吸附无细胞百白破联合疫苗成品内毒素含量检测  

Determination of endotoxin content in final product of diphtheria, tetanus and acellular pertussis combined vaccine, adsorbed

作　　者：卫辰[1] 裴宇盛[2] 晁哲[1] 骆鹏[1] 谭亚军[1] 王丽婵[1] 马霄[1] 侯启明[1] 

机构地区：[1]中国食品药品检定研究院百白破疫苗与毒素室卫生部生物技术产品检定方法及其标准化重点实验室,北京100050 [2]中国食品药品检定研究院药理室,北京100050

出　　处：《中国生物制品学杂志》2015年第4期385-389,共5页Chinese Journal of Biologicals

基　　金：国家863计划"疫苗效果和质量评价新技术研究"(2012AA02A402)

摘　　要：目的 对氢氧化铝佐剂吸附的无细胞百白破联合疫苗（diphtheria,tetanus and acellular pertussis combined vaccine,adsorbed,DTaP）成品中内毒素含量进行测定,确定将内毒素检查方法纳入2015版《中国药典》中百白破类疫苗质量标准的可行性和必要性。方法 参考《中国药典》三部（2010版）附录狱E建立动态浊度法,测定解吸附前后DTaP中细菌内毒素含量,并进行标准曲线的可靠性及干扰试验验证。结果 该方法在0.005-10 EU/ml范围内线性较好,r=-0.997 1。干扰试验样品回收率在50%-200%。实测样品内毒素值从低于检测限至433 EU/ml。不同厂家及同一厂家不同批次产品解吸附前后的内毒素测定值差异显著,CV值在36.9%-89.0%之间。结论 该方法可靠性、线性、回收率、灵敏度均符合要求,为进一步制定DTaP相关质量标准提供参考。现有热原检测方法不能全面反映DTaP成品中内毒素水平,有必要将内毒素检测纳入DTaP质量标准,以更好地控制疫苗安全性。Objective To determine the endotoxin content in final product of adsorbed diphtheria, tetanus and acellular pertussis （DTaP） combined vaccine containing aluminum hydroxide adjuvant as well as the necessity and feasibility of inclusion of test for endotoxin in DPT in Chinese Pharmacopoeia （2015 edition）. Methods A kinetic turbidimetrie method for determination of bacterial endotoxin content in DTaP before and after desorption was developed according to the requirements in appendix X]IE of Chinese Pharmacopoeia （Volume m, 2010 edition）, of which the standard curve and recovery rate were verified. Results The developed method showed good linearity within a concentration range of 0. 005 10 EU/ml （r = -0. 997 1 ）. The recovery rate of samples in interference test was 50%- 200%. The endotoxin content in samples ranged from less than detection limit to 433 EU/ml. The endotoxin contents in products by various manufacturers and those of various batches by the same manufacturer showed significant difference, with CVs of 36. 9 % - 89. 0%. Conclusion The reliability, linearity, recovery rate and sensitivity of developed method met the relevant requirements, which provided a reference for further development of the quality standard of DTaP. The current ppyrogen test could not reflect the eudotoxin level in final product of DTaP completely, so it was necessary to include the test for endotoxin to the quality control standard of DTaP in order to control the safety of vaccine more effectively.

关 键 词：无细胞百白破联合疫苗 内毒素含量 

分 类 号：R378.42[医药卫生—病原生物学]