Highly sensitive and specific high-performance liquid chromatographic analysis of 7-hydroxywarfarin, a marker for human cytochrome P-4502C9 activity.
The formation of 7-hydroxywarfarin in incubations of (S)-warfarin with human liver microsomes reflects their cytochrome P-4502C9 activity. This paper describes a rapid high-performance liquid chromatographic method for the determination of 7-hydroxywarfarin with high sensitivity, selectivity, and a simple sample clean-up procedure. Separation was achieved with a C18 reversed-phase column and quantification by fluorometric detection. The method employs an internal standard resulting in good accuracy and precision. The limit of detection is 150 fmol for 7-hydroxywarfarin.
Lang
D
D
Department of Toxicology and Pharmacology, University of Erlangen-Nürnberg, Germany.
Böcker
R
R
D000925:Anticoagulants / Q000032:analysis / Q000378:metabolism; D001189:Aryl Hydrocarbon Hydroxylases; D002851:Chromatography, High Pressure Liquid / Q000379:methods* / Q000706:statistics & numerical data*; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008862:Microsomes, Liver / Q000201:enzymology; D012680:Sensitivity and Specificity; D039181:Steroid 16-alpha-Hydroxylase; D013250:Steroid Hydroxylases / Q000378:metabolism*; D014859:Warfarin / Q000031:analogs & derivatives* / Q000032:analysis / Q000378:metabolism
D000925:Anticoagulants; D014859:Warfarin; D003577:Cytochrome P-450 Enzyme System; C087837:7-hydroxywarfarin; D013250:Steroid Hydroxylases; D001189:Aryl Hydrocarbon Hydroxylases; D039181:Steroid 16-alpha-Hydroxylase