同步荧光光谱比率法检测过氧亚硝酸根及其清除剂的研究  

Study on Ratiometric Determination of Peroxynitrite and Its Scavenger Based on Synchronous Fluorescence Spectroscopy

作　　者：杜俊鸥[1] 郭祥群[1] 

机构地区：[1]厦门大学化学化工学院化学系,现代分析科学教育部重点实验室,化学生物学福建省重点实验室,福建厦门361005

出　　处：《光谱学与光谱分析》2008年第8期1875-1878,共4页Spectroscopy and Spectral Analysis

基　　金：国家自然科学基金项目(20375032)资助

摘　　要：利用同步荧光光谱建立了一种新的荧光比率法检测过氧亚硝酸根（ONOO-）及评价其清除剂的方法。酪氨酸自身有荧光,在pH 8.5的磷酸缓冲溶液中,酪氨酸与CO2和ONOO-反应生成强荧光的酪氨酸二聚体,利用同步荧光光谱技术,在荧光发射光谱中能同时获得酪氨酸单体（λem=364nm）与二聚体的荧光峰（λem=406nm）,且两峰的荧光强度的比值与ONOO-的浓度呈计量相关。结果表明荧光强度的比值不受实验参数改变的影响,与传统的荧光法相比,有利于扩大检测的线性范围,提高灵敏度。荧光强度的比值与ONOO-的浓度在1.60×10^-7mol·L^-1～6.00×10^-6mol·L^-1范围内呈线性关系,线性相关系数为0.999,检测限为1.84×10^-8mol·L^-1。对浓度为1.00×10^-6mol·L^-1ONOO-平行测定8次,其相对标准偏差为2.4%。利用该法测得抗癌药米托蒽醌的IC50为0.065μg·mL^-1。方法简便易行,试剂廉价易得,体系稳定,可作为一种检测ONOO-及筛选其清除剂的方法。A ratiometrie fluorimetry is proposed for the determination of peroxynitrite （ONOO-） and the evaluation of its scavenger based on synchronous fluorescence spectroscopy. L-tyrosine, an intrinsic fluorescent aminoacid, reacts with peroxynitrite and carbon dioxide, yielding a highly fluorescent dimer of tyrosine in pI-I 8. 5 PBS buffer solution. By synchronous fluorescence scanning, both monomer and dimer emission bands of L-tyrosine appeared at 364 and 406 nm, respectively. The ratio of F406/ F364 is quantitatively related to the concentration of peroxyn/trite, where F406 represents the intensity of synchronous emission band of dimer and F364 represents that of monomer. The method has been showing merit of being insensitive to the changes in experimental parameters and offers a higher sensitivity and a broader responding linear range of the analyte concentration, compared to fluorirnetric method, giving a LOD of 1.84× 10^-8 mol · L^-1 and a linear range of 1.60× 10^-7 mol · L^-1-6. 00× 10^-8 mol · L^-1 for ON（30-, respectively, with a deviation of 2. 4% for 1.00× 10^-6 mol · L^-1 ONOO- （n=8）. A IC50 of 0. 065 μg · mL^-1 for mitoxantrone, an antioxidant and anticancer drug, was also obtained. This method has proved to be simple and speedy. It would be easily used in the determination of ONOO- and its scavenger.

关 键 词：同步荧光 比率法 过氧亚硝酸根 

分 类 号：O657.3[理学—分析化学]