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...

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Bibliographic Details
Published inJournal of chromatography. B, Biomedical applications Vol. 672; no. 2; pp. 305 - 309
Main Authors Lang, D., Böcker, R.
Format Journal Article
LanguageEnglish
Published Netherlands Elsevier B.V 20.10.1995
Subjects
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
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Summary: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 C 18 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.
Bibliography:ObjectType-Article-1
SourceType-Scholarly Journals-1
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ISSN:0378-4347
1572-6495
DOI:10.1016/0378-4347(95)00222-5