Monitoring of Cytochrome P450 2C9 isoform reaction with Diclofenac by In-Capillary System based on Transverse Diffusion of Laminar Flow Profiles
| Authors | |
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| Year of publication | 2010 |
| Type | Article in Proceedings |
| Conference | Book of Abtracts 16th Latin-American Symposium LACE 2010 |
| MU Faculty or unit | |
| Citation | |
| Keywords | EMMA; cytochromes P450; drug metabolism; diclofenac |
| Description | In this study a method for cytochrome P450 2C9 (CYP2C9) reaction with diclofenac as a probe substrate was developed. 30 min incubation time and sandwich layout, where a plug of CYP2C9 is surrounded by two plugs of diclofenac and NADPH mixture, were selected as appropriate incubation conditions for final method within the optimizing. Although bare silica capillary was used for performing the analyses of samples consisting of enzyme preparations, strong rinsing procedure with 1 M H3PO4 and 1 M NaOH applied between following runs provides good method's repeatability with RSD values of 1.52 % and 4.83 % (n = 6) for migration times and peak areas of 4'-hydroxydiclofenac, respectively. Since the methodology of TDLFP allows changing the incubation mixture composition in simple way, the introduced method represents a promising tool for in-capillary inhibition studies of CYP2C9. |
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