A simple method for plasma total vitamin C analysis suitable for routine clinical laboratory use
Open accessAbstract
BACKGROUND: In-hospital hypovitaminosis C is highly prevalent but almost completely unrecognized. Medical awareness of this potentially important disorder is hindered by the inability of most hospital laboratories to determine plasma vitamin C concentrations. The availability of a simple, reliable method for analyzing plasma vitamin C could increase opportunities for routine plasma vitamin C analysis in clinical medicine. METHODS: Plasma vitamin C can be analyzed by high performance liquid chromatography (HPLC) with electrochemical (EC) or ultraviolet (UV) light detection. We modified existing UV-HPLC methods for plasma total vitamin C analysis (the sum of ascorbic and dehydroascorbic acid) to develop a simple, constant-low-pH sample reduction procedure followed by isocratic reverse-phase HPLC separation using a purely aqueous low-pH non-buffered mobile phase. Although EC-HPLC is widely recommended over UV-HPLC for plasma total vitamin C analysis, the two methods have never been directly compared. We formally compared the simplified UV-HPLC method with EC-HPLC in 80 consecutive clinical samples. RESULTS: The simplified UV-HPLC method was less expensive, easier to set up, required fewer reagents and no pH adjustments, and demonstrated greater sample stability than many existing methods for plasma vitamin C analysis. When compared with the gold-standard EC-HPLC method in 80 consecutive clinical samples exhibiting a wide range of plasma vitamin C concentrations, it performed equivalently. CONCLUSION: The easy set up, simplicity and sensitivity of the plasma vitamin C analysis method described here could make it practical in a normally equipped hospital laboratory. Unlike any prior UV-HPLC method for plasma total vitamin C analysis, it was rigorously compared with the gold-standard EC-HPLC method and performed equivalently. Adoption of this method could increase the availability of plasma vitamin C analysis in clinical medicine.
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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 [226]
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References 80 · Cited by 88
- Authors
- Robitaille, L., & Hoffer, L. J.
- Affiliations
- Jewish General Hospital (CA)
- Corresponding author
- Hoffer, L. John
- ORCID
- Hoffer, L. John
- Venue
- Nutrition Journal
- Published
- 2015-12
- Volume
- 15
- Issue
- 1
- Pages
- 40-40
- ISSN
- 1475-2891 (ISSN-L)
- PMC
- PMC4839128
- OpenAlex
- W2336490808
- Cited by
- 88
- References
- 80
- Citations by year
- 2026: 2 · 2025: 14 · 2024: 7 · 2023: 11 · 2022: 12 · 2021: 9 · 2020: 8 · 2019: 9 · 2018: 10 · 2017: 4 · 2016: 1 · 2015: 1
- FWCI
- 5.14
- Type
- journal article
- Language
- en
- Update notices
- correction (2016-05-05)
- Open access
- gold
- doi.org
- Licence
- Locations
- Nutrition Journal (publishedVersion, cc-by)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- Funders
- Lotte and John Hecht Memorial Foundation (None)
- McGill University (None)
- Topics
- Health Sciences › Nursing › Nutrition and Dietetics › Vitamin C and Antioxidants Research
- Health Sciences › Medicine › Biochemistry › Antioxidant Activity and Oxidative Stress
- Physical Sciences › Chemistry › Analytical Chemistry › Analytical Methods in Pharmaceuticals
- Keywords
- plasma vitamin C, vitamin C, dehydroascorbic acid, hypovitaminosis C, sample reduction, method comparison, clinical laboratory assay
- MeSH terms
- Ascorbic Acid, Blood Chemical Analysis, Chromatography, High Pressure Liquid, Humans, Nutritional Status, Vitamins, Reproducibility of Results
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