Clinical Research and Biosciences (I) Pvt. Ltd., Hyderabad, Andhra Pradesh, India.
J Chromatogr B Analyt Technol Biomed Life Sci. 2010 Feb 1;878(3-4):502-8. doi: 10.1016/j.jchromb.2009.12.026. Epub 2010 Jan 4.
A simple, sensitive and reliable method is described for simultaneous quantification of Clopidogrel and its metabolite in human plasma by using HTLC-MS/MS. The analytical procedure involves on-line coupling of extraction with Cyclone P (50 mm x 0.5 mm 50 microm) HTLC column by injecting 15 microL sample and chromatographic separation is performed with Cohesive Propel C18 (5 microm, 3.0 mm x 50 mm), followed by quantification with mass detector in SRM mode using ESI as an interface. The calibration curves were linear over a concentration range of 0.1-8 ng/mL of Clopidogrel and 70 ng/mL to 6 microg/mL of its metabolite using 20 mL human plasma per batch. The total run time of analysis was 7.5 min and the lower limits of quantification were 0.1 ng/mL for Clopidogrel and 70 ng/mL for its metabolite. The method validation was carried out in terms of specificity, sensitivity, linearity, precision, accuracy and stability. The validated method was applied in bioavailability and bioequivalence study.
本文描述了一种简单、灵敏、可靠的 HTLC-MS/MS 法同时测定人血浆中氯吡格雷及其代谢物的浓度。分析过程包括在线萃取与 Cyclone P（50mm×0.5mm，50μm）HTLC 柱的耦联，进样 15μL，采用 Cohesive Propel C18（5μm，3.0mm×50mm）色谱柱进行分离，以电喷雾（ESI）作为接口，采用选择反应监测（SRM）模式进行质谱检测。氯吡格雷的浓度在 0.1-8ng/mL 范围内，其代谢物的浓度在 70ng/mL 至 6μg/mL 范围内呈线性关系，每批 20mL 人血浆。分析总用时 7.5min，氯吡格雷和其代谢物的定量下限分别为 0.1ng/mL 和 70ng/mL。该方法经过特异性、灵敏度、线性、精密度、准确度和稳定性验证后，用于生物利用度和生物等效性研究。
