基于UPLC-MS/MS技术同时测定异环磷酰胺原料药中主成分及有关物质  

Simultaneous determination of principal components and related substances in isocyclophosphamide APIs based on UPLC-MS/MS technique

作　　者：张燕 闫占宽 王辰 曹蕾 李辉辉 刘钦伟 杨春 戚利强 王洪森 ZHANG Yan;YAN Zhankuan;WANG Chen;CAO Lei;LI Huihui;LIU Qinwei;YANG Chun;QI Liqiang;WANG Hongsen(Jiangsu Hengrui Pharmaceutical Co.,Ltd.,Lianyungang 222000,China;Jiangsu Original Drug R&D Co.,Ltd.,Lianyungang 222000,China;School of Pharmacy,Jiangsu Ocean University,Lianyungang 222005,China)

机构地区：[1]江苏恒瑞医药股份有限公司,江苏连云港222000 [2]江苏原创药物研发有限公司,江苏连云港222000 [3]江苏海洋大学药学院,江苏连云港222005

出　　处：《药物评价研究》2023年第12期2535-2539,共5页Drug Evaluation Research

基　　金：江苏省产教融合一流课程《药物分析》建设项目;江苏海洋大学专业学位研究生课程案例库建设项目(YJSZY202102);江苏海洋大学2023年一流课程立项;连云港市政策引导产学研项目(CYX2202)。

摘　　要：目的采用超高效液相色谱质谱联用(UPLC-MS/MS)法同时测定异环磷酰胺原料药中主成分及2种有关物质。方法采用ACQUITY UPLC?HSS T3(100 mm×2.1 mm,1.8μm)色谱柱,以甲醇(A)-5 mmol·L^(-1)碳酸氢铵溶液(B)为流动相进行梯度洗脱,对异环磷酰胺原料药中的异环磷酰胺、杂质G、杂质E进行分离,体积流量为0.3 mL·min^(-1),进样量为10μL,柱温为30℃,质子离子源ESI,正离子模式下以多反应监测方式(MRM)进行定量分析。结果异环磷酰胺、杂质E、杂质G的线性范围分别1.00~50.00、1.00~50.00、0.01~0.5 ng·mL^(-1),线性关系良好,r均大于0.9994;且该方法的仪器精密度、重复性良好,异环磷酰胺的回收率为98.1%~101.7%、RSD为0.40%~1.73%,杂质E的回收率为96.1%~111.1%、RSD为0.67%~2.34%,杂质G的回收率为93.7%~108.3%、RSD为2.2%~7.6%,符合标准。原料药中的异环磷酰胺质量分数为97.1~98.5%,杂质E均未检出,杂质G在1501批次检出量最高,为0.00925%,杂质E和G未超千分之一限度。结论建立的UPLC-MS/MS法灵敏度较高、重复性好,为异环磷酰胺原料药的质量控制提供了一种可行的分析方法,适用于该产品的质量控制。Objective To use liquid chromatography-mass spectrometry(UPLC-MS/MS)to simultaneously determine the principal components and two related substances in ifosfamide API.Methods The ACQUITY UPLC®HSS T3(100 mm×2.1 mm,1.8µm)column was used to perform gradient elution of methanol(A)-5 mmol·L−1 ammonium bicarbonate(B)as the mobile phase,and ifosfamide,impurity G and impurity E in ifosfamide API were separated with a flow rate of 0.3 mL·min^(−1),an injection volume of 10μL,a column temperature of 30℃,and a proton ion source ESI.Quantification in positive ion mode in multi-reaction monitoring mode(MRM).Results The linear ranges of ifosfamide,impurity E and impurity G were 1.00—50.00、1.00—50.00、0.01—0.5 ng·mL^(−1)respectively,with good linear relationships and correlation coefficients greater than 0.994,and the instrument precision and repeatability of the method were good.The recoveries of isocyclophosphamide were 98.1%—101.7%with RSD of 0.40%—1.73%.The recovery of impurity E was 96.1%—111.1%with RSD of 0.67%—2.34%.The recoveries of impurity G were 93.7%—108.3%with RSD of 2.2%—7.6%.The mass fraction of ifosfamide in the raw material was 97.1%—98.5%,and impurity E had not been detected.The detection amount of impurity G in batch 1501 is 0.00925%,and impurities E and G do not exceed the limit of one thousandth.Conclusion The proposed method has high sensitivity and good repeatability,which provides a feasible analysis method for the quality control of ifosfamide API,which is suitable for the quality control of this product.

关 键 词：异环磷酰胺 杂质E 杂质G 有关物质 UPLC-MS/MS 原料药 质量控制 

分 类 号：R932[医药卫生—生药学]