中国中药杂志

2010, v.35(15) 1965-1969

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细胞因子诱导法检测8种注射液内毒素含量
Detection of bacterial endotoxin content in eight kinds of injection by cytokine revulsion

易艳;刘婷;郝然;郭静;李春英;曹春雨;赵雍;高双荣;回连强;梁爱华;
YI Yan,LIU Ting,HAO Ran,GUO Jing,LI Chunying,CAO Chunyu,ZHAO Yong,GAO Shuangrong,HUI Lianqiang,LIANG Aihua(Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China)

摘要(Abstract):

目的:以巨噬细胞系RAW264.7细胞为工具,研究内毒素诱导RAW264.7细胞产生TNF-α的量效关系,对8种注射液的内毒素含量进行检测,探讨该方法用于注射液内毒素检测的可行性及干扰因素。方法:以不同浓度的内毒素诱导RAW264.7细胞产生TNF-α,绘制量效关系曲线,通过对鱼腥草注射液、双黄连粉针剂、双黄连注射液、清开灵注射液、葛根素注射液、香丹注射液、纤溶酶注射液、降纤酶注射液TNF-α含量的检测,计算其内毒素含量。结果:较低量的内毒素就能引起细胞产生TNF-α,且有良好剂量依赖关系,除07081613,810303A2个批号的清开灵注射液内毒素检测值较大外,其余7种注射剂在最大有效稀释倍数(MVD)范围内的内毒素检测结果都小于1.0EU.mL-1。结论:该方法的检测范围广,灵敏度较高,操作简便,且检测到的内毒素具有生物活性,为注射剂的热原检查方法提供了另一技术手段。
Objective:By using RAW 264.7 macrophage cell line,we studied the dose-effect relationship of endotoxin induced RAW 264.7 cells to release TNF-α,and then detected the content of endotoxin in 8 kinds of injections,so that we can investigate the feasibility and the interference factors of the novel test.Method:By using endotoxin of different concentrations to induce RAW 264.7 cells to release TNF-α,we drew the curve of dose-effect relationship between endotoxin and generated TNF-α.Then we detected the content of TNF-α in Yuxingcao,Shuanghuanglian,Qingkailing,Gegensu,Xiangdan,Qianrongmei and Jiangxianmei injections and Shuanghuanglian powder injection,and calculated their content of endotoxin.Result:The endotoxin could induce the cells to release TNF-α in a good dose-dependent manner,even at a very low concentration.In the range of maximum available dilution multiple,the content of endotoxin in the rest 7 kinds of injections was less than 1.0 EU·mL-1 except Qingkailing injection of two batch.Conclusion:Cytokine revulsion has the advantage of wide detection range,high sensitivity,simple operation,and the detected endotoxin is of bioactivity.This method provides another technical mean for pyrogen test of injections.

关键词(KeyWords): 内毒素检测;细胞因子诱导法;注射液
detection of bacterial endotoxin;cytokine revulsion;injections

Abstract:

Keywords:

基金项目(Foundation): 国家自然科学基金项目(90709043);; 科技部国家科技攻关计划项目(2006BAI14B05);; 国家“重大新药创制”科技重大专项(2009ZX09301-005);; 中国中医科学院自选课题团队建设(2008年度)中药注射剂致敏性检测关键技术研究(2009ZX09502-001)

作者(Authors): 易艳;刘婷;郝然;郭静;李春英;曹春雨;赵雍;高双荣;回连强;梁爱华;
YI Yan,LIU Ting,HAO Ran,GUO Jing,LI Chunying,CAO Chunyu,ZHAO Yong,GAO Shuangrong,HUI Lianqiang,LIANG Aihua(Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China)

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