高灵敏催化荧光分析法测定活性氧自由基过氧亚硝酸阴离子  

Highly Sensitive Catalytic Spectrofluorimetric Determination of Peroxynitrite

作　　者：郑琦[1] 邱芬[1] 刘志洪[1] 蔡汝秀[1] 

机构地区：[1]武汉大学化学与分子科学学院

出　　处：《分析化学》2006年第1期26-30,共5页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金(No.20275027);湖北省自然科学基金(No.2004ABA182)资助课题

摘　　要：采用酶催化动力学分析法研究了过氧亚硝酸的氧化行为。提出了一种催化荧光光谱测定过氧亚硝酸的新方法。根据肌红蛋白所具有的模拟酶性质催化过氧亚硝酸氧化酪氨酸。产生一种具有荧光特性的产物酪氨酸二聚体。在实验选定的最佳反应条件下，该产物的荧光强度与过氧亚硝酸阴离子（ONOO^-）的浓度在6．00×10^-7～3．00×10^-5mol／L范围内呈良好的线性关系；方法检出限为8．20×10^-8mol／L。用停流荧光光谱法研究了酶催化反应的动力学行为，探讨了反应的可能机理，认为酪氨酸的二聚体可能是ONOO^-在体内造成损伤的另一标志物。A novel spectrofluorimetdc method for the determination of peroxynitrite was proposed. The method is based on a mimetic enzyme catalyzed reaction with myoglobin as the catalyst and L-tyrosine as the substrate. A new fluorescent substance was produced that might probably be the coupled dimmer of tyrosine, which, instead of nitryl-tyrosine, was likely to be a new marking substance of ONOO^- injury in vivo. Kinetics of the reaction was studied by stopped-flow method and the possible reaction mechanism was also recommended. The optimum conditions of the determination were studied. The proposed method is simple and highly sensitive with a detection limit of 8.20×10^-8mol/L of peroxynitrite. A linear ralationship is obtained over the peroxynitrite concentration range 6.00×10^-7～3.00×10^-5mol/L with a correlation coefficient of 0. 9971.

关 键 词：过氧亚硝酸 荧光光度法 肌红蛋白 酶催化动力学 酪氨酸 

分 类 号：O657.3[理学—分析化学]