胸腺肽高分子量物质检测方法的优化

摘要

目的 优化现行胸腺肽质量标准中控制高分子量(Mw)物质的检测方法,解决该方法存在的对照品Mw大小与洗脱时间矛盾的问题.方法 采用高效液相凝胶过滤色谱法(HPGFC),改变流动相的组成,建立高Mw物质的分析方法,并进行方法学验证.结果 确定三氟乙酸-乙腈-水(0.05∶45∶55)作为流动相的HPGFC分析方法,在该洗脱条件下,对照品的洗脱顺序符合Mw变化的规律,经过方法学验证,改进后的方法更适于检测高Mw物质的含量.结论 优化后的高Mw物质检测方法可用于胸腺肽制剂的质量控制,并为其他生化制剂高Mw蛋白检测提供了方法学参考.%Purpose To optimize the detection methods for high molecular weight component in the current quality control of thymosin and to solve the contradiction between the molecular weight of standard preparation and elution time. Methods High performance gel filtration chromatography ( HPGFC ) was used to analyze the high molecular weight component. The mobile phase was altered to find out the most suitable one. Also the methodology verification was done. Results The analysis of HPGFC was carried out with the mobile phase of trifluoacetic acid-acetonitrile-water(0. 05:45:55 ). Under the condition, the elu-tion order of control article coincided with the variation of molecular weight. After verification of accuracy, repetitiveness and stability, it was found that the improved method was more suitable for detecting the a-mount of high molecular weight component. Conclusion The optimized detecting method of high molecular weight component can be used in the quality control of thymosin preparation and it provides a methodological reference for the detection of other biochemical high molecular weight proteins.