||High-Performance Liquid Chromatographic Assay of Biphenyl Metabolism by Hepatocytes Cultured in an Embryo/Hepatocyte Co-culture Medium.
Mole, M. L. ;
Sanders, L. ;
Oglesby, L. A. ;
||Health Effects Research Lab., Research Triangle Park, NC. ;Northrop Services, Inc./Environmental Sciences, Research Triangle Park, NC.
Biphenyl compounds ;
High pressure liquid chromatography ;
Cytochrome P450 ;
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A high-performance liquid chromatographic method has been modified for the evaluation of both Phase I and II metabolism of biphenyl by hepatocytes maintained in an embryo/hepatocyte co-culture medium. Extracts of the media, before and after hydrolysis of conjugates, are directly injected onto the HPLC and the major hydroxylated metabolites plus unmetabolized biphenyl are detected by fluorescence after separation under gradient or isocratic conditions. The method is almost free of interferences and is relatively simple and rapid. In the case of monohydroxylated derivatives, the minimum media concentrations which can be measured are 7 to 20 nM (0.07 to 0.2 pmol on-column). Recoveries from culture medium to which known amounts of biphenyl and metabolites had been added were quantitative (90-103%) and the reproducibility good (interassay CV < 5%). The assay was applied to cultures of hepatocytes derived from rabbit and from phenobarbital induced and noninduced rat. (Copyright (c) 1988 Academic Press, Inc.)