重组人生长激素原液及注射液中有关物质UPLC分析方法建立  

Establishment of a UPLC method for the determination of related substances in recombinant human growth hormone bulk and injection

作　　者：陆俊杰 李晶[1] 陈莹 李懿 张伟[1] 张慧[1] 吕萍[1] 秦希月 高向东[2] 梁成罡[1] LU Jun-jie;LI Jing;CHEN Ying;LI Yi;ZHANG Wei;ZHANG Hui;Lü Ping;QIN Xi-yue;GAO Xiang-dong;LIANG Cheng-gang(Division of Hormone,National Institutes for Food and Drug Control,NHK Key Laboratory of Research on Quality and Standardization of Biotech Products,Beijing 102629,China;China Pharmaceutical University College of Life Science and Technology,Nanjing 210009,China;Jilin University College of Life Science,Changchun 130012,China)

机构地区：[1]中国食品药品检定研究院激素室国家卫生健康委员会生物技术产品检定方法及其标准化重点实验室,北京102629 [2]中国药科大学生命科学与技术学院,南京210009 [3]吉林大学生命科学学院,长春130012

出　　处：《药物分析杂志》2022年第1期41-50,共10页Chinese Journal of Pharmaceutical Analysis

基　　金：国家科技重大专项课题资助项目(2018ZX09101001-003-004);国家药典委员会药品标准提高课题,重组人生长激素注射液质量标准提高研究(2018S013)。

摘　　要：目的:建立超高效液相色谱法测定重组人生长激素原液及注射液中的有关物质。方法:采用Waters BEH C;300?色谱柱(150 mm×2.1 mm,1.7μm),以12.5 mmol·L^(-1)磷酸氢二钾缓冲溶液-乙腈为流动相,梯度洗脱,流速0.3 mL·min^(-1),检测波长220 nm,柱温45℃。结果:在选定的色谱条件下,能分离出较多有关物质,且主峰与各有关物质峰均能良好分离;质量浓度在0.25~5 mg·mL^(-1)的范围内线性关系良好。精密度、耐用性及稳定性(72 h内)良好。根据质谱定性结果,探索了制备脱酰胺杂质、氧化杂质、脱水杂质等主要有关物质的处理方法。对于重组人生长激素注射液,本方法与现有标准相比,能够弥补我国注册标准方法中检测有关物质数量少,彼此间分离度不高的缺点,同时也克服了欧洲药典需要2种机理不同的独立实验方能完成有关物质控制的问题。结论:本方法适用于测定重组人生长激素原液及注射液中的有关物质。Objective:To establish a UPLC method for determination of related substances in recombinant human growth hormone bulk and injection.Methods:The analysis was conducted on Waters BEH C;300?column(150 mm×2.1 mm,1.7μm).The mobile phases were 12.5 mmol·L^(-1)dipotassium hydrogen phosphate buffer solution and acetonitrile with gradient elution,the flow rate was 0.3 mL·min^(-1),the column temperature was 45℃,and the detection wavelength was 220 nm.Results:Under the selected chromatographic conditions,more related substances could be separated,and the main peak and each peaks of related substances could be well separated.The linear relationship was good in the concentration range of 0.25-5 mg·mL^(-1).Precision,and durability was good.Stability within 72 h was good.According to the qualitative results of mass spectrometry.The method was explored that preparing the main related substances such as deamination impurities,oxidation impurities and dehydration impurities.For recombinant human growth hormone injection,compared with the existing standard,this method could compensate for the shortcomings of the registration standard such as the small number of substances and the low degree of separation between each other.and overcome the problem thatthe European Pharmacopoeia requires two independent experiments with different mechanisms to complete the control of related substances.Conclusion:This method is suitable for the determination of related substances in recombinant human growth hormone bulk and injection.

关 键 词：重组人生长激素原液 重组人生长激素注射液 有关物质 超高效液相色谱法 脱酰胺杂质 氧化杂质 脱水杂质 

分 类 号：R917[医药卫生—药物分析学]