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

作　　者：易艳[1] 刘婷[1] 郝然[1] 郭静[1] 李春英[1] 曹春雨[1] 赵雍[1] 高双荣[1] 回连强[1] 梁爱华[1] 

机构地区：[1]中国中医科学院中药研究所,北京100700

出　　处：《中国中药杂志》2010年第15期1965-1969,共5页China Journal of Chinese Materia Medica

基　　金：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)

摘　　要：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：内毒素检测 细胞因子诱导法 注射液 

分 类 号：R927.2[医药卫生—药学]