Simultaneous determination of MHD and DMD in dog plasma by high-performance liquid chromatography with fluorescence detection and its application to pharmacokinetic studies

摘要

A rapid, reproducible high-performance liquid chromatographic (HPLC) method with fluorescence detection for the simultaneous determination of 3(or 8)-(1-methoxyethyl)-8(or 3)-(1-hydroxyethyl)-deuteroporphyrin IX (MHD) and 3,8-di-(1-methoxyethyl)-deuteroporphyrin IX (DMD) in dog plasma was described. Fluorescein was used as an internal standard. A simple extraction step with ethyl acetate was performed before chromatography on a Diamonsil C-18 column (5 mu m, 4.6 mmx150 mm). The chromatography used 0.02 mol L-1 sodium acetate/tetrahydrofuran (66:34 v/v). The analytical curve was linear over the concentration range 0.025- 2.5 mu g mL(-1). For a 100 mu L dog plasma sample, the limit of determination for both MHD and DMD was 0.025 mu g mL(-1). The recoveries of MHD and DMD were more than 76 and 89, respectively. The intra-assay (within-run) and interassay (between-run) coefficients of variation (precisions) for MHD and DMD were less than 15. This method was found to be suitable for the analysis of biosamples and was successfully applied to pharmacokinetic studies of Deuxemether in dogs.