Development of an In-Capillary Approach to Nanoscale Automated in Vitro Cytochromes P450 Assays

A method to perform nanoscale automated CYP450-based drug metabolism studies using a capillary as it reaction vessel is described. In-capillary assays consumed only similar to 30 nL of recombinant human CYP450 solution. Ultrafast analysis Of substrates and metabolites was achieved off-line by ultrahigh-pressure liquid chromatography coupled to massspectrometry. This approach was successfully applied to qualitative metabolism studies of six major CYP450 isozymes and CYP2D6 inhibition experiments.