摘要
研究了尿嘧啶猝灭伊文思蓝（Evans blue）的荧光猝灭机理，并根据尿嘧啶对伊文思蓝荧光的猝灭作用，建立了测定尿嘧啶的新方法。当用253nm激发时，伊文思蓝的最大发射波长位于387nm，线性回归方程为：c=11．85IO／IF-12．30，相关系数为0．9990，线性范围为0．2～20mg／L，检出限为0．14mg／L，相对标准偏差（RSD）为3．6％。求得伊文思蓝与尿嘧啶的形成常数为K=4．88×10^3L／mol，试验了pH、干扰离子、放置时间、核酸水解产物等对测定的影响。
The mechanism of fluorescence quenching of Evans blue （EB） in the presence of uracil was studied. The maximum emission wavelength of EB was at 387 nm with the excitation wavelength at 253 nm. It was found that the fluorescence quenching of EB was of a static one and the binding constant（K） was 4. 88 × 10^3 L/mol. A linear relationship was found between the relative fluorescence intensity of EB - uracil system and the concentration of uracil. The effects of pH, stability of EB in the presence of uracil, coexistent ions and the presence of hydrolysates of nucleic acid on the determina- tion of uracil were studied. Under optimum conditions, the linear range of the calibration curve for uracil was 0.2 - 20 mg/L with a correlation coefficient of 0. 999 0 and a detection limit of 0. 14 mg/L. The relative standard deviation （RSD） was 3.6%. The analytical results of the samples obtained by this novel method agreed quite well with those obtained by the HPLC method.
出处
《分析测试学报》
CAS
CSCD
北大核心
2008年第8期859-861,865,共4页
Journal of Instrumental Analysis
基金
山东省自然科学基金资助项目(Y2006B31)
关键词
尿嘧啶
伊文思蓝
荧光猝灭法
猝灭机理
uracil
Evans blue
fluorescence quenching
quenching mechanism
