阴离子表面活性剂与酚藏花红的作用及其在核酸荧光法测定中的应用

Reaction of Phenosafranine with Anionic Surfactants and Fluorescence Spectrometric Determination of DNA

对阳离子荧光染料酚藏花红 (PF)在阴离子表面活性剂溶液状态的吸收光谱和荧光光谱进行了研究。结果表明 ,当阴离子表面活性剂十二烷基硫酸钠 (SDS)的浓度在 0 .8× 10 -3 ～ 2 0× 10 -3 mol/L范围内时 ,酚藏花红形成非荧光现场二聚体 ,且在 5 15nm处出现新的特征吸收峰。现场形成的酚藏花红二聚体作为荧光探针的实验表明 ,核酸可使体系的荧光强度有很大的增强 ,且其增强的程度与核酸加入量呈明显的线性关系 ,据此建立了核酸定量分析的新方法。在最优化条件下 ,用于小牛胸腺DNA、鲱鱼精DNA、酵母RNA的定量测定 ,其线性范围分别为 0 0 6～ 18mg/L、0 0 4～ 2 0mg/L和 0 12～ 2 0mg/L ;检测限分别为 2 1、17和36 μg/L。

The fluorescence spectra and visible spectra of a cationic fluorescent dye, phenosafranine(PF), in the solution of anionic surfactants has been studied. A non-fluorescent dye dimer with a new characteristic peak at 515 nm was formed when the concentration of anionic surfactant was in the range of 8.0×10 -4～ 2.0×10 -3 mol/L. The studies on the possibility of using the in situ dimmer of dye as a fluorescence probe showed that the fluorescence intensities increased significantly and varied linearly with the concentration of nucleic acid when appropriate amounts of nucleic acid was added, thus a new method for the determination of nucleic acid was realized. Upon optimal conditions, the linear range for ct-DNA, fs-DNA, and yeast-RNA was 0.06～18 mg/L, 0.04～20 mg/L and 0.12～20 mg/L, respectively; with detect limit being 21 μg/L, 17 μg/L and 36 μg/L, respectively.