动态浊度法检测23价肺炎链球菌多糖疫苗细菌内毒素的方法确认和应用

摘要

目的 对动态浊度法检测23价肺炎链球菌多糖疫苗(23-valent pneumococcal polysaccharide vaccine,PPV23)细菌内毒素进行方法学确认并加以应用.方法 按照《中华人民共和国药典》2010版三部中的动态浊度法操作,配制系列稀释的细菌内毒素标准品,验证动态浊度法的线性、准确度、重复性和中间精密度.根据2012年和2013年连续20批PPV23成品的细菌内毒素检测结果,分析PPV23对动态浊度法的影响.结果 动态浊度法的标准曲线具有可靠性,细菌内毒素标准品的回收率为90％～129％,变异系数为6.2％～12.4％,均符合规定的标准,且操作人员(F=4.80,P=0.06)和时间(F=1.01,P=0.34)变化对检测结果没有影响.2012年和2013年各连续20批PPV23成品的细菌内毒素检测结果保持稳定,细菌内毒素均值分别为0.165和0.355 EU/ml,PPV23对动态浊度法没有影响(F=0.00,P=0.97).结论 动态浊度法有效可行,可用于PPV23细菌内毒素检测.%Objective To validate and apply the turbidimetric-kinetic method for determination of bacterial endotoxin in 23-valent pneumococcal polysaccharide vaccine (PPV23).Methods The bacterial endotoxin standards were diluted serially in accordance with the turbidimetric-kinetic method in Chinese Pharmacopoeia 2010 edition (Volume Ⅲ).Linearity,accuracy,repeatability and intermediate precision of the turbidimetric-kinetic method were validated.According to the determination results of bacterial endotoxins in 20 batches of consecutively manufactured PPV23 in 2012 and 2013,the influence of PPV23 on the turbidimetric-kinetic method was studied.Results The standard curve of the turbidimetric-kinetic method was reliabile.The recoveries of the bacterial endotoxin standard were 90％-129％,and the coefficients of variation were 6.2％-12.4％.Both results met standard requirement.The changes of operator (F=4.80,P=0.06) and time (F=1.01,P=0.34) had no effect on the turbidimetric-kinetic method.The determination results of bacterial endotoxins in 20 batches of consecutively manufactured PPV23 in 2012 and 2013 remained stable,and the average values of bacterial endotoxins in 20 batches of PPV23 in 2012 and 2013 were 0.165 and 0.355 EU/ml,respectively.PPV23 had no influence on the turbidimetric-kinetic method.Conclusion The turbidimetric-kinetic method is effective and feasible and can be used to detect bacterial endotoxin in PPV23.