摘要
伴随纳米科学和纳米技术的迅猛发展,将形貌可控的功能纳米材料引入到电化学传感研究中,实现新颖传感界面的构建和调控以解决环境监测、疾病早期诊断等领域存在的疑难问题,已经成为未来电分析化学发展的必然趋势之一。本论文采用化学法、电沉积法和生物法,制备了形貌独特的金属、非金属及其复合纳米材料,并构置了十三种新型电化学传感器,研究了蛋白质(酶)的直接电化学及其对小分子物质的电催化,建立了测定葡萄糖、过氧化氢、肼和亚硝酸盐的分析新方法。该研究可为蛋白质的直接电化学研究提供借鉴,拓展了电化学传感和纳米材料的研究内容。全文共分五章,作者的主要贡献如下：
1、采用化学法制备了三种功能化石墨烯材料,构置了Nafion/Hb-IL-GE/CCE、 Hb-IL-AuNPs-GE/CCE和Hb-Fe3O4-GE/CCE传感器,研究了Hb直接电化学和电催化行为,建立了测定过氧化氢的分析新方法。研究表明,IL改善了GE在电极表面的固载性能,提高其稳定性,AuNPs改善了GE与蛋白之间的相互作用,Fe304克服了GE自身缺陷,同时提高了目标电极的稳定性和电催化性能。三种传感器中,Ib-Fe3O4-GE/CCE的分析性能最佳,其对H202的电催化还原响应时间<3s,线性范围为1.5×10-6-5.9×10-4mol·L-1,检出限为5×10-7mol·L-1(S/N=3),重复性和稳定性良好。
采用水热法制备了卷发梳状碳CLC,构置了Hb/{CLC-PDDA/AuNPs}3/PDA/ITO传感器,研究了Hb在传感界面上的电化学行为,建立了测定H202的分析新方法。在最佳的实验条件下,该传感器测定H2O2响应时间<2s,线性范围为6.0×10-8～1.6×10-3mol·L-1,检出限为3.0×10-8mol·L-1(S/N=3)。
采用气／液界面反应,制备了纳米四氧化三铁,构置了i-Fe3O4/CCE传感器,建立了肼测定的新方法。与传统方法比较,该方法反应条件温和、无需填充惰性气体保护所制备的Fe304形成三维网状膜结构；该传感器测定N2H4响应时间<2s,线性范围为1.0×10-7～6.0×10-4mol·L-1,灵敏度为152μA-(1.0×10-3mol·L-1)-1·cm-2,检出限为5.0×10-8mol·L-1(S/N=3)。
2、采用模板牺牲电沉积法制备了PtNPs和PdNPs,构置了PtNPs/nanoZnO/GCE和t-PdNPs/GCE传感器,建立了测定N2H4的分析新方法。实验表明,nanoZnO纳米孔结构控制了氢气泡的释放使得形成的PtNPs纳米簇孔径分布均匀,而Ethaline的加入,有效抑制了牺牲模板中气泡的干扰,形成的PdNPs具有高度分散性的特性；与i-Fe3O4/CCE和PtNPs/nanoZnO/GCE相比较,t-PdNPs/GCE测定N2H4的检出限更低,达到3.0×10-8mol·L-1(S/N=3)。
采用超声辅助电沉积制备了Co-GE和Cu-GE,构置了Co-GE/GCE和Cu-GE/GCE传感器,建立了测定肼和葡萄糖的分析新方法。研究表明,络合作用使得石墨烯和纳米钴在电极表面上分步沉积,形成花状的Co-GE异质体,构置的传感器测定N2H4的灵敏度高达562.5μA·(1.0×10-3mol·L-1)-1·cm-2; Ethaline的加入,使得石墨烯和纳米铜在电极上同步沉积,形成了花状Cu-GE复合物,构置的传感器测定葡萄糖的过电位降低到0.3V左右,响应时间<3s,线性范围为5.0×10-6～10-4mol·L-1和9.0×10-4～1.1×10-2mol·L-1,检出限为1.0×10-6mol·L-1(S/N=3)。
将一锅法与电沉积相结合,通过苯胺的电聚合过程将Hb携裹式固载于ITO,构置了Hb-PANI/ZnO-AuNPs/ITO传感器,建立了测定过氧化氢的分析新方法。研究表明,ZnO、AuNPs和PANI的协同作用,使得Hb稳定固载并有效保持原有生物活性,构置的传感器与本论文上述提及的H202电化学生物传感器相比较,电子转移速率极大的提高,ks为4.4s-1。
3、以鸡蛋蛋清为模板制备纳米金,构置了Mb-Cys-AuD/GCE传感器,建立了测定亚硝酸盐的分析新方法。研究表明,蛋清模板使得纳米金形成过程中扩散受阻,从而形成晶枝状结构。构置的传感器优于文献报道,测定NO2-的线性范围为5.0×10-7-4.0×10-4mol·L-1,检出限为3.0×10-7mo1.L-1(S/N=3),KMapp为2.0x10-4mo1.L-1
采用葡萄糖氧化酶催化葡萄糖氧化反应在GOx-CLC/PGE和GOx/DNA-GE/GCE上诱导生成金纳米粒子,构置了两种葡萄糖电化学生物传感器,建立了测定葡萄糖的分析新方法。研究表明,生物催化本身具有的富集作用与CLC、DNA-GE和AuNPs信号放大效应产生协同作用。与文献已有方法相比较,构置的葡萄糖传感器灵敏度高达2.44×104μA-(1.0×10-3mol·L-1)-1,提高了1.2～4000倍,检出限低至3.0×10-8mol·L-1(S/N=3),降低了1-3个数量级。
With the rapid development of nanoscience and nanotechnology, the inctoudition of morphology controllable functionized nanomaterials into the investigation of electrochemical sensing, can achieve the fabrication and control of novel electrochemical sensing interfaces to slove the existing problems in environmental monitoring, early disease diagnosis and so on. It becomes one of the inevitable trends of electroanalytical chemistry's development. In this thesis, chemical method, electrodeposition method and biological method were used to synthetize metal, nonmetal and composite nanomaterials with unique morphology and thirteen novel electrochemical sensors were also fabricated. Electrochemical behavior of redox proteins(enzymes) and the biocatalysis towards small molecular substances were investigated and new electeochemical analytical methods for the determination of H2O2, glucose, N2H4as well as NO2were established. These researches provided guidance for the investigation of the direct electrochemistry of proteins(enzymes) and enriched the research contens of electrochemical sensors and namomaterials. There are five chapters in the thesis, and the main contributions of the author are summarized and presented as follows:
1. Three kinds of fonctionilized graphene(GE):IL-GE, AuNPs-GE. Fe3O4-GE were prepared by chemical reduction method, Nafion/Hb-IL-GE/CCE, Hb-IL-AuNPs-GE/CCE, Hb-Fe3O4-GE/CCE sensors were fabricated for the investigation of direct electrochemistry and electrocatalysis of Hb, thereby, new electeochemical analytical methods for the determination of H2O2were established. IL improved the stripping of GE from the electrode surface, AuNPs enhanced the interaction between GE and protein, Fe3O4not only overcome inherent defect of GE, but also improved stability and electrocatalytic performance. of the proposed electrode. Among them, Hb-Fe3O4-GE/CCE exhibited the best analytical performance for the determination of H2O2, the response time was less than3s and linearity range was1.5×10-6～5.9×10-4mol·L-1with a detection limit of5×10-7mol·L-1(S/N=3). It also showed good repeatability and stability.
By controlling conditions of hydrothermal reactions, a new kind of carbon nanomaterial with comb-Like morphology (CLC) was prepared. Based on this, Hb/{PDDA-CLC/AuNPs}3/PDA/ITO sensor was fabricated by layer-by-layer assembling and employed for the investigation of electrochemistry behaviors of Hb, thereby, a new analysis method was established for the determination of H2O2. Under optimized experimental condition, the linearity range for the determination of H2O2was6.0×10-8～1.6×10-3mol·L-1with a detection limit of3.0×10-8mol·L-1(S/N=3). Furthermore, the biosensor also showed a fast response (within2s) and a high stability.
i-Fe3O4of3D network was successfully prepared via a simple chemical precipitation at the gas/liquid interface with ammonia vapor. Based on this, a novel sensor was successfully fabricated for the determination of N2H4. Comparing with typical preparation method of Fe3O4,the gas/liquid interface reaction can take place without the protection of any inert gas and a film of i-Fe3O4floating at the interface was obtained. The sensor displayed a response time less than2s, a sensitivity of152μA·(1.0×10-3mol·L-1)-1·cm-2and the linearity of1.0×10-7～6.0×10-4mol·L-1with a detection limit of5.0×10-8mol·L-1(S/N=3).
2. With the novel concept that coalition of replacement action and template assisted electrodeposition, PtNPs and PdNPs were prepared by electrodeposition, thereby, a new analysis method was established for the determination of N2H4. The results indicated that nanopore structure of nanoZnO effectively controlled the release of the hydrogen bubble to form PtNPs clusters with uniform pore size distribution, the addition of Ethaline, can restrain the hydrogen bubble on sacrificial templates to form highly decentralized PdNPs. Comparing with i-Fe3O4/CCE and PtNPs/nanoZnO/GCE, for the determination of N2H4, t-PdNPs/GCE had a lower detection limit of3.0×10-8mol·L-1(S/N=3).
Co-GE and Cu-GE were preaprted by ultrasonic assisted eledtrodposition, Co-GE/GCE and Cu-GE/GCE were fabricated, thereby, new electeochemical analytical methods for the determination of N2H4and glucose were established. The results indicated that complexationmade the successive electrodeposition of GE and Co to form hierarchical flower-like Co grown on petalage-like GE. Based on this, the fabricated sensor showed high sensetivity of562.5μA·(1.0×10-3mol·L-1)-1·cm-2for the determination of N2H4; The addition of Ethaline led to GE and CuNPs electrodepose simultaneously and formed flower-like Cu-GE composite. Based on this, the fabricated sensor showed a lower overpotential of0.3V for the determination of glucose. The sensor displayed a response time less than3s, the linearity of5.0×10-6～9.0×10-4mol·L-1and9.0×10-4～1.1×10-2mol·L-1with a detection limit of1.0×10-6mol·L-1(S/N=3).
One-pot electrodepostion was used to wrap Hb onto ITO and Hb-PANI/ZnO-AuNPs/ITO was fabricated for the investigation of direct electrochemistry and electrocatalysis of Hb, thereby, new electeochemical analytical method for the determination of H2O2was established. The results indicated the synergistic effects of ZnO, AuNPs and PANI made Hb keep the original bioactivity effectively. Comparing with H2O2mentioned above, the electron transfer rate was greatly increased, ks was4.4s-1.
3. Gold dendrites(AuD) were synthesized with egg white as the soft template and a novel nitrite(NO2) biosensor was fabricated by assembly of a myoglobin (Mb)-L-cysteamine(Cys)-AuD biological hybrid. The results indicated diffusion limited aggregation processes lead to the formation of complicated shapes qualitatively similar to real dendrites. Compering with literatures, the sensor was high performance.It displayed the linearity of5.0×10-7～4.0×10-4mol·L-1with a detection limit of3.0×10-7mol·L-1(S/N=3), KappMwas2.0×10-4mol·L-1.
Two kinds of glucose biosensors were fabricated by GOx biocatalytical induced deposition and accumulation of AuNPs on the surface of GOx-CLC/PGE and GOx/DNA-GE/GCE. The results indicated the synergistic effects of the advantages of biocatalysis itself, such as the accumulation, amplification effects and the signal amplification of CLC, DNA-GE and AuNPs, led to the high sensitivity and low detection limit of the fabricated biosensors. Compared with other methods, the glucose biosensors showed sensitivity that higher1.2-4000times of2.44×104μA·(1.0×10-3mol·L-1)-1and detection limit that lower one to three orders of magnitude of3.0×10-8mol·L-1(S/N=3).
引文
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