LC-MS/MS法测定血浆中伊拉地平的浓度及其人体药代动力学研究

目的：建立LC-MS/MS法测定人血浆中伊拉地平的浓度，研究中国健康人单剂量口服伊拉地平胶囊后体内药代动力学。方法：采用Waters Xterra^®MS C₁₈色谱柱(2.1 mm×100 mm，3.5 μm)，以乙腈-0.5 mmol·L^-1醋酸铵水溶液为流动相，梯度洗脱(0~1min，20%乙腈；1~3 min，20%→90%乙腈；3~5 min，90%乙腈；5~5.1 min，90%→20%乙腈；5.1~9 min，20%乙腈)，流速0.3 mL·min^-1，柱温40 ℃；负离子方式检测，扫描方式为多反应监测(MRM)，固相萃取法处理血浆样品；9名受试者单剂量口服10 mg伊拉地平胶囊后，以LC-MS/MS法测定血浆中伊拉地平浓度，使用WinNonlin^® 6.3软件对血药浓度数据进行处理，计算药动学参数。结果：血浆中伊拉地平浓度线性范围为10~10000 pg·mL^-1，定量下限为10 pg·mL^-1，批内、批间精密度均小于15%。单剂量口服10 mg伊拉地平胶囊后主要的药动学参数C_max为(4610.21±147.91)pg·mL^-1，T_max为(1.46±0.42)h，t_1/2为(11.65±4.38)h，AUC_0-t(34466±11140)pg·h·mL^-1。结论：本文建立的LC-MS/MS法符合方法学考察要求，适合于伊拉地平胶囊在中国健康受试者体内药代动力学研究。

Objective: To develop an LC-MS/MS method for quantification of isradipine in human plasma and study the in vivopharmacokinetics after administration of isradipine capsules at a single dose to healthy Chinese volunteers. Methods: The isradipine concentrations were determined by LC-MS/MS using a Waters Xterra^® MS C₁₈column(2.1 mm×100 mm,3.5 μm)with a mobile phase of acetonitrile-water(containing 0.5 mmol·mL^-1 ammonium acetate)by gradient elution(0-1 min,20% acetonitrile;1-3 min,20%→90% acetonitrile;3-5 min,90% acetonitrile;5-5.1 min,90%→20% acetonitrile;5.1-9 min,20% acetonitrile)at a flow rate of 0.3 mL·min^-1;the column temperature was 40 ℃ under negative ionization(MRM)mode.Plasma samples were processed by solid phase extraction.After oral administration of isradipine capsules at a single dose to 9 healthy volunteers,the plasma concentrations of isradipine were determined by LC-MS/MS method.The plasma concentrations were determined and the pharmacokinetic parameters were calculated by WinNonlin^® 6.3. Results: The calibration curve was linear within the range of 10-10000 pg·mL^-1.The LLOQ was 10 pg·mL^-1 and RSDs of intra-and inter-day were less than 15%.The pharmacokinetic parameters after administration of 10 mg isradipine capsules at a single dose were as follows:C_maxwas(4610.21±147.91)pg·mL^-1;T_max was(1.46±0.42)h;t_1/2 was(11.65±4.38)h;AUC_0-twas(34466±11140)pg·h·mL^-1. Conclusion: The developed LC-MS/MS method met the requirements of methodology verification and was suitable for the pharmacokinetic study of isradipine in healthy Chinese volunteers.