35-7/DeLeenheer../Liquid Chromatographic Determination of Vitamin C

Published: Journal of Chromatographic Science, Volume 35, Number 7, July 1997, pp. 333–336.

Liquid Chromatographic Determination of Vitamin C in Aquatic Organisms
H.J. Nelis, A.P. De Leenheer, G. Merchie, P. Lavens, and P. Sorgeloos

A reversed-phase ion-pair liquid chromatographic method using electrochemical detection and internal standardization with isoascorbic acid (IAA) is described for the determination of ascorbic acid (AA) and dehydroascorbic acid (DHAA) in aquatic organisms. Several extractants are compared with regard to recovery and stability of AA and DHAA, including 5% metaphosphoric acid (MPA)–0.54mM ethylenediaminetetraacetic acid (EDTA), 1% acetic acid (Hac)–1mM EDTA, 1% Hac–0.1% MPA–1mM EDTA, 2mM homocysteine–1mM EDTA, and water–methanol (70:30, v/v) containing 0.7mM EDTA. The two nonacidic extractants afford insufficient protection of AA (water–methanol–EDTA) or DHAA (homocysteine–EDTA). High ionic strength acidic mixtures (e.g., 5%MPA) may be associated with a negative drift in the retention times of AA and IAA. Hac (1%)–EDTA (1mM) yields lower recovery except when supplemented with MPA. A mixture of 1% Hac–0.1% MPA–1mM EDTA is useful in extracting AA from the brine shrimp Artemia, Brachionus, fish eggs, fish larvae, and postlarvae of shrimp as part of its routine high-performance liquid chromatographic determination in these matrices.

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