本文選題：熒光　切入點(diǎn)：探針　出處：《吉林大學(xué)》2017年博士論文

【摘要】：近年來,熒光探針作為一種優(yōu)秀的化學(xué)檢測(cè)試劑,由于其具有高的靈敏度和選擇性、操作簡單、響應(yīng)迅速等優(yōu)點(diǎn),不僅適合于傳統(tǒng)實(shí)驗(yàn),還可用于生物體內(nèi)研究。尤其是在活體生物成像研究方面,熒光探針幾乎不對(duì)細(xì)胞造成損傷,而且還可以實(shí)現(xiàn)活體的可視化研究,能夠同時(shí)顯示出較高的空間和時(shí)間分辨能力。因此,設(shè)計(jì)和合成新穎的熒光探針,利用它們識(shí)別特定的分析物,具有重要的意義。在本論文中,我們?cè)O(shè)計(jì)合成了基于不同原理的、可用于檢測(cè)Cu~(2+)、Hg~(2+)、Cd~(2+)和ClO~-的三種新型熒光探針,主要研究內(nèi)容如下:(1)設(shè)計(jì)合成了一種對(duì)于Cu~(2+)具有識(shí)別能力的新型羅丹明衍生物(RhS)。該熒光探針可用于測(cè)定飲用水和活體細(xì)胞中Cu~(2+)的濃度,并且具有可逆性。其原理為Cu~(2+)與RhS發(fā)生配位,RhS的螺酰胺環(huán)發(fā)生開環(huán)反應(yīng),導(dǎo)致其熒光發(fā)光強(qiáng)度增加,而其他金屬離子無此性能,因此該探針對(duì)于Cu~(2+)具有較高的選擇性。通過Job's點(diǎn)曲線和質(zhì)譜(MS)確定了RhS和Cu~(2+)配位的化學(xué)計(jì)量比為1:1,該實(shí)驗(yàn)結(jié)果同時(shí)得到了密度泛函(DFT)理論計(jì)算的支持。該探針對(duì)于測(cè)定Cu~(2+)的線性范圍和檢出限分別為0.50~20.00和0.11μmol L-1。飲用水樣品的分析結(jié)果顯示該探針檢測(cè)Cu~(2+)的回收率為100.4~101.2%,此外,探針還顯示了其在活體He La細(xì)胞中用于監(jiān)測(cè)Cu~(2+)的潛力。(2)基于熒光共振能量轉(zhuǎn)移(FRET)機(jī)理,設(shè)計(jì)合成了一種可用于測(cè)定中性水樣品中Hg~(2+)和監(jiān)測(cè)活細(xì)胞內(nèi)Hg~(2+)水平的熒光探針DRh。通過UV-Vis和熒光光譜實(shí)驗(yàn),考察了探針DRh對(duì)于金屬陽離子的選擇性,結(jié)果表明探針DRh能夠?qū)g~(2+)進(jìn)行高選擇性的識(shí)別。通過Job's點(diǎn)曲線和質(zhì)譜(MS)確定了DRh和Hg~(2+)配位的化學(xué)計(jì)量比為1:1,并結(jié)合密度泛函(DFT)和含時(shí)密度泛函(TD-DFT)理論計(jì)算研究了配位和發(fā)光機(jī)理。該探針對(duì)于測(cè)定Hg~(2+)的線性范圍和檢出限分別為0~30.00和0.008μmol L-1。中性水樣品的分析結(jié)果顯示檢測(cè)Hg~(2+)的回收率為104.5~107.9%。此外,該探針還應(yīng)用于活體HeLa細(xì)胞中Hg~(2+)的成像研究。(3)設(shè)計(jì)合成了一種水溶性基于熒光素的增強(qiáng)型熒光探針TF。該探針可用于熒光選擇性的檢測(cè)生理pH條件下的ClO~-和Cd~(2+),熒光增強(qiáng)幅度分別為64倍和4倍,增強(qiáng)原理分別為氧化反應(yīng)和配位反應(yīng)。探針對(duì)于ClO~-和Cd~(2+)的檢出限分別為0.068μmol L-1和2.2μmol L-1。采用密度泛函(DFT)和含時(shí)密度泛函(TD-DFT)理論計(jì)算方法對(duì)ClO~-、Cd~(2+)與TF的結(jié)合方式與反應(yīng)機(jī)理進(jìn)行了研究。此外,該探針還顯示出了在活體細(xì)胞中用于監(jiān)測(cè)ClO~-和Cd~(2+)的潛力。
[Abstract]:In recent years, as a kind of excellent chemical detection reagent, fluorescent probe is not only suitable for traditional experiments, but also can be used in vivo because of its high sensitivity and selectivity, simple operation and rapid response.Especially in the study of in vivo biological imaging, fluorescent probes can hardly damage the cells, but also can realize the visual research in vivo, and can display high spatial and time resolution at the same time.Therefore, it is of great significance to design and synthesize novel fluorescent probes and use them to identify specific analytes.In this thesis, we have designed and synthesized three novel fluorescent probes based on different principles, which can be used for the detection of Cu~(2 and ClO-.The main research contents are as follows: (1) A novel Rhodamine derivative with recognition ability for Cu~(2 was designed and synthesized.The fluorescent probe can be used to determine the concentration of Cu~(2 in drinking water and living cells.The principle is that Cu~(2) reacts with the spiroamide ring of RhS, which leads to the increase of fluorescence intensity, but other metal ions do not, so the probe has a high selectivity for Cu~(2).The stoichiometric ratio of RhS and Cu~(2 is determined to be 1: 1 by Job's point curve and MS. The experimental results are supported by density functional theory.The linear range and detection limit of the probe for the determination of Cu~(2 were 0.50 渭 mol L ~ (-1) and 0.11 渭 mol L ~ (-1), respectively.The analysis of drinking water samples showed that the recovery rate of the probe for detecting Cu~(2 was 100.4 / 101.2. In addition, the probe also showed its potential for monitoring Cu~(2 in He-La cells in vivo, based on the fluorescence resonance energy transfer (FRET) mechanism.A fluorescent probe, DRH, which can be used to determine the level of Hg~(2 in neutral water samples and to monitor Hg~(2 in living cells, has been designed and synthesized.The selectivity of probe DRh to metal cations was investigated by UV-Vis and fluorescence spectra. The results showed that the probe DRh could recognize Hg~(2 with high selectivity.The stoichiometric ratio of DRh and Hg~(2 was determined to be 1: 1 by Job's point curve and MS). The coordination and luminescence mechanism were studied by means of density functional theory (DFT) and time-dependent density functional theory (TD-DFTT).The linear range and detection limit of the probe for the determination of Hg~(2 were 0 ~ 30.00 渭 mol L ~ (-1) and 0.008 渭 mol L ~ (-1), respectively.The analytical results of neutral water samples showed that the recovery of Hg~(2 was 104.5% 107.9%.In addition, the probe was applied to the imaging study of Hg~(2 in living HeLa cells. A water-soluble enhanced fluorescent probe based on fluorescein was designed and synthesized.The probe can be used for the selective fluorescence detection of ClO- and Cd~(2 at physiological pH. The fluorescence enhancement amplitude is 64 times and 4 times, respectively. The enhancement principle is oxidation reaction and coordination reaction, respectively.The detection limits for ClO- and Cd~(2 were 0.068 渭 mol L-1 and 2.2 渭 mol L-1, respectively.The binding mode and the reaction mechanism of ClOOO- (-) and TD-DFT2) with TF were studied by means of density functional theory (DFT) and time-dependent density functional theory (TD-DFTT).In addition, the probe shows the potential for monitoring ClO- and Cd~(2 in living cells.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2017
【分類號(hào)】：O657.3

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