HPLC-MS/MS偶联在线固相萃取技术同时定量测定人血清中6个精神科药物浓度

目的：建立并验证同时测定人血清中氟哌啶醇、氟奋乃静、氟哌噻吨、丁螺环酮、阿普唑仑和米安色林6个精神科药物的在线柱切换LC-MS/MS的灵敏和高选择性方法。方法：40 μL血清样本加入乙腈450 μL（含2 ng·mL^-1的H³-利培酮内标）进行蛋白质沉淀，4℃离心（4 000 r·min^-1）10 min，后取上清20 μL进样，在Waters Xbridge^TM C₈ column（5 mm，2.1 mm×30 mm）固相萃取（solid-phase extraction，SPE）柱，以1%氨水（pH 11）-乙腈溶液（95∶5）为流动相，流速0.5 mL·min^-1，进行初步分离和纯化，在线转移到Xbridge^TM C₁₈（3.5 mm，100 mm×2.1 mm）分析柱（柱温∶室温）进行分离，采用乙腈-1%氨水溶液（pH 11）（70∶30）的流动相，以0.2 mL·min^-1的流速洗脱，串联质谱仪检测。结果：6个精神科药物线性范围覆盖临床有效治疗参考浓度范围和实验室警戒浓度。6个不同来源血清配制高、中、低质控3个水平的基质效应RSD≤15%。所有分析物日内、日间精密度（n=5）在1.1%~8.2%，准确度（n=5）在6.6%~7.6%。结论：该方法灵敏、准确、简单、快速，可用于临床血药浓度检测。

Objective:To develop a sensitive and selective high performance liquid chromatography-tandem mass spectrometry(HPLC-MS/MS)on-line coupled with solid phase extraction for the determination of 6 psychotropic drugs in human serum. Methods:40 μL serum was precipitated by adding 450 μL acetonitrile(containing 2 ng·mL^-1 IS)and then centrifuged(4 000 r·min^-1)at 4℃for 10 min. 20 μL supernatant was injected into the chromatographic system. The analytes were automatically pre-concentrated and pre-purified by Xbridge^TM C₈ solid phase extraction(SPE)cartridge(5 mm,2.1 mm×30 mm)with a mobile phase consisting of acetonitrile and 1% ammonium hydroxide solution(pH 11) (5:95)at a flow rate of 0.5 mL·min^-1,then separated on Xbridge^TM C₁₈ column(3.5 μm, 2.1 mm×100 mm)at room temperature with a mobile phase consisting of acetonitrile and 1% ammonium hydroxide solution(pH 11)(70:30)at a flow rate of 0.2 mL·min^-1,and detected with tandem mass detector. Results:The linear range of 6 psychiatric drugs covered the reference concentration range and laboratory alert concentration of clinical effective treatment. The matrix effects were assessed with the quality control samples,prepared by using the blank serum samples from 6 individual donors,on high,medium and low levels. And the overall relative standard deviation(RSD)for the concentration was not greater than 15%. The intra- and inter-day precisions(n=5)for all analytes were between 1.1% and 8.2%,and the method accuracy(n=5)was between 6.6% and 7.6%. Conclusion:A sensitive, accurate,simple and rapid on-line SPE-HPLC-MS/MS has been developed and validated for the therapeutic drug monitoring(TDM)of 6 psychotropic drugs.