Ascorbic acid determination in serum and aqueous humour by high-performance liquid chromatography

Area of Science:

• Biochemistry
• Analytical Chemistry
• Ophthalmology

Background:

• Ascorbic acid (Vitamin C) plays a crucial role in ocular health.
• Accurate measurement of ascorbic acid in biological fluids is essential for diagnosing and managing certain conditions.
• Existing methods for ascorbic acid determination can be time-consuming or lack sensitivity.

Purpose of the Study:

• To develop a rapid and sensitive method for quantifying ascorbic acid in serum and aqueous humour.
• To establish a reliable analytical technique for clinical applications in ophthalmology.

Main Methods:

• High-performance liquid chromatography (HPLC) using a Supelcosil LC-HN2 analytical column.
• Weak anion exchange chromatography mode for separation.
• UV detection at 254 nm for sensitive quantification.

Main Results:

• Successful separation of ascorbic acid from interfering substances in serum and aqueous humour within 5 minutes.
• Detection limit for ascorbic acid established at 20 pmol.
• Quantification of ascorbic acid concentrations in samples from 10 cataract patients.

Conclusions:

• The developed HPLC method offers a fast and sensitive approach for ascorbic acid determination.
• This method is suitable for clinical analysis of ascorbic acid in ocular fluids and serum.
• The findings provide baseline data for ascorbic acid levels in cataract patients.