人血清中舍曲林、度洛西汀、氟伏沙明、氟西汀和去甲氟西汀的LC-MS/MS方法建立及临床应用

目的 建立人血清中舍曲林、度洛西汀、氟伏沙明、氟西汀和去甲氟西汀浓度同时测定的LC-MS/MS方法,实现临床样本的治疗药物监测。方法 以N-甲基氟西汀和N-甲基度洛西汀为内标,用甲醇-乙腈(1∶1,V∶V)混合溶剂沉淀蛋白;使用ES Sonoma C₁₈(2)色谱柱(2.1 mm×50 mm,3 μm),以0.2%乙腈和50%甲醇水溶液(含6 mmol·L^-1乙酸铵)为流动相,梯度洗脱;使用三重四级杆质谱进行质量分析,采用电喷雾离子源,在正离子模式下以MRM的扫描模式进行定量分析。结果 所建方法经验证,空白基质对分析物和内标无干扰,注射完高浓度样品后各分析物和内标均无残留,基质效应符合文件要求(内标归一化的基质因子变异系数CV%＜15%),处理后的样品24 h内分析稳定、未处理样本4 ℃冰箱存放1周稳定,舍曲林、度洛西汀、氟伏沙明、氟西汀及去甲氟西汀分别在9.22~295.16、10.80~345.60、8.10~259.12、10.11~323.54及5.90~188.70 ng·mL^-1内均具有良好的线性关系,r²＞0.99,方法的准确度高、精密度好。结论 该方法的样本处理简单,样本分析时间短,可实现大量临床样本的快速分析,已成功应用于临床服用舍曲林、度洛西汀、氟伏沙明、或氟西汀的患者血药浓度监测。

Development and Validation of a LC-MS/MS Assay for Therapeutic Drug Monitoring of Sertraline,Duloxetine, Fluvoxamine,Fluoxetine and Norfluoxetine in Serum

OBJECTIVE To develop a simple and reliable liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for therapeutic drug monitoring (TDM) of sertraline, duloxetine, fluvoxamine, fluoxetine and norfluoxetine in serum. METHODS These analytes were extracted by protein precipitation using N-methyl fluoxetine and N-methyl duloxetine as internal standard. The separation was achieved on ES Sonoma C₁₈(2) column (2.1 mm×50 mm, 3 μm) with gradient eluted program using 0.2% formic acid acetonitrile and 50% methanol-water with 6 mmol·L^-1 ammonium acetate as mobile phase. API 4000 triple quadrupole system equipped with an electrospray ionization ion-trap source (ESI) was used for monitoring the analytes and which was operated in the positive ion mode. Analytes were detected in the multiple reaction monitoring (MRM) mode. RESULTS The linear calibration curve of sertraline, duloxetine, fluvoxamine, fluoxetine and norfluoxetine were obtained in the concentration range of 9.22-295.16 (r²=0.997 4), 10.80-345.60 (r²=0.995 5), 8.10-259.12 (r²=0.995 2), 10.11-323.54 (r²=0.996 7), 5.90-188.70 (r²=0.995 7) ng·mL^-1, respectively. The accuracy of intra-batch and inter-batch assays was between 93.44% and 106.20% and the precision CV% was less than 10.00%. The selectivity, matrix effect and carry-over meet the requirements. The raw samples could stay stable for seven days in 4 ℃ condition, and post-preparative stability was validated for 24 h in room temperature and meet the requirement. CONCLUSION A simple and reliable LC-MS/MS method is established and validated. And this method has already been used to monitor the serum concentration of sertraline, duloxetine, fluvoxamine, fluoxetine and norfluoxetine.