核/壳量子点纳米复合物的制备及其可视化荧光识别研究
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摘要
双荧光发射纳米复合材料的制备及其分析应用近年来引起了人们极大的兴趣。量子点（Quantum dots, QDs）具有独特的荧光性质，基于QDs制备的双荧光发射纳米复合材料在荧光分子成像、双荧光比率传感及可视化荧光测定领域具有独特的优势和发展潜力。本文研究了发不同荧光的QDs、不同形貌的金纳米结构的可控制备，在此基础上构建了核/壳QDs纳米复合物，探索了纳米复合物在环境和生物技术领域中的可视化荧光识别。主要内容如下：
1.三聚磷酸钠稳定的CdS QDs的制备及荧光猝灭法测定铁。以三聚磷酸钠(STPP)为稳定剂，在碱性条件下成功合成了分散较均匀、稳定性好、发射峰位可调、半峰宽25nm左右的水溶性CdS量子点。研究了STPP和硫脲的比例、反应时间及pH等条件对CdS QDs发光特性的影响。探讨了痕量Fe2＋离子对CdSQDs荧光的猝灭作用。考察了缓冲溶液、pH、共存离子等因素的影响。在最佳实验条件下,在0.02~1.2M范围内，483nm处F0/F与Fe2＋呈现良好的线性关系，检测限LOD(3/)为6.7nM。用于实际矿泉水样中Fe的测定，回收率95.0%-105.0%。
2. CdSe/CdS@SiQ2纳米复合物的制备及荧光猝灭法测定铜。首先，以STPP作为稳定剂，合成了水溶性CdSe和CdSe/CdS核/壳量子点；然后，通过正硅酸四乙酯(TEOS)水解成功制备了CdSe/CdS@SiQ2核壳/结构的纳米复合物，用荧光光谱、紫外-可见吸收光谱、XRD对其进行了表征。根据Cu~(2~+)会选择性猝灭CdSe/CdS@SiQ2纳米复合物的荧光，建立了CdSe/CdS@SiO_2纳米复合物荧光猝灭法测定水样中Cu~(2~+)的新方法。其线性范围为0.02~0.6μM，检测限为10nM。
3.介孔ZnO/CdS/SiO_2核/壳纳米复合物的制备及可视化荧光测定Ag＋。首先，以STPP为稳定剂，合成ZnO/CdS核/壳QDs，然后直接把ZnO/CdS QDs水溶液加入含有TEOS、氨水的乙醇溶液中，发展了一种新的制备具有介孔结构的核/壳型ZnO/CdS@SiO_2纳米粒子的新方法。Ag~+可以通过介孔与核内ZnO/CdSQDs作用，导致其带边发射猝灭，同时在595nm处的缺陷发光信号增强，在紫外灯的照射下发出红色荧光。根据紫外灯照射下Ag~+致使ZnO/CdS@SiO_2红色荧光增强，我们用同一种量子点的双色荧光的变化实现对Ag~+的可视化、选择性测定，测定下限为0.04μM。根据595nm处Ag~+与ZnO/CdS@SiO_2作用的荧光增强建立了高选择性和高灵敏测定Ag~+的新方法，线性范围为0.03~0.24μM，检测限(3)为3.30nM。
4. CdTe@SiO_2/Au纳米复合物的制备及可视化多色荧光测定Hg~(2~+)。以壳聚糖为稳定剂和还原剂成功合成了表面等离子体共振吸收峰位可调的金纳米盘。在壳聚糖稳定的球状金纳米溶液中加入半胱氨酸，发现球状金纳米粒子可组装成金纳米链。然后，以溶菌酶为稳定剂和还原剂，合成了具有双荧光（蓝、红）发射的金纳米簇(Au CNs)。通过扫描电镜(SEM)、透射电镜(TEM)、X射线衍射(XRD)等表征手段对晶体结构和形貌进行了解析；通过动态光散射、共振散射光谱、吸收光谱和荧光光谱确证Au纳米结构在溶液中形成，并讨论了金纳米结构的生长机理。把具有双荧光（红、蓝）发射性质的Au CNs和具有绿色荧光发射性质的CdTe@SiO_2混合，制备了在同一激发下能发射三色荧光（红、绿、蓝，RGB）的CdTe@SiO_2/Au纳米复合物。研究了Hg~(2~+)对CdTe@SiO_2/Au纳米复合物的RGB荧光的猝灭作用，发现随着Hg~(2~+)浓度的增加纳米复合物呈现由红橙、黄、绿、蓝的颜色变化。由此建立一种可视化多色荧光识别模式，裸眼测定Hg~(2~+)的含量。
Currently, the fabrication and applieations of the composite nanostructures withdual-emission fluorescence is of great interest。Combining the unique fluoreseenceproperties of Quantum dots (QDs), the QDs nanocomposites with dual-emissiondemonstrate great potentials in fluorescence molecular imaging, dual-fluorescenceratio sensor, and visual fluorescence detection. In this thesis, controllable fabricationof QDs with different fluorescence color and Au nanopartieles has been investigated,and then Core/Shell QDs nanocomposites have been faeilely fabrieated, we have alsoinvestigated the visual fluorescent recognition of such nanoeomposites in the field ofbiotechnology and the environment. The main points are as follows:
1. The fabrication of STPP-stabilized CdS QDs and the determination of traceiron by fluorescence quenching method. The uniform dispersion of water-soluble CdSQDs stabilized by STPP was successfully synthesized in an alkaline medium. Theemission peaks of CdS QDs were tunable, and the half peak width is about25nm.The fluorescence spectral properties of CdS QDs at different pH values, thiourea,STPP and reaction time were investigated. A fluorescence quenching method for thedetermination of trace iron (Ⅱ) was developed, and the factors involving pH, buffersolution, and coexistence ion were investigated. Under the optimum conditions, thevalues of F0/F at483nm are linearly proportional to the concentration of Fe~(2~+)from0.02-1.2μM. The detection limit of the developed method was6.7nM for Fe~(2~+).Mineral water samples were analyzed with the recovery of95.0%-105.0%.
2. The fabrication of the CdSe/CdS@SiQ2core/shell nanocomposites and thedetermination of trace Cu~(2~+)by fluorescence quenching method. Water-soluble CdSeand CdSe/CdS core/shell quantum dots were successfully syntheticed in presence ofSTTP as a stabilizer. CdSe/CdS@SiQ2core-shell nano composites were obtained viathe hydrolysis of Tetraethyl silicate (TEOS). The fluorescence spectra,ultraviolet-visible absorption spectroscopy (UV), TEM and XRD were used toquantify the structure and characteristions of CdSe/CdS@SiQ2nanocomplexes. Dueto Cu~(2~+)can cause fluorescent quenching of CdSe/CdS@SiQ2nanocomplexes selectively, we set up a new method for the determination of Cu2~+in water samples.The linear range from0.02to0.6μM and the detection limitation is10nM.
3. The fabrication of the mesoporous ZnO/CdS@SiO_2Core/Shell nanostructureand visual fluorescence detection of Ag~+. A new method for the preparation ofmesoporous ZnO/CdS@SiO_2Core/Shell nanostructure has been developed. Themesoporous silica shells allow Ag~+to enter into the interior of the nanostructures tocontact with ZnO/CdS core, accordingly causes the quenching of its band edgeemission along with a simultaneous enhancement of red emission at around595nm.So, a novel visual fluorescence detection strategy for Ag~+ion is proposed based on acommon Core/Shell Quantum dots nanostructure. Under optimal conditions, theenhanced fluorescence intensity at595nm increased linearly with the concentrationof Ag~+ion ranging from0.03μM to0.24μM with a detection limit (3σ) of3.3nM.
4. The fabrication of the CdTe@SiO_2/Au nanocomposites and visualmulti-colored fluorescent detection of Hg~(2~+). By chitosan as a stabilizer and reducingagent, the tunable surface plasmon resonance absorption peak of gold nanoplate hasbeen successfully fabricated. It was observed that the linear chains of Au nanosphereswere formed by the addition of L-Cysteine in CTS-stabilized Au nanospheressolution. After that, highly fluorescent Au clusters with dual-emission (blue and red)have been synthesized by lysozyme as stabilized and reducing agents. The structureand morphology of the three types of crystals was characterized via scanning electronmicroscopy (SEM), transmission electron microseopy (TEM) and X-ray diffraction(XRD). The formation of Au nano-structure was determined by the use of UV-vis,dynamic light scattering as well as resonance light scattering and fluorescence spectra.The growth mechanism of gold nano-structure was discussed. The CdTe@SiO_2/Aunanocomposite with three-color fluorescence (red, green and blue, RGB) under asingle excitation wavelength was synthesized when the Au CNs with dual-emission(blue and red colors) and CdTe@SiO_2with green fluorescent were mixed. Thethree-color fluorescence quenching of CdTe@SiO_2/Au nanocomposite by addingHg~(2~+)was studied. It was shown that the fluorescent color of the nanocompositechanged from red-organe, yellow and green to blue with increase of the concentrationof Hg~(2~+). Therefore, the visual mode with multi-color fluresence was established and the content of Hg~(2+)was able to evaluate by naked-eyes.
1.三聚磷酸钠稳定的CdS QDs的制备及荧光猝灭法测定铁。以三聚磷酸钠(STPP)为稳定剂，在碱性条件下成功合成了分散较均匀、稳定性好、发射峰位可调、半峰宽25nm左右的水溶性CdS量子点。研究了STPP和硫脲的比例、反应时间及pH等条件对CdS QDs发光特性的影响。探讨了痕量Fe2＋离子对CdSQDs荧光的猝灭作用。考察了缓冲溶液、pH、共存离子等因素的影响。在最佳实验条件下,在0.02~1.2M范围内，483nm处F0/F与Fe2＋呈现良好的线性关系，检测限LOD(3/)为6.7nM。用于实际矿泉水样中Fe的测定，回收率95.0%-105.0%。
2. CdSe/CdS@SiQ2纳米复合物的制备及荧光猝灭法测定铜。首先，以STPP作为稳定剂，合成了水溶性CdSe和CdSe/CdS核/壳量子点；然后，通过正硅酸四乙酯(TEOS)水解成功制备了CdSe/CdS@SiQ2核壳/结构的纳米复合物，用荧光光谱、紫外-可见吸收光谱、XRD对其进行了表征。根据Cu~(2~+)会选择性猝灭CdSe/CdS@SiQ2纳米复合物的荧光，建立了CdSe/CdS@SiO_2纳米复合物荧光猝灭法测定水样中Cu~(2~+)的新方法。其线性范围为0.02~0.6μM，检测限为10nM。
3.介孔ZnO/CdS/SiO_2核/壳纳米复合物的制备及可视化荧光测定Ag＋。首先，以STPP为稳定剂，合成ZnO/CdS核/壳QDs，然后直接把ZnO/CdS QDs水溶液加入含有TEOS、氨水的乙醇溶液中，发展了一种新的制备具有介孔结构的核/壳型ZnO/CdS@SiO_2纳米粒子的新方法。Ag~+可以通过介孔与核内ZnO/CdSQDs作用，导致其带边发射猝灭，同时在595nm处的缺陷发光信号增强，在紫外灯的照射下发出红色荧光。根据紫外灯照射下Ag~+致使ZnO/CdS@SiO_2红色荧光增强，我们用同一种量子点的双色荧光的变化实现对Ag~+的可视化、选择性测定，测定下限为0.04μM。根据595nm处Ag~+与ZnO/CdS@SiO_2作用的荧光增强建立了高选择性和高灵敏测定Ag~+的新方法，线性范围为0.03~0.24μM，检测限(3)为3.30nM。
4. CdTe@SiO_2/Au纳米复合物的制备及可视化多色荧光测定Hg~(2~+)。以壳聚糖为稳定剂和还原剂成功合成了表面等离子体共振吸收峰位可调的金纳米盘。在壳聚糖稳定的球状金纳米溶液中加入半胱氨酸，发现球状金纳米粒子可组装成金纳米链。然后，以溶菌酶为稳定剂和还原剂，合成了具有双荧光（蓝、红）发射的金纳米簇(Au CNs)。通过扫描电镜(SEM)、透射电镜(TEM)、X射线衍射(XRD)等表征手段对晶体结构和形貌进行了解析；通过动态光散射、共振散射光谱、吸收光谱和荧光光谱确证Au纳米结构在溶液中形成，并讨论了金纳米结构的生长机理。把具有双荧光（红、蓝）发射性质的Au CNs和具有绿色荧光发射性质的CdTe@SiO_2混合，制备了在同一激发下能发射三色荧光（红、绿、蓝，RGB）的CdTe@SiO_2/Au纳米复合物。研究了Hg~(2~+)对CdTe@SiO_2/Au纳米复合物的RGB荧光的猝灭作用，发现随着Hg~(2~+)浓度的增加纳米复合物呈现由红橙、黄、绿、蓝的颜色变化。由此建立一种可视化多色荧光识别模式，裸眼测定Hg~(2~+)的含量。
Currently, the fabrication and applieations of the composite nanostructures withdual-emission fluorescence is of great interest。Combining the unique fluoreseenceproperties of Quantum dots (QDs), the QDs nanocomposites with dual-emissiondemonstrate great potentials in fluorescence molecular imaging, dual-fluorescenceratio sensor, and visual fluorescence detection. In this thesis, controllable fabricationof QDs with different fluorescence color and Au nanopartieles has been investigated,and then Core/Shell QDs nanocomposites have been faeilely fabrieated, we have alsoinvestigated the visual fluorescent recognition of such nanoeomposites in the field ofbiotechnology and the environment. The main points are as follows:
1. The fabrication of STPP-stabilized CdS QDs and the determination of traceiron by fluorescence quenching method. The uniform dispersion of water-soluble CdSQDs stabilized by STPP was successfully synthesized in an alkaline medium. Theemission peaks of CdS QDs were tunable, and the half peak width is about25nm.The fluorescence spectral properties of CdS QDs at different pH values, thiourea,STPP and reaction time were investigated. A fluorescence quenching method for thedetermination of trace iron (Ⅱ) was developed, and the factors involving pH, buffersolution, and coexistence ion were investigated. Under the optimum conditions, thevalues of F0/F at483nm are linearly proportional to the concentration of Fe~(2~+)from0.02-1.2μM. The detection limit of the developed method was6.7nM for Fe~(2~+).Mineral water samples were analyzed with the recovery of95.0%-105.0%.
2. The fabrication of the CdSe/CdS@SiQ2core/shell nanocomposites and thedetermination of trace Cu~(2~+)by fluorescence quenching method. Water-soluble CdSeand CdSe/CdS core/shell quantum dots were successfully syntheticed in presence ofSTTP as a stabilizer. CdSe/CdS@SiQ2core-shell nano composites were obtained viathe hydrolysis of Tetraethyl silicate (TEOS). The fluorescence spectra,ultraviolet-visible absorption spectroscopy (UV), TEM and XRD were used toquantify the structure and characteristions of CdSe/CdS@SiQ2nanocomplexes. Dueto Cu~(2~+)can cause fluorescent quenching of CdSe/CdS@SiQ2nanocomplexes selectively, we set up a new method for the determination of Cu2~+in water samples.The linear range from0.02to0.6μM and the detection limitation is10nM.
3. The fabrication of the mesoporous ZnO/CdS@SiO_2Core/Shell nanostructureand visual fluorescence detection of Ag~+. A new method for the preparation ofmesoporous ZnO/CdS@SiO_2Core/Shell nanostructure has been developed. Themesoporous silica shells allow Ag~+to enter into the interior of the nanostructures tocontact with ZnO/CdS core, accordingly causes the quenching of its band edgeemission along with a simultaneous enhancement of red emission at around595nm.So, a novel visual fluorescence detection strategy for Ag~+ion is proposed based on acommon Core/Shell Quantum dots nanostructure. Under optimal conditions, theenhanced fluorescence intensity at595nm increased linearly with the concentrationof Ag~+ion ranging from0.03μM to0.24μM with a detection limit (3σ) of3.3nM.
4. The fabrication of the CdTe@SiO_2/Au nanocomposites and visualmulti-colored fluorescent detection of Hg~(2~+). By chitosan as a stabilizer and reducingagent, the tunable surface plasmon resonance absorption peak of gold nanoplate hasbeen successfully fabricated. It was observed that the linear chains of Au nanosphereswere formed by the addition of L-Cysteine in CTS-stabilized Au nanospheressolution. After that, highly fluorescent Au clusters with dual-emission (blue and red)have been synthesized by lysozyme as stabilized and reducing agents. The structureand morphology of the three types of crystals was characterized via scanning electronmicroscopy (SEM), transmission electron microseopy (TEM) and X-ray diffraction(XRD). The formation of Au nano-structure was determined by the use of UV-vis,dynamic light scattering as well as resonance light scattering and fluorescence spectra.The growth mechanism of gold nano-structure was discussed. The CdTe@SiO_2/Aunanocomposite with three-color fluorescence (red, green and blue, RGB) under asingle excitation wavelength was synthesized when the Au CNs with dual-emission(blue and red colors) and CdTe@SiO_2with green fluorescent were mixed. Thethree-color fluorescence quenching of CdTe@SiO_2/Au nanocomposite by addingHg~(2~+)was studied. It was shown that the fluorescent color of the nanocompositechanged from red-organe, yellow and green to blue with increase of the concentrationof Hg~(2~+). Therefore, the visual mode with multi-color fluresence was established and the content of Hg~(2+)was able to evaluate by naked-eyes.
引文
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