A method for the sequential colorimetric determination of serum triglycerides and cholesterol

Area of Science:

• Clinical Chemistry
• Analytical Chemistry
• Biochemistry

Background:

• Accurate quantification of serum lipids, including triglycerides and cholesterol, is crucial for diagnosing and managing cardiovascular diseases.
• Existing methods for triglyceride and cholesterol determination often require separate analyses, increasing sample volume and processing time.

Purpose of the Study:

• To develop a simplified, sequential spectrophotometric method for the simultaneous determination of triglycerides and cholesterol from a single serum sample.
• To validate the accuracy and reliability of the new method by comparing its results with established individual procedures.

Main Methods:

• A two-stage spectrophotometric procedure involving enzymatic reactions.
• Stage one: Hydrolysis of triglycerides and cholesterol esters, with free fatty acid scavenging for sample clarity.
• Stage two: Enzymatic oxidation of free cholesterol and cholesterol from esters, producing a red chromogen measured at 510 nm.

Main Results:

• The method successfully quantifies both triglycerides and cholesterol sequentially from a single sample.
• The developed spectrophotometric assay produces a distinct red chromogen with maximal absorbance at 510 nm.
• Results showed good correlation with established, separate methods for triglyceride and cholesterol analysis.

Conclusions:

• The proposed spectrophotometric method offers a simple, efficient, and accurate approach for the simultaneous determination of serum triglycerides and cholesterol.
• This technique reduces sample requirements and analysis time, making it a valuable tool in clinical diagnostics.
• The method's good correlation with existing procedures supports its clinical utility and reliability.