HPLC法测定患血浆中氯吡格雷羧酸代谢产物的血药浓度.docVIP

HPLC法测定患者血浆中氯吡格雷羧酸代谢产物的血药浓度 刘丽娜1* , 夏泉2,32，许杜娟 1#,2，3 （1.安徽医科大学药学院， 2.安徽医科大学第一附属医院药剂科 , 3.国家中医药管理局中药化学三级实验室，安徽 合肥 230032） 摘 要：目的 建立一种简易快速的HPLC-UV法测定人血浆中氯吡格雷羧酸代谢产物（CCA）的浓度。 方法 以苯妥英钠为内标，血浆样品经6%高氯酸液-液萃取。Hypersil ODS2 (4.6×250mm,5μm)，流动相为（0.05M磷酸二氢钾，三乙胺调PH至5.7）：乙腈（78:22）。紫外检测羧酸氯吡格雷（CCA）的波长为220nm，流速1.0ml/min,柱温50℃。结果 羧酸氯吡格雷（CCA）血药浓度范围在0.10-8.0μg/ml范围内线性关系良好(r=0.9995);平均回收率为100.75%；分析方法最低检测线0.05μg/ml。日内、日间RSD均3%,冻融稳定性RSD均10%（n=5）。结论 本方法简便，准确，灵敏度高，专属性和稳定性较好，适用于氯吡格雷制剂的临床研究和药动学研究。 关键词 ： 氯吡格雷；氯吡格雷羧酸代谢产物；高效液相色谱法；血药浓度 High-performance liquid chromatographic determination of inactive carboxylic acid metabolite of clopidogrel in human plasma LIU Lina1* ，XIA Quan2,32, XU Dujuan 1#,2,3 ， （1. School of Pharmacy, Anhui Medical University, 2. Department of Pharmacy ,the First Affiliated Hosp-ital of Anhui Medicine University, 3. Third-Grade Pharmaceutical Chemistry Laboratory of State Administration of Traditional Chinese Medicine, Anhui Hefei 230032, China） Abstract ： Objective To establish a simple and rapid method（HPLC-UV）for the determination of carboxylic acid metabolite of clopidogrel (CCA) in human plasma. Method The analytical procedure involves liquid–liquid extraction of the analyte and an internal standard （phenytoin sodium）with chemical reagent-6% perchloric acid. The separation was achieved using Hypersil ODS C18 (4.6×250mm,5μm) analytical column with mobile phase consisting of 0.05M phosphate buffer containing triethylamine（pH 5.7）and acetonitrile（78:22 v/v）at detector wavelength of 220 nm. The flow rate was 1.0ml/min and the column temperature was set at 50℃. Result A good linearity was shown in the concentration range of 0.10–8.0μg/mL（r=0.995）; the average recovery was 100.75%;the LOQ was found to be 0.05 μg/ml. Both the inter-day and intra-day RSD were lower than 3%;the freeze-thaw stability RSD was less the 10%（n=5）. Conclusion The method was convenient, reliable, sensitive which could be applied in the clinical investigation and pharmacokinetic study o