Single-cuvet sequential determination of triglyceride and cholesterol

Area of Science:

• Clinical Chemistry
• Biochemical Assays
• Analytical Chemistry

Background:

• Accurate quantification of serum lipids like triglyceride and cholesterol is crucial for cardiovascular disease risk assessment.
• Existing methods may require separate assays or lack efficiency for routine clinical diagnostics.

Purpose of the Study:

• To develop and validate a sequential kinetic assay for simultaneous determination of serum triglyceride and cholesterol in a single cuvette.
• To assess the method's accuracy, precision, linearity, and interference profile for clinical application.

Main Methods:

• A sequential enzymatic method was employed, assaying triglyceride first using ultraviolet detection with ATP and NADH.
• Cholesterol was subsequently assayed using cholesterol oxidase, peroxidase, and the Trinder reaction, with NADH interference managed by adding pyruvate or glycerol.
• The assay was validated on both random-access and batch analyzers, evaluating within-run and between-run coefficients of variation (CVs).

Main Results:

• The method demonstrated excellent linearity up to 8.0 g/L for both analytes.
• High precision was observed with within-run CVs as low as 1.5% for cholesterol and 4.0% for triglyceride.
• Between-run CVs ranged from 8.1% to 12.5% for cholesterol and 8.9% to 12.8% for triglyceride.
• Minimal interference was noted from bilirubin, hemoglobin, and common drugs.
• Correlation studies with established methods yielded high coefficients (r values 0.985-0.995 for cholesterol, 0.984-0.991 for triglyceride).

Conclusions:

• The described sequential kinetic assay provides a reliable and efficient method for simultaneous measurement of serum triglyceride and cholesterol.
• This approach simplifies lipid profile analysis, offering a valuable tool for clinical diagnostics and cardiovascular risk assessment.