The Synthesis And Properties Of Nitric Oxide Fluorescence Probes

As an informative molecule, Nitric oxide widely exits in the cells of living organisms, and directly involves in the transmission of information in physical activities. Because Nitric oxide research has become more and more necessary and nitric oxide itself does not have fluorescence signals, it is necessary for fluorescence analysis to have the fluorophore through the introduction of reagents and reaction with the fluorophore itself different in optical properties of certain compounds. By changing the optical properties of fluorophores, the qualitative and quantitative determination of nitric oxide is realized. The paper is divided into five chapters on nitric oxide including the principles of design, classification, synthesis and properties of the study. In the first chapter, the paper overviewes the design principles, classification of nitric oxide fluorescence probe, and introduces the research status and research progress.In the second chapter, the amino-fluorescein and 1,4,8,11-tetraaza tetradecane were synthesized to Cu complexes and characterized by IR, NMR, UV-vis and fluorescence spectra. In 0.1 mol/L pH 7.40 Tris-HCl buffer solution, it was carried out:（1） Adding NOC-18 for experimental comparison; （2） NO concentration on the sensitivity of fluorescent probes; （3） Testing the selectivity of nitric oxide probe and the pH of such experiments to test the performance of a fluorescent probe. After testing, whenλex = 465 nm ,we found the fluorescent probe exhibits a maximum absobption on the wavelength of 522 nm and maximum emission wavelength in the pH 4.5; When H₂O₂, ONOO- and OH- were in the presence of testing, Fluorescent probe demonstrated good selectivity and results well.In the third chapter, dichlorofluorescein and o-phenylenediamine were synthesized to lactam and characterized by IR, NMR, UV-vis and fluorescence spectra. Then the lactam reacted with nitric oxide for dichlorofluorescein benzotriazole which can be decomposed into dichlorofluorescein and benzotriazole in aqueous solution. Because benzotriazole has a strong fluorescent itself, it were often used to detect NO concentration, which can qualitative and quantitative. In the 0.1mol/L pH 7.40 PBS buffer solution, it was carried out: （1） Testing of adding nitric oxide releasing agent before and after the change of fluorescence intensity of the reaction; （2） Under testing of changes reaction time in response to nitric oxide changes in the fluorescence intensity ;（3） Testing under different reaction pH changes of fluorescence intensity. After testing, the discovery of the fluorescent probe had a maximum emission wavelength withλex = 521nm, at excitation wavelength withλem = 544nm. And fluorescent probes in H₂O₂, ONOO-and OH- solution, its showing good selectivity and experimental results well.In the fourth chapter, o-phenylenediamine and Eosin Y were synthesized to Eosin Y lactam and characterized by IR, NMR, UV-vis and fluorescence spectra. In 0.1 mol/L pH 7.40 in the PBS buffer solution, it was carried out: （1） Adding nitric oxide releasing agent before and after the change of fluorescence intensity of the reaction; （2） Different nitric oxide concentration on the reaction; （3） Fluorescent probe selection of other experiments. After testing, the discovery of new fluorescent probe were test with a large excitation and emission wavelengths （λex /λem: 553/574 nm）. When the coexistence of NO3-, H₂O₂ and nitric oxide in solution, the lactam showed good selectivity to nitric oxide.In the last chapter, it was summarized for the whole thesis.