Measurement of dexfenfluramide metabolism in rat liver microsomes by gas chromatography-mass spectometry

Victoria S. Haritos, Michael S. Ching, Hany Ghabrial, Jorma T. Ahokas

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Abstract

A specific and useful method was developed for the determination of dexfenfluramine metabolism by microsomal systems utilising CC-MS. The synthesis of two metabolites 1-(3-trifluoromethylphenyl)propan-2-ol('alcohol') and 1-(3-trifluoromethylphenyl)-1,2-propanediol ('diol') via straightforward routes, were confirmed by MS and NMR spectra. The conditions for extraction from alkalinised microsomal mixtures of the metabolites nordexfenfluramine, 1-(3-trifluoromethylphenyl)propan-2-one ('ketone'), alcohol and diol, their conversion to trifluoroacetate derivatives and analysis by GC-MS-SIM are described. Calibration curves were constructed between 48 and 9662 nM and fitted to quadratic equations (r2>0.999). The method precision was good over low (121 nM) medium (2415 nM) and above medium (9662 nM) concentrations for all metabolites; the within- and day-to-day coefficients of variation ranged between 2.5-12.4% and 6.7-17.5%, respectively. The accuracy, measured as bias, was very good both within- and day-to-day (range: -0.4-12.6%, 0.8-18.9%). For most metabolites, the C.V. for the assay and bias increased at 121 nM. Dexfenfluramine metabolism by rat liver microsomes was investigated using the assay method and showed a concentration dependent increase in nordexfenfluramine and ketone metabolites over the substrate range of 5-200 μM.

Original languageEnglish
Pages (from-to)327-336
Number of pages10
JournalJournal of Chromatography B - Biomedical Applications
Volume693
Issue number2
DOIs
Publication statusPublished - 6 Jun 1997
Externally publishedYes

Keywords

  • Dexfenfluramine

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