Simultaneous determination of clopidogrel and its carboxylic acid metabolite by capillary electrophoresis |
| |
Authors: | Hugo Serra Maria do Rosário Bronze Ana Luísa Simplício |
| |
Institution: | 1. IBET/ITQB-UNL, Av. da República–EAN, 2780-157 Oeiras, Portugal;2. Laboratório Medinfar, S.A., Apartado 6645, Venda Nova, 2701-357 Amadora, Portugal;3. iMed, Faculdade de Farmácia da UL, Av. das Forças Armadas, 1649-019 Lisboa, Portugal |
| |
Abstract: | A capillary electrophoresis method was developed and validated for the first time for the analysis of clopidogrel and its carboxylic acid metabolite. Prior to method optimization, the pH dependence of effective mobility of both compounds was determined in order to define the initial pH of the running buffer. The optimized method demonstrated to be selective, and linear in the concentration range of 2–100 μM for both compounds. The method limits of detection and quantification were, respectively, 1.2 and 3.7 μM for clopidogrel and 1.1 and 3.2 μM for the carboxylic acid metabolite. Moreover, method validation demonstrated acceptable results for method repeatability (RSD < 7%), intermediate precision (RSD < 7%) and accuracy (85–96%) and is suitable for the quantitative analysis of clopidogrel and its metabolite in serum samples. The validated method was also applied to the determination of the kinetic parameters of the enzymatic hydrolysis of clopidogrel. An apparent Km of 145 ± 30 μM and Vmax of 0.4, 1.5 and 3.4 μM/min, respectively for the enzyme concentrations 1.0, 2.0 and 4.0 U/ml, were obtained. |
| |
Keywords: | Capillary electrophoresis Clopidogrel Carboxylic acid metabolite Method validation |
本文献已被 ScienceDirect 等数据库收录! |
|