达卡巴嗪原料药中N-亚硝基二甲胺和N-亚硝基二乙胺的UHPLC-APCI-MS/MS方法的建立

目的:采用UHPLC-APCI-MS/MS法测定达卡巴嗪原料药中的N-亚硝基二甲胺(NDMA)和N-亚硝基二乙胺(NDEA),建立控制该药品遗传毒性杂质的方法。方法:采用Kinetex F5色谱柱(3 mm×100 mm,2.6μm),流动相为0.1%甲酸溶液-甲醇,梯度洗脱,流速0.4 mL·min^-1,柱温40℃,质谱离子化方式为APCI,正离子模式,多重反应监测,检测离子对为m/z 75.1/58.0(NDMA),103.0/47.0(NDEA)。结果:NDMA和NDEA在一定范围内线性关系良好(r>0.990),检出限溶液浓度分别为0.03030、0.003787 ng·mL(-1),精密度、稳定性试验满足检测要求,回收率为97~115%,RSD均小于5%。结论:本文建立的方法准确、快速灵敏、专属性强,可用于控制达卡巴嗪原料药中遗传毒性杂质NDMA和NDEA的含量。

Establishment of standard method of NDMA and NDEA in Dacarbazine by UHPLC-APCI-MS/MS

Objective:To establish a method for determination of the genotoxic impurities of NDMA and NDEA in the active pharmaceutical ingredient of dacarbazine by UHPLC-APCI-MS/MS.Methods:The test was performed in Kinetex F5 column(3 mm×100 mm,2.6μm)under the gradient elution of 0.1%formic acid solution(mobile phase A)and methanol(mobile phase B).The flow rate was 0.4 mL·min-1,and the column temperature was 40℃.The ionization mode of mass spectrometry was APCI with positive ion mode,and determination was conducted in MRM scan mode by the external standard method.The ion pairs were m/z 75.1/58.0 for NDMA and m/z 103.0/47.0 for NDEA.Results:The established method for the separation and analysis of NDMA and NDEA was validated.The linear relationship in the linear range was good(r>0.990).The limits of detection were 0.03030 and 0.003787 ng·mL^-1,respectively.The precision and stability results met the criteria,the recovery rate was 97%-115%,and all the relative standard deviations were less than 5%.Conclusion:The established method is accurate,rapid,sensitive and specific.It can be used for the quality control of the genotoxic impurities of NDMA and NDEA in API of Dacarbazine.