基于新型微纳米材料的电化学（生物）传感器的研究
摘要
纳米材料是指三维空间尺度至少有一维处于纳米量级(1-100 nm)的材料，具有表面效应、体积效应、量子尺寸效应和宏观量子隧道效应等。由于纳米材料的这些独特性能，使得其在科学研究的各个领域都有非常广泛的应用前景。本文以具有优良结构的微纳米材料为基础来开发和研制性能优良的电化学及生物传感器。主要内容如下：
1.以聚苯乙烯小球为模板，通过电化学沉积的方法将金沉积到模板空隙中，四氢呋喃去除模板。所得金膜修饰电极具有层状有序的三维孔状结构，比表面积大，孔壁交错贯通，为葡萄糖的氧化提供了大量的反应位点，从而大大提高了其对葡萄糖响应的灵敏度；此外，该传感器可以在较低电位下实现葡萄糖的测定，有效的避免了抗坏血酸等物质的干扰。
2.以聚碳酸酯为模板，通过电化学沉积的方法将铂铅合金沉积到模板的孔隙中去，然后通过有机溶剂去除模板，即得具有微纳米线阵列结构的铂铅合金修饰电极。该修饰电极具有纳米微电极阵列的电化学行为，对葡萄糖展现出良好的电催化响应，可以在pH=7.4的中性磷酸缓冲体系中，在较低的电位下实现葡萄糖的测定。
3.以金纳米粒子/分子筛复合材料为基底，采用半胱胺作为交联剂，通过共价键合的方法将醛基化的葡萄糖氧化酶固定到基底电极上，从而实现葡萄糖生物传感器的构筑。该传感器对葡萄糖响应的线性范围上限可达14 mmol/L，此外，金纳米粒子/分子筛复合材料为酶分子提供了一个非常类似于生物体的微环境，从而可以有效保持生物分子的活性，进而提高了传感器的稳定性。
4.首先将柠檬酸根稳定的金纳米粒子与重氮树脂进行层层自组装，然后通过紫外光照使层间作用力由静电吸引转换为共价作用，从而得到一个具有高度稳定性的层数可控的金纳米粒子多层膜。随后将该金纳米粒子多层膜修饰电极浸入到多巴胺的溶液中，对多巴胺进行吸附，所得的多巴胺修饰电极对抗坏血酸响应灵敏，可以实现抗坏血酸的定量测定。
Nanomaterials, with their size in the range of 1-100 nm, have recently become one of themost exciting forefront fields in material science. Due to their small size, these materialsexhibit quanta-size effect, small-size effect, surface effect and tunneling effect that differfrom both bulk material and the individual atoms from which they comprised. With theseunique properties, they are widely used in the fields of catalysis, optical absorption,medicine, magnetic medium, new materials synthesis and particularly attractive in sensorapplications. Introduction of nanomaterials to the system of electrochemical sensors andbiosensors is of considerable interest, since they can play an important role in improvingthe sensor performance due to their large specific surface area, good conductivity specialcatalytic property and excellent biocompatibility.
In chapter 2, an enzyme-free glucose sensor has been developed using a threedimensionalinverse-opal gold film electrode (3DGFE) obtained by electrochemicaldeposition of gold in the interspaces of polystyrene templates. The gold films werecharacterized by scanning electron microscopy (SEM) and cyclic voltammetry (CV). Theelectrochemical oxidation of glucose in the presence of interferents (ascorbic acid, and so on) at different operating potentials on the 3DGFE has been investigated in detail. A lowoperating potential of -0.30 V was chosen for glucose detection, since the interferencecould be well avoided at this potential, whereas the current response for glucose oxidationwas still sensitive. The amperometric response of the sensor increased with the increase ofglucose concentration with a linear range of 5×10-6 ~ 10-2 mol/L and a detection limit of3.2μmol/L (signal-to-noise ratio of 3). The glucose sensor exhibited a high sensitivity of46.6μA (mmol/L)-1 cm-2, which could be ascribed to the unique surface structure of thethree-dimensionally interconnected porous structured gold films. The sensor with highsensitivity, good selectivity and stability is attractive for the practical glucose detection.
In chapter 3, Pt-Pb nanowire array was directly synthesized by electrochemicaldeposition of Pt-Pb alloy into the pores of macroporous polycarbonate template andsubsequent chemical etching of the template. The morphology and the composition of thePt-Pb nanowires were characterized by scanning electron microscopy (SEM) and X-rayphotoelectron spectroscopy (XPS), respectively. Cyclic voltammetry (CV) was used toevaluate the electrochemical performance of the Pt-Pb nanowire array electrode. Directglucose oxidation at such Pt-Pb nanowire array electrode was investigated detailedly bydiscussing the effect of the structure and materials of the electrode on electrocatalyticoxidation of glucose. As a result, we found that the Pt-Pb nanowire array electrode with athree-dimensional structure exhibited high electrocatalytic activity to glucose oxidation inneutral condition and could be used for the development of nonenzymatic glucose sensor.To effectively avoid the interference coming from ascorbic acid, a negative potential of -0.20 V was chosen for glucose detection, and the sensitivity of the sensor to glucoseoxidation was 11.25μA (mmol/L)-1 cm-2 with linearity up to 11 mmol/L, and a detectionlimit of 8 ?mol/L estimated at a signal-to-noise ratio of 3.
Mesoporous silicas (MPSs) due to their good mechanical, thermal, chemical stability and large surface area have been proven to be promising as biomaterial immobilizationmatrix. It is worth noting that both the proper surface characteristics of the MPSs and goodmatching of the sizes of the enzyme molecules to the pore diameters of the MPSs arenecessary to the immobilization of enzyme. Unless the two factors are satisfied, theimmobilization of enzymes on MPSs will hardly reach. In addition, the poor conductivityof MPSs is also an important drawback of the materials, which might influence theperformance of the biosensor in amperometric detection. Gold nanoparticles (GNPs) havebeen exploited in biomolecule immobilization. Thus, the introduction of GNPs to MPSs isexpected to conquer the drawbacks of MPSs in enzyme immobilization. In chapter 4, goldnanoparticles-mesoporous silica composite (GNPs-MPS) is developed as a novel enzymeimmobilization matrix for biosensor construction. The mesoporous silica SBA-15 ischosen and the GNPs-SBA-15 is formed from AuCl4- adsorbed H2N-SBA-15 by NaBH4reduction. The synthesis process of the composite is monitored by UV-vis spectroscopyand the product is characterized by transmission electron microscopy (TEM) measurement.An amperometric glucose biosensor is built by immobilizing IO4--oxidized-glucoseoxidase (CHO-GOD) on GNPs-MPS modified Au electrode using 2-aminoethanethiol as across-linker. Cyclic voltammetry (CV) and amperometry are employed to investigate thecatalytic behavior of the biosensor to the oxidation of glucose. As a result, the biosensorexhibits an excellent bioelectrocatalytic response to glucose with a fast response time lessthan 7 s, a broad linear range of 0.02～14 mmol/L , high sensitivity of 6.1μA (mmol/L)-1cm-2, as well as good long-term stability and reproducibility. These performances could beascribed to the GNPs-MPS’s features, such as excellent conductivity, large surface areaand good biocompatibility.
The design, fabrication, study and application of nanoparticle-based nanostructuredfilms are currently intensely investigated research areas in materials chemistry. Thepotential utility of such thin films includes catalysis, optics, electrics and biosensors. The electrostatic layer-by-layer assembly of nanoparticles and polyelectrolytes is regarded asone of the most simple and versatile method for the construction of ultrathin organizedmultilayers. One of the key advantages of this approach has proved to be a rapid andexperimentally very convenient way to produce complex layered structures with precisecontrol of layer composition and thickness. However, the main defect of the multilayerfilms from electrostatic force is less stability toward polar solvents or electrolyte aqueoussolution, which limits its application range. However, electrostatic layer-by-layerassembled multilayers containing diazo-resins can be converted to covalent bonding at theinterfaces by UV irradiation. This method combines the simplicity of the ionic selfassemblytechnique and high stability of the covalently attached multilayer films. Inchapter 5, by using an ionic layer-by-layer self-assembly technique, the fabrication ofhighly stable diazo-resins/colloidal gold nanoparticles multilayer films on quartz wafer, Sislide and glassy carbon electrodes (GCEs) was achieved by the UV irradiation of layer-bylayerself-assemble multilayer films consisting of diazo-resins (DAR) and citrate-cappedcolloidal gold nanoparticles. UV irradiation converted the electrostatic interaction intocovalent bonds at the interfaces. These fabricating processes were followed and furtherconfirmed by UV-Vis spectrometry, Fourier transform infrared spectrometer (FTIR),cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Atomicforce microscopy (AFM) shows that these assemblies of colloidal gold nanoparticlesmultilayer films are highly stable and can be kept for a long time, only being removed byphysical scrape. As a biocompatible substrate, the result gold nanoparticles multiplayerfilm can be modified with dopamine, and used for ascorbic acid detection.
1.以聚苯乙烯小球为模板，通过电化学沉积的方法将金沉积到模板空隙中，四氢呋喃去除模板。所得金膜修饰电极具有层状有序的三维孔状结构，比表面积大，孔壁交错贯通，为葡萄糖的氧化提供了大量的反应位点，从而大大提高了其对葡萄糖响应的灵敏度；此外，该传感器可以在较低电位下实现葡萄糖的测定，有效的避免了抗坏血酸等物质的干扰。
2.以聚碳酸酯为模板，通过电化学沉积的方法将铂铅合金沉积到模板的孔隙中去，然后通过有机溶剂去除模板，即得具有微纳米线阵列结构的铂铅合金修饰电极。该修饰电极具有纳米微电极阵列的电化学行为，对葡萄糖展现出良好的电催化响应，可以在pH=7.4的中性磷酸缓冲体系中，在较低的电位下实现葡萄糖的测定。
3.以金纳米粒子/分子筛复合材料为基底，采用半胱胺作为交联剂，通过共价键合的方法将醛基化的葡萄糖氧化酶固定到基底电极上，从而实现葡萄糖生物传感器的构筑。该传感器对葡萄糖响应的线性范围上限可达14 mmol/L，此外，金纳米粒子/分子筛复合材料为酶分子提供了一个非常类似于生物体的微环境，从而可以有效保持生物分子的活性，进而提高了传感器的稳定性。
4.首先将柠檬酸根稳定的金纳米粒子与重氮树脂进行层层自组装，然后通过紫外光照使层间作用力由静电吸引转换为共价作用，从而得到一个具有高度稳定性的层数可控的金纳米粒子多层膜。随后将该金纳米粒子多层膜修饰电极浸入到多巴胺的溶液中，对多巴胺进行吸附，所得的多巴胺修饰电极对抗坏血酸响应灵敏，可以实现抗坏血酸的定量测定。
Nanomaterials, with their size in the range of 1-100 nm, have recently become one of themost exciting forefront fields in material science. Due to their small size, these materialsexhibit quanta-size effect, small-size effect, surface effect and tunneling effect that differfrom both bulk material and the individual atoms from which they comprised. With theseunique properties, they are widely used in the fields of catalysis, optical absorption,medicine, magnetic medium, new materials synthesis and particularly attractive in sensorapplications. Introduction of nanomaterials to the system of electrochemical sensors andbiosensors is of considerable interest, since they can play an important role in improvingthe sensor performance due to their large specific surface area, good conductivity specialcatalytic property and excellent biocompatibility.
In chapter 2, an enzyme-free glucose sensor has been developed using a threedimensionalinverse-opal gold film electrode (3DGFE) obtained by electrochemicaldeposition of gold in the interspaces of polystyrene templates. The gold films werecharacterized by scanning electron microscopy (SEM) and cyclic voltammetry (CV). Theelectrochemical oxidation of glucose in the presence of interferents (ascorbic acid, and so on) at different operating potentials on the 3DGFE has been investigated in detail. A lowoperating potential of -0.30 V was chosen for glucose detection, since the interferencecould be well avoided at this potential, whereas the current response for glucose oxidationwas still sensitive. The amperometric response of the sensor increased with the increase ofglucose concentration with a linear range of 5×10-6 ~ 10-2 mol/L and a detection limit of3.2μmol/L (signal-to-noise ratio of 3). The glucose sensor exhibited a high sensitivity of46.6μA (mmol/L)-1 cm-2, which could be ascribed to the unique surface structure of thethree-dimensionally interconnected porous structured gold films. The sensor with highsensitivity, good selectivity and stability is attractive for the practical glucose detection.
In chapter 3, Pt-Pb nanowire array was directly synthesized by electrochemicaldeposition of Pt-Pb alloy into the pores of macroporous polycarbonate template andsubsequent chemical etching of the template. The morphology and the composition of thePt-Pb nanowires were characterized by scanning electron microscopy (SEM) and X-rayphotoelectron spectroscopy (XPS), respectively. Cyclic voltammetry (CV) was used toevaluate the electrochemical performance of the Pt-Pb nanowire array electrode. Directglucose oxidation at such Pt-Pb nanowire array electrode was investigated detailedly bydiscussing the effect of the structure and materials of the electrode on electrocatalyticoxidation of glucose. As a result, we found that the Pt-Pb nanowire array electrode with athree-dimensional structure exhibited high electrocatalytic activity to glucose oxidation inneutral condition and could be used for the development of nonenzymatic glucose sensor.To effectively avoid the interference coming from ascorbic acid, a negative potential of -0.20 V was chosen for glucose detection, and the sensitivity of the sensor to glucoseoxidation was 11.25μA (mmol/L)-1 cm-2 with linearity up to 11 mmol/L, and a detectionlimit of 8 ?mol/L estimated at a signal-to-noise ratio of 3.
Mesoporous silicas (MPSs) due to their good mechanical, thermal, chemical stability and large surface area have been proven to be promising as biomaterial immobilizationmatrix. It is worth noting that both the proper surface characteristics of the MPSs and goodmatching of the sizes of the enzyme molecules to the pore diameters of the MPSs arenecessary to the immobilization of enzyme. Unless the two factors are satisfied, theimmobilization of enzymes on MPSs will hardly reach. In addition, the poor conductivityof MPSs is also an important drawback of the materials, which might influence theperformance of the biosensor in amperometric detection. Gold nanoparticles (GNPs) havebeen exploited in biomolecule immobilization. Thus, the introduction of GNPs to MPSs isexpected to conquer the drawbacks of MPSs in enzyme immobilization. In chapter 4, goldnanoparticles-mesoporous silica composite (GNPs-MPS) is developed as a novel enzymeimmobilization matrix for biosensor construction. The mesoporous silica SBA-15 ischosen and the GNPs-SBA-15 is formed from AuCl4- adsorbed H2N-SBA-15 by NaBH4reduction. The synthesis process of the composite is monitored by UV-vis spectroscopyand the product is characterized by transmission electron microscopy (TEM) measurement.An amperometric glucose biosensor is built by immobilizing IO4--oxidized-glucoseoxidase (CHO-GOD) on GNPs-MPS modified Au electrode using 2-aminoethanethiol as across-linker. Cyclic voltammetry (CV) and amperometry are employed to investigate thecatalytic behavior of the biosensor to the oxidation of glucose. As a result, the biosensorexhibits an excellent bioelectrocatalytic response to glucose with a fast response time lessthan 7 s, a broad linear range of 0.02～14 mmol/L , high sensitivity of 6.1μA (mmol/L)-1cm-2, as well as good long-term stability and reproducibility. These performances could beascribed to the GNPs-MPS’s features, such as excellent conductivity, large surface areaand good biocompatibility.
The design, fabrication, study and application of nanoparticle-based nanostructuredfilms are currently intensely investigated research areas in materials chemistry. Thepotential utility of such thin films includes catalysis, optics, electrics and biosensors. The electrostatic layer-by-layer assembly of nanoparticles and polyelectrolytes is regarded asone of the most simple and versatile method for the construction of ultrathin organizedmultilayers. One of the key advantages of this approach has proved to be a rapid andexperimentally very convenient way to produce complex layered structures with precisecontrol of layer composition and thickness. However, the main defect of the multilayerfilms from electrostatic force is less stability toward polar solvents or electrolyte aqueoussolution, which limits its application range. However, electrostatic layer-by-layerassembled multilayers containing diazo-resins can be converted to covalent bonding at theinterfaces by UV irradiation. This method combines the simplicity of the ionic selfassemblytechnique and high stability of the covalently attached multilayer films. Inchapter 5, by using an ionic layer-by-layer self-assembly technique, the fabrication ofhighly stable diazo-resins/colloidal gold nanoparticles multilayer films on quartz wafer, Sislide and glassy carbon electrodes (GCEs) was achieved by the UV irradiation of layer-bylayerself-assemble multilayer films consisting of diazo-resins (DAR) and citrate-cappedcolloidal gold nanoparticles. UV irradiation converted the electrostatic interaction intocovalent bonds at the interfaces. These fabricating processes were followed and furtherconfirmed by UV-Vis spectrometry, Fourier transform infrared spectrometer (FTIR),cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Atomicforce microscopy (AFM) shows that these assemblies of colloidal gold nanoparticlesmultilayer films are highly stable and can be kept for a long time, only being removed byphysical scrape. As a biocompatible substrate, the result gold nanoparticles multiplayerfilm can be modified with dopamine, and used for ascorbic acid detection.
引文
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