A simple method for plasma total vitamin C analysis suitable for routine clinical laboratory use

Area of Science:

• Clinical Chemistry
• Analytical Chemistry
• Biochemistry

Background:

• In-hospital hypovitaminosis C is common but often undetected due to limited lab analysis.
• Lack of accessible plasma vitamin C testing hinders clinical recognition and management.
• A simple, reliable assay is crucial for routine clinical use.

Purpose of the Study:

• To develop and validate a simplified UV-HPLC method for plasma total vitamin C analysis.
• To compare the performance of the simplified UV-HPLC method against the established EC-HPLC gold standard.
• To assess the clinical practicality of the new method in hospital laboratories.

Main Methods:

• Modified existing UV-HPLC techniques for plasma vitamin C determination.
• Developed a constant-low-pH sample reduction and isocratic reverse-phase HPLC separation.
• Compared the simplified UV-HPLC method with EC-HPLC using 80 clinical samples.

Main Results:

• The simplified UV-HPLC method is cost-effective, user-friendly, and offers enhanced sample stability.
• It requires fewer reagents and no pH adjustments, simplifying laboratory procedures.
• Performance was equivalent to the gold-standard EC-HPLC across a wide range of plasma vitamin C levels.

Conclusions:

• The described plasma vitamin C analysis method is practical for standard hospital laboratories due to its simplicity and sensitivity.
• This UV-HPLC method, rigorously validated against EC-HPLC, offers equivalent performance.
• Wider adoption can significantly improve the availability of plasma vitamin C testing in clinical settings.