聚酰胺—胺—生物/纳米功能复合物的合成及其在分析化学中的应用
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摘要
新材料的研究与应用一直是分析化学,尤其是电化学生物传感器的一项重要研究课题。纳米粒子的应用,使传感技术得到飞速发展,不同特性的纳米粒子形成的复合纳米材料,不仅具备纳米粒子自身的特性,还因其协同效应产生新的性质。本研究工作致力于多功能纳米复合物的合成与应用的探索。应用独特性质的碳纳米管、聚酰胺-胺树枝形聚合物合成两元和多元新型PAMAM/MWNT-纳米/生物酶复合物,将制备的酶纳米复合材料修饰到电极表面所得生物传感器可在低电位下实现对过氧化氢和葡萄糖的灵敏检测,能用于生物传感界面的构建。制备的生物传感器具有较长的寿命、较高的灵敏度、较低的检测下限以及快的响应速度。具体内容如下:
1合成了酶-PAMAM复合物和CdTe-PAMAM纳米复合物,并以酶的复合物构建生物传感器。
5.0 G PAMAM表面带有64个亲水的氨基,内部呈空穴状,具有类似天然蛋白质结构,具有很好的生物兼容性,具备同时携带多种物质和负载量大的优点,非常适合制备多功能复合物和生物复合物。在第2章中优化了PAMAM的合成条件。在对回收液成分分析的基础上,成功实现了低代PAMAM回收液直接循环利用的实验室模拟绿色合成。合成了10种纯度和得率均在98%以上的PAMAM(0.5 G-5.0 G)产品,并应用红外与元素分析进行表征。
在第3章中,以NaIO4氧化了的葡萄糖氧化酶和辣根过氧化物酶与PAMAM反应,制备了酶-PAMAM环状纳米复合物。该电极对H2O2具有很高的电流响应灵敏度,其灵敏度达360μA·L·mmol-1·cm-2,电极对H2O2的线性响应范围为3.1μM -2.0 mM,最低检测限达0.80μM。响应时间小于2 s。TEM成像显示酶-PAMAM可以通过自聚合形成环状纳米复合物。由于环状复合物中酶处于高度密集状态,固定量非常大,同时PAMAM的空穴结构,可使酶部分楔入其中,使其处于一种适宜的微环境中,能够较好地保持活性,在PAMAM复合物敏感膜中酶的活性比在一般敏感膜中酶的保持时间明显延长。电极对H2O2的电流响应很稳定,在3周时,其电流值为初始电流的91%,10周时,电流响应仍保持最初时的70 %左右。
HRP/GOx-PAMAM复合物生物传感器在3.0×10-6～1.5×10-3 mol L-1葡萄糖的浓度范围,电极呈线性响应,对葡萄糖的响应电流为170 A·L·mmol-1·cm-2,响应时间约2s。
在第4章中,合成了CdTe-PAMAM复合物,并应用TEM成像技术进行表征。PAMAM与量子点之间的能量转移特性,使得体系的荧光大大增强。由于量子点的表面在复合物中得到有效钝化与保护,其耐漂白性能得到显著增强。提出了结构有序CdTe-PAMAM复合物的可控合成方法。
该复合物荧光探针对Hg2+离子具有高的选择性和灵敏度,优化了Hg2+分析条件。在0.0010mol L-1的硝酸介质中,量子点复合物浓度为1.5×10-4mol L-1时,Hg2+离子浓度在5×10-7～1.2×10-5 mol L-1范围内与体系的相对荧光强度呈良好的线性关系。检测下限为9×10-8 mol L-1。量子点复合物浓度为2.0×10-5mol L-1时,Hg2+离子在1.2×10-9～4.0×10-7 mol L-1体系的相对荧光强度呈良好的线性关系,检测下限为5×10-10 mol L-1,并初步应用于实际样品中微量汞的测定。
2合成了PAMAM-MWNT,酶-PAMAM-MWNT复合物和CdTe-PAMAM-MWNT等新型纳米复合物,并应用酶复合物构建电化学生物传感器。
碳纳米管具有非常好的电催化活性、化学稳定性、生物兼容性和优良的能量/电荷转移特性,但它最大的缺陷是几乎不溶于所有溶剂,尤其是水,极大地局限了这种性能优良的纳米材料的研究与应用。
在第5章中,首先将碳纳米管在HNO3-H2SO4混酸中氧化,使碳纳米管壁以及末端生成羧基,将5.0 G PAMAM球型高分子聚合物接枝到碳纳米管上,合成了PAMAM-MWNT复合物,应用红外与荧光等对复合物进行表征。荧光研究发现复合物PAMAM-MWNT之间具有能量转移特性。由于PAMAM表面大量的氨基,PAMAM-MWNT表面带有大量的亲水基团,复合物在水与醇等有机溶剂中具有很好的分散性和稳定性,较好地解决了碳纳米管的溶解性(分散性)问题,扩展了它的应用范围。
在第6章中,将葡萄糖氧化酶和辣根过氧化酶的羟基用NaIO4选择性地氧化成醛基,然后与PAMAM-MWNT结合制备了酶复合物并将其以溶胶-凝胶法修饰到玻碳电极上制成响应双氧水和葡萄糖的生物传感器。由于复合物中酶与MWNTs等的协同作用,电极具有高灵敏度和快速电流响应;GOx和HRP活性中心和GCE表面实现直接电子转移,电极响应不需要电子媒介。工作电位在-0.34 V时,传感器对葡萄糖呈现高选择性,抗坏血酸、尿酸和多巴胺等其他生物分子不产生干扰。电极对葡萄糖的线性响应范围为4.0μM～1.2 mM,检测限为2.5μM,响应时间约为1s。响应灵敏度为2 200 nA mM-1。在PAMAM-MWNT复合物中,由于PAMAM特殊的类似蛋白质结构特性,可为酶提供适宜的微环境,酶的活性保持时间显著延长。
在第7章中,应用Te粉作为碲源,合成了巯基乙酸修饰的CdTe量子点。以巯基乙酸修饰的CdTe与PAMAM-MWNT共价结合,合成了密度可调控的三元复合物。应用TEM成像和红外、紫外可见光谱等表征了量子点复合物。确定了密度可调控的简单可行的合成路线。复合物中量子点的荧光发生强烈猝灭,表明CdTe-PAMAM-MWNT具有很好的能量转移/电荷转移特性,并发现PAMAM具有传递能量/电荷转移的功能。
3在第8章中,应用溶胶-凝胶法将钴(II)卟啉(CoTPP)和四苯硼钾(TPB)修饰到玻碳电极上研制成功对多巴胺和尿酸具有良好的催化活性的电化学传感器,并辅以红外技术研究了多巴胺在电极上的电催化氧化反应机理。利用膜相中TPB负电排斥作用,有效地消除了AA的干扰。建立了一种同时快速、灵敏测定多巴胺和尿酸的新方法。在0.10 mM AA存在时,DA的线性范围在6.0×10-8～1.0×10-5 M,相关系数为99.86%,检测限达2.0×10-8 M;UA的线性范围为1.0×10-7～2.0×10-5 M,相关系数为99.82%,检测限为7.0×10-8 M。
It is very important to develop new materials suitable for chemical and biosensors in the filed of analytical chemistry. Biosensors have been developed rapidly with the use of nanomaterials. Due to the synergic effect of different nanoparticles (NPs), the composites consisted of multi-nanoparticles possess many new properties different from those of original ones. This research aims to develop a simple and effective strategy for synthesizing dualistic and ternary novel enzymes/nanoparticles modified with poly(amidoamine) (PAMAM)/PAMAM-multi-walled carbon nanotubes (MWNTs). Enzymes in PAMAM/PAMAM-MWNT composites hold a high and long-term activity. The biosensors using these composites can be used to determine hydrogen peroxide at low potential with high sensitivity. Glucose oxidase (GOx) can be immobilized on to form biosensors with long lifetime, high sensitivity, low detection limit and fast response. The results of investigation are summarized as follows:
1 Enzymatic PAMAM and CdTe-PAMAM composites were synthesized for the construction of biosensors.
PAMAM (5.0 G with 64 amino groups in one molecule) characterized by inner cavity and with a structure similar to natural protein possesses excellent water-soluble properties. Being biocompatible, it is a promising material for preparing multi-functional composites. In Chapter 2, optimum conditions of synthesizing PAMAM were studied. A new green synthesis of low generation PAMAM have been experimentized by recycling the recovered solution directly based on the analysis of the recovered solutions in laboratory. Ten kinds of PAMAM (0.5 to 5.0 G) with more than 98% of purity and the yield were synthesized and characterized by IR and elemental analysis.
Horseradish peroxidase (HRP)-PAMAM and (GOx-HRP)-PAMAM were prepared and used for the construction of biosensors (Chapter 3). The nano-loop shape enzyme modified PAMAM were observed in TEM images. The research results demonstrated that the HRP-PAMAM composites could effectively immobilize HRP and retain the catalytic activities of enzyme to large extent. The enzyme biosensor retained 91% of its original activity after 3 weeks, and still held 70% of its original activity after 10 weeks. HRP-PAMAM modified electrode provided a linear response to H2O2 over a concentration range of 3.1×10-6 to 2.0×10-3 mol L-1 with a sensitivity of 360 μA·L·mmol-1·cm-2 with a detection limit of 8×10-7 mol L-1. The current response of the sensor is less than 2 s among the calibration range of H2O2.
The (GOx-HRP)-PAMAM modified membrane showed a linear response to glucose over a concentration range of 3.0×10-6 to 1.5×10-3 mol L-1 with a sensitivity of 170μA·L·mmol-1·cm-2 and a response time about 2s.
CdTe-PAMAM composite was prepared and characterized by TEM and UV-Vis spectra (Chapter 4). The simple controllable method for preparing the composite was proposed. Bleach-proof property of quantum dots (QDs) has been greatly improved because the surface of QDs was passivated and protected effectively in the composites. The fluorescence probe of CdTe-PAMAM showed high selectivity and sensitivity to mercuric ion. The relative fluorescence intensity decreased linearly with the mercury ions concentration in the range from 5×10-7 to 1.2×10-5 mol L-1 and the detection limit was 9×10-8 mol L-1 under the condition of using 1.5×10-4 mol L-1 composite. The relative fluorescence intensity decreased linearly with the mercury ions concentration in the range from 1.2×10-9 to 4.0×10-7 mol L-1 with the detection limit of 5×10-10 mol L-1 at 2.0×10-5 mol L-1of the composite. The method was applied to detect trace mercury in real samples with satisfactory results.
2 A novel composite PAMAM-MWNT and ternary enzymatic composites modified with PAMAM-MWNT as well as CdTe quantum dots attached PAMAM- MWNT were synthesized for construction of biosensors.
Carbon nanotubes possess many unique properties such as high electrocatalytic effect, fast electron transfer, chemical stability, excellent compatibility with biomaterals and energy/electron transfer capabilities. A major problem in using CNTs is their poor dispersability in all types of solvents, especially in water.
MWNTs were first oxidized by refluxing in concentrated HNO3 and H2SO4 to produce carboxylated groups on the wall as well as the tips of the MWNTs. By grafting with PAMAM dendrimer, novel carbon nanotube nanocomposites have been successfully prepared and were characterized by infrared (IR) and fluorescence (FL) spectra (Chapter 5). The novel functionalized matrix with plenty amino groups circumvents the troublesome solubility problem of CNTs in solvents, especially in water, greatly expanding the scope of the application of carbon nanotubes. The composites are excellent dispersible and long-term stable in water and in ethanol. The fluorescence research showed the property of energy transfer between PAMAM and MWNTs in the composites.
Carbohydrate groups on the peripheral surface of the GOx and the HRP molecules were oxidized with peridate to carboaldehydes. The GOx and HRP immobilized PAMAM-MWNT based on the functional WMNTs were synthesized (Chapter 6). The activity of enzymes can be retained well in the composites due to the favorable micro-environment of PAMAM. The bi-enzymatic PAMAM-MWNT nanocomposites are highly dispersible in water and show very promising applications in the fabrication of mediator-free bi-enzymatic biosensors for sensitive detection of H2O2 and glucose. The synergic effect of nanocomposite with CNT and high loading of GOx and HRP result in very high sensitivity to glucose with a current response of 2 200 nA mM-1 and fast response (~ 1s). The modified electrode exhibits a wide linear response range for glucose from 4.0μM to 1.2 mM, with a detection limit of 2.5μM. The negative electrode potential of -0.34 V is favorable for glucose detection in real samples without interference caused by other biomolecules.
A novel ternary nanocomposite of CdTe-PAMAM-MWNT was synthesized by covalently linking CdTe QDs onto highly water-soluble MWNTs functionalized with dendritic PAMAM (Chapter 7). A facile method for controlling the density of QDs in the composite has been developed by simply changing the ratio of QDs/PAMAM- MWNT. The fluorescence intensity of the CdTe-PAMAM hybrid was substantially enhanced as compared to that of QDs, and the fluorescence was quenched greatly when QDs reacted with PAMAM-MWNT. The experimentally observed phenomena indicate that electron and energy transfer took place efficiently between QDs, PAMAM and MWNTs in the composite.
3. In Chapter 8, we report the combination of the charge repelling property of tetraphenylborate (TPB) anion and the electro-oxidation catalytic effect of cobalt(II) tetrakis-phenylporphyrin (CoTPP) embedded in a sol gel ceramic film to develop a modified glassy carbon electrode (CoTPP-TPB-SGGCE) for the simultaneous determination of dopamine (DA) and uric acid (UA) determination. The optimized CoTPP-TPB-SGGCE shows excellent sensitivity and selectivity for the DA and UA analysis. As high as 2 000 fold acceptable tolerance of AA for the determination of trace DA and UA are reached. In the presence of 0.10 mM ascorbic acid (AA), the linear concentration range for DA is from 6.0×10-8 to 2.5×10-5 mol L-1 and the detection limit is 2.0×10-8 mol L-1. For UA, the linear concentration range is from 1.0×10-7 to 3.5×10-5 mol L-1 with the detection limit of 7.0×10-8 mol L-1. The results of experiment demonstrated that the novel CoTPP-TPB-SGGCE shows high stability and reliability. For 6.00μM DA and UA, a total of 12 measurements were taken in one week, and the relative standard deviation is 2.05% and 2.68% respectively. No obvious shift of peak current and peak potential is observed over a three-month lifetime test. The response of the sensor is very quick and response time is approximately 1 s.
1合成了酶-PAMAM复合物和CdTe-PAMAM纳米复合物,并以酶的复合物构建生物传感器。
5.0 G PAMAM表面带有64个亲水的氨基,内部呈空穴状,具有类似天然蛋白质结构,具有很好的生物兼容性,具备同时携带多种物质和负载量大的优点,非常适合制备多功能复合物和生物复合物。在第2章中优化了PAMAM的合成条件。在对回收液成分分析的基础上,成功实现了低代PAMAM回收液直接循环利用的实验室模拟绿色合成。合成了10种纯度和得率均在98%以上的PAMAM(0.5 G-5.0 G)产品,并应用红外与元素分析进行表征。
在第3章中,以NaIO4氧化了的葡萄糖氧化酶和辣根过氧化物酶与PAMAM反应,制备了酶-PAMAM环状纳米复合物。该电极对H2O2具有很高的电流响应灵敏度,其灵敏度达360μA·L·mmol-1·cm-2,电极对H2O2的线性响应范围为3.1μM -2.0 mM,最低检测限达0.80μM。响应时间小于2 s。TEM成像显示酶-PAMAM可以通过自聚合形成环状纳米复合物。由于环状复合物中酶处于高度密集状态,固定量非常大,同时PAMAM的空穴结构,可使酶部分楔入其中,使其处于一种适宜的微环境中,能够较好地保持活性,在PAMAM复合物敏感膜中酶的活性比在一般敏感膜中酶的保持时间明显延长。电极对H2O2的电流响应很稳定,在3周时,其电流值为初始电流的91%,10周时,电流响应仍保持最初时的70 %左右。
HRP/GOx-PAMAM复合物生物传感器在3.0×10-6～1.5×10-3 mol L-1葡萄糖的浓度范围,电极呈线性响应,对葡萄糖的响应电流为170 A·L·mmol-1·cm-2,响应时间约2s。
在第4章中,合成了CdTe-PAMAM复合物,并应用TEM成像技术进行表征。PAMAM与量子点之间的能量转移特性,使得体系的荧光大大增强。由于量子点的表面在复合物中得到有效钝化与保护,其耐漂白性能得到显著增强。提出了结构有序CdTe-PAMAM复合物的可控合成方法。
该复合物荧光探针对Hg2+离子具有高的选择性和灵敏度,优化了Hg2+分析条件。在0.0010mol L-1的硝酸介质中,量子点复合物浓度为1.5×10-4mol L-1时,Hg2+离子浓度在5×10-7～1.2×10-5 mol L-1范围内与体系的相对荧光强度呈良好的线性关系。检测下限为9×10-8 mol L-1。量子点复合物浓度为2.0×10-5mol L-1时,Hg2+离子在1.2×10-9～4.0×10-7 mol L-1体系的相对荧光强度呈良好的线性关系,检测下限为5×10-10 mol L-1,并初步应用于实际样品中微量汞的测定。
2合成了PAMAM-MWNT,酶-PAMAM-MWNT复合物和CdTe-PAMAM-MWNT等新型纳米复合物,并应用酶复合物构建电化学生物传感器。
碳纳米管具有非常好的电催化活性、化学稳定性、生物兼容性和优良的能量/电荷转移特性,但它最大的缺陷是几乎不溶于所有溶剂,尤其是水,极大地局限了这种性能优良的纳米材料的研究与应用。
在第5章中,首先将碳纳米管在HNO3-H2SO4混酸中氧化,使碳纳米管壁以及末端生成羧基,将5.0 G PAMAM球型高分子聚合物接枝到碳纳米管上,合成了PAMAM-MWNT复合物,应用红外与荧光等对复合物进行表征。荧光研究发现复合物PAMAM-MWNT之间具有能量转移特性。由于PAMAM表面大量的氨基,PAMAM-MWNT表面带有大量的亲水基团,复合物在水与醇等有机溶剂中具有很好的分散性和稳定性,较好地解决了碳纳米管的溶解性(分散性)问题,扩展了它的应用范围。
在第6章中,将葡萄糖氧化酶和辣根过氧化酶的羟基用NaIO4选择性地氧化成醛基,然后与PAMAM-MWNT结合制备了酶复合物并将其以溶胶-凝胶法修饰到玻碳电极上制成响应双氧水和葡萄糖的生物传感器。由于复合物中酶与MWNTs等的协同作用,电极具有高灵敏度和快速电流响应;GOx和HRP活性中心和GCE表面实现直接电子转移,电极响应不需要电子媒介。工作电位在-0.34 V时,传感器对葡萄糖呈现高选择性,抗坏血酸、尿酸和多巴胺等其他生物分子不产生干扰。电极对葡萄糖的线性响应范围为4.0μM～1.2 mM,检测限为2.5μM,响应时间约为1s。响应灵敏度为2 200 nA mM-1。在PAMAM-MWNT复合物中,由于PAMAM特殊的类似蛋白质结构特性,可为酶提供适宜的微环境,酶的活性保持时间显著延长。
在第7章中,应用Te粉作为碲源,合成了巯基乙酸修饰的CdTe量子点。以巯基乙酸修饰的CdTe与PAMAM-MWNT共价结合,合成了密度可调控的三元复合物。应用TEM成像和红外、紫外可见光谱等表征了量子点复合物。确定了密度可调控的简单可行的合成路线。复合物中量子点的荧光发生强烈猝灭,表明CdTe-PAMAM-MWNT具有很好的能量转移/电荷转移特性,并发现PAMAM具有传递能量/电荷转移的功能。
3在第8章中,应用溶胶-凝胶法将钴(II)卟啉(CoTPP)和四苯硼钾(TPB)修饰到玻碳电极上研制成功对多巴胺和尿酸具有良好的催化活性的电化学传感器,并辅以红外技术研究了多巴胺在电极上的电催化氧化反应机理。利用膜相中TPB负电排斥作用,有效地消除了AA的干扰。建立了一种同时快速、灵敏测定多巴胺和尿酸的新方法。在0.10 mM AA存在时,DA的线性范围在6.0×10-8～1.0×10-5 M,相关系数为99.86%,检测限达2.0×10-8 M;UA的线性范围为1.0×10-7～2.0×10-5 M,相关系数为99.82%,检测限为7.0×10-8 M。
It is very important to develop new materials suitable for chemical and biosensors in the filed of analytical chemistry. Biosensors have been developed rapidly with the use of nanomaterials. Due to the synergic effect of different nanoparticles (NPs), the composites consisted of multi-nanoparticles possess many new properties different from those of original ones. This research aims to develop a simple and effective strategy for synthesizing dualistic and ternary novel enzymes/nanoparticles modified with poly(amidoamine) (PAMAM)/PAMAM-multi-walled carbon nanotubes (MWNTs). Enzymes in PAMAM/PAMAM-MWNT composites hold a high and long-term activity. The biosensors using these composites can be used to determine hydrogen peroxide at low potential with high sensitivity. Glucose oxidase (GOx) can be immobilized on to form biosensors with long lifetime, high sensitivity, low detection limit and fast response. The results of investigation are summarized as follows:
1 Enzymatic PAMAM and CdTe-PAMAM composites were synthesized for the construction of biosensors.
PAMAM (5.0 G with 64 amino groups in one molecule) characterized by inner cavity and with a structure similar to natural protein possesses excellent water-soluble properties. Being biocompatible, it is a promising material for preparing multi-functional composites. In Chapter 2, optimum conditions of synthesizing PAMAM were studied. A new green synthesis of low generation PAMAM have been experimentized by recycling the recovered solution directly based on the analysis of the recovered solutions in laboratory. Ten kinds of PAMAM (0.5 to 5.0 G) with more than 98% of purity and the yield were synthesized and characterized by IR and elemental analysis.
Horseradish peroxidase (HRP)-PAMAM and (GOx-HRP)-PAMAM were prepared and used for the construction of biosensors (Chapter 3). The nano-loop shape enzyme modified PAMAM were observed in TEM images. The research results demonstrated that the HRP-PAMAM composites could effectively immobilize HRP and retain the catalytic activities of enzyme to large extent. The enzyme biosensor retained 91% of its original activity after 3 weeks, and still held 70% of its original activity after 10 weeks. HRP-PAMAM modified electrode provided a linear response to H2O2 over a concentration range of 3.1×10-6 to 2.0×10-3 mol L-1 with a sensitivity of 360 μA·L·mmol-1·cm-2 with a detection limit of 8×10-7 mol L-1. The current response of the sensor is less than 2 s among the calibration range of H2O2.
The (GOx-HRP)-PAMAM modified membrane showed a linear response to glucose over a concentration range of 3.0×10-6 to 1.5×10-3 mol L-1 with a sensitivity of 170μA·L·mmol-1·cm-2 and a response time about 2s.
CdTe-PAMAM composite was prepared and characterized by TEM and UV-Vis spectra (Chapter 4). The simple controllable method for preparing the composite was proposed. Bleach-proof property of quantum dots (QDs) has been greatly improved because the surface of QDs was passivated and protected effectively in the composites. The fluorescence probe of CdTe-PAMAM showed high selectivity and sensitivity to mercuric ion. The relative fluorescence intensity decreased linearly with the mercury ions concentration in the range from 5×10-7 to 1.2×10-5 mol L-1 and the detection limit was 9×10-8 mol L-1 under the condition of using 1.5×10-4 mol L-1 composite. The relative fluorescence intensity decreased linearly with the mercury ions concentration in the range from 1.2×10-9 to 4.0×10-7 mol L-1 with the detection limit of 5×10-10 mol L-1 at 2.0×10-5 mol L-1of the composite. The method was applied to detect trace mercury in real samples with satisfactory results.
2 A novel composite PAMAM-MWNT and ternary enzymatic composites modified with PAMAM-MWNT as well as CdTe quantum dots attached PAMAM- MWNT were synthesized for construction of biosensors.
Carbon nanotubes possess many unique properties such as high electrocatalytic effect, fast electron transfer, chemical stability, excellent compatibility with biomaterals and energy/electron transfer capabilities. A major problem in using CNTs is their poor dispersability in all types of solvents, especially in water.
MWNTs were first oxidized by refluxing in concentrated HNO3 and H2SO4 to produce carboxylated groups on the wall as well as the tips of the MWNTs. By grafting with PAMAM dendrimer, novel carbon nanotube nanocomposites have been successfully prepared and were characterized by infrared (IR) and fluorescence (FL) spectra (Chapter 5). The novel functionalized matrix with plenty amino groups circumvents the troublesome solubility problem of CNTs in solvents, especially in water, greatly expanding the scope of the application of carbon nanotubes. The composites are excellent dispersible and long-term stable in water and in ethanol. The fluorescence research showed the property of energy transfer between PAMAM and MWNTs in the composites.
Carbohydrate groups on the peripheral surface of the GOx and the HRP molecules were oxidized with peridate to carboaldehydes. The GOx and HRP immobilized PAMAM-MWNT based on the functional WMNTs were synthesized (Chapter 6). The activity of enzymes can be retained well in the composites due to the favorable micro-environment of PAMAM. The bi-enzymatic PAMAM-MWNT nanocomposites are highly dispersible in water and show very promising applications in the fabrication of mediator-free bi-enzymatic biosensors for sensitive detection of H2O2 and glucose. The synergic effect of nanocomposite with CNT and high loading of GOx and HRP result in very high sensitivity to glucose with a current response of 2 200 nA mM-1 and fast response (~ 1s). The modified electrode exhibits a wide linear response range for glucose from 4.0μM to 1.2 mM, with a detection limit of 2.5μM. The negative electrode potential of -0.34 V is favorable for glucose detection in real samples without interference caused by other biomolecules.
A novel ternary nanocomposite of CdTe-PAMAM-MWNT was synthesized by covalently linking CdTe QDs onto highly water-soluble MWNTs functionalized with dendritic PAMAM (Chapter 7). A facile method for controlling the density of QDs in the composite has been developed by simply changing the ratio of QDs/PAMAM- MWNT. The fluorescence intensity of the CdTe-PAMAM hybrid was substantially enhanced as compared to that of QDs, and the fluorescence was quenched greatly when QDs reacted with PAMAM-MWNT. The experimentally observed phenomena indicate that electron and energy transfer took place efficiently between QDs, PAMAM and MWNTs in the composite.
3. In Chapter 8, we report the combination of the charge repelling property of tetraphenylborate (TPB) anion and the electro-oxidation catalytic effect of cobalt(II) tetrakis-phenylporphyrin (CoTPP) embedded in a sol gel ceramic film to develop a modified glassy carbon electrode (CoTPP-TPB-SGGCE) for the simultaneous determination of dopamine (DA) and uric acid (UA) determination. The optimized CoTPP-TPB-SGGCE shows excellent sensitivity and selectivity for the DA and UA analysis. As high as 2 000 fold acceptable tolerance of AA for the determination of trace DA and UA are reached. In the presence of 0.10 mM ascorbic acid (AA), the linear concentration range for DA is from 6.0×10-8 to 2.5×10-5 mol L-1 and the detection limit is 2.0×10-8 mol L-1. For UA, the linear concentration range is from 1.0×10-7 to 3.5×10-5 mol L-1 with the detection limit of 7.0×10-8 mol L-1. The results of experiment demonstrated that the novel CoTPP-TPB-SGGCE shows high stability and reliability. For 6.00μM DA and UA, a total of 12 measurements were taken in one week, and the relative standard deviation is 2.05% and 2.68% respectively. No obvious shift of peak current and peak potential is observed over a three-month lifetime test. The response of the sensor is very quick and response time is approximately 1 s.
引文
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