Abstract: Objective To establish an analytical method for the simultaneous determination of 12 antidepressants including moclobemide and bupropion in human blood by ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS). Methods Blood samples were purified through protein precipitation with acetonitrile that contained internal standard (proadifen), then the samples were injected into the UHPLC-MS/MS. The separation was performed on an ACQUITY UPLC HSS T₃ column (2.1 mm×100 mm, 1.8 µm) using gradient elution with a flow rate of 0.35 mL/min. The mobile phase consisted of water solution and acetonitrile solution that both contained 2 mmol/mL ammonium formate and 0.1% formic acid. The analytes were detected using multiple reaction monitoring mode. The electrospray ionization source was performed in positive mode.The internal standard method was used for quantification. Results The correlation coefficients of the 12 antidepressants were better than 0.999 in the corresponding concentration ranges, and the limits of detection ranged from 0.05 ng/mL to 0.2 ng/mL, the limits of quantification ranged from 0.5 ng/mL to 1 ng/mL. At different spiked concentrations (low, medium and high), the accuracy ranged from -7.1% to 11.7%, the intra-day precision ranged from 0.1% to 11.5%, the inter-day precision ranged from 1.9% to 9.4%, the extraction recoveries ranged from 85.7% to 113.5%, and the matrix effect ranged from -18.2% to 10.2%. The stability of each component at different levels of concentration and under different environment conditions ranged from -9.30% to 11.30%. All of the above results meet the requirements for biological sample analysis and forensic toxicology analysis. Conclusion The method is selective, reproducible and precise. The simultaneous determination of 12 antidepressants in human blood can be applied in forensic toxicology practice and it will be of great use in the analysis of the cause of death in unnatural death cases.

Key words: forensic toxicology, ultra high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS), antidepressant, human blood

摘要： 目的 建立超高效液相色谱-串联质谱法同时测定人血中吗氯贝胺、安非他酮等12种抗抑郁药的分析方法。方法 血液样品经乙腈（含内标普罗地芬）沉淀蛋白后进样测定。采用ACQUITY UPLC HSS T₃色谱性（2.1 mm×100 mm，1.8 μm）进行分离，以2 mmol/mL甲酸铵和0.1％甲酸的水溶液-2 mmol/mL甲酸铵和0.1％甲酸的乙腈溶液体系进行梯度洗脱，流速0.35 mL/min。以电喷雾离子源正离子和多反应监测模式进行检测，内标法定量。结果 在相应质量浓度范围内，12种抗抑郁药的相关系数均大于0.999，检出限为0.05~0.2 ng/mL，定量限为0.5~1 ng/mL；在低、中、高不同添加浓度条件下，准确度为-7.1%~11.7%，日内精密度为0.1%~11.5%，日间精密度为1.9%~9.4%，提取回收率为85.7%~113.5%，基质效应为-18.2%~10.2%；各成分在不同添加水平和不同环境下的稳定性为-9.30%~11.30%。综上均符合生物样品分析及法医毒物分析要求。结论 该方法选择性强、重现性好、精密度高，可同时检测人血中12种抗抑郁药，可适用于法医毒物检验鉴定，对非正常死亡案（事）件中死者死因分析具有重要意义。

关键词: 法医毒物学, 超高效液相色谱-串联质谱法, 抗抑郁药, 人血