Clin Chim Acta. 2013 Aug 23;423:32-4. doi: 10.1016/j.cca.2013.04.014. Epub 2013 Apr 23.
Most antipsychotic drugs that are commonly prescribed in the USA are monitored by liquid and gas chromatographic methods. Method performance has been improved using ultra high pressure liquid chromatography coupled to tandem mass spectrometry (LC-MS/MS). A rapid and simple procedure for monitoring haloperidol, thiothixene, fluphenazine, and perphenazine is described here.
Antipsychotic drug concentrations in serum and plasma were determined by LCMS/MS (Waters Acquity UPLC TQD). The instrument is operated with an ESI interface, in multiple reaction monitoring (MRM), and positive ion mode. The resolution of both quadrupoles was maintained at unit mass with a peak width at half height of 0.7amu. Data analysis was performed using the Waters Quanlynx software. Serum or plasma samples were thawed at room temperature and a 100μL aliquot was placed in a tube. Then 300μL of precipitating reagent (acetonitrile-methanol [50:50, volume: volume]) containing the internal standard (0.12ng/μL Imipramine-D3) was added to each tube. The samples were vortexed and centrifuged. The supernatant was transferred to an autosampler vial and 8μL was injected into the UPLC-MS/MS. Utilizing a Waters Acquity UPLC HSS T3 1.8μm, 2.1×50mm column at 25ºC, the analytes were separated using a timed, linear gradient of acetonitrile and water, each having 0.1% formic acid added. The column is eluted into the LC-MS/MS to detect imipramine D3 at transition 284.25>89.10, haloperidol at 376.18>165.06, thiothixene at 444.27>139.24, fluphenazine at 438.27>171.11, and perphenazine at 404.19>143.07. Secondary transitions for each analyte are also monitored for imipramine D3 at 284.25>193.10, haloperidol at 376.18>122.97, thiothixene at 444.27>97.93, fluphenazine at 438.27>143.08, and perphenazine at 404.19>171.11. The run-time is 1.8min per injection with baseline resolved chromatographic separation.
The analytical measurement range was 0.2 to 12.0ng/mL for fluphenazine and perphenazine, and was 1 to 60.0ng/mL for haloperidol and thiothixene. Intra-assay and inter-assay imprecisions (CV) were less than 15% at two concentrations for each analyte.
By utilizing a LC-MS/MS method we combined two previously established analytical assays into one, yielding a 75% time-savings on set-up, and a significantly shortened analytical run-time. These changes reduced the turn-around time for analysis and eliminated interference issues resulting in fewer injections and increased column lifetime.
在美国，大多数常用的抗精神病药物都采用液相色谱和气相色谱方法进行监测。使用超高效液相色谱-串联质谱（LC-MS/MS）技术可以提高方法性能。本文介绍了一种快速简单的方法，用于监测氟哌啶醇、硫利达嗪、氟奋乃静和奋乃静的浓度。
采用 LCMS/MS（Waters Acquity UPLC TQD）测定血清和血浆中的抗精神病药物浓度。仪器采用电喷雾接口，多重反应监测（MRM）和正离子模式操作。两个四极杆的分辨率均保持在单位质量，半峰宽为 0.7amu。数据分析采用 Waters Quanlynx 软件进行。血清或血浆样品在室温下解冻，取 100μL 置于管中。然后向每个管中加入 300μL 沉淀试剂（乙腈-甲醇[50:50，体积：体积]），内含内标（0.12ng/μL 丙咪嗪-D3）。样品涡旋并离心。将上清液转移至自动进样瓶中，用 8μL 进样到 UPLC-MS/MS。采用 Waters Acquity UPLC HSS T3 1.8μm，2.1×50mm 柱，在 25°C 下，使用定时、线性梯度的乙腈和水，每种溶剂均添加 0.1%甲酸进行分离。将柱洗脱至 LC-MS/MS 中，以在过渡 284.25>89.10 处检测丙咪嗪 D3，在 376.18>165.06 处检测氟哌啶醇，在 444.27>139.24 处检测硫利达嗪，在 438.27>171.11 处检测氟奋乃静，在 404.19>143.07 处检测奋乃静。每个分析物的次要转换也在 284.25>193.10 处监测丙咪嗪 D3，在 376.18>122.97 处监测氟哌啶醇，在 444.27>97.93 处监测硫利达嗪，在 438.27>143.08 处监测氟奋乃静，在 404.19>171.11 处监测奋乃静。每个进样的运行时间为 1.8 分钟，具有基线分离的色谱分离。
氟奋乃静和奋乃静的分析测量范围为 0.2 至 12.0ng/mL，氟哌啶醇和硫利达嗪的分析测量范围为 1 至 60.0ng/mL。每个分析物的两个浓度的日内和日间精密度（CV）均小于 15%。
通过使用 LC-MS/MS 方法，我们将两种先前建立的分析方法结合在一起，在设置方面节省了 75%的时间，并显著缩短了分析运行时间。这些变化减少了分析的周转时间，并消除了干扰问题，从而减少了注射次数和增加了柱寿命。
