血红蛋白在裸银电极上的直接电化学及其分析应用

报道了血红蛋白(Hb)在裸银电极上的电化学氧化还原行为。在+0.4～-0.2V(vs.SCE)电位范围内于pH=4.5的0.1mol/LNaAc-HAc底液中,血红蛋白产生一对灵敏的氧化还原峰。峰电位之差△E为0.25V(扫描速度20mV/s).动力学研究表明:电极反应的电子转移数n为0.94,表现电子传递速率常数Ks为0.032.连续电位扫描30min,峰电流变化分别为0.2μA(还原峰)和0.15μA(氧化峰).两峰与血红蛋白浓度在2×10^-7～2×10^-6mol/L和2×10^-6～1.5×10^-5mol/L范围内均呈良好线性关系,已应用于血红蛋白的分析测定。     

Electrodeposited Graphene and Silver Nanoparticles Modified Electrode for Direct Electrochemistry and Electrocatalysis of Hemoglobin

By using a 1‐butylpyridinium hexafluorophosphate based carbon ionic liquid electrode (CILE) as the working electrode, graphene (GR) nanosheets and silver nanoparticles (Ag NPs) were step by step electrodeposited on the surface of the CILE with potentiostatic method. The fabricated Ag/GR/CILE was used as a new platform for protein electrochemistry and hemoglobin (Hb) was immobilized on its surface with chitosan (CTS) as film forming material. In 0.1 mol/L phosphate buffer solution, a pair of well‐defined and quasi‐reversible redox peaks appeared on the CTS/Hb/Ag/GR/CILE with a formal peak potential of ?0.202 V (vs. SCE) and a peak‐to‐peak separation (ΔE_p) of 68 mV, which indicated that direct electrochemistry of Hb was realized on the modified electrode. The results could be attributed to the synergistic effects of Ag NPs and GR nanosheets on the electrode surface, which provided a specific three‐dimensional structure with high conductivity and good biocompatibility. The Hb modified electrode showed excellent electrocatalysis to the reduction of trichloroacetic acid in the concentration range from 0.8 to 22.0 mmol/L with a detection limit of 0.42 mmol/L (3σ). Moreover, the modified electrode exhibited favorable reproducibility, long term stability and accuracy, with potential applications in the third‐generation electrochemical biosensor.     

血红蛋白在碳纳米管修饰碳糊电极上的直接电化学行为

利用吸附法将血红蛋白(Hb)固定在碳纳米管修饰碳糊电极表面,制成稳定的固载Hb碳纳米管修饰电极,研究了Hb在碳纳米管修饰电极上的直接电化学行为.固载Hb的碳纳米管修饰电极在pH=7.0的PBS（磷酸盐缓冲溶液）中有一对相当可逆的循环伏安氧化还原峰,为Hb血红素辅基Fe(Ⅲ)/Fe(Ⅱ)电对的特征峰.式电位为－0.160 V(vs SCE),随扫描速度变化很小.电子转移数为1.021,近似为一个辅基发生电子转移.Hb在碳纳米管修饰电极表面的电子转移常数为0.0816 s－1,远大于亚甲蓝作媒介体时Hb的电子转移反应速率常数.应用于过氧化氢、三氯乙酸和硝基苯等的电催化还原,固定在碳纳米管修饰碳糊电极的血红蛋白表现出稳定且较高的催化活性.     

Direct Electrochemistry and Electrocatalysis of Hemoglobin at PAMAM Dendrimer-MWNTs-Au Nanoparticles Composite Film Modified Glassy Carbon Electrode

The direct electron transfer of hemoglobin at the PAMAM-MWNTs-AuNPs composite film modified glassy carbon electrode was studied. In a phosphate buffer solution(PBS, pH=7.0), the formal potential(E 0' ) of Hb was –0.105 V versus SCE, the electron transfer rate constant was 4.66 s –1 . E 0′ of Hb at the modified electrode was linearly varied in a pH range of 5.0—8.0 with a slope of –49.2 mV/pH. The Hb/PAMAM-MWNTs-AuNPs/GCE gave an ex-cellent electrocatalytic response to the reduction of hydrogen peroxide. The catalytic current increased linearly with H 2 O 2 concentration in a range of 1.0×10 ?6 to 2.2×10 ?3 mol/L. The detection limit was 2.0×10 ?7 mol/L at a signal to noise ratio of 3. The Michaelis-Menten constant(K ma pp ) was 2.95 mmol/L.     

碳糊电极上无机膜固载血红蛋白的直接电化学

报道了用硅溶胶-凝胶(Sol-gel)膜将血红蛋白(Hb)固载于碳糊电极上的直接电化学行为.研究结果表明,Hb-Sol-gel修饰的碳糊电极在pH=7.0的缓冲溶液中于-0.275V(vs.Ag/AgCl)处有一对可逆的循环伏安氧化-还原峰,为Hb血红素辅基Fe(Ⅲ)/Fe(Ⅱ)电对的特征峰.HbFe(Ⅲ)/Fe(Ⅱ)电对的式量电位在pH5.0～11.0范围内与溶液pH值呈线性关系,表明Hb的电化学还原很可能是一个质子伴随着一个电子的电极过程.FTIR光谱证实,Sol-gel膜对Hb的固载没有破坏其天然结构.Hb-Sol-gel修饰的碳糊电极能够催化还原H₂O₂,可望将其用于制作第三代生物传感器.     

细胞色素C在糖及醇修饰金电极上的直接电化学

细胞色素C在生物体内能进行可逆的氧化还原反应，但它知不能直接在金属电极上进行可逆的电化学反应．因此在早期细胞色素（：的电化学研究中，人们往往借助干一类称为媒介体山的分子，这给细胞色素C的电化学反应研究带来很大的不便．IO77年，hill等人间发现在4／一联毗陡存在下     

血红蛋白在离子液体[BMIM]PF6碳糊电极上的直接电化学

制备了离子液体[BMIM]PF6修饰碳糊电极(CILE), 并对其形貌和电化学行为进行了表征. 采用涂布法利用壳聚糖-皂土有机-无机复合膜将血红蛋白(Hb)固定于CILE电极表面, 利用紫外可见光谱、红外光谱和电化学方法等手段对包埋于膜内的Hb的性质进行了表征. 结果表明, Hb在薄膜内保持了其原始构象与生物活性, 循环伏安实验表明, 在pH=7.0的Britton-Robinson (B-R)缓冲液中, Hb表现出一对峰形良好的准可逆氧化还原峰, 为Hb Fe(III)/Fe(II)电对的特征峰, 对其直接电化学行为进行了研究, 求出式电位为-0.352 V(vs SCE), 电子转移数为0.885, 电荷传递系数为0.578, 表观异相电子转移速率常数为0.149 s-1.     

Direct Electrochemistry of Hemoglobin and its Electrocatalysis Based on a Carbon Nanotube Paste Electrode

A new hemoglobin (Hb) and carbon nanotube (CNT) modified carbon paste electrode was fabricated by simply mixing the Hb, CNT with carbon powder and liquid paraffin homogeneously. To prevent the leakage of Hb from the electrode surface, a Nafion film was further applied on the surface of the Hb‐CNT composite paste electrode. The modified electrode was characterized by scanning electron microscopy (SEM) and electrochemical impedance spectroscopy (EIS). Direct electrochemistry of hemoglobin in this paste electrode was easily achieved and a pair of well‐defined quasi‐reversible redox peaks of a heme Fe(III)/Fe(II) couple appeared with a formal potential (E^0′) of ?0.441 V (vs. SCE) in pH 7.0 phosphate buffer solution (PBS). The electrochemical behaviors of Hb in the composite electrode were carefully studied. The fabricated modified bioelectrode showed good electrocatalytic ability for reduction of H₂O₂ and trichloroacetic acid (TCA), which shows potential applications in third generation biosensors.     

A Highly Sensitive and Selective Enzymatic Biosensor Based on Direct Electrochemistry of Hemoglobin at Zinc Oxide Nanoparticles Modified Activated Screen Printed Carbon Electrode

A sensitive enzymatic biosensor has been developed for the detection of hydrogen peroxide (H₂O₂), nitrite ( ) and trichloroacetic acid (TCA) by using hemoglobin (Hb) immobilized on activated screen printed carbon electrode (ASPCE) and zinc oxide (ZnO) composite. A pair of well defined redox peaks is observed with a heterogeneous electron transfer rate constant (K_s) of 5.27 s^?1 for Hb at ASPCE/ZnO. The biosensor exhibits the detection of H₂O₂, TCA and in the concentration range of 0.5–129.5 µmol L^?1, 2.5–72.5 mmol L^?1 and 0.2–674 µmol L^?1 with the detection limit of 0.083 µmol L^?1, 0.12 mmol L^?1 and 0.069 µmol L^?1, respectively.     

L-Cysteine Modified Silver Electrode and Its Application to the Study of the Electrochemistry of Hemoglobin

Abstract

L-cysteine can be modified onto a silver electrode by covalent binding through the sulphur to give very stable and long-lived chemically modified electrodes(CMEs). Hemoglobin(Hb) exhibits an excellent voltammetric response at the resulting CMEs. Linear sweep voltammetric(LSV) measurements of Hb on the CMEs reveal the existence of a linear relationship between the cathodic peak current and the concentration of Hb in the range of 5 X 10^?7 - 1 X 10^?5 mol/L. The detection limit is about 2 X 10^?7 mol/L. The relative standard deviation of results is 4.5% for 10 successive determinations at 2 X 10^?6 mol/L. Other proteins and chemicals present in samples do not interfere in the assay.     

血红蛋白在二氧化铈修饰电极上的直接电化学

将二氧化铈(CeO₂)与酶复合修饰电极, 采用循环伏安法研究了血红蛋白(Hb)在CeO₂修饰的玻碳电极上的电化学行为. 实验表明, 固定在CeO₂材料上的Hb, 不仅能有效地与电极表面进行直接电子转移, 而且能够保持其生物催化活性. 制得的Nafion/CeO₂/Hb/GC修饰电极的电子传递速率k_s为(0.68±0.09) s^－1, 对H₂O₂的检测限为1.013 μmol•L^－1, 重现性和稳定性较好. CeO₂在实验中体现出一定的生物相容性, 起到了促进Hb与电极之间进行直接电子传递的作用. CeO₂修饰电极进行蛋白质直接电化学测定以及酶生物电催化的成功实践, 为稀土氧化物材料在电化学传感领域中的应用开辟了思路.     

马心血红蛋白在氧化铟电极上的直接电子传递反应

马心血红蛋白在氧化铟电极上的直接电子传递反应杨秀娟，菊，陆天虹（中国科学院长春应用化学研究所电分析化学开放实验室，长春，１３００２２）关键词血红蛋白，氧化铟电极，循环伏安法蛋白质与电极之间的电子转移在某种程度上类似于生物体内蛋白质分子之间的电子转移过...     

Application of NiMoO4 Nanorods for the Direct Electrochemistry and Electrocatalysis of Hemoglobin with Carbon Ionic Liquid Electrode

In this paper NiMoO₄ nanorods were synthesized and used to accelerate the direct electron transfer of hemoglobin (Hb). By using an ionic liquid (IL) 1‐butylpyridinium hexafluorophosphate (BPPF₆) modified carbon paste electrode (CILE) as the basic electrode, NiMoO₄ nanorods and Hb composite biomaterial was further cast on the surface of CILE and fixed by chitosan (CTS) to establish a modified electrode denoted as CTS/NiMoO₄‐Hb/CILE. UV‐vis and FT‐IR spectroscopic results showed that Hb in the film retained its native structures without any conformational changes. Electrochemical behaviors of Hb entrapped in the film were carefully investigated by cyclic voltammetry with a pair of well‐defined and quasi‐reversible redox voltammetric peaks appearing in phosphate buffer solution (PBS, pH 3.0), which was attributed to the direct electrochemistry of the electroactive center of Hb heme Fe(III)/Fe(II). The results were ascribed to the specific characteristic of NiMoO₄ nanorods, which accelerated the direct electron transfer rate of Hb with the underlying CILE. The electrochemical parameters of Hb in the composite film were further carefully calculated with the results of the electron transfer number (n) as 1.08, the charge transfer coefficient (α) as 0.39 and the electron‐transfer rate constant (k_s) as 0.82 s^?1. The Hb modified electrode showed good electrocatalytic ability toward the reduction of trichloroacetic acid (TCA) in the concentration range from 0.2 to 26.0 mmol/L with a detection limit of 0.072 mmol/L (3σ), and H₂O₂ in the concentration range from 0.1 to 426.0 µmol/L with a detection limit of 3.16×10^?8 mol/L (3σ).     

血红蛋白在TiO2-石墨烯/离子液体/壳聚糖纳米复合膜修饰电极上的直接电化学

基于TiO2-石墨烯、离子液体和壳聚糖复合膜修饰玻碳电极制备了一种新型的电化学传感器。用循环伏安法研究了血红蛋白在该修饰电极上的直接电化学行为。结果表明,该纳米复合膜能有效地促进血红蛋白在电极上的直接电子转移,保持其生物催化活性。该传感器对H2O2具有良好的催化性能。H2O2的电流响应信号与其浓度在20～860μmol/L范围内呈良好的线性关系,检出限为0.1μmol/L(S/N=3)。传感器具有良好的稳定性和重现性。     

Direct Electrochemistry of Hemoglobin on Single‐Walled Carbon Nanotubes Modified Carbon Ionic Liquid Electrode and Its Electrocatalysis

In this article we report on the fabrication of a carbon ionic liquid electrode (CILE) by using a room temperature ionic liquid of 1‐butyl‐3‐methylimidazolium hexafluorophosphate (BMIMPF₆) as binder. It was further modified by single‐walled carbon nanotubes (SWCNTs) to get a SWCNTs modified CILE denoted as SWCNTs/CILE. The redox protein of hemoglobin (Hb) was further immobilized on the surface of SWCNTs/CILE with the help of Nafion film. UV‐vis and FT‐IR spectra indicated that the immobilized Hb retained its native conformation in the composite film. The direct electrochemistry of Hb on the SWCNTs/CILE was carefully studied in pH 7.0 phosphate buffer solution (PBS). Cyclic voltammetric results indicated that a pair of well‐defined and quasireversible voltammetric peaks of Hb heme Fe(III)/Fe(II) was obtained with the formal potential (E°') at ?0.306 V (vs. SCE). The electrochemical parameters such as the electron transfer coefficient (α), the electron transfer number (n) and the apparent heterogeneous electron transfer rate constant (k_s) were calculated as 0.34, 0.989 and 0.538 s^?1, respectively. The fabricated Hb modified electrode showed good electrocatalytic ability to the reduction of trichloroacetic acid (TCA) in the concentration range from 20.0 to 150.0 mmol/L with the detection limit of 10.0 mmol/L (3σ).     

Immobilization of Hemoglobin on Cobalt Nanoparticles-modified Indium Tin Oxide Electrode:Direct Electrochemistry and Electrocatalytic Activity

A cobalt nanoparticles-attached indium tin oxide(CoNPs/ITO) electrode was applied to the immobilization of hemoglobin(Hb) and an Hb modified CoNPs/ITO electrode(Hb/CoNPs/ITO) was prepared. The direct electron transfer of Hb was shown by the well-behaved voltammetric responses for Hb/CoNPs/ITO electrode and the effects of scan rate and pH value were observed. Based on the catalytic activity of Hb immobilized on the CoNPs/ITO electrode toward the reduction of H₂O₂, a mediator-free H₂O₂ sensor was developed. A linear relationship existed between the catalytic current and the H₂O₂ concentration in a range of 1.0―100.0 μmol/L with a detection limit (S/N=3) of 0.2 μmol/L.     

Direct Electrochemistry of Cytochrome c at Gold Electrode Modified with Fumed Silica

Direct electrochemistry of horse heart cytochrome c (cytc) has been obtained at a gold electrode constructed by self‐assembling fumed silica particles (FSPs) onto a silane monolayer. A pair of well‐defined and nearly symmetrical redox peaks of cytc is obtained at the FSPs film modified gold electrode. Cyclic voltammetry (CV) and tapping‐mode atomic force microscopy (AFM) are used to characterize the FSPs film modified electrode, showing that the FSPs can provide a favorable microenvironment for cytc and facilitate the direct electron transfer between the cytc and the gold electrode, which may propose an approach to realize the direct electrochemistry of other proteins.     

Hemoglobin Modified Carbon Paste Electrode: Direct Electrochemistry and Electrocatalysis

Abstract

A new hemoglobin (Hb) modified carbon paste (CP) electrode was fabricated by simply mixing the hemoglobin with carbon powder and paraffin homogeneously. To prevent the leakage of Hb from the electrode surface, a Nafion film was further applied on the surface of Hb-carbon composite paste electrode. Direct electrochemistry of hemoglobin in the paste electrode was easily achieved, and a pair of well-defined quasi-reversible redox peak of heme Fe(III)/Fe(II) couple appeared with the formal potential (E^0′) as ?0.335 V (vs. Saturated calomel electrode; CE) in pH 7.0 phosphate buffer solution (PBS). The fabricated Hb modified electrode showed good electrocatalytic ability to the reduction of trichloroacetic acid (TCA) and H₂O₂.     

Electrochemistry and Electrocatalysis of Hemoglobin on 1‐Pyrenebutanoic Acid Succinimidyl Ester/Multiwalled Carbon Nanotube and Au Nanoparticle Modified Electrode

A biocompatible nanocomposite film was fabricated for hemoglobin (Hb) molecules immobilization. This film consists of multiwalled carbon nanotubes (MWNTs), 1‐pyrenebutanoic acid, succinimidyl ester (PASE), hemoglobin (Hb) and Au nanoparticles (AuNPs). Herein, PASE molecules physically adsorbed onto MWNTs, and its groups then covalently bond with Hb. AuNPs were then linked with Hb/PASE/MWNTs via electrostatic adsorption force. UV‐visible adsorption spectra, Fourier transform infrared spectra, scanning electron microscope and electrochemical impedance spectroscopy have characterized the film. Cyclic voltammetry (CV) scans showed that in the film Hb has well‐defined redox reaction, with the formal potential (E°) at about ?0.27 V (vs. Ag/AgCl). Herein, differential pulse voltammetry (DPV) was employed to electrochemically detect the Hb in the film with a detection limit of 9.3×10^?8 M. The proposed method was also succeeded for Hb detection in clinical blood samples. Furthermore, the Hb in the film showed good electrocatalytic activities to the reduction of H₂O₂, TCA, NaNO₂ and O₂.     

血红蛋白直接电化学的界面设计与传感研究进展

血红蛋白为人体红细胞中的一种主要蛋白质,是血液中运输氧气的主要物质。由于其类酶的性质、确定的结构以及分布的广泛性,长期以来一直是氧化还原蛋白质直接电化学以及生物传感研究的理想模型。近年来,受益于材料科学与信息技术的快速发展,血红蛋白直接电化学的界面设计以及生物传感已成为当前的研究热点。为此,本文对近年来这方面的研究进展进行综述。通过介绍血红蛋白直接电化学界面设计的基本方法以及生物传感的原理和研究现状,以探索今后的发展趋势。