基因重组鲎试剂检测促肝细胞生长素注射液中细菌内毒素含量适用性研究

Suitability Study of Recombinant Limulus Amebocyte Lysate for the Determination of Bacterial Endotoxins in Hepatocyte Growth-Promoting Factors Injection

目的 探讨基因重组鲎试剂用于测定促肝细胞生长素注射液中细菌内毒素含量的适用性。方法 分别使用基因重组鲎试剂和天然美洲鲎试剂建立细菌内毒素标准曲线,采用供试品干扰试验测定供试品溶液中外加内毒素的回收比,采用动态显色法测定供试品中细菌内毒素含量,并比较2种鲎试剂检测结果的差异。结果 2种鲎试剂细菌内毒素浓度在0.008~5 EU/mL范围内与反应时间线性关系较好(|r|≥0.998)。供试品在稀释2倍或20倍时对反应无干扰,细菌内毒素回收比均在50%~200%之间,26批供试品的细菌内毒素含量均小于限值(37.5 EU/mL)。2种鲎试剂细菌内毒素含量检测结果无显著差异(P> 0.05)。结论 该研究中建立了该制剂细胞内毒素的定量检测方法,准确性和灵敏度好,可用于该产品生产、流通环节细菌内毒素的质量控制。

Objective To investigate the suitability of recombinant limulus amebocyte lysate(LAL)for the determination of bacterial endotoxins in Hepatocyte Growth-Promoting Factors Injection.Methods The bacterial endotoxin standard curves were established by the recombinant LAL and LAL,respectively.The recovery rate of endotoxins added to the test solution was determined by the interference test of the test sample,the content of bacterial endotoxins in test sample was determined by the kinetic chromogenic assay,and the content differences determined by recombinant LAL and LAL were compared.Results The concentration of bacterial endotoxins determined by recombinant LAL and LAL in the range of 0.008-5 EU/mL showed a good linear relationship with reaction time(|r|≥0.998).There was no interference when the test sample was diluted by two or twenty multiples,and the recovery rate of bacterial endotoxins was in the range of 50%to 200%.The content of bacterial endotoxins in 26 batches of test samples was lower than the limit(37.5 EU/mL).There was no significant difference in the content of bacterial endotoxins determined by recombinant LAL and LAL(P>0.05).Conclusion The established quantitative determination method for bacterial endotoxin in Hepatocyte Growth-Promoting Factors Injection is accurate and sensitive,which can be used for the quality control of bacterial endotoxin in the production and circulation of this product.