重组C因子法检测泰拉霉素注射液细菌内毒素的方法学研究

Investigation on the Applicability of Bacterial Endotoxins Test Using Recombinant Factor C in Tulathromycin Injection for Detection of Bacterial Endotoxin

关键词:  重组C因子  泰拉霉素注射液  细菌内毒素

Key words:  recombinant factor C  tulathromycin injection  bacterial endotoxin

摘要:

建立泰拉霉素注射液的重组C因子细菌内毒素检测方法,并根据《中国兽药典》一部(2020年版)9101兽药质量分析方法验证指导原则的规定,对该方法的线性范围、准确度、精密性、专属性、检测限及耐用性进行验证,与动态显色法进行比较。重组C因子法在内毒素标准品为0.005～5 EU/mL范围内与logδRFU均呈良好的线性关系,r均＞0.980；回收率结果均在50%～200%的范围内,变异系数均小于10%,为弱变异性；重组C因子不与 β-葡聚糖发生反应,动态显色法检测β-葡聚糖浓度为0.47EU/mL；检测限均为 0.005EU/mL；斜率、截距和实测值不受增益值改变的影响。重组C因子法具有良好的准确度、精密性、专属性及耐用性,可应用于兽医专用注射液细菌内毒素的检查。

Abstract:

A recombinant C-factor bacterial endotoxin assay was established for tulathromycin injection, and the linear range, accuracy, precision, specificity, detection limit and durability of the method were validated according to the ＜9101＞ Guidelines for the Verification of Veterinary Medicine Quality Analysis Methods in Chinese Veterinary Pharmacopoeia（Volume Ⅰ，2020 edition）, and compared with the dynamic colorimetric method. The recombinant C-factor method showed good linear relationship with logδRFU in the range of 0.005-5 EU/mL for endotoxin standards, with r＞0.980; the recoveries were in the range of 50%-200%, and the coefficients of variation were less than 10%, which were weakly variable; the recombinant C-factor did not react with β-glucan, and the dynamic colorimetric method detected the concentration of β-glucan at 0.47 EU/mL; the detection limits were 0.005 EU/mL; the slopes, intercepts and measured values were not affected by the change of gain values. The recombinant C-factor method has good accuracy, precision, specificity and durability, and can be applied to the examination of bacterial endotoxin in veterinary special injection.