一些金属离子和阴离子光学传感器的构建及其分析应用研究
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摘要
荧光探针的研究在光学传感器的发展中一直占据重要地位。在过去的十几年中,有关光学探针的研究得到了飞速发展,各种有机染料、共轭聚合物、半导体量子点和贵金属纳米微粒作为荧光探针已大量用于各种分析之中,这些探针具有快速、灵敏、荧光效率高等优点,但是仍然存在一些不足,比如,有机染料和共轭聚合物的光稳定性有待提高,而且多数都涉及到复杂的化学合成；半导体量子点的毒性大。近年来,介于分子和纳米材料之间的金属纳米簇受到广泛关注,主要原因是其尺寸接近于费米波长,具有类似于分子的荧光特性,并且合成方法简单、良好的生物相容性等。因此,金属纳米簇作为荧光探针有诱人的发展前景,目前的研究主要集中在贵金属纳米簇,尤其是金及银纳米簇,尽管它们在分析化学中的应用已有大量报道,但是如何制备高稳定性及高量子产率的纳米簇仍然是一个瓶颈,而且有关铜纳米簇的研究较少。因此,研究合成高稳定性及高量子产率铜纳米簇的方法,并探索其作为荧光探针识别金属离子和阴离子的工作具有重要意义。为此,本论文以铜纳米簇荧光探针为重点,开展了：1)、基于小分子的荧光探针识别Cu2+的研究；2)、基于功能化银纳米的光散射探针同时测定Pb2+和半胱氨酸的研究；3)、利用简单方法合成了荧光铜纳米簇,探索了其作为荧光探针的可行性。根据以上结果,构建了新的测定金属离子和阴离子的光学传感器,并成功将其用于环境分析和生化分析之中。全文共六章,主要内容如下：
第一章：文献综述和选题依据。简要介绍了1)近年来基于金属纳米材料的光学传感器的构建及其分析应用；2)金属纳米簇的合成、荧光性质、分析应用及其存在的问题。根据文献分析结果,提出本论文的选题依据。
第二章：开展了基于小分子荧光性质的方法识别Cu2+的研究,实验发现在碱性条件下,Cu2+对过氧化氢氧化对甲基苯酚具有高效的催化能力,甲基苯酚的氧化产物2,2'-二羟基-5,5'-二甲基联苯具有强的荧光。通过甲基苯酚-过氧化氢体系的荧光光谱表明Cu2+具有催化能力。据此,建立了一种简单、灵敏和选择性好的测定Cu2+的荧光分析方法。在优化条件下,测定Cu2+的线性范围为0.3μM-50μM,检出限为10nM。对7μM的Cu2+重复测定15次,其相对标准偏差为2.1%。将上述方法应用于湖水,江水和游泳池水中微量Cu2+的测定,并获得了满意结果,并且测定结果与原子吸收方法测定结果很吻合。
第三章：开展了基于功能化银纳米的光散射探针同时测定pb2+和半胱氨酸的研究,其基本原理是：二硫代氨基甲酸盐有强的金属亲和力,当pb2+与二硫代氨基甲酸盐功能化的Ag NPs作用时,促使DTC-Ag NPs团聚,从而使Ag NPs的共振光散射信号增强,可用于识别pb2+。加入半胱氨酸时,由于半胱氨酸与pb2+形成更强的Pb-S键,使得pb2+从DTC-Ag NPs表面脱离下来,导致DTC-Ag NPs重新分散在水溶液中,致使Ag NPs的共振光散射信号降低,可用于识别半胱氨酸。通过直接将二硫化碳和二乙醇胺作为前体分子在超声条件下充分混合即可原位合成二硫代氨基甲酸盐。研究了溶液的pH.DTC-Ag NPs的浓度和离子强度对共振光散射体系的影响。在优化条件下,功能化银纳米探针测定pb2+的线性范围为O.01μM-60μM,检出限可达4nM。本方法具有选择性好、线性响应范围宽、灵敏度高等特点。用于河水和自来水中pb2+的测定,获得满意结果。同时研究了基于功能化银纳米光散射探针用于检测半胱氨酸的可行性。
第四章：提出了室温下一步法简单、方便制备酒石酸稳定的水溶性荧光铜纳米簇(Cu NCs)的合成新方法,并且用UV-Vis吸收光谱、高分辨透射电镜、动力学光散射及FT-IR等对其进行了表征,其荧光量子产率为2.2%。加入酒石酸作为稳定剂是为了防止Cu NCs的聚集和氧化,并且增加其水溶性和稳定性,研究表明即使在1M NaCl的高离子强度溶液中,Cu NCs的荧光强度也保持不变。进一步研究表明,经过简单的酒石酸表面修饰的铜纳米簇能够高选择性识别A13+,其它常见的金属共存离子对Al3+的识别影响很小。其机理可能是A13+与Cu NCs表面的羧基和羟基作用,导致铜纳米簇的荧光增强并且波长发生蓝移；详细研究了各因素对Cu NCs识别Al3+的影响,获得了优化条件,在此条件下,本法对A13+的检出限可达12.5nM。另外,由于F-与Al3+具有更强的亲和力,当有F-存在时,使得Al3+脱离酒石酸稳定的Cu NCs表面,释放Cu NCs,伴随着荧光强度的降低和波长的红移。详细研究了各因素对Cu NCs识别F-的影响,并且探讨了本法用于传感检测F-的可行性。以上结果表明,本文合成的酒石酸-Cu NCs可作为荧光探针用于同时测定A13+和F-本方法具有简单、快速、灵敏高、选择性好。
第五章：为得到高荧光量子产率的铜纳米簇,提出了一步法合成单宁酸(TA)功能化的铜纳米簇(TA-Cu NCs),通过紫外可见光谱、红外光谱(FT-IR)、荧光光谱、TEM和XPS等手段对其进行一系列表征。TA-Cu NCs在激发波长为360nm,发射波长430nm处具有强的荧光,其量子产量为14%。TA-Cu NCs在0.3M的NaCl水溶液中仍然稳定存在,并且其荧光强度不受pH影响。通过电子传递的作用,Fe3+能使TA-Cu NCs的荧光发生猝灭,然而其他金属离子并没有这种效果。换言之,Fe2+或是H202单独加入都不能改变TA-Cu NCs的荧光,但是如果二者混合就能导致TA-Cu NCs的荧光猝灭。基于此,TA-Cu NCs能够用于构建区分Fe3+、Fe2+以及它们的混合物的分子逻辑门系统。这个简单的化学传感器能够快速、灵敏地和选择性地用于测定Fe3+,测定Fe3+的线性范围为10nM-10μM,检出下限低至10nM。同时,将本法用于实际水样中Fe3+和Fe2+的测定,获得满意结果。为进
一步探索高荧光量子产率的TA-Cu NCs在活体细胞成像中的应用潜力,用CCK-8方法评价了TA-Cu NCs对A549细胞的毒性,结果表明,即使使用10μM TA-Cu NCs,对A549细胞的成活率(100%)几乎无影响,表明其细胞毒性低,有良好的生物相容性,可用于活细胞成像,检测细胞中的Fe3+。
第六章：开展了利用TA-Cu NCs作为荧光开关探针识别磷酸根的研究。其基本原理是：磷酸根与TA-Cu NCs竞争性结合Eu3+。Eu3+存在下,Eu3+能够通过与相邻TA-Cu NCs表面的羧基和羟基结合导致其发生团聚,使TA-Cu NCs的荧光猝灭；当加入磷酸根时,由于Eu3+与磷酸根的亲和力大于与TA-Cu NCs羧基的结合力,磷酸根能将Eu3+从TA-Cu NCs的表面拉下来,使得TA-Cu NCs的荧光恢复。因此,借助Eu3+, TA-Cu NCs可作为荧光开关探针识别磷酸根。通过优化各种实验条件后,得出TA-Cu NCs荧光探针检测磷酸根的线性范围为0.07～80μM,检出限可达9.6nM。基于TA-Cu NCs与Eu3+联合建立了选择性好,灵敏度高的测定磷酸根的荧光法,并成功用于实际水样中磷酸根的测定。
The fluorescent probes continue to retain an important position in the development of optical sensor. In the past decades, it has been a rapid development for the research on optical probe. Various fluorescent probes, such as organic dyes, conjugated polymers, semiconductor quantum dots, and noble metal nanoparticles, have been extensively used for various analyzes. They promise advantages of rapidity, sensitivity, high quantum yield, and so on. However, these fluorescent probes still suffers from one or more major deficiencies, such as, poor light stability of organic dyes and conjugated polymers, complex chemical synthesis, high environmental toxicity of semiconductor quantum dots. Recently metal nanoclusters, which provide the missing link between atomic and nanoparticle behavior in metals, have drawn the most attention, because of their molecule-like fluorescence properties due to their sizes compared to the Fermi wavelength of electrons, simple synthesis methods, and good biocompatibility. To date, most of previous work concentrated on the noble mental nanoclusters, especially Au NCs and Ag NCs. Although various synthetic methods have been proposed for the preparation of noble metal NCs, the obtained fluorescent Au NCs and Ag NCs have some drawbacks including low quantum yield and stability. Also, compared to extensive studies on Au NCs and Ag NCs, Cu NCs were much less studied. Hence, it remains a great significance for the development of a simple and facile strategy for the synthesis of water soluble, extremely stable, and highly quantum efficient fluorescent Cu NCs and the application of the resulted Cu NCs in the detection of mental ions and anions. Thus, this thesis focused on the fluorescent probes of copper nanoclusters, and carry out the following work:1) The research of small molecule as fluorescent probes for sensing of Cu2+;2) The research of functional Ag nanoparticles (NPs) as resonance light scattering (RLS) probes for simultaneous sensing of Pb2+and cysteine;3) Facile preparation of fluorescent Cu nanoclusters (NCs) and the research of the suitability of Cu NCs as fluorescent probes.
On this basis, the development of some metal ions and anions optical sensors were successfully used for environmental analysis and biochemical analysis. The thesis is divided into six chapters:
Chapter1:It introduces literature review and the reasons why to select this subject. In brief,1) The development of the optical sensors based on metal nanomaterials and analytical applications in recent years;2) Synthesis of metal nanoclusters, fluorescence properties, analytical applications and some problems. According to the results of literature analysis, we presented in the reasons why to select this subject on this thesis.
Chapter2:The research about small molecule as fluorescent probes for sensing of Cu2+is developed. It was found that Cu2+shows great catalytic effect on p-cresol oxidation by hydrogen peroxide under alkaline condition, leading to an intense fluorescence signal due to fast formation of2,2'-dihydroxy-5,5'-dimethylbiphenyl, an oxidation product of p-cresol. Investigation on the fluorescence spectra of the p-cresol-hydrogen peroxide system demonstrated the catalytic behavior of Cu2+. On this basis, a very simple, sensitive, and selective fluorescent method was established for the determination of trace copper in this study. Under the optimal conditions, the proposed system could respond down to1.0x10-8mol L-1of Cu2+with a linear calibration range from3.0×l0-7mol L-1to5.0×5mol L-1. The relative standard deviation (RSD) is2.1%for7.0x10-6mol L-1Cu2+(n=15). The proposed method was successfully applied to determine trace Cu2+in lake, river and swimming pool water samples with satisfactory results. The results given by the proposed method are in good agreement with those given by atomic absorption spectroscopy method.
Chapter3:We investigated the suitability of functional Ag nanoparticles (NPs) as resonance light scattering (RLS) probes for simultaneous sensing of Pb2+and cysteine. This strategy was based on the fact that Pb2+could induce the aggregation of DTC-Ag NPs due to strong metal affinity of DTC along with the enhanced RLS signal. Meanwhile, due to the strong binding preference of cysteine toward Pb2+by forming Pb2+-S bond, Pb2+was removed from the surface of the DTC-Ag NPs, leading to redispersion of DTC-Ag NPs, along with the decreased RLS signal. The DTC capping ligands are generated by a very simple in situ method through reaction of carbon disulfide with diethanolamine as primary precursor molecules under ultrasonic irradation. After optimizing some experimental conditions (including pH value of solution, concentration of DTC-Ag NPs, and ion strength), a very simple and facile sensing system has been developed for detection of Pb2+in water based on RLS technology. The proposed system promises excellent selectivity, wide linear response range and high sensitivity for Pb2+. The linear response ranges for Pb2+was from0.01μM to60μM. The limit of detection (S/N=3?) for Pb2+was as low as4nM. The proposed method was successfully used to detect Pb2+in river and tap water samples, indicating the potential of this new, sensitive and selective method in water quality monitoring. The possibility of the proposed system for sensing of cysteine was also investigated.
Chapter4:Facile preparation of water soluble and fluorescent Cu nanoclusters (NCs) stabilized by tartaric acid at room temperature is described and characterized using UV-Vis absorption spectrum, high-resolution transmission electron microscopy, dynamic light scattering and Fourier transform infrared spectroscopy (FT-IR), giving a quantum yield (QY)2.2%against the reference of quinine sulfate. The tartaric acid layer was introduced to prevent the Cu NCs from aggregation or oxidation, increase their water solubility and was found to be highly stable even in1M NaCl. Additionally, this simple surface optimization resulted in surprisingly high efficiency of selective Al3+sensing and other coexistence ions have no obvious effects on the sensing of Al3+. The sensing mechanism to be hypothesized is the interaction between Al3+and carboxyl and hydroxyl groups on the Cu surface along with the enhanced fluorescence signal and blue shift wavelength. Under optical condition, the limit of detection (LOD) for Al3+was as low as12.5nM. Meanwhile, due to the strong binding preference of fluoride toward Al3+, Al3+was removed from the surface of the tartaric acid-Cu NCs, resulting in the disconnection of the interaction between Al3+ions and tartaric acid-Cu NCs, along with the decreased fluorescence signal of tartaric acid-Cu NCs and red shift wavelength. The possibility of the proposed system for sensing of fluoride was also investigated. This result revealed a simple and practical strategy for Al3+and fluoride detection using fluorescent Cu NCs as sensor probe. The method is simple, rapid, sensitive and selective.
Chapter5:To obtain Cu nanoclusters (NCs) with high quantum yield, a simple, one-step facile route for preparation of water soluble and fluorescent Cu NCs stabilized by tannic acid (TA) as a model of small molecule is described. The as-prepared TA capped Cu NCs (TA-CuNCs) are characterized by UV-vis spectroscopy, Fourier transform infrared (FT-IR) spectroscopy, luminescence, transmission electron microscopy (TEM), and X-ray photoelectron spectroscopy (XPS). The Cu NCs show luminescence properties having excitation and emission maxima at360nm and430nm, respectively, with a quantum yield of about14%. The TA-Cu NCs are very stable even in0.3M NaCl, and their luminescent properties show pH independent. The fluorescence (FL) of the TA-Cu NCs is quenched by Fe3+through an electron transfer mechanism, but not by other metal ions. Furthermore, the FL of the TA-Cu NCs shows no changes with the addition of Fe2+or H2O2individually, but it is quenched significantly by their mixture. On this basis, TA-Cu NCs have been demonstrated to build the combination logic gate system to discriminate Fe3+, Fe2+and their mixture. This facile chemosensor can offer a rapid, reliable, sensitive, and selective sensing of Fe3+ions with detection limit as low as10nM and a dynamic range from10nM to10μM. The detection results for ferrous ions in water samples obtained by this method agreed well with that by atomic absorbance spectrometry, suggesting the potential application of this sensing system. Moreover, to demonstrate the potential application of the prepared TA-Cu NCs fluorescent probe with high QY for in cell imaging, the cytotoxicity of TA-Cu NCs was evaluated by CCK-8assays, in which no significant cytotoxic effects have been observed in the presence of TA-Cu NCs, and almost100%of A549cells are alive even in the presence of10μM TA-Cu NCs, indicating it is low toxicity for cell, good biocompatibility and suitable for imaging studies. We demonstrate the feasibility of using TA-Cu NCs for determination of Fe3+in living cells.
Chapter6:We have employed tannic acid stabilized copper clusters (TA-Cu NCs) as a novel swithable fluorescence sensor for the detection of the phosphate (Pi) in solution, which was based on the competition between Pi and TA—Cu NCs for the interaction with the Europium ions (Eu+). First, Eu+could induce the aggregation of TA-Cu NCs due to coordination to the oxygen-donor atoms of the carboxylate groups and hydroxyl on the Cu NCs surfaces along with the decrease in the FL intensity of the Cu NCs. Because Eu3+displays a higher affinity for the oxygen-donor atoms in Pi than for the carboxylate groups and hydroxyl on the TA-Cu NCs surface, there is a competition for Eu3+between the oxygen-donor atoms from Pi and those from the carboxylate groups on the Cu NCs surface, which effectively diminishing its quenching efficiency, and the FL intensity of Cu NCs is recovered. Under optical condition, a linear relationship (R2=0.9963) was observed with Pi concentrations from0.07to80μM and the detection limit at an S/N ratio of3for Pi was9.6nM. Because fluorescence quenching of the TA-Cu NCs by Eu3+is inhibited by Pi selectivity, we have obtained a highly sensitive and selective assay for Pi. Herein, a novel switchable phosphate sensing system with highly sensitivity and selectivity is designed using stable and fluorescent Cu NCs that bear surface groups of carboxy groups, combined with Eu3+Moreover, we demonstrate the feasibility of using TA-Cu NCs for determination of Pi in environmental smples.
第一章：文献综述和选题依据。简要介绍了1)近年来基于金属纳米材料的光学传感器的构建及其分析应用；2)金属纳米簇的合成、荧光性质、分析应用及其存在的问题。根据文献分析结果,提出本论文的选题依据。
第二章：开展了基于小分子荧光性质的方法识别Cu2+的研究,实验发现在碱性条件下,Cu2+对过氧化氢氧化对甲基苯酚具有高效的催化能力,甲基苯酚的氧化产物2,2'-二羟基-5,5'-二甲基联苯具有强的荧光。通过甲基苯酚-过氧化氢体系的荧光光谱表明Cu2+具有催化能力。据此,建立了一种简单、灵敏和选择性好的测定Cu2+的荧光分析方法。在优化条件下,测定Cu2+的线性范围为0.3μM-50μM,检出限为10nM。对7μM的Cu2+重复测定15次,其相对标准偏差为2.1%。将上述方法应用于湖水,江水和游泳池水中微量Cu2+的测定,并获得了满意结果,并且测定结果与原子吸收方法测定结果很吻合。
第三章：开展了基于功能化银纳米的光散射探针同时测定pb2+和半胱氨酸的研究,其基本原理是：二硫代氨基甲酸盐有强的金属亲和力,当pb2+与二硫代氨基甲酸盐功能化的Ag NPs作用时,促使DTC-Ag NPs团聚,从而使Ag NPs的共振光散射信号增强,可用于识别pb2+。加入半胱氨酸时,由于半胱氨酸与pb2+形成更强的Pb-S键,使得pb2+从DTC-Ag NPs表面脱离下来,导致DTC-Ag NPs重新分散在水溶液中,致使Ag NPs的共振光散射信号降低,可用于识别半胱氨酸。通过直接将二硫化碳和二乙醇胺作为前体分子在超声条件下充分混合即可原位合成二硫代氨基甲酸盐。研究了溶液的pH.DTC-Ag NPs的浓度和离子强度对共振光散射体系的影响。在优化条件下,功能化银纳米探针测定pb2+的线性范围为O.01μM-60μM,检出限可达4nM。本方法具有选择性好、线性响应范围宽、灵敏度高等特点。用于河水和自来水中pb2+的测定,获得满意结果。同时研究了基于功能化银纳米光散射探针用于检测半胱氨酸的可行性。
第四章：提出了室温下一步法简单、方便制备酒石酸稳定的水溶性荧光铜纳米簇(Cu NCs)的合成新方法,并且用UV-Vis吸收光谱、高分辨透射电镜、动力学光散射及FT-IR等对其进行了表征,其荧光量子产率为2.2%。加入酒石酸作为稳定剂是为了防止Cu NCs的聚集和氧化,并且增加其水溶性和稳定性,研究表明即使在1M NaCl的高离子强度溶液中,Cu NCs的荧光强度也保持不变。进一步研究表明,经过简单的酒石酸表面修饰的铜纳米簇能够高选择性识别A13+,其它常见的金属共存离子对Al3+的识别影响很小。其机理可能是A13+与Cu NCs表面的羧基和羟基作用,导致铜纳米簇的荧光增强并且波长发生蓝移；详细研究了各因素对Cu NCs识别Al3+的影响,获得了优化条件,在此条件下,本法对A13+的检出限可达12.5nM。另外,由于F-与Al3+具有更强的亲和力,当有F-存在时,使得Al3+脱离酒石酸稳定的Cu NCs表面,释放Cu NCs,伴随着荧光强度的降低和波长的红移。详细研究了各因素对Cu NCs识别F-的影响,并且探讨了本法用于传感检测F-的可行性。以上结果表明,本文合成的酒石酸-Cu NCs可作为荧光探针用于同时测定A13+和F-本方法具有简单、快速、灵敏高、选择性好。
第五章：为得到高荧光量子产率的铜纳米簇,提出了一步法合成单宁酸(TA)功能化的铜纳米簇(TA-Cu NCs),通过紫外可见光谱、红外光谱(FT-IR)、荧光光谱、TEM和XPS等手段对其进行一系列表征。TA-Cu NCs在激发波长为360nm,发射波长430nm处具有强的荧光,其量子产量为14%。TA-Cu NCs在0.3M的NaCl水溶液中仍然稳定存在,并且其荧光强度不受pH影响。通过电子传递的作用,Fe3+能使TA-Cu NCs的荧光发生猝灭,然而其他金属离子并没有这种效果。换言之,Fe2+或是H202单独加入都不能改变TA-Cu NCs的荧光,但是如果二者混合就能导致TA-Cu NCs的荧光猝灭。基于此,TA-Cu NCs能够用于构建区分Fe3+、Fe2+以及它们的混合物的分子逻辑门系统。这个简单的化学传感器能够快速、灵敏地和选择性地用于测定Fe3+,测定Fe3+的线性范围为10nM-10μM,检出下限低至10nM。同时,将本法用于实际水样中Fe3+和Fe2+的测定,获得满意结果。为进
一步探索高荧光量子产率的TA-Cu NCs在活体细胞成像中的应用潜力,用CCK-8方法评价了TA-Cu NCs对A549细胞的毒性,结果表明,即使使用10μM TA-Cu NCs,对A549细胞的成活率(100%)几乎无影响,表明其细胞毒性低,有良好的生物相容性,可用于活细胞成像,检测细胞中的Fe3+。
第六章：开展了利用TA-Cu NCs作为荧光开关探针识别磷酸根的研究。其基本原理是：磷酸根与TA-Cu NCs竞争性结合Eu3+。Eu3+存在下,Eu3+能够通过与相邻TA-Cu NCs表面的羧基和羟基结合导致其发生团聚,使TA-Cu NCs的荧光猝灭；当加入磷酸根时,由于Eu3+与磷酸根的亲和力大于与TA-Cu NCs羧基的结合力,磷酸根能将Eu3+从TA-Cu NCs的表面拉下来,使得TA-Cu NCs的荧光恢复。因此,借助Eu3+, TA-Cu NCs可作为荧光开关探针识别磷酸根。通过优化各种实验条件后,得出TA-Cu NCs荧光探针检测磷酸根的线性范围为0.07～80μM,检出限可达9.6nM。基于TA-Cu NCs与Eu3+联合建立了选择性好,灵敏度高的测定磷酸根的荧光法,并成功用于实际水样中磷酸根的测定。
The fluorescent probes continue to retain an important position in the development of optical sensor. In the past decades, it has been a rapid development for the research on optical probe. Various fluorescent probes, such as organic dyes, conjugated polymers, semiconductor quantum dots, and noble metal nanoparticles, have been extensively used for various analyzes. They promise advantages of rapidity, sensitivity, high quantum yield, and so on. However, these fluorescent probes still suffers from one or more major deficiencies, such as, poor light stability of organic dyes and conjugated polymers, complex chemical synthesis, high environmental toxicity of semiconductor quantum dots. Recently metal nanoclusters, which provide the missing link between atomic and nanoparticle behavior in metals, have drawn the most attention, because of their molecule-like fluorescence properties due to their sizes compared to the Fermi wavelength of electrons, simple synthesis methods, and good biocompatibility. To date, most of previous work concentrated on the noble mental nanoclusters, especially Au NCs and Ag NCs. Although various synthetic methods have been proposed for the preparation of noble metal NCs, the obtained fluorescent Au NCs and Ag NCs have some drawbacks including low quantum yield and stability. Also, compared to extensive studies on Au NCs and Ag NCs, Cu NCs were much less studied. Hence, it remains a great significance for the development of a simple and facile strategy for the synthesis of water soluble, extremely stable, and highly quantum efficient fluorescent Cu NCs and the application of the resulted Cu NCs in the detection of mental ions and anions. Thus, this thesis focused on the fluorescent probes of copper nanoclusters, and carry out the following work:1) The research of small molecule as fluorescent probes for sensing of Cu2+;2) The research of functional Ag nanoparticles (NPs) as resonance light scattering (RLS) probes for simultaneous sensing of Pb2+and cysteine;3) Facile preparation of fluorescent Cu nanoclusters (NCs) and the research of the suitability of Cu NCs as fluorescent probes.
On this basis, the development of some metal ions and anions optical sensors were successfully used for environmental analysis and biochemical analysis. The thesis is divided into six chapters:
Chapter1:It introduces literature review and the reasons why to select this subject. In brief,1) The development of the optical sensors based on metal nanomaterials and analytical applications in recent years;2) Synthesis of metal nanoclusters, fluorescence properties, analytical applications and some problems. According to the results of literature analysis, we presented in the reasons why to select this subject on this thesis.
Chapter2:The research about small molecule as fluorescent probes for sensing of Cu2+is developed. It was found that Cu2+shows great catalytic effect on p-cresol oxidation by hydrogen peroxide under alkaline condition, leading to an intense fluorescence signal due to fast formation of2,2'-dihydroxy-5,5'-dimethylbiphenyl, an oxidation product of p-cresol. Investigation on the fluorescence spectra of the p-cresol-hydrogen peroxide system demonstrated the catalytic behavior of Cu2+. On this basis, a very simple, sensitive, and selective fluorescent method was established for the determination of trace copper in this study. Under the optimal conditions, the proposed system could respond down to1.0x10-8mol L-1of Cu2+with a linear calibration range from3.0×l0-7mol L-1to5.0×5mol L-1. The relative standard deviation (RSD) is2.1%for7.0x10-6mol L-1Cu2+(n=15). The proposed method was successfully applied to determine trace Cu2+in lake, river and swimming pool water samples with satisfactory results. The results given by the proposed method are in good agreement with those given by atomic absorption spectroscopy method.
Chapter3:We investigated the suitability of functional Ag nanoparticles (NPs) as resonance light scattering (RLS) probes for simultaneous sensing of Pb2+and cysteine. This strategy was based on the fact that Pb2+could induce the aggregation of DTC-Ag NPs due to strong metal affinity of DTC along with the enhanced RLS signal. Meanwhile, due to the strong binding preference of cysteine toward Pb2+by forming Pb2+-S bond, Pb2+was removed from the surface of the DTC-Ag NPs, leading to redispersion of DTC-Ag NPs, along with the decreased RLS signal. The DTC capping ligands are generated by a very simple in situ method through reaction of carbon disulfide with diethanolamine as primary precursor molecules under ultrasonic irradation. After optimizing some experimental conditions (including pH value of solution, concentration of DTC-Ag NPs, and ion strength), a very simple and facile sensing system has been developed for detection of Pb2+in water based on RLS technology. The proposed system promises excellent selectivity, wide linear response range and high sensitivity for Pb2+. The linear response ranges for Pb2+was from0.01μM to60μM. The limit of detection (S/N=3?) for Pb2+was as low as4nM. The proposed method was successfully used to detect Pb2+in river and tap water samples, indicating the potential of this new, sensitive and selective method in water quality monitoring. The possibility of the proposed system for sensing of cysteine was also investigated.
Chapter4:Facile preparation of water soluble and fluorescent Cu nanoclusters (NCs) stabilized by tartaric acid at room temperature is described and characterized using UV-Vis absorption spectrum, high-resolution transmission electron microscopy, dynamic light scattering and Fourier transform infrared spectroscopy (FT-IR), giving a quantum yield (QY)2.2%against the reference of quinine sulfate. The tartaric acid layer was introduced to prevent the Cu NCs from aggregation or oxidation, increase their water solubility and was found to be highly stable even in1M NaCl. Additionally, this simple surface optimization resulted in surprisingly high efficiency of selective Al3+sensing and other coexistence ions have no obvious effects on the sensing of Al3+. The sensing mechanism to be hypothesized is the interaction between Al3+and carboxyl and hydroxyl groups on the Cu surface along with the enhanced fluorescence signal and blue shift wavelength. Under optical condition, the limit of detection (LOD) for Al3+was as low as12.5nM. Meanwhile, due to the strong binding preference of fluoride toward Al3+, Al3+was removed from the surface of the tartaric acid-Cu NCs, resulting in the disconnection of the interaction between Al3+ions and tartaric acid-Cu NCs, along with the decreased fluorescence signal of tartaric acid-Cu NCs and red shift wavelength. The possibility of the proposed system for sensing of fluoride was also investigated. This result revealed a simple and practical strategy for Al3+and fluoride detection using fluorescent Cu NCs as sensor probe. The method is simple, rapid, sensitive and selective.
Chapter5:To obtain Cu nanoclusters (NCs) with high quantum yield, a simple, one-step facile route for preparation of water soluble and fluorescent Cu NCs stabilized by tannic acid (TA) as a model of small molecule is described. The as-prepared TA capped Cu NCs (TA-CuNCs) are characterized by UV-vis spectroscopy, Fourier transform infrared (FT-IR) spectroscopy, luminescence, transmission electron microscopy (TEM), and X-ray photoelectron spectroscopy (XPS). The Cu NCs show luminescence properties having excitation and emission maxima at360nm and430nm, respectively, with a quantum yield of about14%. The TA-Cu NCs are very stable even in0.3M NaCl, and their luminescent properties show pH independent. The fluorescence (FL) of the TA-Cu NCs is quenched by Fe3+through an electron transfer mechanism, but not by other metal ions. Furthermore, the FL of the TA-Cu NCs shows no changes with the addition of Fe2+or H2O2individually, but it is quenched significantly by their mixture. On this basis, TA-Cu NCs have been demonstrated to build the combination logic gate system to discriminate Fe3+, Fe2+and their mixture. This facile chemosensor can offer a rapid, reliable, sensitive, and selective sensing of Fe3+ions with detection limit as low as10nM and a dynamic range from10nM to10μM. The detection results for ferrous ions in water samples obtained by this method agreed well with that by atomic absorbance spectrometry, suggesting the potential application of this sensing system. Moreover, to demonstrate the potential application of the prepared TA-Cu NCs fluorescent probe with high QY for in cell imaging, the cytotoxicity of TA-Cu NCs was evaluated by CCK-8assays, in which no significant cytotoxic effects have been observed in the presence of TA-Cu NCs, and almost100%of A549cells are alive even in the presence of10μM TA-Cu NCs, indicating it is low toxicity for cell, good biocompatibility and suitable for imaging studies. We demonstrate the feasibility of using TA-Cu NCs for determination of Fe3+in living cells.
Chapter6:We have employed tannic acid stabilized copper clusters (TA-Cu NCs) as a novel swithable fluorescence sensor for the detection of the phosphate (Pi) in solution, which was based on the competition between Pi and TA—Cu NCs for the interaction with the Europium ions (Eu+). First, Eu+could induce the aggregation of TA-Cu NCs due to coordination to the oxygen-donor atoms of the carboxylate groups and hydroxyl on the Cu NCs surfaces along with the decrease in the FL intensity of the Cu NCs. Because Eu3+displays a higher affinity for the oxygen-donor atoms in Pi than for the carboxylate groups and hydroxyl on the TA-Cu NCs surface, there is a competition for Eu3+between the oxygen-donor atoms from Pi and those from the carboxylate groups on the Cu NCs surface, which effectively diminishing its quenching efficiency, and the FL intensity of Cu NCs is recovered. Under optical condition, a linear relationship (R2=0.9963) was observed with Pi concentrations from0.07to80μM and the detection limit at an S/N ratio of3for Pi was9.6nM. Because fluorescence quenching of the TA-Cu NCs by Eu3+is inhibited by Pi selectivity, we have obtained a highly sensitive and selective assay for Pi. Herein, a novel switchable phosphate sensing system with highly sensitivity and selectivity is designed using stable and fluorescent Cu NCs that bear surface groups of carboxy groups, combined with Eu3+Moreover, we demonstrate the feasibility of using TA-Cu NCs for determination of Pi in environmental smples.
引文
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