Development of an on-line high performance liquid chromatography detection system for human cytochrome P450 1A2 inhibitors in extracts of natural products.
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.
Jeurissen
Suzanne M F
SM
Laboratory of Organic Chemistry, Wageningen University, Dreijenplein 8, 6703 HB Wageningen, The Netherlands. suzanne.jeurissen@wur.nl
Claassen
Frank W
FW
Havlik
Jaroslav
J
Bouwmans
Evelien E
EE
Cnubben
Nicole H P
NH
Sudhölter
Ernst J R
EJ
Rietjens
Ivonne M C M
IM
van Beek
Teris A
TA
D001688:Biological Products / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D065609:Cytochrome P-450 CYP1A2 Inhibitors; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000032:analysis* / Q000737:chemistry / Q000494:pharmacology*; D006801:Humans; D020128:Inhibitory Concentration 50; D020901:Kava; D000432:Methanol; D018649:Ocimum basilicum; D009862:Online Systems; D010088:Oxidoreductases / Q000378:metabolism; D010936:Plant Extracts / Q000737:chemistry*; D013050:Spectrometry, Fluorescence; D013056:Spectrophotometry, Ultraviolet
D001688:Biological Products; D065609:Cytochrome P-450 CYP1A2 Inhibitors; D004791:Enzyme Inhibitors; D010936:Plant Extracts; D003577:Cytochrome P-450 Enzyme System; D010088:Oxidoreductases; C076307:methoxyresorufin-O-demethylase; D000432:Methanol