摘要
在pH4.56的BR缓冲溶液中,DNA能使硫堇的光散射增强,荧光发生猝灭。本实验基于硫堇的三维光谱讨论了其发光属性,并通过在普通荧光光度计上单次扫描同时获得的539nm处的光散射强度（I539）与630nm处的荧光强度（F630）之比,建立了测定痕量DNA的光散射/荧光比率法。当硫堇的浓度为1.0×10^-4mol/L时,方法的线性范围为0.1~1.6mg/L,检测限为（3σ）为12.0μg/L。将所建立的方法用于DNA合成样品的测定,回收率为94.9%~107.0%,相对标准偏差小于3.0%。
DNA could quench the fluorescence of thionine in BR buffer at pH 4.56, and result in light scattering enhancement. In this contribution, the emission nature of thionine was discussed based on the investigations of its three-dimensional spectra, and a determination method of trace amount of DNA was further devel- oped with a common spectrofluorometer to measure the light emission signals through one-time scanning by which a ratio of the intensity of light scattering at 539 nm with that of fluorescence at 630 nm could be available. It was found that this method has the linear range from 0.1 to 1.6 mg/L with the detection of limit （3σ） of 12..0 μg/L for 1.0 ×10 ^-4mol/L Thionine. The method has been applied for the detection of DNA in artificial samples with the recovery from 94.9% to 107.0% with the RSD lower than 3.0%.
出处
《分析化学》
SCIE
EI
CAS
CSCD
北大核心
2008年第4期529-532,共4页
Chinese Journal of Analytical Chemistry
基金
国家自然科学基金(No.20425517)资助项目
关键词
硫堇
脱氧核糖核酸(DNA)
光散射/荧光比率
三维光谱
Thionine, deoxyribonucleic acid （ DNA）, light scattering-fluorescence ratio, three-dimensionalspectra
