摘要
目的:建立适用于人凝血因子Ⅷ效价测定的生色底物法。方法:用生色底物法(包括动力学法和终点法)进行人凝血因子Ⅷ的效价测定,采用量反应平行线法进行效价计算,并对该方法进行方法学验证。采用经验证的方法对来自7家企业的21批人凝血因子Ⅷ样品进行效价测定,并与现行标准的一期法结果进行比较。结果:效价测定范围为标示量的64%~156%;以效价测定值的对数对理论值的对数作线性回归,动力学法的相关系数r为0.998;相对偏倚为-0.2%~1.0%,其90%置信区间为(-2.3%,2.2%);中间精密度为1.8%~2.1%,其95%置信上限范围在3.2%~3.9%;终点法的相关系数r为0.998;相对偏倚为-0.6%~1.2%,其90%置信区间为(-1.7%,2.6%);中间精密度为1.5%~2.4%,其95%置信上限范围在2.7%~4.4%;不同批次的试剂盒和不同分析人员对测定结果的影响无统计学意义,方法耐用性良好。21批样品生色底物法量反应平行线测定法测得结果与现行标准结果一致性良好。结论:建立的生色底物法可用于人凝血因子Ⅷ效价的测定,引入的量反应平行线模型可增加试验数据可靠性判断,并有效控制测定方法的误差,从而更好地控制人凝血因子Ⅷ质量。
Objective:To establish chromogenic substrate assay for biological activity determination of human coagulation factorⅧ.Methods:The biological activity of human coagulation factorⅧwas determined by chromogenic substrate assay,the results were calculated by parallel-line model,and then the method was verified.Finally,21 batches of human coagulation factorⅧfrom seven manufacturers were investigated by using the method and the results were compared with those calculated by one-stage assay with standard curve model.Results:The determination range was between 64%and 156%of the potency stated on the label.In kinetic method,the correlation coefficient between measured and labelled values was 0.998.The relative bias were ranged from-0.2%to 1.0%with 90%confidence intervals from-2.3%to 2.2%.The intermediate precision was 1.8%-2.1%with 95%upper confidence limit from 3.2%to 3.9%.In end-point method,the correlation coefficient between measured and labelled values was 0.998.The relative bias were ranged from-0.6%to 1.2%with 90%confidence intervals from-1.7%to 2.6%.The intermediate precision was 1.5%-2.4%with 95%upper confidence limit from 2.7%to 4.4%.The effects of different batch numbers and analysts on the determination results were not statistically significant,and the method showed good durability.The results of 21 batches measured by chromogenic substrate assay with parallel-line model were in good agreement with those obtained by one-stage assay with standard curve model.Conclusion:The established chromogenic substrate method can be used for the biological activity determination of human coagulation factorⅧ.The parallel-line model introduced can increase the reliability of the data and effectively control the error of the determination method,so as to better control the quality of human coagulation factorⅧ.
作者
史卓维
王鸣人
段徐华
邵泓
陈钢
Shi Zhuowei;Wang Mingren;Duan Xuhua;Shao Hong;Chen Gang(Shanghai Institute for Food and Drug Control,NMPA Key Laboratory for Quality Control of Therapeutic Monoclonal Antiodies,Shanghai 201203,China;Institutes of Biomedical Sciences,Fudan University)
出处
《中国药师》
CAS
2022年第7期1296-1301,共6页
China Pharmacist
关键词
人凝血因子Ⅷ
效价测定
生色底物法
量反应平行线测定法
Human coagulation factorⅧ
Biological activity determination
Chromogenic substrate assay
Parallel-line model
