Detection of bacterial endotoxin content in eight kinds of injection by cytokine revulsion

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.