The Affiliated Brain Hospital of Guangzhou Medical University (Guangzhou Huiai Hospital), 36 Mingxin Road, Guangzhou 510370, China.
The Affiliated Brain Hospital of Guangzhou Medical University (Guangzhou Huiai Hospital), 36 Mingxin Road, Guangzhou 510370, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2018 Aug 15;1092:506-514. doi: 10.1016/j.jchromb.2018.05.026. Epub 2018 May 18.
Adjunctive therapy with olanzapine and fluoxetine has been shown to be beneficial in treatment-resistant depression and the depressive phase of bipolar disorder. Consensus guidelines issued by the Arbeitsgemeinschaft für Neuropsychopharmakologie und Pharmakopsychiatrie strongly recommend that patients taking olanzapine undergo therapeutic drug monitoring (TDM), and suggest that TDM is useful for patients taking fluoxetine. The aim of the current study was to develop and validate a sensitive, practical, and robust liquid chromatography-tandem mass spectrometry method (LC-MS/MS) for simultaneous determination of olanzapine, fluoxetine, and norfluoxetine in human plasma for routine TDM. Simple liquid-liquid extraction using ethyl acetate was used to extract olanzapine, fluoxetine, and norfluoxetine from 200 μL of pre-basified human plasma. Analytes were separated on an Agilent Eclipse Plus C column (4.6 × 100 mm, 5 μm) eluted with a mobile phase consisting of methanol:20 mM ammonium formate buffer (82.5:17.5, v/v), and then quantified using an electrospray ionization source operated in positive ion multiple reaction monitoring mode. The linear range for the analytes was 0.2-25 ng/mL, covering the vast majority of levels encountered in real-life samples. A weighting factor of 1/x best fit the calibration curves. The mean internal standard-normalized matrix effects for all analytes were 99.5%-110%. The extraction recoveries were 75%-85% for olanzapine and olanzapine‑d, and 58%-69% for fluoxetine, norfluoxetine, and their deuterated internal standards. Accuracy and precision values also met the acceptance criteria. The stability assessments showed that QC samples containing the three analytes were stable for at least 1 d at room temperature, 21 d at -70 °C, and through three freeze-thaw cycles. Post-preparation storage for 2 d in the autosampler did not cause obvious degradation of the investigated compounds. This validated high performance LC-MS/MS method was successfully applied to a pharmacokinetic study in healthy male volunteers.
奥氮平联合氟西汀辅助治疗已被证明对治疗抵抗性抑郁症和双相情感障碍的抑郁期有效。德国神经精神药理学和精神药理学工作组发布的共识指南强烈建议服用奥氮平的患者进行治疗药物监测（TDM），并建议 TDM 对服用氟西汀的患者有用。本研究旨在开发和验证一种灵敏、实用和强大的液相色谱-串联质谱法（LC-MS/MS），用于常规 TDM 同时测定人血浆中的奥氮平、氟西汀和去甲氟西汀。使用乙酸乙酯进行简单的液-液萃取，从 200µL 预碱化的人血浆中提取奥氮平、氟西汀和去甲氟西汀。分析物在 Agilent Eclipse Plus C 柱（4.6×100mm，5μm）上分离，流动相由甲醇：20mM 甲酸铵缓冲液（82.5:17.5，v/v）组成，然后使用电喷雾电离源在正离子多反应监测模式下进行定量。分析物的线性范围为 0.2-25ng/mL，涵盖了实际样本中遇到的绝大多数水平。校准曲线的最佳权重因子为 1/x。所有分析物的内标归一化基质效应均值为 99.5%-110%。奥氮平和奥氮平-d 的提取回收率为 75%-85%，氟西汀、去甲氟西汀及其氘代内标为 58%-69%。准确度和精密度值也符合验收标准。稳定性评估表明，含三种分析物的 QC 样品在室温下至少稳定 1d、-70°C 下至少稳定 21d、通过三次冻融循环稳定。在自动进样器中储存 2d 后，未引起所研究化合物的明显降解。该验证后的高效液相色谱-串联质谱法成功应用于健康男性志愿者的药代动力学研究。
