非水溶性样品用的细菌内毒素标准品的研制  

Development of bacterial endotoxin standard for insoluble samples

作　　者：裴宇盛[1] 陈晨[1] 赵小燕 高华[1] 蔡彤[1] PEI Yu-sheng;CHEN Chen;ZHAO Xiao-yan;GAO Hua;CAI Tong(National Institutes for Food and Drug Control, Beijing 102629, China)

机构地区：[1]中国食品药品检定研究院,北京102629

出　　处：《中国药理学通报》2022年第6期944-948,共5页Chinese Pharmacological Bulletin

基　　金：国家科技重大专项(No 2018ZX09101001);国家药典委员会药品标准制修订研究课题(No 2020Y007)。

摘　　要：目的为非水溶性样品细菌内毒素检测建立一批内毒素标准物质。方法将细菌内毒素原料通过分装冻干(不添加赋形剂)制备了候选品A和B。两批候选品采用3种方法(实验室个数:凝胶法13个、动态浊度法9个、动态显色法5个)进行协作标定。结果经统计分析,3种方法标定候选品A的效价几何平均值分别为680.1 EU、827.0 EU、800.8 EU,RSD分别为22.4%、16.2%、16.7%,3种方法的方差分析P值为0.067,差异无显著性,候选品A的效价加权平均值为774.0 EU(95%置信区间为721.0~831.0,FL%为7.10)。3种方法标定候选品B的效价几何平均值分别为1640.6 EU、1828.6 EU、3224.8 EU,RSD分别为33.9%、47.0%、54.4%,3种方法的方差分析P值为0.030,差异具有显著性,用卡方检验对权重进行校正,得到候选品B的校正加权平均效价值为1822.7 EU(95%置信区间为1548.7~2145.2,FL%为16.4)。根据结果淘汰候选品B。结论经国家标准物质委员会批准,候选品A成为我国首批细菌内毒素国家标准品(非水溶性样品专用),效价为每支700 EU。Aim To establish a batch of endotoxin standard for bacterial endotoxin detection of insoluble samples.Methods Candidate A and candidate B were prepared by freeze-drying bacterial endotoxin without excipient.The two batches of candidates were calibrated by three methods,including 13 laboratories for gel method,9 laboratories for kinetic-turbidimetric assay and 5 laboratories for kinetic chromogenic assay.Results After statistical analysis,the geometric mean values of gel method,kinetic-turbidimetric assay and kinetic chromogenic assay calibration of candidate A were 680.1 EU,827.0 EU and 800.8 EU,with RSD of 22.4%,16.2%and 16.7%,respectively.The P value of variance analysis of calibration results of the three methods was 0.067,showing no significant difference.The weighted mean of potency was 774.0 EU(95%confidence interval 721.0~831.0,FL%7.10).The geometric mean values of the calibration of candidate B by gel method,kinetic-turbidimetric assay and kinetic chromogenic assay method were 1640.6 EU,1828.6 EU and 3224.8 EU,with RSD of 33.9%,47.0%and 54.4%,respectively.The P value of variance analysis of the calibration results of the three methods was 0.030,showing significant difference.Chi-square test was used to correct the weight of each method,and weighted average of the results of the three methods was used to obtain a corrected weighted average efficiency value of 1822.7 EU(95%confidence interval 1548.7~2145.2,FL%16.4).Candidate B was eliminated based on the results.Conclusion Candidate A has become the first batch of national standard bacterial endotoxin(for insoluble samples only)approved by National Standard Substance Committee of China,and the potency is 700 EU.

关 键 词：细菌内毒素 标准物质 非水溶性样品 协作标定 凝胶法 光度测定法 

分 类 号：R378[医药卫生—病原生物学] R927.1[医药卫生—基础医学]