高等学校化学学报

高等学校化学学报

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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

蒋治良, 李建福, 梁爱惠, 李纪顺, 汤亚芳, 王素梅, 张南南   

  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
  • 收稿日期:2007-12-05 修回日期:1900-01-01 出版日期:2008-10-10 发布日期:2008-10-10
  • 通讯作者: 蒋治良

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

JIANG Zhi-Liang*, LI Jian-Fu, LIANG Ai-Hui, LI Ji-Shun, TANG Ya-Fang, WANG Su-Mei, ZHANG Nan-Nan   

  1. Department of Material and Chemical Engineering, Key Laboratory of New Processing Technology for Nonferrous Metals and Materials of Education Ministry, Guilin University of Technology, Guilin 541004, China
  • Received:2007-12-05 Revised:1900-01-01 Online:2008-10-10 Published:2008-10-10
  • Contact: JIANG Zhi-Liang

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被引次数

2

摘要： 在pH 4.8 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8.3×10-12～4.17×10-10 g/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; 3.6×10-12 g/mL HRP和5.4×10-12 g/mL HRP. 该法用于辣根过氧化酶(HRP)的测定, 结果满意.

关键词: 邻苯二胺, 辣根过氧化酶, 过氧化氢, 瑞利散射光谱法

Abstract: 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 coefficient and the detection limit were ΔI445 nm=2.23c+11, ΔI484 nm=1.47c+4.8; 0.9982, 0.9919; 3.6×10-12 g/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.

Key words: o-Phenylenediamine, Horseradish peroxidase, Hydrogen peroxide, Rayleigh scattering assay

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