Syntheses And Applications Of Rhodamine, Fluorescein, Nile Red And Nile Blue Derivatives

Based on rhodamine, fluorescein, nile red and nile blue in this thesis, respectively, rhodamine derivatives were designed and synthesized in order to modificate their fluorescence quantum yield, the large Stokes shift and incorporate reactive groups. Fluorescein derivatives were designed and synthesized in order to modificate pH-sensitive range, light-sensitive group, binding specificity and the large Stokes shift. Nile red and nile blue derivatives were designed and synthesized in order to modificate fluorescence quantum yield and water solubility. Then fluorescence properities of these derivatives were investigated by UV-vis spectra and fluorescence spectra, and the better fluorescent probes were selected and applied in the preparation of fluorescent microspheres,the determination of nitrite in water and the detection of polyhydroxyalkanoates of bacteria.Allyl rhodamine B, allyl fluorescein, allyl nile red, asymmetrical rhodafluor and pyrrolidinyl rhodamine S were designed and synthesized. And then they were used in the preparation of polystyrene-based fluorescent microspheres by the dispersion copolymerization and absorption method. These fluorescent microspheres were characterized by environmental scanning electron microscopy, laser scanning confocal microscopy, fluorescence spectrophotometry and fourier transform infrared spectrometer. They exhibited good dispersion and stable and high fluorescence intensity. Spectral properties of copolymerization fluorescent microspheres in tetrahydrofuran indicated these individual characteristics of labels should be maintained in the fluorescent microspheres. The differences of the fluorescent spectra between five fluorescent microspheres and their corresponding parent labels in ethanol had been investigated. Furthermore, copolymerization fluorescent microspheres were functionalized with amino groups, and the amount of amino groups was determined.A series of asymmetrical rhodamine derivatives were prepared and selected the better fluorescent probes. They were N steeled asymmetrical rhodamine containing 7-hydroxy-1,2,3,4-tetrahydroquinoline, which had a high fluorescence quantum yield and large Stokes shift. They can react with nitrite in an acidic medium to form a nitroso product, which had much lower fluorescence. Therefore, the method for trace nitrites was developed, which was simple, sensitive and selective. This method was applied to the determination of nitrite in tap water and lake water with satisfactory results.Two kinds of xanthamide fluorescein derivatives with active groups were prepared, which can be used in biological analysis. The results of their fluorescent properties showed that they can offer a wide variety of absorption and emission maximums. And they had excellent binding specificity and photostability and large Stokes shift. In addition, they can be employed as a biological fluorescence tag in acidic, neutral and alkaline environments.Rhodamine derivatives with chlorine, bromine and carboxyl group on the bottom ring were prepared. On this basis, the complicated rhodamine derivatives with reactive group can be designed from them, which would be suitable for biological detection.1-(Fluorescein-5-yl)-2,4,6-trimethylpyridinium perchlorate with positive charge was prepared, it can combine with biomolecules with negative charge through electrostatic attraction. It would be applied in biological applications by the fluorescence analysis.Nile red and nile blue derivatives with carboxyl or amino or hydroxyl group were synthesized, which were used to tag biological molecules. The dicarboxylic acid nile red had the better yield than that was reported, nile blue derivatives were found to be clear pH-dependent and could be used as acid-base indicators to measure intracellular pH. In addition, the dicarboxylic acid nile red and nile blue with good solubility in aqueous media were used to develop a simple, quick, safe, highly sensitive staining method to detect polyhydroxyalkanoates of bacteria, which was confirmed by fluorescence images of polyhydroxyalkanoates granules of bacteria.