Rapid high-performance liquid chromatographic method for Vitamin C determination in human milk versus an enzymatic method
Abstract
Vitamin C is an antioxidant that can be considered a possible biomarker of oxidative stability in human milk. A high-performance liquid chromatographic method was developed and validated for determining the total Vitamin C (ascorbic acid and dehydroascorbic acid) and ascorbic acid levels in human milk. This method was then compared with an enzymatic method (a Colorimetric technique) for quantifying ascorbic acid levels. Repeatability and reproducibility were acceptable for all methods. However, the high-performance liquid chromatography (HPLC) technique provided more satisfactory results than the enzymatic method due to this last method detected 37% less ascorbic acid and does not determine the total Vitamin C because of the enzymatic method cannot reduce the dehydroascorbic acid (DHA) to ascorbic acid. Furthermore, the HPLC method has the added advantages that it requires less reagents and material, and is simpler and less time consuming than the enzymatic method. In conclusion, the drawbacks of this enzymatic method would justify its substitution for a HPLC method.
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L-ascorbic acid and dehydroascorbic acid can degrade during sample preparation for HPLC/CE analysis.
“Since the target analytes can degrade during the sample preparation process, stabilizers and/or chelators need to be added to the sample [223,226,228].”
From Vitamin C-Sources, Physiological Role, Kinetics, Deficiency, Use, Toxicity, and Determination (2021) · cites this paper as [223]
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Vitamin C-Sources, Physiological Role, Kinetics, Deficiency, Use, Toxicity, and Determination
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- Authors
- ROMEUNADAL, M., MORERAPONS, S., CASTELLOTE, A., & LOPEZSABATER, M.
- Affiliations
- Universitat de Barcelona (ES)
- ORCID
- CASTELLOTE, A
- Venue
- Journal of Chromatography B
- Publisher
- Elsevier BV
- Published
- 2006-01-02
- Volume
- 830
- Issue
- 1
- Pages
- 41-46
- ISSN
- 1570-0232 (ISSN-L)
- OpenAlex
- W2092805468
- Cited by
- 109
- References
- 28
- Citations by year
- 2026: 3 · 2025: 2 · 2024: 5 · 2023: 4 · 2022: 5 · 2021: 8 · 2020: 5 · 2019: 2 · 2018: 11 · 2017: 7 · 2016: 4 · 2015: 9 · 2014: 5 · 2013: 4 · 2012: 7
- FWCI
- 4.26
- Type
- journal article
- Language
- en
- Open access
- closed
- Locations
- Journal of Chromatography B (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Generalitat de Catalunya
- Topics
- Health Sciences › Nursing › Nutrition and Dietetics › Infant Nutrition and Health
- Health Sciences › Nursing › Nutrition and Dietetics › Vitamin C and Antioxidants Research
- Physical Sciences › Chemistry › Analytical Chemistry › Analytical Methods in Pharmaceuticals
- Keywords
- vitamin C, breast milk, dehydroascorbic acid, HPLC method validation, enzyme activity assay, colorimetric assay, method comparison
- MeSH terms
- Ascorbic Acid, Chromatography, High Pressure Liquid, Enzymes, Humans, Milk, Human, Spectrophotometry, Ultraviolet
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