A simple fluorometric determination of vitamin C

Area of Science:

• Analytical Chemistry
• Biochemistry

Background:

• Accurate quantification of vitamin C (ascorbic acid and dehydroascorbic acid) is crucial for nutritional and health studies.
• Existing methods may lack specificity or sensitivity for complex biological matrices.

Purpose of the Study:

• To develop a simple, accurate, and sensitive fluorometric method for vitamin C determination.
• To optimize extraction and separation techniques for analyzing vitamin C in natural products.

Main Methods:

• Utilized 4,5-dimethyl-o-phenylenediamine (DMPD) as a fluorescent reagent for vitamin C.
• Investigated isobutanol and n-butanol for extracting fluorometric derivatives.
• Employed High-Performance Liquid Chromatography (HPLC) for separating vitamin C derivatives from interfering compounds.

Main Results:

• DMPD reacted with dehydroascorbic acid to form a stable, fluorescent product (Ex; 360 nm, Em; 440 nm).
• Isobutanol extraction demonstrated superior specificity compared to n-butanol by minimizing co-extraction of keto acids.
• The developed method showed improved sensitivity and accuracy for vitamin C determination in natural products.

Conclusions:

• The DMPD-based fluorometric method, coupled with isobutanol extraction and HPLC, provides a robust approach for vitamin C analysis.
• This method enhances the reliability of vitamin C quantification in diverse natural product samples.