UHPLC-MS/MS法测定瑞戈非尼中两种痕量基因毒性杂质

Determination of two trace genotoxic impurities in Regorafenib by UHPLC-MS/MS

目的建立超高效液相色谱-串联质谱(UHPLC-MS/MS)方法测定瑞戈非尼中两类基因毒性杂质[3-氟-4-氨基苯酚(SM2)、3-三氟甲基-4-氯异氰酸苯酯(SM3)]的含量。方法采用色谱柱Agilent RRHD C 18(2.1 mm×100 mm,1.8μm),流动相为0.05%甲酸水-甲醇,梯度洗脱,流速为0.3 mL·min^(-1),柱温为35℃;采用Agilent 1290-6470A液质联用仪和电喷雾离子源(AJS ESI)源检测,正负离子模式采集数据。结果本方法的专属性、线性与范围、定量限与检测限、准确度、精密度及稳定性均符合《中国药典》的验证指标。结论本方法操作简便,结果可靠,用于两批瑞戈非尼中潜在毒性杂质(3-氟-4-氨基苯酚、3-三氟甲基-4-氯异氰酸苯酯)的测定。两批样品中均未检测到SM2,而基因毒性杂质SM3的含量分别为13.6×10^(-6)和41.4×10^(-6)。

Objective To establish an UHPLC-MS/MS analytical method for the determination of genotoxic impurities in Regorafenib.Methods The method was achieved on a Agilent RRHD C 18 column(2.1 mm×100 mm,1.8μm)utilizing a mobile phase of 0.05%Formic acid water(A)-Methanol(B)with gradient elution at the flow rate of 0.3 mL·min 0x0E䥺Symbolm@@0x0F 1.The temperature of column was set at 35℃.The Agilent 1290-6470A LC-MS was used to detect(AJS ESI source,in Multiple Reaction Monitoring mode).Results The specificity,linearity and range,LOQ and LOD,accuracy,precision and stability of the method were all in accordance with the validation of Chinese Pharmacopoeia.Conclusion The proposed method was successfully applied to determine the trace-level PGIs in two batches of regorafenib.The contents of SM2 were not observed in the two batches of samples,whereas the contents of SM3 were determined to be 13.6×10^(-6) and 41.4×10^(-6),respectively.