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Cholesterol Oxidase Method
Structure of Cholesterol
Serum/Plasma Cholesterol
Biomedical Importance of Cholesterol
Sources of Serum Cholesterol
Serum Cholesterol Estimation
• Fluorometric
• Spectrophotometric
Chemical method
Biochemical/enzymatic method
Enzymatic Method
for Cholesterol Estimation
History
1967---Tonks
1974—Alian et al.
Principle
Cholesterol esterase
Cholesterol Esters ---------------------------→ Cholesterol + Fatty Acids
Cholesterol oxidase
Cholesterol + O2 ------------------------→ Cholesten-3-one + H2O2
Peroxidase
2 H2O2 + 4-aminoantipyrine + Phenol -----------→ Quinoneimine + 4 H 2O
Reaction Scheme
Requirements: Equipments
Incubator
Requirements: Equipments
Centrifuge Machine
Requirements: Equipments
Colorimeter
Requirements: Reagents
• Preparation of Reagents
• Preparation of Standards
• Preparation of Blank
• Preparation of Serum
Method
Parameters of Analysis
• Cross Contamination
• Interferences in the Measurement
• Influence of Drugs on Clinical Laboratory
Calculation
Presentation of results
Interpretation of results
Performance Characteristics
• Reportable range
• Precision
Standard Reference