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  本文選題：熒光探針　切入點：5-氨基熒光素　出處：《遵義醫(yī)學(xué)院》2017年碩士論文　論文類型：學(xué)位論文

【摘要】：目的:構(gòu)建基于熒光素的NO型熒光探針,并對其識別NO的性能進(jìn)行研究。方法:以4-硝基鄰二甲苯為起始原料,經(jīng)氧化、脫水、加成及還原等反應(yīng)制備5-氨基熒光素(L_1),再將其與4-(2-氨乙基)嗎啉反應(yīng)制備5-氨基熒光素嗎啉衍生物(L_2)。采用~1H-NMR,~(13)C-NMR,HR-MS,FT-IR及單晶衍射等現(xiàn)代分析技術(shù)對L_1和L_2的組成和結(jié)構(gòu)進(jìn)行表征。在Tris-HCl緩沖溶液中利用紫外-可見吸收光譜法和熒光分光光度法考察L_1和L_2識別NO的性能和機(jī)理。結(jié)果:經(jīng)~1H-NMR,~(13)C-NMR,HR-MS,FT-IR及單晶衍射確證L_1和L_2均為預(yù)期目標(biāo)化合物。它們均有選擇性識別NO的性能。L_1是以熒光增強(qiáng)脫氨基化對NO進(jìn)行識別,其熒光強(qiáng)度與一氧化氮供體NOC-18在0-8.0×10~(-5) mol/L濃度范圍內(nèi)呈現(xiàn)良好的線性關(guān)系;研究范圍內(nèi)的活性氧和氮物種中Cl O-對L_1識別NO稍有干擾,而抗壞血酸和脫氫抗壞血酸對L_1識別NO有較大干擾;L_1識別NO機(jī)理經(jīng)~1H-NMR與HRMS證實為脫氨基作用。細(xì)胞毒性實驗發(fā)現(xiàn):L_1對Raw 264.7細(xì)胞具有低毒性,并能在Raw 264.7細(xì)胞中實現(xiàn)對NO的熒光成像。L_2是以熒光猝滅脫氨基機(jī)理進(jìn)行熒光猝滅識別NO。L_2自身在紫外燈下呈藍(lán)色的熒光,在L_2與NO作用后呈現(xiàn)的是熒光猝滅的現(xiàn)象,并且其猝滅作用在研究范圍內(nèi)的活性氧和氮物種中受NO_3~-的影響。細(xì)胞毒性實驗顯示:L_2對Raw 264.7細(xì)胞具有低毒性。結(jié)論:構(gòu)建的兩個5-氨基熒光素衍生物L(fēng)_1和L_2均通過脫氨基反應(yīng)實現(xiàn)對NO的熒光探測,L_1為熒光增強(qiáng)型NO熒光探針,L_2為熒光猝滅型NO熒光探針。
[Abstract]:Objective: to construct no fluorescent probe based on fluorescein and study its ability to recognize no. Methods: 4-nitro-o-xylene was oxidized and dehydrated. 5-aminofluorescein L _ (1) was prepared by addition and reduction reactions, and then synthesized by reaction of 5-aminofluorescein morpholine with 4-( 2-aminoethyl) morpholine. The composition and structure of L1 and L2 were analyzed by modern analytical techniques such as 1H-NMR-13C-NMR-MSFT-IR, single crystal diffraction and other modern analytical techniques, such as the addition and reduction of 5-aminofluorescein and the synthesis of 5-aminofluorescein morpholine derivatives. The performance and mechanism of no recognition by L _ (1) and L _ (2) in Tris-HCl buffer solution were investigated by UV-Vis absorption spectrometry and fluorescence spectrophotometry. Results: 1H-NMR-13C-NMR-HR-MSFT-IR and single crystal diffraction confirmed that L _ 1 and L _ S _ 2 were expected target compounds. They all have the ability of selective recognition of no. LS-1 is used to recognize no by fluorescence enhanced deamination. The fluorescence intensity showed a good linear relationship with the concentration of nitric oxide donor NOC-18 in the range of 0-8.0 脳 10 ~ (-5) mol/L. However, ascorbic acid and dehydroascorbic acid have great interference on the recognition of no in L1. The mechanism of no recognition in L1 was confirmed by 1H-NMR and HRMS. The fluorescence imaging of no in Raw 264.7 cells. LST2 is a fluorescence quenching mechanism to recognize the blue fluorescence of NO.L_2 under ultraviolet lamp, and the phenomenon of fluorescence quenching after the interaction between L2 and no. The cytotoxicity test showed that the two 5-aminofluorescein derivatives L1 and L2 were of low toxicity to Raw 264.7 cells. Conclusion: both of the constructed 5-aminofluorescein derivatives L1 and L2 are common. Fluorescence detection of no by over deamination reaction: L1 is a fluorescence enhanced no fluorescence probe and L2 is a fluorescence quenching no fluorescence probe.
【學(xué)位授予單位】：遵義醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：O657.3

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本文編號：1601874