Development of a High Sensitivity Method for the Analysis of Clopidogrel and Clopidogrel Carboxylic Acid Metabolite in Human K_2EDTA Plasma Using UHPLC-MS-MS

摘要

Bioanalytical methods often involve the quantification of a parent compound to determine the pharmacokinetic properties of a potential drug. In some cases, it may also be necessary to quantify a metabolism product in addition to the parent molecule as these may be either active or toxic. Simultaneous determination can be challenging because of differences in chemical properties, such as acidity, basicity, and polarity, which can significantly increase the difficulty in developing both the extraction method as well as suitable chromatography. Developing chromatographic conditions involves not only separating the parent from the metabolite, but also effectively separating both compounds from potential coeluted contaminants such as phospholipids that may affect ionization efficiency and cause suppression or enhancement. The stabilities of the parent and metabolite also must be considered, as any interconversion between the two during the bioanalytical procedure may lead to variability within the results. In this article, we discuss the method development process required for the accurate quantification of both clopidogrel and its acid metabolite with a lower limit of quantification (LLOQ) of 1 pg/mL in human plasma.