High-performance liquid chromatographic assay of biphenyl metabolism by hepatocytes cultured in an embryo/hepatocyte co-culture medium
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.
- Research Organization:
- Environmental Protection Agency, Research Triangle Park, NC (USA). Health Effects Research Lab.
- OSTI ID:
- 6919061
- Report Number(s):
- PB-90-185216/XAB; EPA--600/J-88/485
- Country of Publication:
- United States
- Language:
- English
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Related Subjects
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.
ANIMAL CELLS
ANIMALS
AROMATICS
BIOASSAY
BIPHENYL
CHROMATOGRAPHY
EMBRYOS
FLUORESCENCE
HYDROCARBONS
LIQUID COLUMN CHROMATOGRAPHY
LIVER CELLS
LUMINESCENCE
MAMMALS
METABOLISM
METABOLITES
ORGANIC COMPOUNDS
RABBITS
RATS
RODENTS
SEPARATION PROCESSES
SOMATIC CELLS
TABLES
TOXICITY
VERTEBRATES