LC-MS-MS法同时测定人血浆中阿戈美拉汀及其2种代谢产物的浓度

目的建立同时测定人血浆中阿戈美拉汀及其代谢产物7-去甲阿戈美拉汀和3-羟基-阿戈美拉汀浓度的方法。方法采用LC-MS/MS法(液相色谱串联质谱)进行检测,以非那西汀为内标、Phenomenex ODS-3为色谱柱,A(0.2%甲酸铵+100%水)-B(甲醇)=30∶70为流动相进行色谱分离。采用ESI(电喷雾电离源)和MRM(多反应监测)的方式进行正电荷检测,用作定量分析的阿戈美拉汀、7-去甲-阿戈美拉汀、3-羟基-阿戈美拉汀和非那西汀的离子反应对m/z分别为244.1→185.1、230.1→171.1、260.1→201.1、180.1→110.1。结果阿戈美拉汀、7-去甲-阿戈美拉汀、3-羟基-阿戈美拉汀分别在0.045 7~100 ng/m L、0.137 2~300 ng/m L、0.457 2~1000 ng/m L范围内线性关系良好;最低检测限分别为0.045 7、0.137 2、0.457 2 ng/m L;所有待测样本的日内及日间精密度和准确度均符合相关生物样品分析的要求,且RSD<10%。在稳定性试验中,血浆样品在室温放置4 h、在4℃放置24 h、经历3次冷冻-解冻循环及在-80℃中放置3个月等条件下均可保持稳定。结论该方法具有操作简便、检测快速及灵敏性好等特点,能够用于临床相关药动学研究。

Simultaneous determination of agomelatine and two of its metabolites in human plasma by LC-MS/MS

Objective To establish a method for the simultaneous determination of agomelatine,7-desmethyl-agomelatine and 3-hydroxy-agomelatine in human plasma.Methods The LC-MS/MS method was used with phenacetin as internal standard,and the determination was performed on Phenomenex ODS-3 column with 0.2% ammonium formate+100% water-methanol (30∶70) as the mobile phase.Agomelatine (244.1→185.1),7-desmethyl-agomelatine (260.1→201.1) and 3-hydroxy-agomelatine (180.1→110.1) were detected by electrospray ionization (ESI) with positive ion in MRM mode quantitatively.Results The linear range was good within 0.045 7~100 ng/mL of agomelatine,0.137 2~300 ng/mL of 7-desmethyl-agomelatine and 0.457 2~1 000 ng/mL of 3-hydroxy-agomelatine,respectively,and the lowest limits of quantitation were 0.045 7,0.137 2 and 0.457 2 ng/mL,respectively.The intra-day and inter-day precision and accuracy were all in line with the requirements of biological sample analysis.The plasma samples were stable after 3 freeze-thaw cycles and being stored for 4 h at room temperature and 90 d at-80 ℃.Conclusion The method is rapid,sensitive and repeatable,which can be used for the determination of plasma concentration and pharmacokinetic study.