Development of an on-line high performance liquid chromatography detection system for human cytochrome P450 1A2 inhibitors in extracts of natural products

S.M.F. Jeurissen, F.W. Claassen, J. Havlik, E. Bouwmans, N.H.P. Cnubben, E.J.R. Sudhölter, I.M.C.M. Rietjens, T.A. van Beek

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Abstract

An on-line HPLC screening method for detection of inhibitors of human cytochrome P450 1A2 in extracts was developed. HPLC separation of extracts is connected to a continuous methoxyresorufin-O-demethylation (MROD) assay in which recombinant human P450 1A2 converts methoxyresorufin to its fluorescent metabolite resorufin. The system was tested with three P450 1A2 inhibitors, for which minimum detectable amounts (MDA) ranging from 0.7 to 9.5 ng were obtained. Analysis of a kava kava and a basil extract showed that the on-line system is applicable to complex mixtures, since in both extracts, peaks with P450 1A2 inhibiting activity were observed
Original languageEnglish
Pages (from-to)81-89
JournalJournal of Chromatography. A, Including electrophoresis and other separation methods
Volume1141
Issue number1
DOIs
Publication statusPublished - 2007

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Keywords

  • tandem mass-spectrometry
  • biochemical detection
  • catalytic activity
  • coupled online
  • enzymes
  • methyleugenol
  • metabolism
  • bioactivation
  • fluorescence
  • activation

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