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Stereoselective determination of p ‐hydroxyphenyl‐phenylhydantoin enantiomers in rat liver microsomal incubates by reversed‐phase high‐performance liquid chromatography using β‐cyclodextrin as chiral mobile phase additives
Author(s) -
Yao T. W.,
Zeng S.
Publication year - 2001
Publication title -
biomedical chromatography
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.4
H-Index - 65
eISSN - 1099-0801
pISSN - 0269-3879
DOI - 10.1002/bmc.50
Subject(s) - chemistry , chromatography , enantiomer , stereoselectivity , ammonium acetate , glucuronidation , high performance liquid chromatography , microsome , cyclodextrin , urea , phase (matter) , in vitro , organic chemistry , biochemistry , catalysis
An analytical method was developed for determination of p ‐hydroxyphenylphenylhydantoin enantiomers in rat liver microsome by using reversed‐phase high‐performance liquid chromatography. A 50 mm C 8 column was used as the analytical column. The mobile phase was made up of 8.8 mmol/L β‐cyclodextrin, 0.25 mol/L urea and 0.05 mol/L ammonium acetate in water. The assay was linear from 2.05 to 410.0 µmol/L for each enantiomer. The limits of detection and of quantitation for the method were 0.90 and 2.05 µmol/L for each enantiomer, respectively. The analytical method afforded average recoveries of 93.59 ± 2.75%and 94.72 ± 1.78% for S ‐ and R‐p ‐hydroxyphenylphenylhydantoin, respectively. The method allowed study of the in vitro glucuronidation of p ‐hydroxyphenylphenylhydantoin in rat liver microsomal incubates. The stereoselectivity of p ‐hydroxyphenylphenylhydantoin phase II metabolism was observed. Copyright © 2001 John Wiley & Sons, Ltd.