Qin Ting, Qin Feng, Li Ning, Lu Shan, Liu Wei, Li Famei
Department of Analytical Chemistry, Shenyang Pharmaceutical University, People's Republic of China.
Biomed Chromatogr. 2010 Mar;24(3):268-73. doi: 10.1002/bmc.1283.
Dipyridamole is a classic platelet inhibitor which has been a key medicine in clinical therapy of thrombosis and cerebrovascular disease. A rapid, selective and convenient method using high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) was developed for determination of dipyridamole in human plasma. After protein precipitation of 200 microL plasma with methanol, dipyridamole and diazepam (internal standard) were chromatographed on an Ultimate XB-C(18) (50 x 2.1 mm i.d, 3 microm) column with the mobile phase consisting of methanol-ammonium acetate (5 mM; 80 : 20, v/v) at a flow rate of 0.25 mL/min. The detection was performed on a triple quadrupole tandem mass spectrometer by multiple reaction monitoring mode via positive eletrospray ionization source (ESI(+)). The retention times of dipyridamole and diazepam were 1.4 and 1.2 min, respectively. The method was validated over a concentration range of 0.0180-4.50 microg/mL (r(2) > or = 0.99) with a lower limit of quantitation (LLOQ) of 0.0180 microg/mL for dipyridamole. The intra- and inter-day precisions (RSD) of the assay at all three QC levels were 1.6-12.7% with an accuracy (RE) of -4.3-1.9%, which meets the requirements of the FDA guidance. The HPLC-MS/MS method herein described was proved to be suitable for pharmacokinetic study of sustained-release dipyridamole tablet in volunteers after oral administration.
双嘧达莫是一种经典的血小板抑制剂，一直是血栓形成和脑血管疾病临床治疗中的关键药物。建立了一种快速、选择性且简便的高效液相色谱 - 串联质谱法（HPLC-MS/MS）用于测定人血浆中的双嘧达莫。用甲醇对200微升血浆进行蛋白沉淀后，双嘧达莫和地西泮（内标）在Ultimate XB-C(18)（50×2.1毫米内径，3微米）柱上进行色谱分离，流动相由甲醇 - 醋酸铵（5 mM；80 : 20，v/v）组成，流速为0.25毫升/分钟。通过正电喷雾电离源（ESI(+)）在三重四极杆串联质谱仪上采用多反应监测模式进行检测。双嘧达莫和地西泮的保留时间分别为1.4分钟和1.2分钟。该方法在0.0180 - 4.50微克/毫升的浓度范围内得到验证（r(2)≥0.99），双嘧达莫的定量下限（LLOQ）为0.0180微克/毫升。在所有三个质量控制水平下，该测定的日内和日间精密度（RSD）为1.6 - 12.7%，准确度（RE）为 -4.3 - 1.9%，符合FDA指南的要求。本文所述的HPLC-MS/MS方法被证明适用于志愿者口服双嘧达莫缓释片后的药代动力学研究。
