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目的:建立测定人血浆中托吡酯浓度的HPLC-MS-MS方法。方法:采用液液萃取法,以甲基叔丁基醚为提取溶剂;HPLC采用Hypersil ODS C18色谱柱(150 mm×4.6 mm,5μm),流动相为5 mmol·L-1醋酸铵溶液-甲醇(25∶75),流速0.5 mL·min-1;质谱采用ESI源,负离子检测模式,定量分析离子为托吡酯m/z338.1[M-H]-→78.0,双氯芬酸钠m/z294.1[M-H]-→249.7。结果:人血浆中托吡酯的标准曲线范围为10.00~3 600.00 ng.mL-1(n=7),r>0.999;日内和日间精密度均小于8.9%;提取回收率(低、中、高)分别为(75.5±3.1)%,(77.0±1.6)%和(72.8±1.5)%,内标提取回收率为(76.1±3.5)%;基质效应(低、中、高)分别为(99.2±5.6)%,(98.9±4.2)%和(99.4±3.8)%,内标基质效应为(100.4±1.3)%。结论:该方法灵敏、快速、专属性强,可用于人血浆中托吡酯浓度的测定。
Objective: To establish a HPLC-MS-MS method for the determination of topiramate in human plasma. Methods: Liquid-liquid extraction was used with methyl tert-butyl ether as extraction solvent. The HPLC was carried out on a Hypersil ODS C18 column (150 mm × 4.6 mm, 5 μm) with a mobile phase of 5 mmol·L -1 ammonium acetate solution-methanol (25:75) at a flow rate of 0.5 mL · min-1. The ESI-MS and negative ion detection methods were used for the mass spectrometry. Quantitative analysis of the topiramate m / z 338.1 [MH] - → 78.0, diclofenac sodium m / z 294.1 [MH] - → 249.7. Results: The range of standard curve of topiramate in human plasma ranged from 10.00 to 3600.00 ng.mL-1 (n = 7), r> 0.999. The intra- and inter-day precision was less than 8.9%. The extraction recovery rate (low, middle and high ) Were (75.5 ± 3.1)%, (77.0 ± 1.6)% and (72.8 ± 1.5)% respectively, and the recovery rate of internal standard was (76.1 ± 3.5)%. The matrix effects (low, middle and high) ± 5.6%, (98.9 ± 4.2)% and (99.4 ± 3.8)% respectively, and the internal standard matrix effect was (100.4 ± 1.3)%. Conclusion: The method is sensitive, rapid and specific and can be used to determine the concentration of topiramate in human plasma.