HPLC-MS/MS法测定人血清中伏硫西汀的浓度

Determination of vortioxetine in human serum by HPLC-MS/MS

目的采用高效液相色谱-串联质谱法(HPLC-MS/MS)建立快速测定人血清中伏硫西汀质量浓度的方法。方法以乙腈为沉淀剂,经蛋白沉淀法处理,进高效液相串联质谱系统(HPLC-MS/MS)分析,色谱柱:Shim-pack VP-ODS柱(2.0 mm×150 mm,5μm),流动相:甲醇-5 mmoL/L甲酸铵缓冲溶液(体积比75∶25),流速:0.4 mL/min,柱温:35℃,进样量:1μL,采用电喷雾离子源、多反应监测模式,定量分析离子对为m/z 299.15→m/z 150.10(伏硫西汀)、m/z 307.25→m/z 153.10(伏硫西汀-D8)。结果伏硫西汀在2~200 ng/mL质量浓度范围内线性关系良好,伏硫西汀批间和批内精密度在0.83%~4.35%之间,准确度在95.48%~103.53%之间,提取回收率在91.93%~97.26%之间。血清质控样品在室温放置24 h、-70℃冻存7 d及14 d、冻融3次后稳定性均在85%~115%之间。结论建立的快速测定人血清中伏硫西汀质量浓度的方法处理简便、灵敏度高、专属性强、检测时间短,适用于临床伏硫西汀的血药浓度监测及药动学研究。

Objective To establish a rapid method for the determination of Vortioxetine in human serum by high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS).Methods The serum was extracted by protein precipitation with acetonitrile as precipitant.The separation was performed on the Shim-pack VP-ODS(2.0 mm×150 mm,5μm)with multiple reaction monitoring(MRM),and methanol:5 mmol/L ammonium formate(75:25)as mobile phase.The flow rate was 0.4 mL/min,the column temperature was 35℃,and the sample injection volume was 1μL.Electrospray ionization(ESI)was used for acquisition and detection in multi reaction monitoring mode.The transitions for quantifying were m/z 299.15→m/z 150.10(Vortioxetine),m/z 307.25→m/z 153.10(Vortioxetine-D8).Results The linear range was 2-200 ng/mL for Vortioxetine.The within-run and between-run precisions were within a range of 0.83%~4.35%,the accuracy was varied between 95.48%~103.53%and the extraction recovery was ranged from 91.93%to 97.26%.The stability under different conditions(24 h at room temperature;7 days and 14 days at-70℃;three freeze-thaw cycles)was ranged from 85%to 115%.Conclusion The rapid,simple,sensitive and specific method was established for determination of Vortioxetine,which is suitable for clinical blood concentration monitoring and pharmacokinetic study of Vortioxetine.