Abstract：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.
王鸣人, 段徐华, 王灿, 郑璐侠, 邵泓, 陈钢. 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[J]. 中国药学杂志, 2020, 55(20): 1711-1716.
WANG Ming-ren, DUAN Xu-hua, WANG Can, ZHENG Lu-xia, SHAO Hong, CHEN Gang. Application of HL60-IL6 Assay in Potency Estimation of Lipopolysaccharide Working Standard Reference. Chinese Pharmaceutical Journal, 2020, 55(20): 1711-1716.
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