Highly sensitive and specific high-performance liquid chromatographic analysis of 7-hydroxywarfarin, a marker for human cytochrome P-4502C9 activity

J Chromatogr B Biomed Appl. 1995 Oct 20;672(2):305-9. doi: 10.1016/0378-4347(95)00222-5.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Anticoagulants / analysis
  • Anticoagulants / metabolism
  • Aryl Hydrocarbon Hydroxylases*
  • Chromatography, High Pressure Liquid / methods*
  • Chromatography, High Pressure Liquid / statistics & numerical data*
  • Cytochrome P-450 Enzyme System / metabolism*
  • Humans
  • Kinetics
  • Microsomes, Liver / enzymology
  • Sensitivity and Specificity
  • Steroid 16-alpha-Hydroxylase*
  • Steroid Hydroxylases / metabolism*
  • Warfarin / analogs & derivatives*
  • Warfarin / analysis
  • Warfarin / metabolism


  • Anticoagulants
  • Warfarin
  • Cytochrome P-450 Enzyme System
  • 7-hydroxywarfarin
  • Steroid Hydroxylases
  • Aryl Hydrocarbon Hydroxylases
  • Steroid 16-alpha-Hydroxylase