基于多级孔纳米材料的电化学酶传感器的制备及应用研究

发布时间：2018-03-14 11:19

本文选题：电化学酶传感器　切入点：乙酰胆碱酯酶　出处：《吉林大学》2017年硕士论文　论文类型：学位论文

【摘要】：随着科学技术的不断进步,电化学酶传感器由于具备高灵敏度和选择性,快速响应,低成本等良好的性能,成为一类检测分析的工具,其应用包括临床,制药,食品,环境和军事等。其中,乙酰胆碱酯酶(ACh E)电化学传感器更是被广泛应用于有机磷和氨基甲酸酯类农药的检测。而构建电化学酶传感器关键的步骤就是对电极表面的修饰。多级孔生物活性玻璃材料在骨缺损修复和药物缓释领域已得到很好的研究,但作为修饰剂在电化学分析检测中的应用较少。本文设计了三种多级孔生物活性玻璃材料修饰的ACh E电化学传感器,分别运用物理吸附法和共价键合法将ACh E固定在多级孔生物活性玻璃材料上。通过扫描电镜、透射电镜、X射线衍射光谱、比表面、傅里叶红外光谱、热重分析、X射线光电子能谱等表征手段,分别对三种材料进行性质研究。在对三种电化学酶传感器的性能进行了研究和优化后,完成了对有机磷农药模型马拉硫磷和二嗪磷的检测。1.以益母草花粉为生物模板的多级孔生物活性玻璃的制备及性质探究利用生物模板法和水热法,选用天然益母草花粉和非离子嵌段共聚物P123(EO20PO70EO20)作为双模板制备多级孔生物活性玻璃(HMBG)。HMBG不仅具有良好的生物相容性和稳定性,而且较大的比表面积可以促进生物分子在材料表面的吸附和孔道内的传输。在多级孔生物活性玻璃微球上嫁接具有氨基末端基团的聚酰胺-胺(PAMAM)树状聚合物,得到活性位点更多的复合多级孔纳米载体(HMBG-PAMAM)。2.物理吸附法固定乙酰胆碱酯酶的电化学传感器的制备及应用通过物理吸附法将ACh E分别固定在HMBG和HMBG-PAMAM两种载体上,再分别修饰碳糊电极制备了HMBG-ACh E和HMBG-PAMAM-ACh E电化学传感器。经过实验条件优化,以马拉硫磷为农药模型,结果显示在马拉硫磷浓度为0.02 ppb~50 ppb和50 ppb~600 ppb的范围内,农药浓度与电流抑制率成一定线性关系,HMBG-ACh E电化学传感器的检测限为0.0135 ppb(S/N=3),HMBG-PAMAM-ACh E电化学传感器的检测限为0.0192 ppb(S/N=3)。3.戊二醛共价键合法固定乙酰胆碱酯酶的电化学传感器的制备及应用通过共价键合法将ACh E的氨基与HMBG上接枝的氨基利用双功能试剂戊二醛(GA)共价键合,得到的材料修饰碳糊电极制备一种HMBG-GA-ACh E电化学传感器。经过实验条件优化,以马拉硫磷和二嗪磷为农药模型,结果显示在马拉硫磷和二嗪磷浓度为0.02 ppb~10 ppb和10 ppb~600 ppb的范围内,农药浓度与电流抑制率成一定线性关系,HMBG-GA-ACh E电化学传感器对马拉硫磷和二嗪磷的检测限分别为0.0156 ppb(S/N=3)和0.0172 ppb(S/N=3)。对三种电化学酶传感器的研究结果表明多级孔生物活性玻璃可以作为固定乙酰胆碱酯酶的载体,可以较好的保持酶的活性。三种乙酰胆碱酯酶电化学传感器灵敏度高,检测范围广,检测限低,并且可以实现对实际样品的检测。
[Abstract]:With the development of science and technology, electrochemical enzyme sensors have become a kind of tool for detection and analysis because of their high sensitivity and selectivity, fast response, low cost and so on. Their applications include clinical, pharmaceutical, food, etc. Environment, military, etc. The electrochemical sensor of acetylcholinesterase (ache) has been widely used in the detection of organophosphorus and carbamate pesticides. The key step to construct the electrochemical enzyme sensor is to modify the electrode surface. Vitreous materials have been well studied in the field of bone defect repair and drug delivery. However, there are few applications as modifiers in electrochemical analysis. In this paper, three kinds of multiporous bioactive glass modified ACh E electrochemical sensors are designed. ACh E was immobilized on multiporous bioactive glass by physical adsorption method and covalent bonding method, respectively. By scanning electron microscope, transmission electron microscope, X-ray diffraction spectrum, specific surface, Fourier transform infrared spectroscopy, The properties of three kinds of materials were studied by means of thermogravimetric analysis and X-ray photoelectron spectroscopy. After the performance of three kinds of electrochemical enzyme sensors were studied and optimized, The detection of malathion and diazinophos in organophosphorus pesticide model was completed. 1. Preparation and properties of multiporous bioactive glass with motherwort pollen as biological template. Using natural Leonurus pollen and Nonionic block copolymer P123O20PO70EO20 as double templates to prepare multiporous bioactive glass, HMBGG. HMBG has good biocompatibility and stability. Moreover, the larger specific surface area can promote the adsorption of biomolecules on the surface of materials and the transport of biomolecules in the pores. The polyamide-amine polyamines (PAMAM) dendritic polymers with amino terminal groups are grafted on multilevel porous bioactive glass microspheres. HMBG-PAMAMN. 2. Preparation and Application of Electrochemical biosensor for immobilization of acetylcholinesterase by physical adsorption method ACh E was immobilized on HMBG and HMBG-PAMAM respectively by physical adsorption. The electrochemical sensors of HMBG-ACh E and HMBG-PAMAM-ACh E were prepared by modified carbon paste electrode respectively. Using malathion as the pesticide model, the experimental conditions were optimized. The results showed that the concentration of malathion was 0.02 ppb~50 ppb and 50 ppb~600 ppb, respectively. The detection limit of HMBG-ACHE electrochemical sensor is 0.0135 ppbS- / Na-3HMBG-PAMAM-ACHE. The detection limit of HMBG-PAMAM-ACHE electrochemical sensor is 0.0192 ppb ~ (-1) 路s / N ~ (3) N ~ (3) N ~ (3). Glutaraldehyde covalent bond method for immobilized acetylcholinesterase. The preparation and application of glutaraldehyde (Glutaraldehyde), a bifunctional reagent, covalently bonded the amino groups of ACh E with the amino groups grafted on HMBG by covalent bonding method. A HMBG-GA-ACh E electrochemical sensor was prepared by modified carbon paste electrode. After optimized experimental conditions, malathion and diazinophos were used as pesticide models. The results showed that the concentration of malathion and diazinophos was 0.02 ppb~10 ppb and 10 ppb~600 ppb, respectively. The detection limits of malathion and diazinphosphos by HMBG-GA-ACHE electrochemical sensor were 0.0156 ppbS- / Na-3 and 0.0172 ppbS- / Na-3, respectively. The results of the study on three kinds of electrochemical enzyme sensors showed that the multipore bioactivity was higher than that of the HMBG-GA-ACHE electrochemical sensor. Glass can be used as a carrier of acetylcholinesterase. Three kinds of acetylcholinesterase electrochemical sensors have high sensitivity, wide detection range, low detection limit, and can be used to detect actual samples.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：TB383.1;O657.1

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