磷脂化重组人铜锌超氧化物歧化酶活性测定方法的建立

目的：建立磷脂化重组人铜锌超氧化物歧化酶（PC-SOD）的活性测定方法，并对方法进行方法学验证。方法：采用经典的氮蓝四唑（NBT）还原法，SOD可竞争结合负氧离子，抑制显色，通过与活性标准品比较抑制显色的程度，计算样品的活性。对方法的专属性、线性与范围、回收率、精密度进行验证。结果：方法符合专属性要求；SOD线性范围0.23~500 μg·mL^-1，PC-SOD线性范围0.46~1000 μg·mL^-1，R²≥0.99；回收率测定结果介于80%~120%，符合活性测定方法对于回收率的要求；SOD原液活性测定结果为（5298±549）U·mg^-1，PC-SOD原液测定结果为（2765±474）U·mg^-1，PC-SOD成品测定结果为（2595±230）U·mg^-1，RSD≤20%，符合一般生物制品活性检测精密度要求。结论：建立并验证了反映药效的PC-SOD活性测定方法，可用于PC-SOD的质量控制和一致性分析。

Objective: To establish and verify a method for PC-SOD activity assay. Methods: Using the NBT deoxidation method,SOD were competitively bound to negative oxygen ions,which inhibited coloration.The samples' activity was calculated through the comparison of the coloration degree between the samples and the reference material.Then the specificity,linear range,recovery,and precision of the method were verified. Results: The method met the specificity requirement.The linear ranges of SOD and PC-SOD were 0.23-500 μg·mL^-1 and 0.46-1000 μg·mL^-1,R²≥0.99.The recovery was within 80%-120%,which met the recovery requirement in activity assay of biological products.The activity assay results of SOD bulk,PC-SOD bulk and PC-SOD product were (5298±549) U·mg^-1,(2765±474) U·mg^-1 and (2595±230) U·mg^-1,respectively;RSD≤20%. Conclusion: The method for PC-SOD activity assay is established and verified,which can be used for the quality control and conformity analysis of PC-SOD.

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