Unit of Pharmacogenetics and Clinical Psychopharmacology, Center for Psychiatric Neurosciences, Department of Psychiatry, Lausanne University Hospital, Hospital of Cery, 1008 Prilly-Lausanne, Switzerland.
J Chromatogr A. 2013 May 31;1292:160-72. doi: 10.1016/j.chroma.2012.12.071. Epub 2013 Jan 9.
A sensitive and selective ultra-high performance liquid chromatography (UHPLC) tandem mass spectrometry (MS/MS) method was developed for the fast quantification of ten psychotropic drugs and metabolites in human plasma for the needs of our laboratory (amisulpride, asenapine, desmethyl-mirtazapine, iloperidone, mirtazapine, norquetiapine, olanzapine, paliperidone, quetiapine and risperidone). Stable isotope-labeled internal standards were used for all analytes, to compensate for the global method variability, including extraction and ionization variations. Sample preparation was performed by generic protein precipitation with acetonitrile. Chromatographic separation was achieved in less than 3.0min on an Acquity UPLC BEH Shield RP18 column (2.1mm×50mm; 1.7μm), using a gradient elution of 10mM ammonium formate buffer pH 3.0 and acetonitrile at a flow rate of 0.4ml/min. The compounds were quantified on a tandem quadrupole mass spectrometer operating in positive electrospray ionization mode, using multiple reaction monitoring. The method was fully validated according to the latest recommendations of international guidelines. Eight point calibration curves were used to cover a large concentration range 0.5-200ng/ml for asenapine, desmethyl-mirtazapine, iloperidone, mirtazapine, olanzapine, paliperidone and risperidone, and 1-1500ng/ml for amisulpride, norquetiapine and quetiapine. Good quantitative performances were achieved in terms of trueness (93.1-111.2%), repeatability (1.3-8.6%) and intermediate precision (1.8-11.5%). Internal standard-normalized matrix effects ranged between 95 and 105%, with a variability never exceeding 6%. The accuracy profiles (total error) were included in the acceptance limits of ±30% for biological samples. This method is therefore suitable for both therapeutic drug monitoring and pharmacokinetic studies.
建立了一种灵敏、选择性的超高效液相色谱（UHPLC）串联质谱（MS/MS）方法，用于快速定量测定人血浆中的十种精神药物及其代谢物，以满足我们实验室的需求（氨磺必利、阿塞那平、去甲米氮平、依匹哌唑、米氮平、诺喹酮、奥氮平、帕利哌酮、喹硫平利培酮）。所有分析物均使用稳定同位素标记的内标，以补偿包括提取和离子化变化在内的全局方法变异性。样品制备采用通用的乙腈沉淀蛋白法。在 Acquity UPLC BEH Shield RP18 柱（2.1mm×50mm；1.7μm）上，采用 10mM 甲酸铵缓冲液 pH 3.0 和乙腈的梯度洗脱，流速为 0.4ml/min，在 3.0min 内完成色谱分离。采用正电喷雾电离模式的串联四极杆质谱仪进行化合物定量分析，采用多重反应监测。该方法根据国际指南的最新建议进行了全面验证。使用 8 个点的校准曲线，覆盖了较大的浓度范围，0.5-200ng/ml 用于阿塞那平、去甲米氮平、依匹哌唑、米氮平、奥氮平、帕利哌酮和利培酮，1-1500ng/ml 用于氨磺必利、诺喹酮和喹硫平。在准确度（93.1-111.2%）、重复性（1.3-8.6%）和中间精密度（1.8-11.5%）方面均取得了良好的定量性能。内标归一化基质效应在 95-105%之间，变化率从不超过 6%。准确度谱（总误差）包含在生物样品的±30%接受限内。因此，该方法适用于治疗药物监测和药代动力学研究。
