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

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0.47 EU/mL;检测限均为0.005 EU/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。

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(VolumeⅠ,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 withβ-glucan,and the dynamic colorimetric method detected the concentration ofβ-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.