Simultaneous determination of clopidogrel and its carboxylic acid metabolite by capillary electrophoresis

publication · 11 years ago
by Hugo Serra, Ana Luisa Simplicio, Maria do Rosário Bronze (IBET/ITQB, Laboratório Medinfar, iMed)
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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 2–100mM for both compounds. The method limits of detection and quantification were, respectively, 1.2 and 3.7mMfor clopidogrel and 1.1 and 3.2mMfor 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±30mMand Vmax of 0.4, 1.5 and 3.4mM/min, respectively for the enzyme concentrations 1.0, 2.0 and 4.0 U/ml, were obtained.
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