水溶性双功能荧光探针合成与应用研究
发布时间：2018-10-08 19:30
【摘要】：近年来,荧光检测法因其具有操作简便、灵敏度高、实时快速分析等优点,广泛应用于环境检测分析和生物成像等领域。其中,具有良好水溶性的多功能荧光探针能够高选择性检测复杂体系中两种或以上分析物已成为研究热点。因此,本论文中我们设计合成了三种新颖的水溶性双功能荧光探针,分别实现了对铜离子(Cu~(2+))和硫离子(S~(2-))、四氢呋喃中水含量和pH值、次氯酸根(ClO~-)和温度的快速、灵敏、选择性的荧光检测。本论文的主要内容和结果如下:1.基于荧光能量共振转移(FRET)原理,我们将异硫氰酸荧光素(FITC)共价键连接到支化聚乙烯亚胺(b-PEI)分子上,制得了一种水溶性双功能荧光聚合探针(PF)。PF能够选择性地络合Cu~(2+)生成PF-Cu~(2+)络合物,PF体系荧光减弱,这是由于荧光素(给体)和PF-Cu~(2+)络合物(受体)之间发生了有效地FRET。当向上述PF-Cu~(2+)体系中加入S~(2-),S~(2-)和Cu~(2+)结合形成稳定的CuS沉淀物,PF体系荧光恢复,实现了对Cu~(2+)和S~(2-)高选择性检测。此外,该探针PF被成功应用于自来水和人血清中Cu~(2+)和S~(2-)的高灵敏度荧光检测。2.以对苯二胺为原料,采用水热法,我们合成了一种水溶性双功能红色荧光碳量子点(R-Cdots)。随着pH的增大(2.0~9.0),R-Cdots在610 nm处的荧光逐渐增大,并且在5.0~9.0之间,荧光强度值和pH有较好的线性关系,表明R-Cdots能够灵敏地检测复杂生物样品的pH。此外,R-Cdots具有较强的溶剂极性依赖性,其荧光发射波长随溶剂极性增大而发生红移(~80 nm)。随着四氢呋喃中的水含量增加,其荧光发射波长发生红移且荧光强度减弱(绿色变为红色),检测限低至0.04%(体积分数),实现了对四氢呋喃中水含量的荧光定量检测。3.我们将异硫氰酸罗丹明B通过共价键的方式连接到荧光硅纳米粒子表面氨基上,制得一种水溶性双功能比率型荧光纳米探针(DRFS)。当向DRFS溶液中加入ClO~-,硅纳米粒子的荧光(445 nm)减弱,而罗丹明B的荧光(576 nm)基本保持不变,从而实现了对ClO~-高选择性荧光检测。DRFS不仅可以测定人血清中ClO~-的浓度,而且还能够对细胞中外源性和内源性的ClO~-进行荧光成像。另外,DRFS也可作为一种温度荧光传感器。随着温度的升高,罗丹明B的荧光(576 nm)逐渐下降,而硅纳米粒子的荧光(445 nm)基本保持不变,并且在20℃~60℃之间,荧光强度比值(I576/I445)和温度有较好的线性关系,表明DRFS能够定量监测生物体系中的温度变化。
[Abstract]:In recent years, fluorescence detection method has been widely used in the field of environmental detection and biological imaging because of its advantages of simple operation, high sensitivity, real-time rapid analysis and so on. Among them, multifunctional fluorescent probes with good water solubility can detect two or more analytes in complex systems with high selectivity. Therefore, we have designed and synthesized three novel water-soluble bifunctional fluorescent probes in this thesis. We have achieved the sensitivity and rapidity of water content and pH value, hypochlorite (ClO~-) and temperature of copper ion (Cu~ (2) and sulfur ion (S ~ (2), tetrahydrofuran), respectively. Selective fluorescence detection. The main contents and results of this thesis are as follows: 1. Based on the principle of fluorescence energy resonance transfer (FRET), we connect the covalent bond of fluorescein isothiocyanate (FITC) to the branched polyvinyleneimine (b-PEI) molecule. A water-soluble bifunctional fluorescent polymerization probe (PF) 路PF was prepared, which can selectively complexate Cu~ (2) to form PF-Cu~ (2) complex. The fluorescence of PF-Cu~ (2) complex / PF system is weakened, which is due to the effective FRET. between fluorescein (donor) and PF-Cu~ (2) complex (receptor). When S2- and Cu~ (2) were added to the PF-Cu~ (2) system to form a stable CuS precipitate, the fluorescence recovery was achieved, and the high selectivity detection of Cu~ (2) and S2- was realized. In addition, the probe PF was successfully applied to the detection of Cu~ (2) and S2- in tap water and human serum. A water-soluble bifunctional red fluorescent carbon quantum dot (R-Cdots) was synthesized from p-phenylenediamine by hydrothermal method. With the increase of pH (2.0 ~ 9.0), the fluorescence of R-Cdots at 610 nm increases gradually, and the fluorescence intensity has a good linear relationship with pH in the range of 5.0 ~ 9.0, which indicates that R-Cdots can detect pH. of complex biological samples sensitively. In addition, R-Cdots have strong solvent polarity dependence, and their fluorescence emission wavelengths shift red with the increase of solvent polarity (80 nm). With the increase of water content in tetrahydrofuran, the fluorescence emission wavelength shifts red and the fluorescence intensity decreases (green to red), the detection limit is as low as 0.04% (volume fraction), and the fluorescence quantitative detection of water content in tetrahydrofuran is realized. We have connected Rhodamine isothiocyanate B to the surface amino of fluorescent silicon nanoparticles by covalent bonding to prepare a water-soluble bifunctional fluorescent nano-probe (DRFS). When the fluorescence (445 nm) of ClO~-, silicon nanoparticles was weakened in DRFS solution, but the fluorescence of Rhodamine B (576 nm) remained unchanged, the highly selective fluorescence detection of ClO~- could not only determine the concentration of ClO~- in human serum. Moreover, fluorescent imaging of exogenous and endogenous ClO~- in cells is also possible. In addition, DRFS can also be used as a temperature fluorescence sensor. With the increase of temperature, the fluorescence of rhodamine B (576 nm) decreased gradually, while the fluorescence of silicon nanoparticles (445 nm) remained unchanged, and the fluorescence intensity ratio (I576/I445) had a good linear relationship with temperature between 20 鈩，
本文编号：2258002
[Abstract]:In recent years, fluorescence detection method has been widely used in the field of environmental detection and biological imaging because of its advantages of simple operation, high sensitivity, real-time rapid analysis and so on. Among them, multifunctional fluorescent probes with good water solubility can detect two or more analytes in complex systems with high selectivity. Therefore, we have designed and synthesized three novel water-soluble bifunctional fluorescent probes in this thesis. We have achieved the sensitivity and rapidity of water content and pH value, hypochlorite (ClO~-) and temperature of copper ion (Cu~ (2) and sulfur ion (S ~ (2), tetrahydrofuran), respectively. Selective fluorescence detection. The main contents and results of this thesis are as follows: 1. Based on the principle of fluorescence energy resonance transfer (FRET), we connect the covalent bond of fluorescein isothiocyanate (FITC) to the branched polyvinyleneimine (b-PEI) molecule. A water-soluble bifunctional fluorescent polymerization probe (PF) 路PF was prepared, which can selectively complexate Cu~ (2) to form PF-Cu~ (2) complex. The fluorescence of PF-Cu~ (2) complex / PF system is weakened, which is due to the effective FRET. between fluorescein (donor) and PF-Cu~ (2) complex (receptor). When S2- and Cu~ (2) were added to the PF-Cu~ (2) system to form a stable CuS precipitate, the fluorescence recovery was achieved, and the high selectivity detection of Cu~ (2) and S2- was realized. In addition, the probe PF was successfully applied to the detection of Cu~ (2) and S2- in tap water and human serum. A water-soluble bifunctional red fluorescent carbon quantum dot (R-Cdots) was synthesized from p-phenylenediamine by hydrothermal method. With the increase of pH (2.0 ~ 9.0), the fluorescence of R-Cdots at 610 nm increases gradually, and the fluorescence intensity has a good linear relationship with pH in the range of 5.0 ~ 9.0, which indicates that R-Cdots can detect pH. of complex biological samples sensitively. In addition, R-Cdots have strong solvent polarity dependence, and their fluorescence emission wavelengths shift red with the increase of solvent polarity (80 nm). With the increase of water content in tetrahydrofuran, the fluorescence emission wavelength shifts red and the fluorescence intensity decreases (green to red), the detection limit is as low as 0.04% (volume fraction), and the fluorescence quantitative detection of water content in tetrahydrofuran is realized. We have connected Rhodamine isothiocyanate B to the surface amino of fluorescent silicon nanoparticles by covalent bonding to prepare a water-soluble bifunctional fluorescent nano-probe (DRFS). When the fluorescence (445 nm) of ClO~-, silicon nanoparticles was weakened in DRFS solution, but the fluorescence of Rhodamine B (576 nm) remained unchanged, the highly selective fluorescence detection of ClO~- could not only determine the concentration of ClO~- in human serum. Moreover, fluorescent imaging of exogenous and endogenous ClO~- in cells is also possible. In addition, DRFS can also be used as a temperature fluorescence sensor. With the increase of temperature, the fluorescence of rhodamine B (576 nm) decreased gradually, while the fluorescence of silicon nanoparticles (445 nm) remained unchanged, and the fluorescence intensity ratio (I576/I445) had a good linear relationship with temperature between 20 鈩，
本文编号：2258002
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