HPLC法测定患者血浆中氯吡格雷的羧酸代谢产物浓度

Concentration Determination of Carboxylic Acid Metabolite of Clopidogrel in Human Plasma by HPLC

目的：建立一种简易快速的方法测定人血浆中氯吡格雷代谢产物羧酸氯吡格雷（CCA）的浓度。方法：血浆样品经6％高氯酸液．液萃取后采用高效液相色谱一紫外（HPLC．uv）法进样测定，内标为苯妥英钠，色谱柱为Hypersi10DSC，流动相为0．05mol／L磷酸二氢钾（三乙胺调pH至5．7）．乙腈（78：22），紫外检测波长为220μm，流速为1．0ml／min，柱温为50℃。结果：CCA血药浓度在O．10～8．0μg／ml范围内线性关系良好（r=0．9995），分析方法最低检测限为0．05μg／ml；方法回收率为99．7％～100．2％，提取回收率〉75％；日内、日间RSD均小于3％，冻融稳定性RSD均小于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，适用于氯吡格雷的临床研究和药动学研究。

OBJECTIVE： To establish a simple and rapid method for the determination of carboxylic acid metabolite of clopido grel （CCA） in human plasma. METHODS： After liquid-liquid extraction with 6% perchloric acid, plasma sample was determined by HPLC-UV using phenytoin sodium as internal standard. The separation was performed on Hypersil ODS C column with mobile phase consisting of 0.05 mol/L monopotassium phosphate （pH adjusted to 5.7 using triethylamine）-acetonitrile （78：22） at the flow rate of 1.0 ml/min. UV detection wavelength was set at 220 nm, and the column temperature was 50℃. RESULTS： The linear range of CCA were 0.10-8.0 pg/ml（r=0.999 5）; the method recovery was 99.7%-100.2%, the extraction recovery was 〉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 than 10% （n=5）. CONCLUSIONS： The method is convenient, accurate, sensitive, specific and stable, which could be applied for clinical study and pharmacokinetic study of clopidogrel.