HPLC-MS/MS法同时测定人血浆中6种抗抑郁药的浓度

目的:建立HPLC-MS/MS同时测定人血浆中西酞普兰、米氮平、曲唑酮、氟伏沙明、氯米帕明、文拉法辛血药浓度的方法。方法:血浆经含内标(伏立康唑)乙腈沉淀蛋白后进样测定,采用Kinetex C18柱(50 mm×2.1 mm,2.6μm)进行色谱分离,以乙腈-0.1%甲酸水为流动相,梯度洗脱,流速0.4 mL·min……-1,柱温40℃,进样量2μL;采用电喷雾离子源,正离子模式,多反应监测进行测定。结果:西酞普兰和米氮平质量浓度范围为4~400 ng·mLmL-1(r>0.9986),曲唑酮、氟伏沙明、氯米帕明和文拉法辛质量浓度范围为10~1000 ng·mL-1(r>0.9984);所有分析物日内、日间精密度(n=6)RSD<11.52%,准确度RE在-3.89%~8.14%,符合生物样品分析要求。氯米帕明在高脂血基质及溶血基质中有明显基质效应,其余待测物无明显基质效应。结论:经方法学考察,该方法灵敏、简便、准确、快速,可用于6种抗抑郁药的临床血药浓度检测。

Simultaneous determination of six antidepressants in human plasma by HPLC-MS/MS

OBJECTIVE To establish a HPLC-MS/MS method for simultaneous determination of citalopram, mirtazapine, trazodone, fluvoxamine, clomipramine and venlafaxine in human plasma. METHODS Plasma was precipitated by internal standard (voriconazole) acetonitrile. The protein was separated by Kinetex C₁₈ column (50 mm×2.1 mm, 2.6 μm). Acetonitrile-0.1% formic acid water was used as mobile phase. Gradient elution was performed at a flow rate of 0.4 mL·min^-1, column temperature of 40℃, and sample size of 2 μL. Electrospray ionization source, positive ion mode and multiple reaction monitoring were used for determination. RESULTS The concentration of citalopram and mirtazapine ranged from 4 to 400 ng·mL^-1 (r > 0.998 6), and the concentration of trazodone, fluvoxamine, clomipramine and venlafaxine ranged from 10 to 1 000 ng·mL^-1 (r > 0.998 4); the intra-day and inter-day precision of all analytes (n=6) RSD < 11.52%, and the accuracy RE ranged from -3.89% to 8.14%, which met the requirements of biological sample analysis. Clomipramine has a significant matrix effect in the hyperlipidemia matrix and the hemolysis matrix, while the other substances had no significant matrix effect.CONCLUSION This method is sensitive, simple, accurate and rapid, and can be used for the detection of clinical blood concentration of six antidepressants.