Abstract
This paper reports the development of a reliable method for the separation and determination of p-hydroxyphenylpyruvic acid by high-performance liquid chromatography. An aqueous solution of p-hydroxyphenylpyruvic acid was injected into a Cosmosil C 18-5 column with a mobile phase consisting of acetonitrile-acetic acid-water (18:1:81, v/v/v) with detection at 283 nm. Three peaks were observed on the chromatogram, and were identified as the keto-enol tautomers and the decomposition product p-hydroxybenzaldehyde. The enol form and p-hydroxybenzaldehyde were determined without derivatization and the keto form as its oxime from their peak heights by comparison with those of known amounts of the enol form, p-hydroxybenzaldehyde and the oxime standard of the keto form.
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