Development of an on-line high performance liquid chromatography detection system for human cytochrome P450 1A2 inhibitors in extracts of natural products
article
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.
Topics
Biomedical ResearchMethoxyresorufin-O-demethylaseMixture analysisOn-line detectionP450 1A2 inhibitorsBioassayCytologyEnzyme inhibitionFluorescenceMetabolitesScreeningMethoxyresorufin-O-demethylaseMixture analysisOn-line detectionP450 1A2 inhibitorsHigh performance liquid chromatographyBasil extractCytochrome P450 1A2Cytochrome P450 1A2 inhibitorCytochrome P450 inhibitorFluvoxamineFurafyllineKava extractMethoxyresorufinNaringeninNatural productPlant extractPropranololResorufinResorufin derivativeTacrineAssayBasilDemethylationFluorescenceHigh performance liquid chromatographyMetaboliteMethoxyresorufin O demethylation assayMinimum detectable amountOnline analysisParameterPriority journalScreeningBiological ProductsChromatography, High Pressure LiquidCytochrome P-450 CYP1A2Cytochrome P-450 Enzyme SystemEnzyme InhibitorsHumansInhibitory Concentration 50KavaMethanolOcimum basilicumOnline SystemsOxidoreductasesPlant ExtractsSpectrometry, FluorescenceSpectrophotometry, UltravioletOcimum basilicumPiper methysticum
TNO Identifier
239849
ISSN
00219673
Source
Journal of Chromatography A, 1141(1), pp. 81-89.
Pages
81-89
Files
To receive the publication files, please send an e-mail request to TNO Repository.