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Simultaneous and stereospecific analysis of warfarin oxidative metabolism using 2D LC/Q-TOF
Siji Joseph1, Murali Subramanian2, Smriti Khera3
1Life Science Center, Agilent Technologies, Inc., Bangalore 560048, India.
Background:
Warfarin is a widely used racemic anticoagulant with narrow therapeutic range and wide interindividual response to treatment. This is due to the extensive and differential clearance of R- and S-warfarin with the involvement of several polymorphic CYP450 enzymes resulting in the formation of several stereoisomeric oxidative metabolites.
Results:
A stereospecific 2DLC/Q-TOF method was developed for the simultaneous identification and quantitation of hydroxylated warfarin metabolites from a single sample analysis. Using this method metabolites from rat microsomal and plated hepatocyte incubations with R-, S- and (R/S)-warfarin were estimated.
Conclusion:
Multiheart cutting with high resolution MS and MS/MS analysis is suggested as a viable approach for achiral-chiral separation of metabolites of warfarin and other chiral or prochiral drugs.
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