發(fā)布時間：2018-04-04 12:42

  本文選題：熒光探針　切入點：激發(fā)態(tài)分子內(nèi)質(zhì)子轉(zhuǎn)移　出處：《浙江大學》2017年博士論文

【摘要】：熒光探針檢測具有靈敏度高、選擇性好、響應迅速、檢測范圍寬、操作簡單方便等優(yōu)點,尤其是其無可比擬的實時性這一特點,結(jié)合熒光成像技術,熒光探針可以用于活細胞及活體中生物分子及生物過程的原位實時無損傷監(jiān)控,因而在有機化學、高分子化學、材料化學、生命科學和醫(yī)藥等各個領域引起了廣泛的關注和研究。本論文對探針分子進行了詳細的描述和研究,通過合理的設計,簡單、成熟的合成,詳細、全面的表征,探索了一系列探針分子的性質(zhì)及其在小分子檢測方面的應用。結(jié)合文獻報道和本課題組之前的工作,我們先是基于激發(fā)態(tài)分子內(nèi)質(zhì)子轉(zhuǎn)移(ESIPT)機理和氟離子與硅反應引起硅-氧鍵斷裂的反應設計合成了兩個熒光探針SiOPI和SiOPIC。兩個探針分子展現(xiàn)出了對氟離子快速而優(yōu)異的靈敏度和選擇性,尤其是SiOPI分子,有效屏蔽了氫氧根的干擾,這一點是氟離子檢測領域長久以來的難點之一。通過1HNMR、29SiNMR和ESI-MS對檢測過程的跟蹤,我們推測出了 SiOPI與氟離子的兩階段反應機理,很好的解釋了在氟離子滴定時其光物理性質(zhì)的變化過程,與我們觀察到的現(xiàn)象完美契合。這兩個階段包括硅-氧鍵斷裂并形成t-BuPh2SiF2-的過程,以及之后過量氟離子存在時去質(zhì)子化的過程。當SiOPI的硅-氧鍵斷裂生成HOPI分子,其ESIPT機理便被激活,可以觀察到一個明顯的熒光紅移過程。因此這兩種探針分子均可用于氟離子檢測,特別是SiOPI分子,展現(xiàn)出了很好的比率熒光的特性。比率熒光探針的選擇性和靈敏度要明顯高于普通探針分子,且受到外部環(huán)境的影響更小,測量數(shù)據(jù)更加準確。接著,我們介紹了兩個基于ESIPT機理和光誘導電子轉(zhuǎn)移(PET)機理設計合成的過氧化氫探針分子P1和P2,通過對其性能研究,我們發(fā)現(xiàn)這兩個探針可用于在堿性環(huán)境中快速檢測過氧化氫。P1的發(fā)色團HPNO的熒光發(fā)射光譜因其ESIPT機理而受到溶劑性質(zhì)的顯著影響。在二氯甲烷中,HPNO表現(xiàn)出雙波長發(fā)射;當溶劑極性上升,在質(zhì)子性溶劑如甲醇、DMF中其酮式異構(gòu)體的發(fā)射峰減弱,烯醇式的熒光發(fā)射占主導作用;而其余三個分子在不同溶劑中則基本不受影響。接著,我們又研究了 P1和P2的發(fā)色團HPNO和B-HBO在堿性環(huán)境中去質(zhì)子化的熒光性質(zhì)。在體系中加入NaOH后,二者均發(fā)生熒光發(fā)射強度的改變,并分別伴隨有紅移或藍移,B-HBO甚至可以觀察到溶液顏色的變化,表明該分子在pH比率檢測方面有著潛在的應用。而P1和P2則表現(xiàn)出了對堿性條件良好的耐受性,其吸收和熒光發(fā)射光譜基本沒有變化。隨后我們研究了 P1和P2在堿性環(huán)境下對過氧化氫的傳感,發(fā)現(xiàn)他們均能很好的檢測過氧化氫。P1在堿性環(huán)境下對過氧化氫的響應表現(xiàn)為比率熒光改變,而P2則表現(xiàn)為熒光猝滅。結(jié)合文獻報道及我們的分子模擬計算結(jié)果,我們推測P2發(fā)生了 PET過程導致其熒光猝滅。兩個探針均顯示出了對過氧化氫優(yōu)異的敏感性和選擇性。此外,其明顯優(yōu)于其他探針分子的響應速率,為過氧化氫檢測提供了新的解決方案。與P1相比,P2展現(xiàn)出了更好地靈敏度,而且是有報道以來的第一例BODIPY用于過氧化氫檢測的熒光探針分子。文章針對氟離子和過氧化氫檢測的特點,設計合成了一些探針分子,并以此為基礎對這些化合物進行了大量的表征工作,并對其在相應分子檢測方面的應用性進行了研究。這些工作不僅豐富了有機熒光化合物的種類,更為有機熒光探針分子在分子傳感領域的應用提供了新的思路和解決方案。
[Abstract]:Fluorescence detection with high sensitivity, good selectivity, fast response, wide detection range, has the advantages of simple and convenient operation, especially the characteristics of the real-time fluorescence imaging technology combined with There is nothing comparable to this, and the fluorescent probe can be used for in situ biological molecules and biological processes of living cells and in vivo in real-time monitoring without damage, so in organic chemistry, polymer chemistry, materials chemistry, life sciences and medicine has aroused widespread concern and research. This paper makes a detailed study on the probe molecule, through reasonable design, simple and mature synthesis, detailed, comprehensive characterization, explore the properties of a series of probe molecules and its application in small molecule detection. Combined with the literature and our previous work, we first excited state intramolecular proton transfer (ESIPT) mechanism based on silicon and fluorine ion and reaction of lead Reaction design of silicon oxygen bond rupture of the synthesized two fluorescent probes SiOPI and SiOPIC. two probe molecules show for fluoride fast and excellent sensitivity and selectivity, especially SiOPI molecules, effectively shielding the hydroxyl interference, this point is one of the difficulties in the field of fluorine ion detection. Through the long long 1HNMR, 29SiNMR and ESI-MS on the detection process of tracking, we conclude that the two stage of the reaction mechanism of SiOPI and fluoride, a good explanation of the changes in the photophysical properties of fluorine ion titration, and we observed the phenomenon of perfect fit. These two stages including silicon oxygen bond rupture and the formation process of t-BuPh2SiF2-, and then there is an excess of fluoride deprotonation. When SiOPI silicon oxygen bond cleavage to generate HOPI molecules, the ESIPT mechanism is activated, can be observed an obvious fluorescence redshift. Therefore, these two kinds of probe molecules can be used for the detection of fluoride ion, especially SiOPI molecules, showing a good ratio of fluorescence. The ratio of fluorescence probe selectivity and sensitivity was higher than that of ordinary probe molecules, and influenced by the external environment of the smaller, more accurate measurement data. Then, we introduce two based on the mechanism of ESIPT and photoinduced electron transfer (PET) mechanism design and synthesis of hydrogen peroxide probe molecules P1 and P2, the performance of research, we find that the two probe can be used for spectral properties affected by the solvent due to its mechanism of ESIPT fluorescence detection of hydrogen peroxide.P1 chromophore HPNO fast in alkaline environment in dichloromethane. In HPNO, showed a dual wavelength emission; when the polarity of the solvent increased in protic solvents such as methanol, DMF emission peak of the keto isomer decreased, fluorescence emission accounted for the enol A leading role; while the remaining three molecules in different solvents is basically unaffected. Then, we also studied the fluorescence properties of P1 and P2 HPNO and B-HBO chromophore deprotonated in the alkaline environment. Adding NaOH in the system, two occurred in the fluorescence emission intensity change, and were accompanied by red shift or a blue shift, B-HBO can even observed color change, show that the molecules have potential applications in pH ratio detection. P1 and P2 showed good tolerance to alkaline conditions, the absorption and fluorescence emission spectra did not change. Then we study the P1 and P2 in alkaline environment the hydrogen peroxide sensor, found that they were good.P1 detection of hydrogen peroxide in alkaline environment in response to hydrogen peroxide showed fluorescence and P2 ratio change, there is fluorescence quenching. Combined with the literature and our points The sub simulation results, we speculate that P2 PET process resulted in the fluorescence quenching. The two probes showed excellent sensitivity and selectivity to hydrogen peroxide. In addition, the response rate was significantly better than the other probe molecules, which provides a new solution for hydrogen peroxide detection. Compared with P1, P2 showed better the sensitivity, and it is the first case of BODIPY reported since the fluorescent probe molecules for hydrogen peroxide detection. According to the characteristics of fluorine ion and hydrogen peroxide detection, some probes were designed and synthesized, and as a basis for the characterization of many of these compounds, and the corresponding molecular detection applications were studied the work not only enriches the types of organic fluorescent compounds, it provides new ideas and organic fluorescent probes used in molecular sensing fields Solution.

【學位授予單位】：浙江大學
【學位級別】：博士
【學位授予年份】：2017
【分類號】：O657.3

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本文編號：1710000