title: CYP2C19-mediated (S)-mephenytoin 4'-hydroxylation assayed by high-performance liquid chromatography with radiometric detection.
abstract: A reverse-phase, high-performance liquid chromatography method is described for the quantification of 4'-hydroxymephenytoin formed enzymatically from 14C-labeled (S)-mephenytoin following incubation with cDNA-expressed CYP2C19 or human liver microsomes. Analytical separation is achieved using a C18 column developed with a gradient from 10 to 100% methanol, with detection using a scintillation detector. This method is applicable to enzymatic studies for determination of CYP2C19-catalyzed (S)-mephenytoin 4'-hydroxylation activity.
authors.affiliation: Discovery Labware, BD Biosciences, Woburn, MA, USA.
authors.forename: Charles L
authors.initials: CL
authors.lastname: Crespi
authors.forename: Thomas K H
authors.initials: TK
authors.lastname: Chang
authors.forename: David J
authors.initials: DJ
authors.lastname: Waxman
mesh_terms: D001189:Aryl Hydrocarbon Hydroxylases / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D065731:Cytochrome P-450 CYP2C19; D006801:Humans; D008617:Mephenytoin / Q000378:metabolism*; D008862:Microsomes, Liver / Q000201:enzymology; D006899:Mixed Function Oxygenases / Q000378:metabolism*; D011874:Radiometry / Q000379:methods*
chemical_list: D006899:Mixed Function Oxygenases; D001189:Aryl Hydrocarbon Hydroxylases; C045793:CYP2C19 protein, human; D065731:Cytochrome P-450 CYP2C19; D008617:Mephenytoin