Quantitative Analysis of Phenyl-Acetic Acid inProteus MirabilisCultures by High Performance Liquid Chromatography

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Abstract
A method was developed for the analysis of phenylacetic acid produced by Proteus mirabilis during a 10-day incubation period. A daily sample was acidified and extracted with petroleum ether for 24 hours in a continuous extraction apparatus. The extract of each day was analyzed by reverse-phase liquid chromatography using UV detection at 254 nm. A 10-μl aliquot of the concentrated extract was injected onto an octadecylsilane column. The mobile phase consisted of 20% methanol and 80% sodium acetate buffer, 0.01 M, pH 4.2. This method may be used for screening other microbial fermentations for the production of similar lipophilic acids.