基质效应对注射用重组人凝血因子Ⅷ效价检测方法的影响

Matrix effect on the determination of Potency in Recombinant Coagulation Factor Ⅷ for injection

目的 考察不同基质对注射用重组人凝血因子Ⅷ(rFⅧ)效价检测方法的影响。方法 采用两种不同检测基质分别建立注射用重组人凝血因子Ⅷ效价检测方法,并对两种基质效应的影响进行了方法验证,包括专属性、线性、准确性/重复性和中间精密度。结果 专属性:高浓度两种基质加标回收率分别为112%,110%,低浓度两种基质加标回收率分别为104%,109%。线性:在0.125~1.000 IU/mL范围内,标准品/样品的浓度和凝固时间之间的相关系数r均>0.99。准确性/重复性:两种基质回收率分别为104%、102%,RSD分别为2.4%、1.9%。中间精密度:两种基质人员因素P值分别为0.72、0.23,日期因素P值分别为0.79、0.85,RSD12次分别为4.2%、3.0%。两种基质检测结果比对:6批重组人凝血因子Ⅷ(rFⅧ)检测结果差异均≤5%,两种检测基质的检测结果无明显差异。结论 两种检测基质均具有良好的专属性、线性、重复性、准确性及中间精密度,适用于重组人凝血因子Ⅷ(rFⅧ)产品的效价检测。

Objective To investigate the matrix effect on the determination of potency in Recombinant Human Coagulation Factor Ⅷ for Injection(rFⅧ). Methods Two different detection matrices were used to establish two methods for detecting the potency in Recombinant Human Coagulation Factor Ⅷ for Injection. And the matrix effect on the determination of potency was determined, including specificity, linearity, repeatability, accuracy and intermediate precision. Results As to the specificity, the recoveries of the two substrates at high vs low concentration level were 112% and 110% vs 104% and 109%,respectively. As to the linearity, in the range of(0.125-1.000) IU/mL, the correlation coefficient between concentration and coagulation time of standard/sample was higher than 0.99. As to the accuracy/repeatability, the recoveries of two matrices was 104% and 102%, and RSD was 2.4% and 1.9%. As to the intermediate precision, personnel factor of two matrices was 0.72 and 0.23, date factor was 0.79 and 0.85, and RSD(for 12 times) was 4.2% and 3.0%. Comparison of two matrices was as follows: Deviation in test results of 6 batches of rFⅧ was all lower than 5%. There was no significant difference between two matrices. Conclusion The two matrices for potency detection show good performance including specificity, linearity, repeatability, accuracy, and intermediate precision. They are suitable for the determination of potency in rFⅧ products.