CYP2C19-mediated (S)-mephenytoin 4'-hydroxylation assayed by high-performance liquid chromatography with radiometric detection.

Crespi CL1, Chang TK, Waxman DJ.
Source: Methods Mol Biol
Publication Date: (2006)
Issue: 320: 115-119
Cells used in publication:
Hepatocyte, mouse
Species: mouse
Tissue Origin: liver
Hepatocyte, rat
Species: rat
Tissue Origin: liver
Hepatocyte, human
Species: human
Tissue Origin: liver
Hepatocyte, Cynomolgus
Species: monkey
Tissue Origin: liver
Hepatocyte, Rhesus
Species: monkey
Tissue Origin: liver
Hepatocyte, canine
Species: canine
Tissue Origin:


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.