LC-MS/MS法同时测定人血浆中氟西汀、去甲氟西汀、帕罗西汀和氟伏沙明的质量浓度

Simultaneous determination of fluoxetine,norfluoxetine,paroxetine and fluvoxamine concentration in human plasma by LC-MS/MS

目的建立一种简单、快速的能同时测定血浆中氟西汀、去甲氟西汀、帕罗西汀和氟伏沙明质量浓度的LC-MS/MS方法。方法血浆经乙腈沉淀后,上清液直接进样分析。色谱柱为Hypersil GOLD aQ,流动相为乙腈-1 mL·L^-1甲酸溶液,梯度洗脱。正离子检测,扫描方式为选择性反应监测,氟西汀、去甲氟西汀、帕罗西汀、氟伏沙明和内标罗红霉素的检测离子对分别为m/z 310.2→44.4,296.1→134.1,330.2→192.1,319.2→259.1和837.7→679.3。结果血浆中氟西汀、去甲氟西汀、帕罗西汀和氟伏沙明的质量浓度在1~1000 ng·mL^-1范围内线性关系良好,最低定量下限为1 ng·mL^-1。氟西汀、去甲氟西汀、帕罗西汀和氟伏沙明的日内和日间精密度RSD值均小于15%,日内和日间相对误差在-15%~15%之间,药物的提取回收率均大于90%,无显著基质效应。药物在反复冻融3次、室温下保存12 h和-80℃保存1个月的条件下稳定。该方法成功应用于抑郁症患者的氟西汀、去甲氟西汀、帕罗西汀和氟伏沙明血药质量浓度监测。结论建立的LC-MS/MS分析方法简便、灵敏、准确,可用于临床血药质量浓度的监测。

Objective To establish an LC-MS/MS method for simultaneous determination of fluoxetine,norfluoxetine,paroxetine and fluvoxamine concentration in human plasma.Methods Plasma samples were deproteinized by adding acetonitrile,and the supernatant was directly analyzed by LC-MS/MS.A Hypersil GOLD aQ column with a mobile phase composed of acetonitrile-1 mL·L^-1 formic acid in gradient mode was utilized to separate the analytes,and detection was carried out in positive and selective reaction monitoring(SRM)mode.The SRM transitions monitored for fluoxetine,norfluoxetine,paroxetine and fluvoxamine and roxithromycin as internal standard were m/z 310.2→44.4,296.1→134.1,330.2→192.1,319.2→259.1 and 837.7→679.3,respectively.Results The proposed method was linear over the range of 1 to 1000 ng·mL^-1 for fluoxetine,norfluoxetine,paroxetine and fluvoxamine,and the lower limit of quantification(LLOQ)was 1 ng·mL^-1 for all of them.The intra-day and inter-day precision were within 15%,and relative error for fluoxetine,norfluoxetine,paroxetine and fluvoxamine were between-15%to 15%,respectively.The recoveries were more than 90%for all of them,and no evident matrix effect was observed.The samples remained stable after freeze-thaw cycles 3 times and preserving at ambient temperature for 12 h and at-80℃for 1 month.The proposed method was successfully used to therapeutic drug monitoring of fluoxetine,norfluoxetine,paroxetine and fluvoxamine in depression patients.Conclusion A simple,sensitive and accurate LC-MS/MS method was established for simultaneous analysis of fluoxetine,norfluoxetine,paroxetine and fluvoxamine in human plasma,and was applied to therapeutic drug monitoring.