (PHENOL FREE)
INTENDED USE
Cholesterol Reagent Set (Phenol Free) can be used for the quantitative determination of total cholesterol in human 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 520nm and a surfactant to facilitate the completion of reaction.
PRINCIPLE
Cholesterol Reagent Set (Phenol Free)
Cholesterol esters are hydrolyzed to produce cholesterol. Hydrogen peroxide is then produced from the oxidation of cholesterol-bycholesterol 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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