摘要
目的检测生脉注射液不同制备工艺中间体含残留蛋白质含量。方法以残留蛋白质为检测目标,采用透析法获取生脉注射液不同制备工艺中间体中大分子杂质,Bradford法测定各大分子杂质中残留蛋白质含量,结合SDS-PAGE电泳分析各残留蛋白质分子质量。结果在0～8μg线性范围内,牛血清白蛋白标准品回归方程为:Y=0.0474X-0.003(r=0.9974);不同工艺对提取液中残留蛋白的影响较大,红参不同工艺提取液含残留蛋白浓度较高,分别为399、353μg/mL;麦冬、五味子提取液残留蛋白含量相对较低;经对SDS-PAGE电泳结果分析,各不同工艺提取液中间体含蛋白质分子质量主要集中于6～11万。结论生脉注射液中间体提取工艺工程中可能有蛋白质残留,应加强残留蛋白质检测及生脉注射液的工艺改进;Bradford法结合SDS-PAGE电泳分析生脉注射液中的残留蛋白质,快速、准确、敏感性及检测结果均较好,可为工业生产提供科学数据参考。
Objective To detect the content of residual protein in the intermediates of Shengmai injection.Method Large molecular impurities in the intermediates of Shengmai injection were obtained by dialysis method;the content of the residual protein was determined by Bradford assay;and the molecular weight of the residual protein was analyzed by SDS-PAGE electrophoresis.Results The method was proved to be linear in the range of 0～8μg with the linear equation was：Y = 0.047 4X-0.003 （r = 0.9974）.The concentration of residual protein was influenced by different technology.The content of residual protein in Radix Ginseng Rubra by different extraction technology was higher than that in Fructus Schisandrae and Raidix Ophiopogonis intermediates.The residual protein＇s molecular weight was concentrated in the 6～11 million by analysis of SDS-PAGE electrophoresis.Conclusion Shengmai injection＇ intermediates of different extraction technology may be contain residual protein,determination of residual protein should be strengthened and the extraction technology need to be improved.The proposed Bradford unifies SDS-PAGE electrophoresis method was rapid,sensitive and accurate,and can be used for improving the manufacture process and for routine quality control of drug products.
出处
《中国医药指南》
2010年第19期57-59,共3页
Guide of China Medicine
