Coupled β-cyclodextrin and reverse-phase high-performance liquid chromatography for assessing biphenyl hydroxylase activity in hepatic 9000g supernatant
✍ Scribed by Dix E. Weaver; Robert B.L. Van Lier
- Publisher
- Elsevier Science
- Year
- 1986
- Tongue
- English
- Weight
- 415 KB
- Volume
- 154
- Category
- Article
- ISSN
- 0003-2697
No coin nor oath required. For personal study only.
✦ Synopsis
Coupled fi-cyclodextrin bonded-phase and reverse-phase high-performance liquid chromatography with fluorescence detection has been employed to detect the major hydroxylated metabolites of biphenyl following in virro incubation with hepatic 9000~ supernatant.
The method requires only 0.3 mg of protein and its sensitivity was as low as 0.36 nmol metabolite formed/mg protein/ h (0.32 pmol injected) for 2-. 3-. and 4-hydroxybiphenyl.
Microsomes need not be purified and no organic extraction or derivatization was required.
The method was employed successfully with samples from rats and mice treated with Aroclor, a-naphthoflavone, or phenobarbital; from monkeys dosed with Aroclor;
and from untreated dogs.