基于多級(jí)孔納米材料的電化學(xué)酶?jìng)鞲衅鞯闹苽浼皯?yīng)用研究
發(fā)布時(shí)間：2018-03-14 11:19
本文選題：電化學(xué)酶?jìng)鞲衅?/strong> 切入點(diǎn)：乙酰膽堿酯酶 出處：《吉林大學(xué)》2017年碩士論文 論文類(lèi)型：學(xué)位論文
【摘要】：隨著科學(xué)技術(shù)的不斷進(jìn)步,電化學(xué)酶?jìng)鞲衅饔捎诰邆涓哽`敏度和選擇性,快速響應(yīng),低成本等良好的性能,成為一類(lèi)檢測(cè)分析的工具,其應(yīng)用包括臨床,制藥,食品,環(huán)境和軍事等。其中,乙酰膽堿酯酶(ACh E)電化學(xué)傳感器更是被廣泛應(yīng)用于有機(jī)磷和氨基甲酸酯類(lèi)農(nóng)藥的檢測(cè)。而構(gòu)建電化學(xué)酶?jìng)鞲衅麝P(guān)鍵的步驟就是對(duì)電極表面的修飾。多級(jí)孔生物活性玻璃材料在骨缺損修復(fù)和藥物緩釋領(lǐng)域已得到很好的研究,但作為修飾劑在電化學(xué)分析檢測(cè)中的應(yīng)用較少。本文設(shè)計(jì)了三種多級(jí)孔生物活性玻璃材料修飾的ACh E電化學(xué)傳感器,分別運(yùn)用物理吸附法和共價(jià)鍵合法將ACh E固定在多級(jí)孔生物活性玻璃材料上。通過(guò)掃描電鏡、透射電鏡、X射線衍射光譜、比表面、傅里葉紅外光譜、熱重分析、X射線光電子能譜等表征手段,分別對(duì)三種材料進(jìn)行性質(zhì)研究。在對(duì)三種電化學(xué)酶?jìng)鞲衅鞯男阅苓M(jìn)行了研究和優(yōu)化后,完成了對(duì)有機(jī)磷農(nóng)藥模型馬拉硫磷和二嗪磷的檢測(cè)。1.以益母草花粉為生物模板的多級(jí)孔生物活性玻璃的制備及性質(zhì)探究利用生物模板法和水熱法,選用天然益母草花粉和非離子嵌段共聚物P123(EO20PO70EO20)作為雙模板制備多級(jí)孔生物活性玻璃(HMBG)。HMBG不僅具有良好的生物相容性和穩(wěn)定性,而且較大的比表面積可以促進(jìn)生物分子在材料表面的吸附和孔道內(nèi)的傳輸。在多級(jí)孔生物活性玻璃微球上嫁接具有氨基末端基團(tuán)的聚酰胺-胺(PAMAM)樹(shù)狀聚合物,得到活性位點(diǎn)更多的復(fù)合多級(jí)孔納米載體(HMBG-PAMAM)。2.物理吸附法固定乙酰膽堿酯酶的電化學(xué)傳感器的制備及應(yīng)用通過(guò)物理吸附法將ACh E分別固定在HMBG和HMBG-PAMAM兩種載體上,再分別修飾碳糊電極制備了HMBG-ACh E和HMBG-PAMAM-ACh E電化學(xué)傳感器。經(jīng)過(guò)實(shí)驗(yàn)條件優(yōu)化,以馬拉硫磷為農(nóng)藥模型,結(jié)果顯示在馬拉硫磷濃度為0.02 ppb~50 ppb和50 ppb~600 ppb的范圍內(nèi),農(nóng)藥濃度與電流抑制率成一定線性關(guān)系,HMBG-ACh E電化學(xué)傳感器的檢測(cè)限為0.0135 ppb(S/N=3),HMBG-PAMAM-ACh E電化學(xué)傳感器的檢測(cè)限為0.0192 ppb(S/N=3)。3.戊二醛共價(jià)鍵合法固定乙酰膽堿酯酶的電化學(xué)傳感器的制備及應(yīng)用通過(guò)共價(jià)鍵合法將ACh E的氨基與HMBG上接枝的氨基利用雙功能試劑戊二醛(GA)共價(jià)鍵合,得到的材料修飾碳糊電極制備一種HMBG-GA-ACh E電化學(xué)傳感器。經(jīng)過(guò)實(shí)驗(yàn)條件優(yōu)化,以馬拉硫磷和二嗪磷為農(nóng)藥模型,結(jié)果顯示在馬拉硫磷和二嗪磷濃度為0.02 ppb~10 ppb和10 ppb~600 ppb的范圍內(nèi),農(nóng)藥濃度與電流抑制率成一定線性關(guān)系,HMBG-GA-ACh E電化學(xué)傳感器對(duì)馬拉硫磷和二嗪磷的檢測(cè)限分別為0.0156 ppb(S/N=3)和0.0172 ppb(S/N=3)。對(duì)三種電化學(xué)酶?jìng)鞲衅鞯难芯拷Y(jié)果表明多級(jí)孔生物活性玻璃可以作為固定乙酰膽堿酯酶的載體,可以較好的保持酶的活性。三種乙酰膽堿酯酶電化學(xué)傳感器靈敏度高,檢測(cè)范圍廣,檢測(cè)限低,并且可以實(shí)現(xiàn)對(duì)實(shí)際樣品的檢測(cè)。
[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.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2017
【分類(lèi)號(hào)】：TB383.1;O657.1
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