廖丹;杨烨;李青耘;谢姗姗;王静;

• 1:成都生物制品研究所有限责任公司

摘要(Abstract)：

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。

关键词(KeyWords)： 过氧化氢酶;酶活性;分光光度法

Abstract:

Keywords:

基金项目(Foundation):

作者(Author): 廖丹;杨烨;李青耘;谢姗姗;王静;

Email:

DOI: 10.13200/j.cnki.cjb.000861

参考文献(References)：

• [1]Andisi VF,Hinojosa CA,de Jong A,et al.Pneumococcal gene complex involved in resistance to extracellular oxidative stress[J].Infect Immun,2012,80(3):1037-1049.
• [2]Wang HF,Zhan HJ.Research progress of determination catalase activity[J].Sci Technol Inno Herald,2009,19(1):7-8.(in Chinese)王华芳,展海军.过氧化氢酶活性测定方法的研究进展[J].科技创新导报,2009,19(1):7-8.
• [3]Krause S.Validation of analytical methods for biopharmaceuticals-A guide to risk-based validation and implementation strategies[M].PDA Bethesda,MD,USA:DHI Publishing,LLC,2007:33-67.
• [4]Center for drug evaluation,CFDA.General principles of method validation for biological products[S].GPH1-1.2005.(in Chinese)国家食品药品监督管理局药品审评中心.生物制品质量控制分析方法验证技术审评一般原则[S].GPH1-1.2005.
• [5]Bekdeser B,zyürek M,GüclüK,et al.Development of a new catalase activity assay for biological samples using optical CUPRAC sensor[J].Spectrochim Acta A Mol Biomol Spectrosc,2014,132(11):485-490.
• [6]Teke M.Development of a new biosensor for determination of catalase activity[J].Prep Biochem Biotechnol,2014,44(6):608-616.
• [7]Standardization Administration of the People's Republic of China.Inspection of grains and oils-Determination of hydrogen peroxidase activity of grain and oilseeds[S].GB/T 5522-2008.Beijing:China Standardization Press,2008.(in Chinese)中华人民共和国国家标准化管理委员会.粮油检验粮食、油料的过氧化氢酶活动度的测定[S].GB/T 5522-2008.北京:中国标准出版社,2008.
• [8]Zhou DD,Wu WW,Yang FL,et al.Comparison of two methods for determination of catalase activity[J].Acta Agricul Jiangxi,2009,21(1):118-120.(in Chinese)周丹丹,吴文卫,杨逢乐,等.两种过氧化氢酶活性测定方法的比较[J].江西农业学报,2009,21(1):118-120.
• [9]Li Y,Schellhorn HE.Rapid kinetic microassay for catalase activity[J].J Biomol Tech,2007,18(4):185-187.
• [10]Iwase T,Tajima A,Sugimoto S,et al.A simple assay for measuring catalase activity:a visual approach[J].Sci Rep,2013,3(10):3081-3084.
• [11]Shivakumar A,Nagaraja P,Chamaraja NA,et al.Determination of catalase activity using chromogenic probe involving iso-nicotinicacidhydrazide and pyrocatechol[J].J Biotechnol,2011,155(4):406-411.
• [12]Pinto PC,Costa AD,Lima JL,et al.Automated evaluation of the effect of ionic liquids on catalase activity[J].Chemosphere,2011,82(11):1620-1628.