超痕量HRP的邻苯二胺微粒瑞利散射光谱分析新方法及其应用  

Rayleigh Scattering Spectral Determination of Ultratrace Horseradish Peroxidase Based on o-Phenylenediamine Particles and Its Application

作　　者：蒋治良[1] 李建福[1] 梁爱惠[1] 李纪顺[1] 汤亚芳[1] 王素梅[1] 张南南[1] 

机构地区：[1]桂林工学院材料与化学工程系,有色金属及材料加工新技术教育部重点实验室,桂林541004

出　　处：《高等学校化学学报》2008年第10期1973-1976,共4页Chemical Journal of Chinese Universities

基　　金：国家自然科学基金(批准号:20365001,20865002);广西自然科学基金(批准号:0728213);桂林工学院科研启动基金资助

摘　　要：在pH4.8的乙酸盐缓冲溶液中,邻苯二胺（OPD）形成粒径约380 nm的微粒,在392,420,445,484和507 nm处有5个较强的Rayleigh散射峰.辣根过氧化酶（HRP）催化H2O2氧化邻苯二胺生成黄色的2,3-二氨基吩嗪产物,反应体系在420,445和484 nm处的Rayleigh散射光信号显著减弱.在最佳条件下,HRP浓度在8.3×10^-12～4.17×10^-10g/mL范围内均与445和484 nm处的Rayleigh散射强度的降低值呈线性关系,其回归方程、相关系数、检出限（3σ）分别为ΔI445 nm=2.23c＋11,ΔI484 nm=1.47c＋4.8;0.9982,0.9919;3.6×10^-12g/mL HRP和5.4×10^-12g/mL HRP.该法用于辣根过氧化酶（HRP）的测定,结果满意.In pH 4. 8 acetate buffer solution, o-phenylenediamine aggregated itself to particles in size of 380 nm, that exhibited five Rayleigh scattering peaks at 392, 420, 445, 484 and 507 nm. Horseradish peroxidase （HRP） has a strong catalytic effect on the H2O2 oxidation of o-phenylenediamine to 2,3-diaminophenazine, that caused significantly decreasing of the Rayleigh scattering intensity at 420, 445 and 484 nm. Under the optimal condition, the concentration of HRP in the range of 8.3 × 10^-12--4.17 × 10^-10 g/mL was all proportional to the decreased Rayleigh scattering intensity at 445 and 484 nm. Its regression equation, correlation co- efficient and the detection limit were ΔI445 nm=2.23c＋11,ΔI484 nm=1.47c＋4.8;0.9982,0.9919;3.6×10^-12g/mL , and 5.4 × 10^-12 g/mL HRP, respectively. A new enzyme catalytic-Rayleigh scattering spectral method was developed for assay of HRP, with a high sensitivity and selectivity, simplicity and rapidity. This assay was applied to the determination of HRP with satisfactory results.

关 键 词：邻苯二胺 辣根过氧化酶 过氧化氢 瑞利散射光谱法 

分 类 号：O656[理学—分析化学]