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Issue 11, 2019

Determination of cellular vitamin C dynamics by HPLC-DAD

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Abstract

A redox-sensitive inter-conversion between ascorbic acid (ASC) and its oxidized form dehydroascorbic acid (DHA) in the intracellular environment has been of exceptional interest to recent metabolomics and pharmaceutical research. We developed a chromatographic protocol to instantly determine these vitamers with each identity from cellular extracts, without any labeling and pretreatments. Owing to its simplicity, one can readily continue the assay for hours, an otherwise difficult to cover timescale at which the intracellular DHA–ASC conversion comes into play. The method was validated for the analysis of pancreatic cancer cells, to our knowledge the first-ever study on a nucleated cell type, to trace in detail their kinetics of glucose transporter-dependent DHA uptake and, simultaneously, that for the intracellular ASC conversion. The simplest of all the relevant techniques and yet with the unique ability to provide each vitamer identity on a high-throughput basis, this method should offer the most practical option for VC-involved physiological and pharmaceutical studies including high-dose VC cancer therapy.

Graphical abstract: Determination of cellular vitamin C dynamics by HPLC-DAD

Supplementary files

Article information


Submitted
19 Nov 2018
Accepted
26 Feb 2019
First published
19 Mar 2019

This article is Open Access

Analyst, 2019,144, 3483-3487
Article type
Communication

Determination of cellular vitamin C dynamics by HPLC-DAD

T. Miyazawa, A. Matsumoto and Y. Miyahara, Analyst, 2019, 144, 3483 DOI: 10.1039/C8AN02240B

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