Dehydroascorbic acid quantification in human plasma: Simultaneous direct measurement of the ascorbic acid/dehydroascorbic acid couple by UPLC/MS-MS.

Autor:	Violet PC; Molecular and Clinical Nutrition Section, Digestive Diseases Branch, Intramural Research Program, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, USA, 20892., Munyan N; Molecular and Clinical Nutrition Section, Digestive Diseases Branch, Intramural Research Program, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, USA, 20892., Luecke HF; Intramural Research Division, National Institute of Environmental Health Sciences, National Institutes of Health, Durham, NC, USA, 27709., Wang Y; Molecular and Clinical Nutrition Section, Digestive Diseases Branch, Intramural Research Program, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, USA, 20892., Lloyd J; Laboratory of Bioorganic Chemistry, Intramural Research Program, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, USA, 20892., Patra K; Molecular and Clinical Nutrition Section, Digestive Diseases Branch, Intramural Research Program, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, USA, 20892., Blakeslee K; Principal Technical Support Specialist, Waters Mid Atlantic District Office, Waters Corporation, Columbia, MD, 21046, USA., Ebenuwa IC; Molecular and Clinical Nutrition Section, Digestive Diseases Branch, Intramural Research Program, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, USA, 20892., Levine M; Molecular and Clinical Nutrition Section, Digestive Diseases Branch, Intramural Research Program, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, USA, 20892. Electronic address: markl@mail.nih.gov.
Jazyk:	angličtina
Zdroj:	Redox biology [Redox Biol] 2024 Dec; Vol. 78, pp. 103425. Date of Electronic Publication: 2024 Nov 15.
DOI:	10.1016/j.redox.2024.103425
Abstrakt:	Ascorbic acid (AA, vitamin C) and dehydroascorbic acid (DHA) constitute a biological couple. No technique can accurately, independently, and simultaneously quantify both members of the couple in animal and human samples, thereby constraining advances in physiology and pathophysiology. Here we describe a new UPLC/MS/MS method to measure both compounds directly and independently in human plasma. Lower limits of quantification were 16 nM, with linear coefficients >0.99 over a 100-fold concentration range. The method was stable and reproducible with <10 % injection-to-injection variation. Use of isotopic labeled internal standards for both compounds ensured precision and accuracy. Plasma preparation required only 2 steps. In plasma samples from 14 anonymized subjects who met criteria for blood donation, mean concentrations were 6±2 μmol/L (mean ± SD) and 56 ± 14 μmol/L for DHA and AA respectively, with (DHA)/(AA + DHA) ratio of 9.8 %. This method represents a pioneering approach to measuring the AA/DHA couple in human plasma. Competing Interests: Declaration of Competing Interest None reported. (Published by Elsevier B.V.)
Databáze:	MEDLINE
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