Wang Na, Xu Fengguo, Zhang Zunjian, Yang Cheng, Sun Xiuhong, Li Jinheng
Key Laboratory of Drug Quality Control and Pharmacovigilance (China Pharmaceutical University), Ministry of Education, Nanjing 210009, People's Republic of China.
Biomed Chromatogr. 2008 Feb;22(2):149-56. doi: 10.1002/bmc.909.
A sensitive, rapid and simple high-performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS) method for simultaneous determination of dipyridamole and salicylic acid in human plasma has been developed and validated. After the addition of diazepam and rosiglitazone as internal standard (IS), plasma samples were prepared by liquid-liquid extraction followed by an isocratic elution with methanol:2 mM ammonium acetate buffer (pH 4.25; 70/30, v/v) on a Shimadzu VP-ODS C(18) column (5 microm, 150 x 2.0 mm I.D.). Detection was performed on a quadrupole mass spectrometer with ESI interface operating in the positive-ion mode for dipyridamole and negative-ion mode for salicylic acid. Calibration curves were linear (r(2) > 0.99) over the concentration range 10-2500 ng/mL for dipyridamole and 30-4000 ng/mL for salicylic acid with acceptable accuracy and precision, respectively. The intra- and inter-batch precisions were less than 15% of the relative standard deviation. The limits of detection of dipyridamole and salicylic acid were 1 and 15 ng/mL, respectively. The validated HPLC-ESI-MS method was successfully applied to a preliminary pharmacokinetic study of fixed-dose combination of sustained-release dipyridamole/aspirin in Chinese healthy male volunteers.
已开发并验证了一种灵敏、快速且简便的高效液相色谱-电喷雾电离-质谱法（HPLC-ESI-MS），用于同时测定人血浆中的双嘧达莫和水杨酸。加入地西泮和罗格列酮作为内标（IS）后，血浆样品通过液-液萃取制备，然后在岛津VP-ODS C(18)柱（5微米，150×2.0毫米内径）上用甲醇:2 mM醋酸铵缓冲液（pH 4.25；70/30，v/v）进行等度洗脱。在四极杆质谱仪上进行检测，电喷雾接口在正离子模式下用于双嘧达莫，负离子模式下用于水杨酸。双嘧达莫在10 - 2500 ng/mL浓度范围内校准曲线呈线性（r(2) > 0.99），水杨酸在30 - 4000 ng/mL浓度范围内校准曲线呈线性，分别具有可接受的准确度和精密度。批内和批间精密度均小于相对标准偏差的15%。双嘧达莫和水杨酸的检测限分别为1和15 ng/mL。经过验证的HPLC-ESI-MS方法成功应用于中国健康男性志愿者中缓释双嘧达莫/阿司匹林固定剂量组合的初步药代动力学研究。
