基于2,6-二-(2-吡啶基)-4-吡啶甲酸的紫外法检测辣根过氧化物酶浓度

Determination of Horseradish Peroxidase Concentration Based on 2,6-Di-(2-pyridyl)-4-pyridyl Carboxylic Acid by UV Method

摘要: 本文首次提出采用2,6-二-(2-吡啶基)-4-吡啶甲酸(配体L)-H₂O₂-HRP测定HRP酶浓度的新体系。利用紫外考查HRP催化H₂O₂氧化配体L体系的光谱变化。在pH = 7.2，25℃下，配体L浓度为6 × 10⁻⁶ mol/L，H2O2浓度为1 × 10⁻⁴ mol/L时，回归方程为A = −0.0034x + 1.072，线性相关系数为0.9909，该体系的检测范围可达10⁻⁸~10⁻⁹ mol/L，灵敏度较高，操作简便，抗干扰能力强，稳定性好，可用作检测游离HRP酶的新方法，具有实际的应用前景。

Abstract: A new system for the determination of horseradish peroxidase concentration by 2,6-di-(2-py- ridyl)^-4-picolinic acid (ligand L)-H₂O₂-HRP was proposed for the first time. The spectral changes of ligand L system catalyzed by HRP for H₂O₂ oxidation were investigated by UV method. At pH = 7.2 and 25˚C, the ligand L concentration is 6 × 10⁻⁶ mol/L, H₂O₂ concentration is 1 × 10⁻⁴ mol/L, the regression equation is A = −0.0034x + 1.072 and the linear correlation coefficient is 0.9909. The detection range of the system can reach 10⁻⁸~10⁻⁹ mol/L. This method has high sensitivity, simple operation, strong anti-interference ability and good stability. It can be used as a new method for the detection of free HRP enzyme and has practical application prospects.