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Determination of vitamin C (ascorbic and dehydroascorbic acids) in foods and feeds
Summary
This study presents a novel method for quantifying ascorbic acid (ASC) using anionic Sephadex chromatography and a new colorimetric reagent, 4-Nitro-1,2-Phenylenediamine (NPD). The technique allows for specific and semi-automated measurement of ASC in extracts.
Area of Science:
- Analytical Chemistry
- Biochemistry
Background:
- Ascorbic acid (ASC) quantification is crucial in various biological and food science applications.
- Existing methods may lack specificity or require complex procedures.
Purpose of the Study:
- To develop a specific and efficient method for the separation and quantification of ascorbic acid.
- To introduce a novel colorimetric assay for dehydroascorbic acid (DASC).
Main Methods:
- Separation of ascorbic acid (ASC) from extract solutions using anionic Sephadex chromatography.
- On-column oxidation of ASC to dehydroascorbic acid (DASC) using p-benzoquinone.
- Colorimetric detection of DASC using 4-Nitro-1,2-Phenylenediamine (NPD) and measurement of optical extinction at 375 nm.
Main Results:
- The developed method demonstrates high specificity for ascorbic acid detection.
- The chromatographic purification process can be semi-automated for higher throughput.
- The assay allows for the quantification of DASC, with potential for ASC reduction prior to analysis.
Conclusions:
- This novel method offers a specific and efficient approach for ascorbic acid analysis.
- The use of anionic Sephadex chromatography combined with NPD colorimetry provides a reliable quantification technique.
- The semi-automated nature of the purification enhances its applicability in routine analysis.