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TOTAL CHOLESTEROL Assay Kit
Kit is an enzymatic, colorimetric assay designed for the quantitative determination of total cholesterol. Cat.No: OttoBC135
Product Description
Cholesterol is a steroid with a secondary hydroxyl group in the C3 position. It is syn‑ thesized in many types of tissue, but particularly in the liver and intestinal wall. Approximately three quarters of cholesterol is newly synthesized and a quarter originates from dietary intake. Cholesterol assays are used for screening for athero‑ sclerotic risk and in the diagnosis and treatment of disorders involving elevated cholesterol levels as well as lipid and lipoprotein metabolic disorders. Cholesterol analysis was first reported by Liebermann in 1885 followed by Burchard in 1889. In the Liebermann‑Burchard reaction, cholesterol forms a bluegreen dye from polymeric unsaturated carbohydrates in an acetic acid/acetic anhydride/ concentrated sulfuric acid medium. The Abell and Kendall method is specific for cholesterol, but is technically complex and requires the use of corrosive reagents. In 1974, Roeschlau and Allain described the first fully enzymatic method. This method is based on the determination of A 4 cholestenone after enzymatic cleavage of the cholesterol ester by cholesterol esterase, conversion of cholesterol by cholesterol oxidase and subsequent measurement by the Trinder reaction of the hydrogen peroxide formed. Optimization of ester cleavage (>99.5%) allows standardization using primary and secondary standards and a direct comparison with the CDC and NIST reference methods. The Analyticon cholesterol assay meets the 1992 National Institutes of Health (NIH) goal of less than or equal to 3% for both precision and bias.
Contents
Cholesterol is converted by oxygen with the aid of cholesterol oxidase to A4 ‑ Cholestenone and hydrogen peroxide. Hydrogen peroxide created forms a red dyestuff by reacting with 4‑aminoantipyrine and phenol under the catalytic action of peroxidase. The color intensity is directly proportional to the concentration of cholesterol and can be determined photometrically.
Cholesterol ester + H2O Cholesterol + fatty acid
Cholesterol + O2Cholesten‑3‑on + H2O2
H2O2 + phenol + 4‑aminoantipyrinequinoneimine dye+4 H2O
Referanslar
1. Abell L. et al. Standard Methods in Clinical Chemistry 1958; 26:2.
2. Allain C.C. et al. Clin Chem 1974;20:470
3. Burchard H. Beitrâge zur Kenntnis der Cholesterine. Dissertation Rostock 1889.
4. Cohn J.S., McNamara J.R., Schaefer E.J.. Lipoprotein Cholesterol Concentrations in the plasma of Human Subjects as Measured in the fed and Fasted States. Clin. Chem 1988;34:2456‑2459
5. Glick M.R., Ryder K.W., Jackson SA.. Graphical Comparisons of Interferences in Clinical Chemistry Instrumentation. Clin Chem 1986;32:470‑474
6. Greinling H., Gressner A.M. eds. Lehrbuch der Klinischen Chemie und Pathobiochemie. 3rd ed. Stuttgart/New York: Schattauer, 1995
7. Liebermann C. Ber Dtsch chem Ges 1885; 18:1803
8. Pisani T., Gebski C.P., Leary E.T. et al. Accurate Direct Determination of Lowdensity Lipoprotein Cholesterol Using an Immunoseparation Reagent and Enzymatic Cholesterol Assay. Arch. Pathol Lab Med 1995;119:1127
9. Recommendations for Improving Cholsterol Measurement: A Report from the Laboratory Standardization Panel of the National Cholesterol Education Program. NIH Publication No. 90‑2964, February 1990
10. Roeschlau P. et al. Z Klein Chem Klein Biochem 1974;12:226
11. Second Report of the Expert Panel on Detection, Evaluation and Treatment of High Blood Cholesterol in Adults. NIH Publikation No. 93‑3096, September 1993
12. Siedel J. Hagele E.O., Ziegenhorn J. et al. Clin Chem 1983;29:1075.
13. Study Group, European Atherosclerosis Society. Strategies for the prevention of coronary heart disease: A policy statement of the European Atherosclerosis Society. European Heart Journal 1987;8:77
14. Thomas, L., Labor und Diagnose, 5th. ed. (1998)
15. Tietz N.W. eds. Clinical Guide to Laboratory Tests, 3.Auaflage.Philadelphia, Pa: W.B. Saunders Company, 1995:130‑131 Wiebe D.A., Bernert J.T. Clin Chem. 1984;30:35
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