Determination of Blood cholesterol
Principle:
Cholesterol forms a colored complex with acetic anhydride and concentrated Sulfuric acid.
Reagents:
Reagent I: Cholesterol standard 200 mg/dl
Reagent II: Acetic anhydride 3.5 mol/l
Acetic acid 5 mol/l
Additional reagent: Sulfuric acid conc.
Sample preparation: Serum, heparinized plasma or EDTA plasma.
Procedure:
Wave length: 578 nm.
Incubation temperature: 20-25°c
Pipette into dry test tubes:
|
|
Blank
|
Standard
|
Sample
|
|
Dist.water
|
0.1 ml
|
|
|
|
Reagent I
|
|
0.1 ml
|
|
|
Sample
|
|
|
0.1 ml
|
|
Reagent II
|
2.5 ml
|
2.5 ml
|
2.5 ml
|
Mix carefully and incubate in water bath at 20-25 °c for 5 min. Add 0.5 ml conc. sulfuric acid by the inside wall of the tube and mix thoroughly and let to stand for 10 min. Read the absorbance of the sample (A sample) and Standard (A standard) against blank.
Calculation:
Cholesterol conc. in mg/dl = A sample / A standard X 200
Interpretation:
Normal value: 150-250mg/dl.
Hypercholesterolemia:
1-Diabetes mellitus.
2-Obstructive jaundice.
3-Liver cirrhosis.
4-Nephrotic syndrome.
5-Hypoparathyroidism and Xanthomatosis.
Hypocholesterolemia:
1- Hyperthyroidism and parathyroidism.
2-Hemolytic anaemia, pernicious anaemia and malabsorption syndrome.
