以香豆素为母体专一性检测生物硫醇及二氧化硫衍生物的荧光探针的设计合成及应用研究

发布时间：2018-04-11 12:02

本文选题：荧光探针 + 香豆素　；参考：《西北大学》2017年硕士论文

【摘要】：谷胱甘肽(GSH)与生物体中的氧化还原平衡状态、机体正常的生命活动息息相关;SO_2作为一种大气污染物,通过影响植物的光合作用进而影响植物的生长发育,其衍生物也被广泛的用作食品添加剂,经常被人们摄入体内。因此构建一种可定性定量,且专一性检测谷胱甘肽(GSH)及S02衍生物的检测方法是非常重要的。香豆素广泛存在于高等植物中,不但容易合成而且易于修饰,因具有高的荧光量子产率和稳定的光学性质而备受关注。本文基于荧光分析法,以香豆素为荧光团分别设计合成了两种专一性检测谷胱甘肽(GSH)及SO_2衍生物的荧光探针。主要内容如下:(1)以香豆素作为荧光团专一性检测谷胱甘肽(GSH)的荧光探针的合成及应用研究。根据文献报道,本文合成了专一性检测谷胱甘肽(GSH)的荧光探针A。从探针A的分子结构可以看出,其7-位存在推电子的N,N-二乙胺基,苯环3-位所连接的烯烃链上既存在吸电子的醛基又存在活泼的卤素C1原子,所以整个探针分子结构存在分子内电荷转移(intramolecular charge transfer,ICT)和卤素氯产生的重原子效应(the heavy atom effect),使其在550 nm处有较弱的荧光发射峰,当向探针A的溶液中加入谷胱甘肽(GSH)后,谷胱甘肽(GSH)中亲核性的氨基会进攻探针A上的醛基,由于生成的中间产物存在亲核性的巯基,巯基就会接着与其临近的氯原子发生亲核取代反应。上述反应不但破坏了探针A分子的ICT效应同时也打破了氯原子的重原子效应,故使溶液的荧光发射波长由550 nm蓝移到500 nm,反应体系在500 nm处的荧光强度与GSH的浓度在0-10 μM范围内具有良好的线性关系,该方法对GSH的检测非常灵敏,检出限为68.7 nM。(2)以苯并吡喃嗡为识别基团的HSO_3~-荧光探针的设计及应用研究。由于苯并吡喃嗡染料特别容易受到亲核试剂的进攻,从而打破了染料原来的π共轭结构,使分子内电荷重新排布,体系的发射波长发生变化。基于苯并吡喃嗡易受到亲核试剂进攻这一原理,本文设计合成了检测HSO_3~-的荧光探针B。探针B中的苯并吡喃嗡部分与香豆素荧光团之间存在分子内的光诱导电子转移过程(photo-induce electron transfer,PET),导致自身荧光很弱,当向探针B的溶液中加入亲核性的HSO_3~-时,HSO_3~-与苯并吡喃嗡中C-4原子发生亲核加成反应,使分子内电荷重新排布,PET效应消失,因此反应体系在510nm处的荧光信号显著升高。同时,上述反应使溶液的颜色也发生了显著地变化由蓝色变为黄色。探针B在30 s内就可以完成对HSO_3~-的检测,反应体系在510 nm处的荧光强度与HSO_3~-的浓度在0.2-7.5 μM范围内具有良好的线性关系,该方法对HSO_3~-的检出限为42nM,同时也成功的应用探针B检测了糖溶液和细胞中的HSO_3~-。
[Abstract]:Glutathione GSH (GSH) is closely related to the redox equilibrium in organism and the normal life activity of organism. As an atmospheric pollutant, GSH affects the growth and development of plants by affecting the photosynthesis of plants.Its derivatives are also widely used as food additives and are often ingested in the body.Therefore, it is very important to construct a qualitative, quantitative and specific method for the detection of glutathione glutathione (GSH) and S02 derivatives.Coumarin is widely found in higher plants, which is not only easy to synthesize but also easy to modify, and has attracted much attention due to its high fluorescence quantum yield and stable optical properties.Based on fluorescence analysis, two specific fluorescent probes for the detection of glutathione (GSH) and SO_2 derivatives were designed and synthesized using coumarin as fluorescence group.The main contents are as follows: 1) Synthesis and application of coumarin as fluorescent probe for glutathione glutathione glutathione (GSH) detection.A fluorescence probe A was synthesized for the specific detection of glutathione glutathione (GSH).From the molecular structure of probe A, it can be seen that there is a N-N- diethylamine group at the 7- site, and the alkene chain connected by the 3-site benzene ring has both an electron absorbing group and an active halogen C1 atom.So the whole molecular structure of the probe is characterized by intramolecular charge transfer charge transfer (ICTI) and the heavy atom effect produced by halogen chlorine, making it have a weaker fluorescence emission peak at 550nm, when glutathione glutathione is added to the solution of probe A).Nucleophilic amino groups in glutathione glutathione (GSH) attack the aldehyde group on probe A. due to the existence of nucleophilic sulfhydryl group in the generated intermediate product, the thiol group then reacts with its adjacent chlorine atom to produce a nucleophilic substitution reaction.The above reactions not only destroy the ICT effect of probe A, but also break the heavy atom effect of chlorine atom.Therefore, the fluorescence emission wavelength of the solution is shifted from 550 nm blue to 500 nm. The fluorescence intensity of the reaction system at 500nm has a good linear relationship with the concentration of GSH in the range of 0-10 渭 M. the method is very sensitive to the detection of GSH.The detection limit is 68.7 NM. 2) the design and application of HSOSP-fluorescence probe with benzopyran as the recognition group.Because the benzopyran dyes are especially vulnerable to the attack of nucleophilic reagents, the original 蟺 conjugate structure of the dyes is broken, the intramolecular charge is rearranged and the emission wavelength of the system changes.Based on the principle that benzopyran is easily attacked by nucleophilic reagents, a fluorescent probe B for the detection of HSO _ 3H _ 3- is designed and synthesized in this paper.The photo-induced electron transfer process between the benzopyran humming part of probe B and the coumarin fluorescence group leads to a very weak self-fluorescence.At the same time, the color of the solution changed significantly from blue to yellow.Probe B can detect HSO3- within 30 seconds. The fluorescence intensity of the reaction system at 510 nm has a good linear relationship with the concentration of HSO3- in the range of 0.2-7.5 渭 M.The detection limit of HSO3- for HSO3- was 42 nM.The probe B was also successfully used to detect HSOS3- in sugar solution and cells.
【学位授予单位】：西北大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：O657.3

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