Separative determination of ascorbic acid and erythorbic acid by high-performance liquid chromatography.

Abstract

High-performance liquid chromatography (HPLC) was applied to the determination of ascorbic acid (AsA) and erythorbic acid (ErA). The apparatus was a Shimadzu model LC-2P Liquid Chromatograph equipped with a UV detector set at 254 nm. The separation was achieved on a LiChrosorb-NH2 column which was pre-treated with 0.1 M ammonium monophosphate solution using a mixture of acetonitrile, acetic acid and water (87 : 2 : 11, vol/vol) as an eluant. The HPLC method has the followimg advantages: AsA and ErA are quantitated after being distinctly separated, analysis time per sample is short and AsA or ErA levels as low as 1.0 .times. 10-2 .mu.g are detectable. Recovery experiments with dehydro-AsA and dehydro-ErA, involving reduction with H2S, gave satisfactory results.