磷脂化重组人铜锌超氧化物歧化酶铜、锌含量测定方法的建立

目的:建立重组人铜锌超氧化物歧化酶( rhCu/Zn-SOD)和磷脂化重组人铜锌超氧化物歧化酶(PC-SOD)中铜、锌含量的测定方法。方法:采用原子吸收光谱法检测,经考察选择稀硝酸溶液作为稀释液,并进行了方法的验证。结果:用稀硝酸溶液稀释样品效果最佳,回收率为98.3%~101.8%,检测限为铜 0.004 8 μg·mL^-1,锌0.005 3 μg·mL^-1,其标准曲线相关系数为0.99以上,精密度的RSD为0.2%~1.2%。测定5批样品的铜、锌含量,结果与理论值一致。结论:建立并验证了 rhCu/Zn-SOD和PC-SOD中铜、锌含量的测定方法,该方法操作简便,精密度和灵敏度符合生物制品理化测定的要求,可用于 rhCu/Zn-SOD和PC-SOD中铜、锌含量的质量控制。

Objective:To establish a method for determination of copper and zinc in recombinant human Cu/Zn Superoxide Dismutase(rhCu/Zn-SOD)and lecithinized recombinant human Cu/Zn superoxide dismutase(PC-SOD).Methods:Atomic absorption method was used and the samples were diluted by nitric acid.In addition,the method was validated.Results:The best method of handling was nitric acid,and the recovery was 98.3%-101.8%.The detection limit of the method was 0.004 8 μg·mL^-1 for copper and 0.005 3 μg·mL^-1 for zinc.The regression coefficient > 0.99,the RSD of precision was 0.2%-1.2%.The copper and zinc content of 5 batch of samples was analyzed by the established method,and the obtained results were accordance with the theoretical value.Conclusion:The method for copper and zinc determination in rhCu/Zn-SOD and PC-SOD was established and validated,which can be used for copper and zinc content quality control for rhCu/Zn-SOD and PC-SOD since the precision and detection limit of the method met the requirement for physical and chemical assay of biological products.