Quantitative Assay Of Cholesterol In Blood
What is the definition of cholesterol?
Cholesterol is a steroid containing a tier 2 hydroxyl group at the C3 position. It is synthesized in different tissue types, but usually concentrates in the liver and intestinal wall. About 80% endogenous and 20% are derived from food.
Linear range: the range of values for which the measurement is accurate.
Limits of measurement: minimum concentration that a measurement can measure
Principles
The cholesterol ester enzyme digestion method is cleaved by the cholesterol esterase enzyme, which produces cholesterol and free fatty acids. Cholesterol oxidase catalyzes the oxidation of cholesterol to cholest-4-en-3-one and hydrogen peroxide. In the presence of peroxidase, hydrogen peroxide acts on the oxidative coupling between 4 - aminophenazone and phenol to produce a red quinone-imine
Reaction equation:
Cholesterol esters + H20 CE cholesterol + RCOOH
Cholesterol + O2 CHOD cholest - 4 - en - 3 - one + H2O2
2H2O2 + 4- AAP + phenol POD quinone - imine dye + 4H20
The color intensity of the dye is directly proportional to the cholesterol concentration. Color intensity is determined by measuring the increase in absorbance.
.Laboratory equipment
Biochemical testing machine Biosytem A25. Centrifuge speed of 4000 rpm
Reagents
Chemical testing firm
0.9% NaCl diluent solution
Safe
Serum, plasma, whole blood at high risk of infection, carrying out operations on patient samples in accordance with the procedures
Wear full protective clothing such as gown, gloves, masks ..
When chemicals, specimens are stuck on gloves, they must be changed immediately; If in eyes, rinse immediately under emergency eyewash
Samples after testing must be treated as infectious waste
Interpret and report the results.
CHOL blood: <200mg / dL Warning: Factors affecting Jaundice: conjugated bilirubin concentration> 274 µmol / l or 16 mg / dl.
Unconjugated bilirubin concentration> 239 µmol / l or 14 mg / dl.
Hemolysis: hemoglobin concentration> 435 olmol / l or 700 mg / dl.
.Lipemia:> 2000 mg / dl. There is a correlation between turbidity and triglycerides concentration.
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