A highly sensitive high-performance liquid chromatography method for the estimation of ascorbic and dehydroascorbic acid in tissues, biological fluids, and foods

doi:10.1016/0003-2697(87)90206-5

Analytical Biochemistry

Volume 165, Issue 1, 15 August 1987, Pages 102-107

https://doi.org/10.1016/0003-2697(87)90206-5 Get rights and content

Abstract

A highly sensitive procedure for determining ascorbic acid (AA) and dehydroascorbic acid (DHAA) by high-performance liquid chromatography with electrochemical detection in biological fluids, tissues, and foods is described. AA is separated in a C18 reverse-phase column after extraction from the sample with metaphosphoric acid. An aliquot of 20 μl of diluted extract is injected into the column for the estimation of AA. DHAA is indirectly estimated by converting it to AA after reduction with dl-homocysteine at pH 7.0–7.2 for 30 min at 25°C. After dilution, a 20-μl aliquot is injected into the column to obtain total vitamin C (AA + DHAA). The concentration of DHAA is calculated by subtraction. AA can be reproducibly quantified at concentrations as low as $50 pg 20 μ l$ of sample extract. The method described here used a specially designed mobile phase, gave greater stability and a noiseless baseline, and increased substantially the sensitivity and precision. The procedure is rapid, analysis being completed within 10 min after sample preparation, and has been successfully applied to biological fluids, tissues, and foods.

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^☆: Bureau of Nutritional Sciences Publication No. 245.

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A highly sensitive high-performance liquid chromatography method for the estimation of ascorbic and dehydroascorbic acid in tissues, biological fluids, and foods☆

Abstract

J. Biol. Chem

Anal. Biochem

Amer. J. Clin. Nutr

Metabolism

J. Assoc. Off. Agric. Chem

J. Assoc. Off. Anal. Chem