UPLC-MS/MS法检测雷迪帕韦原料药中潜在基因毒性杂质残留量

摘要

Objective: To establish a UPLC-MS/MS method for the determination of the residual quantity of potential genetoxic impurity 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide (EDC) in ledipasvir.Methods: The chromatographic conditions were as follows: an Agilent ZORBAX SB-C18 (75 mm×4.6 mm, 3.5 μm) column was used with methanol-0.1% formic acid (50∶50) as the mobile phase.The flow rate was 0.3 ml·min-1.The column temperature was 30 ℃ and the injection volume was 5 μl.The mass spectrometry conditions were as follows: electrospray ionization source (ESI), multistage reaction monitoring (MRM), positive ion scanning mode, ion spray voltage of 2 500 V, ion source temperature of 500 ℃, atomization gas of 379.0 kPa, auxiliary gas of 275.6 kPa, curtain gas of 137.8 kPa, collision gas of 41.3 kPa, ion collision energy of 15 V and scan time of 100 ms.The MRM ion pair for quantitative analysis was m/z→156.2/86.1.Results: The linear range of EDC was 0.03-2.25 μg·ml-1 (r=0.999 0).The limit of detection was 0.03 ng and the limit of quantitation was 0.08 ng.The average recovery was 98.3%(RSD=5.7%, n=9).Conclusion: The method is simple, rapid and accurate, and can be used for the quality control of EDC in ledipasvir.%目的: 建立超高效液相色谱-质谱联用法(UPLC-MS/MS)检测雷迪帕韦原料药中潜在基因毒性杂质1-(3-二甲氨基丙基)-3-乙基碳二亚胺(EDC)的残留量.方法: 色谱条件:采用Agilent ZORBAX SB-C18色谱柱(75 mm×4.6 mm,3.5 μm),流动相为甲醇-0.1%甲酸溶液(50∶50),流速为0.3 ml·min-1,柱温为30 ℃,进样量为5 μl.质谱条件:采用电喷雾离子源(ESI),多级反应监测(MRM),正离子扫描模式,离子喷雾电压为2 500 V,离子源温度为500 ℃,雾化气为379.0 kPa,辅助气为275.6 kPa,气帘气为137.8 kPa,碰撞气为41.3 kPa,离子碰撞能为15 V,扫描时间为100 ms,用于定量分析的MRM离子对为m/z→156.2/86.1.结果: EDC的线性范围为0.03~2.25 μg·ml-1(r=0.999 0);检测限为0.03 ng,定量限为0.08 ng;平均回收率为98.3%(RSD=5.7%,n=9).结论: 本方法操作简便、快捷、结果准确,可用于雷迪帕韦原料药中潜在基因毒性杂质EDC的质量控制.