Cholestrol(CHOD / POD METHOD)
INTENDED USE
This reagent is intended for the in-vitro quantitative determination of cholesterol in human serum.
PRINCIPLE
Cholesterol esters are hydrolyzed by cholesterol esterase to free cholesterol and fatty acids.
Free cholesterol is oxidized by cholesterol oxidase to cholestenone and hydrogen peroxide.
Hydrogen peroxide reacts with 4-aminoantipyrine and phenol in presence of peroxidase to form a red coloured quinoneimine dye.
REACTION
Cholesterol Ester + H₂O → Cholesterol + Fatty acids
Cholesterol + O₂ → Cholestenone + H₂O₂
H₂O₂ + Phenol + 4-aminoantipyrine → Red quinoneimine
REAGENTS
-
Cholesterol Enzyme Reagent
-
Cholesterol Standard (200 mg/dL)
SPECIMEN :- Serum / Plasma
PROCEDURE
| Tube | Reagent | Sample | Standard |
|---|---|---|---|
| Blank | 1.0 ml | – | – |
| Standard | 1.0 ml | – | 10 µl |
| Test | 1.0 ml | 10 µl | – |
Mix well, incubate at 37°C for 5 minutes.
Read absorbance at 505 nm against reagent blank.
CALCULATION
Cholesterol (mg/dL) =
NORMAL VALUE
-
Total Cholesterol: Less than 200 mg/dL
CLINICAL SIGNIFICANCE
Cholesterol is the main lipid found in blood. It is an essential structural component of cell membranes and is involved in the synthesis of steroid hormones and bile acids.
Elevated cholesterol levels are associated with atherosclerosis and increased risk of coronary heart disease.
STORAGE
Store reagent at 2–8°C. Do not freeze.