HPLC-UV method for the simultaneous determinations of ascorbic acid and dehydroascorbic acid in human plasma
Open accessAbstract
A high performance liquid chromatography (HPLC) paired with UV-vis detection method to determine ascorbic acid and its oxidation product, dehydroascorbic acid, in human plasma was developed. Ascorbic acid in human plasma was extracted and stabilized using 10% metaphosphoric acid, and was analyzed by a Symmetry C18 column with 5 mM Hexadecyltrimethylammonium bromide and 50 mM KH2PO4 solution as the mobile phase (1.0 mL/min flow rate). Isoascorbic acid served as the internal standard and ultraviolet detector wavelength was 254 nm and 265 nm. Dehydroascorbic acid concentration was calculated from the differences in ascorbic acid concentration before and after reduction by dithiothreitol reagent. Quantification for ascorbic acid in human plasma was linear from 1–100 μg/mL. The interand intra-day precisions and accuracy were determined and the results were found to be within ±15%. This method was successfully applied to a human pharmacokinetic study of ascorbic acid as well as dehydroascorbic acid after oral administration of 4,000 mg vitamin C tablets to healthy Korean volunteers.
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Vitamin C absorption is relatively slow, as maximal plasma levels are reached after 2-4 h.
“In general, vitamin C absorption is relatively slow, as maximal plasma levels are reached after 2–4 h [6,8,40,58].”
From Vitamin C-Sources, Physiological Role, Kinetics, Deficiency, Use, Toxicity, and Determination (2021) · cites this paper as [58]
The Results (Pharmacokinetic study) report that plasma ascorbic acid reached its maximum at 2.67 ± 0.58 hours after a 4,000 mg oral dose, supporting the 2–4-hour timing. The study involved only three healthy volunteers and does not establish that vitamin C absorption is generally or relatively slow. (LLM, conf=0.86)
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- Authors
- Kim, Y., & Kim, M.-G.
- Affiliations
- Jeonbuk National University Hospital (KR)
- Venue
- Translational and Clinical Pharmacology
- Publisher
- XMLink
- Published
- 2016
- Volume
- 24
- Issue
- 1
- Pages
- 37
- ISSN
- 2289-0882 (ISSN-L)
- 2383-5427
- OpenAlex
- W2346983855
- Cited by
- 33
- References
- 21
- Citations by year
- 2026: 3 · 2025: 6 · 2024: 3 · 2023: 2 · 2022: 9 · 2021: 5 · 2020: 1 · 2019: 1 · 2018: 3
- FWCI
- 1.39
- Type
- journal article
- Language
- en
- Open access
- diamond
- doi.org
- Licence
- Locations
- Translational and Clinical Pharmacology (publishedVersion, cc-by-nc)
- Funders
- Chonbuk National University Hospital
- Topics
- Health Sciences › Nursing › Nutrition and Dietetics › Vitamin C and Antioxidants Research
- Physical Sciences › Chemistry › Analytical Chemistry › Analytical Methods in Pharmaceuticals
- Physical Sciences › Chemistry › Spectroscopy › Analytical Chemistry and Chromatography
- Keywords
- vitamin C, dehydroascorbic acid, human plasma, pharmacokinetics, HPLC-UV method, dithiothreitol reduction, simultaneous determination, oral administration
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