Title
Development of an on-line high performance liquid chromatography detection system for human cytochrome P450 1A2 inhibitors in extracts of natural products
Author
Jeurissen, S.M.F.
Claassen, F.W.
Havlik, J.
Bouwmans, E.E.
Cnubben, N.H.P.
Sudhölter, E.J.R.
Rietjens, I.M.C.M.
van Beek, T.A.
TNO Kwaliteit van Leven
Publication year
2007
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. © 2006 Elsevier B.V. All rights reserved.
Subject
Biomedical Research
Methoxyresorufin-O-demethylase
Mixture analysis
On-line detection
P450 1A2 inhibitors
Bioassay
Cytology
Enzyme inhibition
Fluorescence
Metabolites
Screening
Methoxyresorufin-O-demethylase
Mixture analysis
On-line detection
P450 1A2 inhibitors
High performance liquid chromatography
basil extract
cytochrome P450 1A2
cytochrome P450 1A2 inhibitor
cytochrome P450 inhibitor
fluvoxamine
furafylline
kava extract
methoxyresorufin
naringenin
natural product
plant extract
propranolol
resorufin
resorufin derivative
tacrine
article
assay
basil
demethylation
fluorescence
high performance liquid chromatography
metabolite
methoxyresorufin O demethylation assay
minimum detectable amount
online analysis
parameter
priority journal
screening
Biological Products
Chromatography, High Pressure Liquid
Cytochrome P-450 CYP1A2
Cytochrome P-450 Enzyme System
Enzyme Inhibitors
Humans
Inhibitory Concentration 50
Kava
Methanol
Ocimum basilicum
Online Systems
Oxidoreductases
Plant Extracts
Spectrometry, Fluorescence
Spectrophotometry, Ultraviolet
Ocimum basilicum
Piper methysticum
To reference this document use:
http://resolver.tudelft.nl/uuid:2f6d86ef-a466-4e8b-b5a6-8af5151ecaf8
DOI
https://doi.org/10.1016/j.chroma.2006.12.007
TNO identifier
239849
ISSN
0021-9673
Source
Journal of Chromatography A, 1141 (1), 81-89
Document type
article