Determination of CYP2C9-catalyzed diclofenac 4'-hydroxylation by high-performance liquid chromatography.
A reverse-phase, high-performance liquid chromatography method is described for quantification of diclofenac 4'-hydroxylation catalyzed by human liver microsomes or cDNA-expressed CYP2C9. Analytical separation is achieved using a C18 column developed with a gradient of 30% acetonitrile and 2 mM perchloric acid in water to 100% methanol, with detection at 280 nm. This method is applicable to enzymatic studies for determination of CYP2C9-catalyzed diclofenac 4'-hydroxylation activity.
Crespi
Charles L
CL
Discovery Labware, BD Biosciences, Woburn, MA, USA.
Chang
Thomas K H
TK
Waxman
David J
DJ
D001189:Aryl Hydrocarbon Hydroxylases / Q000378:metabolism*; D002384:Catalysis; D002851:Chromatography, High Pressure Liquid; D065729:Cytochrome P-450 CYP2C9; D004008:Diclofenac / Q000378:metabolism*; D006801:Humans; D006900:Hydroxylation; D008862:Microsomes, Liver / Q000201:enzymology*; D013056:Spectrophotometry, Ultraviolet
D004008:Diclofenac; C450260:CYP2C9 protein, human; D065729:Cytochrome P-450 CYP2C9; D001189:Aryl Hydrocarbon Hydroxylases