UPLC-QDa质谱检测器法检测米拉贝隆中硝基苯类带基因毒警示结构杂质的含量  

Determination of notrobenzenes impurities with alerting structures in mirabegron by UPLC-QDa mass spectrometry method

作　　者：秦永香 许红霞 谌宗永 郭鑫 刘智 孙跃军 王明娟 袁伟锋 孟旻 马昂 刘宇晶 QIN Yong-xiang;XU Hong-xia;SHEN Zong-yong;GUO Xin;LIU Zhi;SUN Yue-jun;WANG Ming-juan;YUAN Wei-feng;MENG Min;MA Ang;LIU Yu-jing(Beijing Sun-novo Pharmaceutical Research Co.,Ltd.,Beijing 102200,China)

机构地区：[1]北京阳光诺和药物研究股份有限公司,北京102200

出　　处：《药物分析杂志》2022年第10期1792-1800,共9页Chinese Journal of Pharmaceutical Analysis

基　　金：国家药品监督管理局化学药品杂质谱研究重点实验室开放课题(NMPA-KLIPCD-2020-06);北京市科委、中关村管委会专项课题(Z211100003421042)。

摘　　要：目的:建立可监控米拉贝隆原料中残留风险最大的4个硝基苯类带基因毒警示结构杂质(MLB-1、MLB-2、MLB-1-IM01、MLB-2-IM01)含量的方法。方法:采用UPLC-QDa质谱检测器系统,XBridge C_(18)柱(150 mm×4.6 mm,3.5μm),以乙腈-水-三氟乙酸(400∶600∶0.5)为流动相,流速0.8 mL·min^(-1),柱温35℃,进样体积2μL;QDa检测器的参数设置:电喷雾离子化-正电离模式(ESI~+),多反应监测模式(MRM),探头温度为600℃,采集速率10点·s^(-1),气流量20 L·min^(-1);采集时间15 min。结果:4个目标硝基苯类杂质在0.03~0.6μg·mL^(-1)浓度范围内(相当于主成分的0.0003%~0.006%水平;4个目标杂质限度为0.003%,总量限度为0.01%)线性关系均良好(r≥0.9973),回收率介于91.8%~119.1%,检测限和定量限分别低于0.1和0.25μg·mL^(-1)(相当于0.0025%水平),能较好地满足目标杂质的检测灵敏度要求。采用拟定工艺生产的多批米拉贝隆样品中均未检出上述4个目标杂质(总量<0.001%,远低于限度规定的0.01%)。结论:该方法灵敏度高,准确性好,在拟定限度范围内呈线性响应,可用于检测米拉贝隆原料中潜在硝基苯类基因毒杂质的残留量监控。Objective:To establish an original method for determination of four nitrobenzenes with alerting structure(MLB-1,MLB-2,MLB-1-IM01 and MLB-2-IM01)most likely residue in mirabegron drug substance.Methods:The Waters Acquity UPLC-QDa mass detector system was applied,with the XBridge C_(18)column(150 mm×4.6 mm,3.5μm)maintaining at 35℃.Mobile phase consisted of 0.05%trifluoroacetic acid in acetonitrile-water(400∶600,v/v)at a flow rate of 0.8 mL·min^(-1).The injection volumes were 2μL.The conditions of QDa detector were as follows:electrospray ionization mode positive(ESI+),multi-reaction monitoring(MRM)mode,probe temperature at 600℃,flow rate of nitrogen at 20 L·min^(-1),with the acquisition rate of 10 points·s^(-1)for 15 min.Results:Four nitrobenzenes all showed good linearity(r≥0.9973)in the range of 0.03-0.6μg·mL^(-1)(equivalent to 0.0003%-0.006%of mirabegron,meanwhile the single and total limit for the four nitrobenzenes was 0.003%and 0.01%,respectively).Their recoveries were between 91.8%-119.1%,and their limits of detection and quantitation were all below 0.1μg·mL^(-1)and 0.25μg·mL^(-1)(0.0025%),respectively,fulfilling the sensitivity requirements for detection of the target impurities.None of the four nitrobenzenes was detected in mirabegron produced with the proposed synthesis and purification process(total amount<0.001%,much lower than the limit of 0.01%).Conclusion:The original method established is sensitive,accurate and shows linear response in the range covering the proposed limit,demonstrating its suitability for trace control of nitrobenzenes in mirabegron drug substances.

关 键 词：超高效液相色谱 QDa检测器 米拉贝隆 硝基苯类 警示结构 基因毒杂质 风险评估 

分 类 号：R917[医药卫生—药物分析学]