High-performance liquid chromatography analysis of CYP2C8-catalyzed paclitaxel 6alpha-hydroxylation.
Paclitaxel (Taxol) is a naturally occurring member of the taxane family of antitumor drugs, which act by stabilizing microtubules. Paclitaxel is inactivated in human liver by a cytochrome P450 (P450)-catalyzed 6alpha-hydroxylation reaction. A reverse-phase, high-performance liquid chromatographic assay is described for the analysis of paclitaxel 6alpha-hydroxylation catalyzed by human liver microsomes or cDNA-expressed P450 enzyme CYP2C8. Analytical separations are achieved using a C18 column with a linear gradient of 10-100% methanol, with detection at 230 nm. This method is applicable to enzymatic studies for determination of CYP2C8-catalyzed paclitaxel 6alpha-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 / Q000379:methods*; D065727:Cytochrome P-450 CYP2C8; D006801:Humans; D006900:Hydroxylation; D008862:Microsomes, Liver / Q000201:enzymology; D017239:Paclitaxel / Q000378:metabolism*; D013056:Spectrophotometry, Ultraviolet
D001189:Aryl Hydrocarbon Hydroxylases; C450259:CYP2C8 protein, human; D065727:Cytochrome P-450 CYP2C8; D017239:Paclitaxel