以1-萘胺为底物伏安法测定辣根过氧化物酶

Voltammetric Determination of Horseradish Peroxidase with 1-Naphthylamine as Substrate

摘要: 提出了1-萘胺-过氧化氢-辣根过氧化物酶(HRP)伏安酶联免疫分析体系应用于HRP的伏安法测定.在pH 7.0的KH2PO4-Na2HPO4缓冲溶液中,HRP催化过氧化氢氧化1-萘胺生成电活性产物,在极谱仪上进行二阶导数伏安线性扫描,该产物在滴汞电极上发生还原反应,于-0.43 V(vs.SCE)左右产生一灵敏的还原峰.HRP浓度在3.0×10-7~5.0×10-5g·L-1范围内与线性扫描峰电流的二阶导数值(ΔI″p)呈线性关系,检出限为2.5×10-7g·L-1.

Abstract: A new system of immunoassay,composing of horseradish peroxidase (HRP),hydrogen peroxide and 1-naphthyl amine,was proposed for the voltammetric determination of HRP.In a phosphate buffer of pH 7.0,the oxidation of 1-naphthylamin by H2O2 was catalyzed by the presence of HRP,producing an electroactive compound which gave a well-defined sensitive reduction peak at the potential -0.43 V (vs.SCE).Under optimum enzyme-catalytic conditions,linear relationship between values of ΔI″p and concentration of HRP was obtained in the range of 3.0×10-7-5.0×10-5g·L-1 of HRP with a detection limit of 2.5×10-7g·L-1.