LC-MS/MS法测定人血浆中尼莫地平浓度及其药动学研究

摘要

目的:建立LC-MS/MS法测定人静脉滴注尼莫地平脂质微球注射液后血浆中尼莫地平浓度的方法,并将该方法应用于尼莫地平脂质微球注射液在健康人体内的药动学研究. 方法: 健康受试者单剂量静脉注射尼莫地平脂质微球注射液1. 0 mg·h-1 ,给药前抽取空白血样4 ml,给药后5, 10, 20, 30, 35 45 min, 1, 1. 5, 2, 3, 4, 5, 6 , 8 h分别采集静脉血4 ml. 含药血浆冻存于--80℃冰箱中待分析. 色谱柱:Agilent TC-C18(4. 6 mm′150 mm, 5 mm),流动相:甲醇与4 mmol·L-1醋酸铵缓冲液(含0. 4%甲酸)体积比为80:20,流速为0. 5 ml·min-1 ,硝苯地平为内标,采用电喷雾离子源,以多反应监测( MRM)方式进行正离子检测. 用于定量分析的离子分别为m/z 419. 2→343. 2(尼莫地平),m/z 347. 1→254. 1(硝苯地平,内标). 结果:尼莫地平的线性范围在0. 15~30 ng·ml-1 ,定量下限为0. 15 ng·ml-1 ,方法提取回收率为79. 7% ~81. 5%,相对回收率为92. 6% ~97. 6%,日内、日间RSD均<10%. 结论:该法准确、灵敏、重现性好,适用于注射用尼莫地平脂质微球注射液人体药动学研究.%Objective: To establish an LC-MS/MS method for the determination of nimodipine in human plasma and apply the method in pharmacokinetic study of nimodipine lipid microspheres for injection. Methods: A single dose of nimodipine lipid micro-spheres for injection was given to 6 healthy volunteers through iv administration. The blood samples ( approximately 4 ml) were drawn in 0 h (pre-dose) and in 5, 10, 20, 30, 35 and 45 minutes, 1, 1. 5, 2, 3, 4, 5, 6 and 8 h after the administration. All of the blood samples were immediately centrifuged and stored at -20°C until the analysis. The separation was performed on an Agilent TC-C18 col-umn (4. 6 mm × 150 mm, 5 μm). Methanol and 4 mmol·L-1 ammonium acetate (80: 20) containing 0. 4% formic acid was used as the mobile phase at a flow rate of 0. 5 ml·min-1. Nifedipine was used as the internal standard. An electrospary ionization (ESI) source was applied and operated in a positive ion mode. Multiple reaction monitoring (MRM) mode with m/z 419. 2→343. 2 and m/z 347. 1→254. 1 was used to quantify nimodipine and nifedipine, respectively. Results:The linear calibration curve of nimodipine was within the range of 0. 15-30 ng·ml-1 . The lower limit of quantification of nimodipine was 0. 15 ng·ml-1 . The extraction recoveries of nimodipine were within the range of 79. 7%-81. 5%, the relative recoveries of nimodipine were 92. 6%-97. 6%, and the intra-and in-ter-day RSDs were less than 10%. Conclusion:The method is sensitive and reproducible, and the results are reliable and accurate. It is applicable in pharmacokinetic study of nimodipine lipid microspheres for injection.