LC-MS/MS法测定富马酸替诺福韦酯中杂质9-丙烯基腺嘌呤的含量

Quantification of 9-propenyladenine in tenofovir disoproxil fumarate by LC-MS/MS

目的建立测定富马酸替诺福韦酯中杂质9-丙烯基腺嘌呤含量的LC-MS/MS法。方法采用Thermo BDS(100 mm×4.6 mm,2.4μm)色谱柱,以甲醇-水(体积比为60∶40)为流动相,流速为0.3 m L·min^(-1)。选用ESI源以多反应监测(MRM)方式进行正离子检测,用于定量分析的离子反应为m/z 176.2→m/z 136.3。结果 9-丙烯基腺嘌呤在0.20~20μg·L^(-1)内线性关系良好,定量限为0.20μg·L^(-1),日内、日间精密度(RSD)均小于11.4%,准确度(RE)在3.7%以内,平均加样回收率为103%(n=6)。结论应用此方法测定了3批原料药中杂质9-丙烯基腺嘌呤的含量,结果均满足世界卫生组织(WHO)对其限量(不超过5 mg·L^(-1))检查的要求。

Objective To develop an LC-MS/MS method for the quantification of 9-propenyladenine（ impurity K） in tenofovir disoproxil fumarate pharmaceutical product. Methods Isocratic chromatographic separation was performed on a Thermo BDS C18 column（ 100 mm×4.6 mm,2.4 m）. The mobile phase consisted of methanol-water（ V∶V = 60∶40） at a flowrate of 0.3 m L·min^（-1）. ESI source was applied and operated in the positive ion mode. M RM mode with the transition of m/z 176.2 m/z→136.3 was used to quantify the impurity K. Results The method was liner in the concentration range from 0.20 to 20 μg·L^（-1）. The limit of quantification was 0.20 μg·L^（-1）. The intra-and inter-day precision values were both below11.4%,and accuracy was within 3.7% in all quality control samples. The average recovery was 103%（ n=6）. Conclusions The method provides a sensitive and specific means for the determination of impurity K in tenofovir disoproxil fumarate pharmaceutical substances,which meets the requirement of that in WHO（ a limit of not more than 5 mg·L^（-1） for the mutagenic impurity 9-propenyladenine,impurity K）.