细菌内毒素国家标准品原料的制备及其理化性质和生物学活性的分析  

Preparation and Properties of Bacterial Endotoxin National Standard Material

作　　者：于婷[1] 蔡彤[1] 张国来[1] 高华[1] 李波[1] 

机构地区：[1]中国药品生物制品检定所,北京100050

出　　处：《中国药学杂志》2010年第5期388-391,共4页Chinese Pharmaceutical Journal

摘　　要：目的制备第7批细菌内毒素国家标准品的原料。方法常规方法培养和收获细菌,超声破碎菌液,热酚水法制备内毒素粗品,充分透析去酚后,浓缩,取上清,超速离心(4℃,105000×g,4h),最后冻干。采用该方法共制备得到3批内毒素原料候选品,对其进行理化分析和生物学活性测定后,确定其中一批作为内毒素国家标准品原料。同时与细菌内毒素国际标准品、中国细菌内毒素国家标准品(NS,Lot:981)和日本细菌内毒素国家标准品进行一系列比较。结果3批原料候选品中,3号内毒素原料的生物学活性最好,杂质含量最低。结论3批内毒素原料的生物学活性以及理化性质基本一致,表明采用的提取方法稳定、重现性好。通过生物学活性和杂质含量的比较,结果表明,3号内毒素原料可以作为制备新一批细菌内毒素国家标准品的原料。OBJECTIVE To prepare the 7th bacterial endotoxin national standard material.METHODS Escherichia coli O11B4 was cultured and harvested by routine method,bacterial liquid was broken by supersound,then crude endotoxin material was extracted by hot phenol-water.After sufficient dialysis to remove all traces of phenol,endotoxin was concentrated.The supernatant was chosen and purified by ultracentrifge at 105 000×g at 4 ℃ for 4 h.Finally the sediment was freeze-dried.Three batches of endotoxin material candidate were prepared by this method,and one of the batches was ascertained to be bacterial endotoxin national standard material by physical chemical and biological activity assay.A series of comparison were carried out between bacterial endotoxin international standard,national standard（the 6th batch）and Japanese national standard.RESULTS No.3 endotoxin material showed the best biological activity and lowest impurity content.CONCLUSION Three batches of endotoxin material almost have the same biological activity,physical and chemical character.The extract method has good stability and reproducibility.Compared by biological activity and impurity content,the No.3 endotoxin material can be used for preparing new batch of bacterical endotoxin national standard material.

关 键 词：内毒素 细菌内毒素国家标准品 热酚水法 

分 类 号：R917[医药卫生—药物分析学]