發(fā)布時間：2018-05-28 06:35

  本文選題：二氧化鈦納米管陣列 + 銀納米顆粒�。� 參考：《東南大學(xué)》2015年博士論文

【摘要】：電化學(xué)酶生物傳感器是一種將電化學(xué)分析方法與酶生物技術(shù)相結(jié)合的生物傳感器,已在臨床檢測、環(huán)境監(jiān)測以及食品、制藥等領(lǐng)域表現(xiàn)出廣闊的應(yīng)用前景。目前,在酶生物傳感器的制備和應(yīng)用過程中,所面臨的最大問題就是酶電極的生物電催化活性較低,且穩(wěn)定性較差,容易失活。然而,此類問題與酶電極制備過程中所采用的酶固定化方法、以及酶固定化所選用載體材料密切相關(guān)。此外,酶電極的生物電催化活性不僅與載體材料的親水性密切相關(guān),而且與載體材料的導(dǎo)電性也密切相關(guān)�；诖�,本論文從新型二氧化鈦(Ti02)納米載體材料用于酶電極的構(gòu)建以及酶的高效固定化方法等方面進(jìn)行了探索性的研究。本論文研究工作主要是基于TiO2 NTAs酶電極的制備及其應(yīng)用研究。設(shè)計(jì)合成了4種不同TiO2 NTAs基載體材料：二氧化鈦納米管陣列(TiO2 NTAs),二氧化鈦納米管陣列/銀納米顆粒(TiO2 NTAs/AgNPs),二氧化鈦納米管陣列/還原氧化石墨烯/銀納米顆粒(TiO2 NTAs/r-GO/AgNPs),氮摻雜二氧化鈦納米管陣列(N-TiO2 NTAs)�；谝陨纤苽涞腡iO2 NTAs基載體材料構(gòu)建了一系列具有較高生物電催化活性的TiO2 NTAs基酶電極,并研究了所制備酶電極在生物電催化領(lǐng)域中的應(yīng)用。本論文研究的具體工作包括以下幾個方面：1、TiO2 NTAs/GOx酶電極的制備及其生物電催化性能研究。TiO2 NTAs具有良好的生物相容性和良好的親水性,并且具有較大的比表面積和較強(qiáng)的吸附能力：此外,高度有序的TiO2納米陣列可以為電子傳輸提供單向通道,有利于電子傳導(dǎo)。因而,TiO2 NTAs是一種理想的酶電極載體材料。探索研究了以TiO2 NTAs為載體材料,在其表面進(jìn)行葡萄糖氧化酶(GOx)的固定化,以制備TiO2 NTAs/GOx酶電極。提出了一種可直接將GOx固定化于TiO2 NTAs表面以制備酶電極的新的酶固定化方法,即改進(jìn)交聯(lián)法。與傳統(tǒng)的交聯(lián)法相比,該種改進(jìn)交聯(lián)法可以直接將酶固定化于載體材料的表面,避免酶的活性中心在固定化過程中被包埋,因此可以有效提升酶電極的生物電催化活性。研究表明,基于TiO2 NTAs/GOx酶電極的生物傳感器對葡萄糖濃度響應(yīng)的線性范圍為0.05-0.65mM,其檢測靈敏度為199.61 μmM-1 cm-2。因此,TiO2 NTAs/GOx酶電極具有良好的生物電催化活性,可以作為葡萄糖生物傳感器用于葡萄糖的檢測。2、TiO2 NTAs/AgNPs/GOx酶電極的制備及其生物電催化性能研究。銀不僅是導(dǎo)電性最好的貴金屬材料,而且具有良好的生物相容性,基于AgNPs修飾酶生物傳感器具有較強(qiáng)的檢測靈敏度。探索研究了以TiO2 NTAs/AgNPs為載體材料的酶電極,通過提高載體材料電子傳遞能力來提升酶電極生物電催化活性。本研究采用了化學(xué)沉積法將AgNPs沉積于TiO2 NTAs表面,得到了TiO2 NTAs/AgNPs,單個銀顆粒的尺寸大小在30到100 nm范圍之間。然后,采用改進(jìn)交聯(lián)法將GOx固定化于TiO2 NTAs/AgNPs載體材料表面制備得到了TiO2 NTAs/AgNPs/GOx酶電極,并研究的該酶電極在生物電催化領(lǐng)域中的應(yīng)用。研究表明,基于TiO2 NTAs/AgNPs/GOx酶電極的生物傳感器對葡萄糖濃度響應(yīng)的線性范圍為0.05-0.65 mM,其檢測靈敏度為207.43 μA mM-1。與TiO2 NTAs/GOx酶電極的檢測靈敏度(199.61 μAmM-1cm-2)相比較為接近或者有輕微的提升,但重要的是TiO2 NTAs/AgNPs電極電子傳遞能力的提升,使得其在酶生物燃料電池領(lǐng)域中的應(yīng)用成為了可能。基于TiO2 NTAs/AgNPs/GOx酶電極的生物燃料電池的開路電壓為0.202 V,短路電流為0.197 mA cm-2最大輸出功率密度為8.66μW cm-2。因此,TiO2 NTAs/AgNPs/GOx酶電極的生物電催化活性高于TiO2 NTAs/GOx酶電極,能夠一定程度上提高葡萄糖生物傳感器的檢測靈敏度性能,并且可以應(yīng)用于構(gòu)建酶生物燃料電池。3、TiO2 NTAs/r-GO/AgNPs/GOx酶電極的制備及其生物電催化性能研究。石墨烯與納米顆粒之間能夠形成的較強(qiáng)的范德華力,可以有效防止其表面所沉積的納米顆粒的團(tuán)聚,在石墨烯的表面進(jìn)行納米顆粒的沉積可以得到較小尺寸的納米顆粒。此外,r-GO表面富含親水性基團(tuán),這些親水性基團(tuán)有助于酶電極構(gòu)建過程中提升酶的固定化效率。為了解決TiO2 NTAs/AgNPs載體材料中AgNPs的團(tuán)聚現(xiàn)象、粒徑大小不一和分布不太均勻等問題,探索研究了以TiO2 NTAs/r-GO/AgNPs為載體材料的酶電極,通過提高表面修飾AgNPs 5分散性能來提升酶電極生物電催化活性。采用化學(xué)沉積法將AgNPs沉積于r-GO修飾的TiO2 NTAs表面,得到了TiO2 NTAs/r-GO/AgNPs。然后,采用改進(jìn)交聯(lián)法將GOx固定化于TiO2 NTAs/r-GO/AgNPs載體材料表面制備得到了TiO2 NTAs/r-GO/AgNPs/GOx酶電極,并研究了該酶電極在生物電催化領(lǐng)域中的應(yīng)用。研究表明,與TiO2 NTAs/AgNPs相比, TiO2 NTAs/r-GO/AgNPs表面AgNPs的覆蓋密度更大,AgNPs顆粒尺寸又小又均勻,單個銀顆粒的尺寸大小在20到30 nm范圍之間。基于TiO2NTAs/r-GO/AgNPs/GOx酶電極的生物傳感器對葡萄糖濃度響應(yīng)的線性范圍為0.05-0.3mM,其檢測靈敏度為257.79 μAmM-1,比較TiO2 NTAs/AgNPs/GOx酶電極的檢測靈敏度(207.43 μA mM-1 cm-2)有明顯的提升；基于TiO2 NTAs/r-GO/AgNPs/GOx酶電極的生物燃料電池的開路電壓為0.225 V,短路電流為0.232 mA cm-2,最大輸出功率密度為13.45 μW cm-2,比較基于TiO2 NTAs/AgNPs/GOx酶電極的生物燃料電池(8.66 μW cm-2)有一定程度的提升。因此,TiO2 NTAs/r-GO/AgNPs/GOx酶電極的生物電催化活性優(yōu)于TiO2 NTAs/AgNPs/GOx酶電極,可以進(jìn)一步提高葡萄糖生物傳感器的檢測靈敏度和酶生物燃料電池的最大輸出功率密度。4、N-TiO2 NTAs/GOx酶電極的制備及其生物電催化性能研究。氮元素?fù)诫s改性Ti02不僅具有較好的親水性,而且與Ti02相比具有較高的導(dǎo)電性。為了既能提升TiO2 NTAs載體材料的電子傳遞能力,又能保持其親水性,采用氮元素?fù)诫s改性TiO2 NTAs制備得到了N-TiO2 NTAs。TiO2和N-TiO2材料的接觸角分別為39°和44°,因此,它們具有相近而優(yōu)良的親水性；N-TiO2 NTAs電荷轉(zhuǎn)移電阻(Rct)為500.2 ohm,明顯低于TiO2 NTAs的Rct (6431 ohm),因此,N-TiO2 NTAs與TiO2 NTAs相比具有更好的電子傳遞能力。采用改進(jìn)交聯(lián)法將GOx固定化于N-TiO2 NTAs載體材料表面制備了N-TiO2 NTAs/GOx酶電極,并研究了該酶電極在生物電催化領(lǐng)域中的應(yīng)用。研究表明,基于N-TiO2 NTAs/GOx酶電極的生物傳感器對葡萄糖濃度響應(yīng)的線性范圍為0.05-0.85 mM,其檢測靈敏度為733.17 μA mM-1 cm-2,比較TiO2 NTAs/GOx酶電極的檢測靈敏度(199.61μA mM-1 cm2)有非常顯著的提升。此外,基于N-TiO2 NTAs/GOx酶電極的生物燃料電池的最大輸出功率密度為23.92 μW cm-2,比較基于TiO2 NTAs/GOx酶電極(5.38 μW cm-2)和TiO2 NTAs/r-GO/AgNPs/GOx酶電極(13.45 μW cm-2)的生物燃料電池都有明顯的提升。因此,N-TiO2 NTAs/GOx酶電極的生物電催化活性明顯優(yōu)于TiO2 NTAs/GOx和Ti02NTAs/r-GO/AgNPs/GOx酶電極,可以顯著提高葡萄糖生物傳感器的檢測靈敏度和酶生物燃料電池的最大輸出功率密度。
[Abstract]:Electrochemical enzyme biosensor is a kind of biosensor which combines electrochemical analysis method with enzyme biological technology. It has been widely used in clinical detection, environmental monitoring, food, pharmaceutical and other fields. At present, the biggest problem in the preparation and application of enzyme biosensor is the production of enzyme electrode. In addition, the bioelectrocatalytic activity of the enzyme electrode is not only closely related to the hydrophilicity of the carrier material, but also is closely related to the hydrophilic property of the carrier material, and the conductivity of the enzyme electrode. Based on this, this thesis has studied the construction of the new titanium dioxide (Ti02) nanoscale material for the construction of the enzyme electrode and the efficient immobilization of the enzyme. The main work of this thesis is based on the preparation and application of the TiO2 NTAs enzyme electrode. 4 different TiO2 NTAs bases are designed and synthesized. Carrier materials: titanium dioxide nanotube array (TiO2 NTAs), titanium dioxide nanotube array / silver nanoparticles (TiO2 NTAs/AgNPs), titanium dioxide nanotube array / reduced graphite oxide / silver nanoparticles (TiO2 NTAs/r-GO/AgNPs), nitrogen doped titanium dioxide nanotube array (N-TiO2 NTAs). Based on the TiO2 NTAs base material prepared above A series of TiO2 NTAs based enzyme electrodes with high bioelectrocatalytic activity were constructed and the application of the enzyme electrode in the field of bioelectrocatalysis was studied. The specific work of this study included the following aspects: 1, the preparation of the TiO2 NTAs/GOx enzyme electrode and the study on the bioelectrocatalytic performance of.TiO2 NTAs. In addition, highly ordered TiO2 nanowire arrays can provide unidirectional channels for electronic transmission and facilitate electron conduction. Therefore, TiO2 NTAs is an ideal material for the carrier of enzyme electrode. TiO2 NTAs as the carrier material is explored and studied. The immobilized glucose oxidase (GOx) is immobilized on the surface to prepare the TiO2 NTAs/GOx enzyme electrode. A new enzyme immobilization method, which can directly immobilize GOx on the surface of TiO2 NTAs to prepare the enzyme electrode, is proposed, which is improved crosslinking method. Compared with the traditional crosslinking method, the improved crosslinking method can directly immobilize the enzyme in the table of carrier material. In order to avoid the enzyme activity center in the immobilized process, it can effectively enhance the bioelectrocatalytic activity of the enzyme electrode. The study shows that the linear range of the response of biosensor based on TiO2 NTAs/GOx enzyme electrode is 0.05-0.65mM, and its sensitivity is 199.61 mM-1 cm-2., therefore, TiO2 NTAs/GOx enzyme electrode It has good bioelectrocatalytic activity and can be used as a glucose biosensor for the determination of.2, TiO2 NTAs/AgNPs/GOx enzyme electrode and its bioelectrocatalytic performance. Silver is not only the best metal material with the best conductivity, but also has good biocompatibility, and the biosensor based on AgNPs modifier has a better effect. The enzyme electrode with TiO2 NTAs/AgNPs as the carrier material is explored and studied. The bioelectrocatalytic activity of the enzyme electrode is enhanced by increasing the electron transfer ability of the carrier material. The chemical deposition method is used to deposit AgNPs on the surface of TiO2 NTAs, and the TiO2 NTAs/ AgNPs is obtained. The size of the single silver particles is from 30 to 100 n. M ranges between them. Then, the modified crosslinking method is used to obtain the TiO2 NTAs/AgNPs/GOx enzyme electrode on the surface of GOx immobilized on the TiO2 NTAs/AgNPs carrier material, and the application of the enzyme electrode in the field of bioelectrocatalysis is studied. The study shows that the linear model of the response of the biosensor based on the TiO2 NTAs/AgNPs/GOx enzyme electrode to the glucose concentration response is shown. The detection sensitivity is 0.05-0.65 mM, and its detection sensitivity is 207.43 mu A mM-1. and the detection sensitivity of TiO2 NTAs/GOx electrode (199.61 mu AmM-1cm-2) is close or slightly elevated, but it is important to improve the electron transfer ability of TiO2 NTAs/AgNPs electrode, so that its application in the field of enzyme biofuel battery has become possible. The open circuit voltage of the biofuel battery of the TiO2 NTAs/AgNPs/GOx enzyme electrode is 0.202 V, the short circuit current is 0.197 mA cm-2 and the maximum output power density is 8.66 u W cm-2., so the bioelectrocatalytic activity of the TiO2 NTAs/AgNPs/GOx enzyme electrode is higher than the TiO2 NTAs/GOx enzyme electrode, and the detection sensitivity of the glucose biosensor can be improved to a certain extent. Degree performance and can be applied to the preparation of.3, TiO2 NTAs/r-GO/AgNPs/GOx enzyme electrode and its bioelectrocatalytic performance. The strong Fan Dehua force formed between graphene and nanoparticles can effectively prevent the agglomeration of nanoparticles deposited on the surface of the graphene, and make nano particles on the surface of graphene. In addition, the r-GO surface is rich in hydrophilic groups. These hydrophilic groups are helpful to the immobilization efficiency of the enzyme during the construction of the enzyme electrode. In order to solve the problem of the aggregation of AgNPs in the carrier material of the TiO2 NTAs/AgNPs carrier, the size of the particle size is small and the distribution is not too uniform and the distribution is not too uniform. The enzyme electrode with TiO2 NTAs/r-GO/AgNPs as the carrier material was investigated by increasing the dispersion properties of the surface modified AgNPs 5 to improve the bioelectrocatalytic activity of the enzyme electrode. AgNPs was deposited on the r-GO modified TiO2 NTAs surface by chemical deposition, and TiO2 NTAs/r-GO/AgNPs. was obtained, and the GOx was immobilized to TiO2 NTAs/r-GO/A by the improved cross linking method. The TiO2 NTAs/r-GO/AgNPs/GOx enzyme electrode was prepared on the surface of the gNPs carrier material, and the application of the enzyme electrode in the field of bioelectrocatalysis was studied. Compared with TiO2 NTAs/AgNPs, the density of AgNPs on the surface of TiO2 NTAs/r-GO/AgNPs was larger, the size of AgNPs particles was small and uniform, and the size of single silver particles was 20 to 30 n. Between the M range, the linear range of the biosensor based on the TiO2NTAs/r-GO/AgNPs/GOx enzyme electrode response to glucose concentration is 0.05-0.3mM, and its detection sensitivity is 257.79 mu AmM-1, and the detection sensitivity of the TiO2 NTAs/AgNPs/GOx electrode (207.43 mu A mM-1 cm-2) has obvious lifting; based on the TiO2 NTAs/r-GO/AgNPs/GOx enzyme electrode. The open circuit voltage of the fuel cell is 0.225 V, the short-circuit current is 0.232 mA cm-2, the maximum output power density is 13.45 W cm-2, and the biofuel battery based on the TiO2 NTAs/AgNPs/GOx enzyme electrode has a certain degree of improvement. Therefore, the bioelectrocatalytic activity of the TiO2 NTAs/r-GO/AgNPs/GOx enzyme electrode is better than that of the TiO2. The GOx enzyme electrode can further improve the sensitivity of the glucose biosensor and the maximum output power density of the enzyme biofuel battery.4, the preparation of the N-TiO2 NTAs/GOx enzyme electrode and its bioelectrocatalytic performance. The nitrogen doped modified Ti02 not only has better hydrophilicity, but also has higher conductivity compared with Ti02. In order to improve the electronic transfer ability of TiO2 NTAs carrier material and keep its hydrophilic property, the contact angles of N-TiO2 NTAs.TiO2 and N-TiO2 materials are 39 and 44 degrees respectively by nitrogen doped modified TiO2 NTAs. Therefore, they have similar and excellent hydrophilicity, and N-TiO2 NTAs charge transfer resistance (Rct) is 500.2 ohm, It is obviously lower than Rct (6431 Ohm) of TiO2 NTAs. Therefore, N-TiO2 NTAs has better electron transfer ability than TiO2 NTAs. The N-TiO2 electrodes are prepared by the modified crosslinking method on the surface of N-TiO2 NTAs carrier material, and the application of the enzyme electrode in the field of bioelectrocatalysis is studied. The linear range of the biosensor on the TAs/GOx enzyme electrode is 0.05-0.85 mM in response to glucose concentration, and its detection sensitivity is 733.17 A mM-1 cm-2, and the detection sensitivity of the TiO2 NTAs/GOx enzyme electrode (199.61 mu A mM-1 cm2) has a very significant improvement. The rate density is 23.92 W cm-2, compared with the TiO2 NTAs/GOx enzyme electrode (5.38 W cm-2) and the TiO2 NTAs/r-GO/AgNPs/GOx enzyme electrode (13.45 mu W cm-2), the biofuel battery has obviously improved. Therefore, the bioelectrocatalytic activity of the N-TiO2 NTAs/GOx enzyme electrode is obviously superior to that of the enzyme electrode and the enzyme electrode. The detection sensitivity of glucose biosensor and the maximum output power density of enzyme biofuel cell were increased.
【學(xué)位授予單位】：東南大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2015
【分類號】：O657.1;TB383.1
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本文編號：1945685