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

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Analytical technologies in the biomedical and life sciences, 2010-06, Vol.878 (19), p.1480-1486
• Main Authors: Serra, Hugo, Bronze, Maria do Rosário, Simplício, Ana Luísa
• Format: Article
• Language: English
• Subjects: Analysis; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Capillary electrophoresis; Carboxylic acid metabolite; Carboxylic Acids - analysis; Carboxylic Acids - chemistry; Carboxylic Acids - metabolism; Clopidogrel; Electrophoresis, Capillary - methods; Fundamental and applied biological sciences. Psychology; General pharmacology; Hydrogen-Ion Concentration; Least-Squares Analysis; Medical sciences; Method validation; Pharmacology. Drug treatments; Reproducibility of Results; Sensitivity and Specificity; Sodium Dodecyl Sulfate; Temperature; Ticlopidine - analogs & derivatives; Ticlopidine - analysis; Ticlopidine - chemistry; Ticlopidine - metabolism
• ISSN: 1570-0232; 1873-376X
• DOI: 10.1016/j.jchromb.2010.03.044

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 K m of 145 ± 30 μM and V max 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.