测定过氧化氢酶活性的分光光度法的建立及验证

Development and verification of spectrophotometry for determination of catalase activity

目的建立测定23价肺炎球菌多糖疫苗的关键原辅材料—过氧化氢酶活性的方法,并进行验证。方法分别采用高锰酸钾滴定法和紫外分光光度法测定过氧化氢酶活性,并对紫外分光光度法进行重复性、日间精密度、不同人员间精密度、耐用性、线性验证。结果两批样品重复测定6次结果的RSD均〈5%;两批样品两名检测人员3日内的检测结果以及两批样品不同工作日间的检测结果差异均无统计学意义（P〉0-05）,样品日间和人员间检测结果的RSD均〈5%;在20-30℃温度范围内该方法耐用性良好,在p H 7.0-7.5范围内耐用性良好;该方法在样品浓度为0.2-1.6μl/ml之间的线性关系良好。结论分光光度法重复性、日间精密度、不同人员间精密度良好,且操作简便,结果可靠,适用于过氧化氢酶的质量控制。

Objective To develop and verify a method for determination of activity of catalase, a key raw material of 23- valent pneumococcal polysaccharide vaccine. Methods Catalase activity was determined by potassium permanganate titration and ultraviolet spectrophotometry respectively, and the latter was verified for reproducibility, precisions on various working days and by various personnel, redness and linearity. Results All the RSDs of six test results of two batches of samples were less than 5%. No significant differences were observed in the test results of two batches of samples by two personnel within 3 d and those on various working days （P 〉 0. 05 ）. Both the RSDs of test results on various working days and by various personnel were less than 5%. The method showed good redness at 20 - 30 ℃ and pH 7. 0 - 7. 5, and a good linearity within a concentration range of 0. 2 - 1. 6 μl / ml. Conclusion Spectrophotometry showed good reproducibility as well as good precisions on various working days and by various personnel, which was simple, reliable and was suitable for the quality control of catalase.