HL60-IL6单核细胞活化反应测定法在细菌内毒素工作标准品效价测定中的应用  

Application of HL60-IL6 Assay in Potency Estimation of Lipopolysaccharide Working Standard Reference

作　　者：王鸣人 段徐华[1] 王灿[1] 郑璐侠[1] 邵泓[1] 陈钢[1] WANG Ming-ren;DUAN Xu-hua;WANG Can;ZHENG Lu-xia;SHAO Hong;CHEN Gang(Shanghai Institute for Food and Drug Control,Shanghai 201203,China)

机构地区：[1]上海市食品药品检验所,上海201203

出　　处：《中国药学杂志》2020年第20期1711-1716,共6页Chinese Pharmaceutical Journal

基　　金：药品医疗器械审评审批制度改革专项资助(ZG2016-4-05)。

摘　　要：目的将HL60-IL6单核细胞活化反应测定法应用于细菌内毒素工作标准品的效价测定中。方法将细菌内毒素国家标准品作为标准品,细菌内毒素工作标准品作为待标品,按《美国药典》<1033>生物检定方法验证指导原则进行方法学验证,验证指标包括专属性、相对准确度、精密度、线性和范围。采用经验证的HL60-IL6单核细胞活化反应测定法对细菌内毒素工作标准品进行效价测定,并比较与其标示量的一致性。结果专属性考察结果符合可接受标准。相对偏差的置信区间均在-11%~12%内,直线回归方程斜率在0.8~1.25内,中间精密度均低于20%,线性相关系数为0.967,大于0.95,均符合可接受标准。相对效价水平范围为0.64~1.56。采用经验证的HL60-IL6法进行7次效价测定结果的平均值为每支81.9 EU,置信区间为每支75.04~89.36 EU,与标示值(每支80 EU)一致。结论采用HL60-IL6法进行内毒素工作标准品效价测定的结果与其标示量一致,该法可用于细菌内毒素工作标准品的效价测定。OBJECTIVE To apply HL60-IL6 assay in the potency estimation of lipopolysaccharide(LPS)working standard reference.METHODS Using LPS national standard reference as reference and LPS working standard reference as sample,the HL60-IL6 assay was firstly validated according to USP<1033>.The parameters including relative accuracy,specificity,intermediate precision(IP),linearity and range were verified.Once the assay was validated it was then applied to the potency estimation of LPS working standard reference,and the result was compared with that from conventional bacterial endotoxin test.RESULTS Specificity was established for LPS working standard reference.The confidential interval of relative bias was<12%and the slope ratio of linearity relationship between known and measured relative potency was between 0.8-1.25.The IP was<20%.The linearity was 0.967,over 0.95.All of the parameters conformed with the acceptance criteria.Based on the acceptance for IP equal to 20%and for relative potency equal to 12%,the range of the bioassay was 0.64 to 1.56.After validation,the HL60-IL6 assay was used for potency estimation of LPS working reference standard.The potency was estimated as 81.9 EU·vial-1 and its confidential interval was 75.04-89.36 EU·vial-1,which was consistent with the labeled value(80 EU·vial-1).CONCLUSION The HL60-IL6 assay can be applied to the potency estimation of LPS working reference standard.

关 键 词：HL60-IL6法 细菌内毒素标准品 效价测定 生物检定方法验证 《美国药典》 

分 类 号：R917[医药卫生—药物分析学]