基于Fe(Ⅲ)和四甲基联苯胺显色反应分析碱性磷酸酶活力  

Alkaline phosphatase activity analysis based on the chromogenic reaction of iron( Ⅲ)-3,3’,5,5’-tetramethylbenzidine

作　　者：赵灵芝[1,2] 赵柳 张小清[1] 苗延青[1] ZHAO Lingzhi;ZHAO Liu;ZHANG Xiaoqing;MIAO Yanqing(Department of Pharmacy,Xi’an Medical College,Xi’an 710021;Key Laboratory of Analytical Chemistry for Life Science of Shaanxi Province,School of Chemistry and Chemical Engineering,Shaanxi Normal University,Xi’an 710062;Shaanxi Agricultural Inspection and Testing Center,Xi’an 710003)

机构地区：[1]西安医学院药学院,西安710021 [2]生命分析重点实验室,陕西师范大学化学化工学院,西安710062 [3]陕西省农业检验检测中心,西安710003

出　　处：《分析试验室》2020年第5期502-506,共5页Chinese Journal of Analysis Laboratory

基　　金：国家自然科学基金(81701014);陕西省科技厅项目(2018JM2027);陕西省教育厅项目(18JK0655);国家自然科学基金培育项目(2018GJFY13);西安市未央区科学计划项目(201846);西安医学院药学省级重点学科建设项目(2016YXXK15)资助。

摘　　要：基于酶底物和产物不同的氧化还原性质,建立了Fe(Ⅲ)和四甲基联苯胺(TMB)显色反应检测抗坏血酸和碱性磷酸酶(ALP)活力的比色法。在最佳条件下,吸光度与ALP活力的对数在0.5~200 U/L范围内呈现良好依赖性,检测限为0.05 U/L。方法已用于ALP抑制剂Na3VO4抑制率评估和监测兔血浆样品中内源性ALP水平,在临床应用和现场检测显示出巨大应用潜力。Fe3+-3,3’,5,5’-tetramethylbenzidine( TMB) was developing for detecting ascorbic acid( AA) and alkaline phosphatase( ALP) activity which was based on the different redox properties of ALP substrate AA-P and product AA. Under optimal conditions,UV-vis adsorbance exhibited a good quantitative relationship with LogcALPin the range of 0. 5-200 U/L with a detection limit of 0. 05 U/L. The method has been successfully applied to detect ALP inhibitor Na3VO4 and the endogenous ALP level of rabbit plasma samples. This method was simple and fast without the preparation of nanomaterials,holding great potential in clinical applications and in the point-of-care testing.

关 键 词：碱性磷酸酶(ALP) 抗坏血酸 比色法 3 3’ 5 5’-四甲基联苯胺 铁(Ⅲ) 

分 类 号：O657.3[理学—分析化学]