核酸水解产物中腺嘌呤的荧光猝灭法测定及机理研究  

Studies on the Determination of Adenine in DNA Hydrolyzate Based on the Fluorescence Quenching of Evans Blue and Its Mechanism

作　　者：张敏[1] 赵虎[1] 吕庆銮[1] 岳宁宁[1] 张苗[1] 王怀友[1] 

机构地区：[1]山东师范大学化学化工与材料科学学院,山东济南250014

出　　处：《分析测试学报》2008年第3期263-265,269,共4页Journal of Instrumental Analysis

基　　金：山东省自然科学基金资助项目(Y2006B31)

摘　　要：研究了腺嘌呤猝灭伊文思蓝（Evans blue）的荧光猝灭机理并建立了测定腺嘌呤的新方法。当用253am激发，伊文思蓝的最大发射波长位于387nm，腺嘌呤对伊文思蓝的荧光具有猝灭作用。线性回归方程为：p（mg／L）：1.836×10^3I^-1F-9.110，相关系数为0.9997，线性范围为0.20-20.0mg／L，检出限为0.18mg／L，相对标准偏差（RSD）为5.6％。试验了pH、干扰离子、放置时间等对测定的影响，该方法可用于核酸水解产物中腺嘌呤的测定。The mechanism of fluorescence quenching of Evans blue （EB） with adenine was studied and a novel method was thus developed for the determination of adenine in DNA hydrolyzate. The re- suits showed that the wavelength of maximum fluorescence emission of EB was at 387 nm when excited with a wavelength of 253 nm, and that the EB fluorescence was quenched in the presence of adenine. The effects of pH, Stability of EB blue in the presence of adenine, foreign ions and the presence of hydrolyzates of nucleic acid on the determination of adenine were examined. The linear range of the calibration curve for adenine was 0.20-20.0 mg/L with a correlation coefficient of 0.9997. The detection limit was 0.18 mg/L and the relative standard derivation was 5.6% . The proposed method has been applied to the determination of adenine in DNA hydrolyzates, the results with those obtained by the HPLC method. agreed quite well with those obtained by the HPLC method.

关 键 词：腺嘌呤 伊文思蓝 荧光猝灭法 猝灭机理 脱氧核糖核酸 

分 类 号：O657.3[理学—分析化学] Q522[理学—化学]