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BACKGROUND
Cholesterol is found in bile, blood and brain tissue and is one type of steroid that contains a secondary hydroxyl group in C position. It acts as a precursor to vitamin D, acids and steroids. Synthesis of cholesterol occurs in various tissues but mostly in intestinal walls and liver. The amount of total cholesterol present in serum is vital for diagnosing hyper-cholestereasmias.1
So, the objective of this test is to estimate the total cholesterol in Plasma.
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REQUIREMENTS
Reagents: Cholesterol standard – 200 mg/dl
Pipes buffer: pH-6.9
Phenol – 90 mmol/l
Cholesterol oxidase – 26 mmol/l
Cholesterol esterase – 500 µ/l
Peroxidase – 500 µ/l
4-Aminoantipyirine – 0.4 mmol/l
Apparatus: Test tubes
Bunsen burner
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PROCEDURE
Prepare three test tubes and label it as blank, standard and sample. Then in each test tube add constituents like this:
Blank: Add 1 ml of R2 reagent and 10 microliter distilled water
Standard: 1 ml R2 reagent and 10 microliter of R reagent.
Sample: 1 ml of R2 reagent and 10 microlitres of serum.
Blend and incubate for 5 minutes at 37ºC. Measure the specimen absorbance and standard against reagent blank at 545 nm.2,3
Total Cholesterol = (Absorbance of specimen/Absorbance of standard) x [Standard]
Expected value:
Risk classification Total Cholesterol
Desirable 200 mg/dl (5.2 mmol/l)
Borderline high 200-239 mg/dl (5.2-6.2 mmol/l)
High 240 mg/dl (6.2 mmol/l)
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CONCLUSION
An increase in cholesterol level is dangerous for health.
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REFERENCES
- Miller, N. E., F. Hammett, S. Saltissi, S. Rao, H. van Zeller, J. Coltart, and B. Lewis. 1981. Relation of angiographically defined coronary artery disease to plasma lipoprotein subfractions and apolipoproteins. Br. Med. J. 282: 1741–1744.
- Calabresi, L., G. Franceschini, M. Sirtori, G. Gianfranceschi, P. Werba, and C. R. Sirtori. 1990. Influence of serum triglycerides on the HDL pattern in normal subjects and patients with coronary artery disease. Atherosclerosis. 84: 41–48.
- Syvanne, M., M. Ahola, S. Lahdenpera, J. Kahri, T. Kuusi, K. S. Virtanen, and M. R. Taskinen. 1995. High density lipoprotein subfractions in non-insulin-dependent diabetes mellitus and coronary artery disease. J. Lipid Res. 36: 573–582.
