This paper describes the high-performance liquid chromatographic separation of UV-absorbing bile alcohol derivatives. Bile alcohols were treated with 3 alpha-hydroxysteroid dehydrogenase to form the corresponding 3-keto bile alcohols. The 3-keto bile alcohols produced were converted to the 2,4-dinitrophenylhydrazone derivatives, separated using a Nova-Pak Phenyl column, and monitored at 364 nm. The separation of stereoisomers related to the configuration of hydroxyl groups on the side chain of the bile alcohols, which was not achieved by gas chromatography, could also be accomplished.
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