Immobilization and Bioelectrochemistry of Hemoglobin Based on Carrageenan and Room Temperature Ionic Liquid Composite Film

A novel biopolymer/room‐temperature ionic liquid composite film based on carrageenan, room temperature ionic liquid (IL) [1‐butyl‐3‐methylimidazolium tetra?uoroborate ([BMIM]BF₄)] was explored for immobilization of hemoglobin (Hb) and construction of biosensor. Direct electrochemistry and electrocatalytic behaviors of Hb entrapped in the IL‐carrageenan composite ?lm on the surface of glassy carbon electrode (GCE) were investigated. UV‐vis spectroscopy demonstrated that Hb in the IL‐carrageenan composite ?lm could retain its native secondary structure. A pair of well‐de?ned redox peaks of Hb was obtained at the Hb‐IL‐carrageenan composite ?lm modi?ed electrode through direct electron transfer between the protein and the underlying electrode. The heterogeneous electron transfer rate constant (k_s) was 2.02 s^?1, indicating great facilitation of the electron transfer between Hb and IL‐carrageenan composite film modi?ed electrode. The modi?ed electrode showed excellent electrocatalytic activity toward reduction of hydrogen peroxide with a linear range of 5.0×10^?6 to 1.5×10^?4 mol/L and the detection limit was 2.12×10^?7 mol/L (S/N=3). The apparent Michaelis‐Menten constant K_M^app for hydrogen peroxide was estimated to be 0.02 mmol/L, indicating that the biosensor possessed high af?nity to hydrogen peroxide. In addition, the proposed biosensor showed good reproducibility and stability.     

Hydrogen Peroxide Biosensor Based on Immobilization of Hemoglobin on Au@Ag Nanoparticles Modified Carbon Ionic Liquid Electrode

A hydrogen peroxide (H₂O₂) biosensor based on the combination of Au@Ag core‐shell nanoparticles with a hemoglobin‐chitosan‐1‐butyl‐3‐methyl‐imidazolium tetrafluoroborate (Hb‐CHIT‐BMIM×BF₄) composite film was prepared. UV‐vis spectroscopy and transmission electron microscopy confirmed a core‐shell nanostructure of Au@Ag nanoparticle was successfully obtained. Cyclic voltammetric results showed a pair of well‐defined redox peaks appeared with the formal potential (E^O′) of ‐0.301 V (versus Ag/AgCl reference electrode) and the peak‐to‐peak separation (ΔE_p) was 84 mV in 0.1 M phosphate buffer solutions. Due to the synergetic effect of Au@Ag core‐shell nanoparticles and Hb‐CHIT‐BMIM×BF₄, the biosensor exhibited good electrocatalytic activity to the reduction of H₂O₂ in a linear range from 1.0 × 10^?6 to 1.0 × 10^?3 M with a detection limit of 4 × 10^?7 M (S/N = 3). The apparent Michaelis‐Menten constant (K_M) was estimated to be 4.4 × 10^?4 M, showing its high affinity. Thus, the study proved that the combination of Au@Ag core‐shell nanoparticles and Hb‐CHIT‐BMIM×BF₄ is able to open up new opportunities for the design of enzymatic biosensors.     

室温离子液体六氟磷酸正丁基吡啶修饰碳糊电极的制备与表征

以室温离子液体(RTIL)六氟磷酸正丁基吡啶(BPPF6)代替传统固体石蜡为粘合剂与石墨粉相混合制备了一种新型的离子液体修饰碳糊电极(RTIL/CPE)。优化出制备电极时石墨与BPPF6的比例为3∶1(w/w),采用扫描电子显微镜对其表面形貌进行了表征,以铁氰化钾为电化学探针对RTIL/CPE的电化学行为进行了研究,并与传统石蜡碳糊电极(CPE)进行了比较。结果表明由于BPPF6具有较高的导电性,使RTIL/CPE比CPE具有更高的导电效率,铁氰化钾在电极上的可逆性变好,ΔEp值为64mV,峰电流响应增加3.5倍,电极过程由吸附控制变为扩散控制,根据计时库仑法求解出铁氰化钾的扩散系数为1.39×10-4cm2/s。     

Electrocatalysis Oxidation of GMP Based on Layered Double Hydroxide Functionalized with Anionic Surfactant and Room Temperature Ionic Liquid Modified Glassy Carbon Electrode

The electrocatalysis oxidation of guanosine‐5′‐monophosphate (GMP) was investigated on Mg‐Al layered double hydroxide (LDH) functionalized with sodium dodecyl sulfate (SDS) and room temperature ionic liquid (RTIL) modified glass carbon electrode (GCE). The cyclic voltammogram of GMP on the modified electrode (RTIL/ LDH‐SDS/GCE) exhibited a well defined anodic peak at 1.091 V in 0.2 mol·L^?1 pH 4.4 acetate buffer solution. The GMP oxidation was enhanced in the presence of anionic surfactant in the ?lms. The results suggest that the surfactant molecules intercalate the LDH layers to preconcentrate GMP molecules and the RTIL showed good ionic conductivity. The experimental parameters were optimized, the kinetic parameters were investigated and the probable oxidation mechanism was proposed. Under the optimized conditions, the oxidation peak current was proportional to GMP concentration in the range from 5.0×10^?7 to 1.0×10^?4 mol·L^?1 with the correlation coefficient of 0.9987 and the detection limit was 1.0×10^?7 mol·L^?1. The RTIL/LDH‐SDS/GCE showed a good electrochemical response to the oxidation of GMP and would be developed into a new biosensor.     

甲烷氧化菌素功能化金纳米层层自组装修饰电极上过氧化氢的催化还原

借助巯基试剂,在纳米金颗粒表面修饰生物活性物质Mb,制备保持有Mb生物活性的功能化金纳米巯基乙胺-Au NPs-Mb.采用UV-Vis、FTIR光谱和投射电镜表征其结构,该纳米颗粒分布均匀且粒径均一,并显著改善了金纳米颗粒团聚现象.以Mb功能化金纳米为基元,采用单层自组装及层层自组装方式将其修饰到裸金电极表面.各Mb或Mb-Cu电极的电化学测试并未借助电子传递媒介.配位Cu~(2+)后,修饰有Mb的单层及层层自组装修饰的催化还原能力均显著提升.其中Cu~(2+)配位的{巯基乙胺-Au NPs-Mb}3/Au修饰电极作为一种新型H2O2生物传感器,响应时间大约为2 s,米氏常数KappM为0.787 mmol/L,表现出了较强的还原H2O2的催化活性,且稳定性较好.     

基于硫堇/碳纳米管修饰电极的新型过氧化氢电化学传感器

基于碳纳米管(CNTs)和硫堇(Th)的协同效应,将辣根过氧化物酶(HRP)通过戊二醛(GA)交联作用固定在硫堇(Th)/CNTs修饰电极上,构造了一种新型酶电极(HRP/GA-Th/CNTs/GC)。CNTs静电吸附正电荷的Th,而Th不仅可以促进电极和酶的氧化还原活性中心之间的电子传递,而且能使CNTs氨基(—NH2)功能化,从而利于HRP的固定。基于HRP/GA-Th/CNTs/GC电极的过氧化氢传感器具有较好的传感性能,且检出限低(0.3μmol.L-1)、响应时间短(5 s内)、抗干扰能力强。     

Electrodeposited Silver Nanoparticles on Carbon Ionic Liquid Electrode for Electrocatalytic Sensing of Hydrogen Peroxide

Silver nanoparticles (narrowly dispersed in diameter) were electrodeposited on carbon ionic liquid electrode (CILE) surface using a two‐step potentiostatic method. Potentiostatic double pulse technique was used as a suitable and simple method for controlling the size and morphologies of silver nanoparticles electrodeposited on CILE. The obtained silver nanoparticles deposited on CILE surface showed excellent electrocatalytic activity (low overpotential of ?0.35 V vs. Ag/AgCl) towards reduction of hydrogen peroxide. A linear dynamic range of 2–200 μM with an experimental detection limit of 0.7 μM (S/N=3) and reproducibility of 4.1% (n=5) make the constructed sensor suitable for peroxide determination in aqueous solutions.     

An Unmediated Hydrogen Peroxide Sensor Based on a Hemoglobin-sds Film Modified Electrode

ABSTRACT

An unmediated hydrogen peroxide sensor is designed in this paper by employing a hemoglobin-SDS film modified electrode. Hemoglobin exhibits direct (unmediated) electrochemistry at the modified electrode. The protein also shows elegant catalytic activity towards the electrochemical reduction of hydrogen peroxide. Consequently, a prototype hydrogen peroxide sensor is prepared. Under optimum conditions, this sensor provides a linear response over the hydrogen peroxide concentrations in the range of 1×10^-5～1×10^-4 mol/L. The detection limit was 2×10^-6 mol/L The relative standard deviation was 4.2% for 6 successive determinations of the hydrogen peroxide at 1×10^-5 mol/L. This configuration is shown to be sensitive, stable and easily fabricated. It might be useful in the biological and industrial fields.     

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 and Electrocatalysis of Hemoglobin Based on Nafion‐Room Temperature Ionic Liquids‐Multiwalled Carbon Nanotubes Composite Film

A robust and effective composite film combined the benefits of Nafion, room temperature ionic liquid (RTIL) and multi‐wall carbon nanotubes (MWNTs) was prepared. Hemoglobin (Hb) was successfully immobilized on glassy carbon electrode surface by entrapping in the composite film. Direct electrochemistry and electrocatalysis of immobilized Hb were investigated in detail. A pair of well‐defined and quasi‐reversible redox peaks of Hb was obtained in 0.10 mol·L^?1 pH 7.0 phosphate buffer solution (PBS), indicating that the Nafion‐RTIL‐MWNTs film showed an obvious promotion for the direct electron transfer between Hb and the underlying electrode. The immobilized Hb exhibited an excellent electrocatalytic activity towards the reduction of H₂O₂. The catalysis current was linear to H₂O₂ concentration in the range of 2.0×10^?6 to 2.5×10^?4 mol·L^?1, with a detection limit of 8.0×10^?7 mol·L^?1 (S/N=3). The apparent Michaelis‐Menten constant (K_m^app) was calculated to be 0.34 mmol·L^?1. Moreover, the modified electrode displayed a good stability and reproducibility. Based on the composite film, a third‐generation reagentless biosensor could be constructed for the determination of H₂O₂.     

血红蛋白在壳聚糖修饰碳纳米管上的电化学特性及对过氧化氢的电催化分析

用壳聚糖对多壁碳纳米管进行修饰,构建了一种用于固定血红蛋白的新型复合材料,并研究了血红蛋白在该碳纳米管上的电化学性质及其对过氧化氢的电催化活性.扫描电镜结果表明,壳聚糖修饰的多壁碳纳米管呈单一的纳米管状,并能均匀分散在玻碳电极表面.紫外光谱分析表明血红蛋白在该复合膜内能很好地保持其原有的二级结构.将该材料固定在玻碳电极上后,血红蛋白能成功地实现其直接电化学.根据峰电位差随着扫描的变化,计算得到血红蛋白在壳聚糖修饰的碳纳米管膜上的电荷转移系数为0.57,表观电子转移速率常数为7.02 s-1.同时,该电极对过氧化氢显示出良好的催化性能,电流响应信号与H2O2浓度在1.0×10-6 ～1.5×10-3 mol/L间呈线性关系,检出限为5.0×10-7 mol/L.修饰电极显示了良好的稳定性.     

基于杂多酸的负载离子液相催化在醇氧化中的应用

采用溶胶-凝胶法制备的离子液体功能化的二氧化硅对于杂多酸是一种可行的载体,三种商用杂多酸成功的负载在离子液体功能化的二氧化硅上作为醇氧化的催化剂.利用FTIR、XRD、XPS对该种负载离子液相催化剂进行了表征.基于磷钨酸的负载离子液相催化剂活性最好,高产率(大部分＞93%)的得到了相应的羰基化合物.且该催化剂经过简单的...     

Amperometric Biosensor for Hydrogen Peroxide Based on Electrodeposited Sub-micrometer Gold Modified Glassy Carbon Electrode

IntroductionAmperometricbiosensorofhydrogenperoxideisofpracticalimportancebecauseofitswideapplicationsinchemical,biological,clinical,environmentalandmanyotherfields.Forimprovementofsensor抯quality,vari-ouskindsofchemicalmodificationmethodshavebeendevelopedforreducingredoxoverpotentialsofH2O2atelectrodesurfaces,increasingthedetectionsensitivity,linearrange,stabilityandlivetime.Ithasbeenshownthattheuseofsub-micrometersizedmetalparticlessuchasPt-blackcansignificantlyimprovethequalityofthebiosens…     

一种基于碳纳米管的安培型过氧化氢生物传感器

利用硫堇(th ion ine,Th i)作为介体结合多壁碳纳米管(MWNTs)、壳聚糖(ch itosan,CH IT)、辣根过氧化酶(HRP)的混合包埋物制作过氧化氢(H2O2)生物传感器。研究结果表明所得传感器对H2O2具有明显的增敏效应,线性范围为0.03~5.5 mmol/L,相关系数为0.9995;检出限为19μmol/L(S/N=3),具有良好的稳定性及工作寿命。     

辣根过氧化物酶修饰电极在离子液体[EMIM]BF4中的直接电化学

将壳聚糖(Chi)-辣根过氧化物酶(HRP)-多壁碳纳米管(MWCNTs)的复合物修饰在玻碳电极(GCE)表面, 制备了HRP修饰电极(Chi-HRP-MWCNTs/GCE), 并将其用于在亲水性离子液体1-乙基-3-甲基咪唑四氟硼酸([EMIM]BF4)中HRP的直接电化学研究. 紫外可见光谱和红外光谱表明, HRP在复合物内保持了其原始构象. 电化学研究表明, 该修饰电极在[EMIM]BF4中的循环伏安图上出现了一对峰形良好、几乎对称的氧化还原峰, 式量电位为－0.247 V (vs. Ag/AgCl), 说明包埋在Chi-MWCNTs中的HRP与电极之间发生了直接电子传递; HRP在电极表面直接电子转移的速率常数ks为3.12 s－1; 在65 ℃的[EMIM]BF4中HRP仍然保持其活性; HRP修饰电极对过氧化氢的还原具有电催化作用, 其表观米氏常数Km为5.6×10－5 mol•L－1, 催化电流与过氧化氢浓度在5.0×10－7～5.0×10－5 mol•L－1范围内呈线性关系, 检出限为2.0×10－7 mol•L－1. 该研究为非水相生物传感器的构制提供了一种新途径.     

Reduced Graphene Oxide|Carbon Ceramic Electrode Modified with CdS‐Hemoglobin as a Sensitive Hydrogen Peroxide Biosensor

A sensitive hydrogen peroxide (H₂O₂) biosensor was developed based on a reduced graphene oxide|carbon ceramic electrode (RGO|CCE) modified with cadmium sulfide‐hemoglobin (CdS‐Hb). The electron transfer kinetics of Hb were promoted due to the synergetic function of RGO and CdS nanoparticles. The transfer coefficient (α) and the heterogeneous electron transfer rate constant (k_s) were calculated to be 0.54 and 2.6 s^?1, respectively, indicating a great facilitation achieved in the electron transfer between Hb and the electrode surface. The biosensor showed a good linear response to the reduction of H₂O₂ over the concentration range of 2–240 µM with a detection limit of 0.24 µM (S/N=3) and a sensitivity of 1.056 µA µM^?1 cm^?2. The high surface coverage of the CdS‐Hb modified RGO|CCE (1.04×10^?8 mol cm^?2) and a smaller value of the apparent Michaelis? Menten constant (0.24 mM) confirmed excellent loading of Hb and high affinity of the biosensor for hydrogen peroxide.     

Electrochemical Determination of Ascorbic Acid in Room Temperature Ionic Liquid BPPF6 Modified Carbon Paste Electrode

A room temperature ionic liquid N‐butylpyridinium hexafluorophosphate (BPPF₆) was used as a binder to make an ionic liquid modified carbon paste electrode (IL‐CPE), which showed good characteristics such as simple preparation procedure, fast electrochemical response and good conductivity. The electrochemical oxidation of ascorbic acid (AA) on the new IL‐CPE was carefully studied. The oxidation peak potential of AA on the IL‐CPE appeared at 109 mV (vs. SCE), which was about 338 mV decrease of the overpotential compared to that obtained on the traditional carbon paste electrode (CPE) and the oxidation peak current was increased for about four times. The electrochemical parameters of AA on the IL‐CPE were calculated with the charge transfer coefficient (α) and the electrode reaction rate constant (k_s) as 0.87 and 0.800 s^?1, respectively. Based on the relationship of the oxidation peak current and the concentration of AA a sensitive analytical method was established with cyclic voltammetry. The linear range for AA determination was in the range from 1.0×10^?5 to 3.0×10^?3 mol/L with the linear regression equation as I_p (μA)=?2.52–0.064C (μmol/L) (n=13, γ=0.9942) and the detection limit was calculated as 8.0×10^?6 mol/L (3σ). The proposed method was free of the interferences of coexisting substances such as dopamine (DA) and amino acids etc., and successfully applied to the vitamin C tablets determination.     

Fabrication and Electrochemical Behavior of Hemoglobin Modified Carbon Ionic Liquid Electrode

A new hemoglobin (Hb) and room temperature ionic liquid modified carbon paste electrode was constructed by mixing Hb with 1‐butyl‐3‐methylimidazolium hexafluorophosphate (BMIMPF₆) and graphite powder together. The Hb modified carbon ionic liquid electrode (Hb‐CILE) was further characterized by FT‐IR spectra, scanning electron microscopy (SEM) and electrochemical impedance spectroscopy (EIS). Hb in the carbon ionic liquid electrode remained its natural structure and showed good direct electrochemical behaviors. A pair of well‐defined quasireversible redox peaks appeared with the apparent standard potential (E′) as ?0.334 (vs. SCE) in pH 7.0 phosphate buffer solution (PBS). The electrochemical parameters such as the electron transfer number (n), the electron transfer coefficient (α) and the heterogeneous electron transfer kinetic constant (k_s) of the electrode reaction were calculated with the results as 1.2, 0.465 and 0.434 s^?1, respectively. The fabricated Hb‐CILE exhibited excellent electrocatalytic activity to the reduction of H₂O₂. The calibration range for H₂O₂ quantitation was between 8.0×10^?6 mol/L and 2.8×10^?4 mol/L with the linear regression equation as I_ss (μA)=0.12 C (μmol/L)+0.73 (n=18, γ=0.997) and the detection limit as 1.0×10^?6 mol/L (3σ). The apparent Michaelis–Menten constant (K_M^app) of Hb in the modified electrode was estimated to be 1.103 mmol/L. The surface of this electrochemical sensor can be renewed by a simple polishing step and showed good reproducibility.     

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.     

基于室温离子液体的活化石墨烯粉末超级电容储能性能

室温离子液体(RTILs)具有电压窗口高等优点,被认为是实现超级电容高性能储能的绿色电解液。但是,离子液体的电导率低、粘度高,使得其储能性能不佳。本文探究了溶剂效应对离子液体超级电容储能性能的影响。以石墨烯粉末为活性材料,选取1-丁基-3-甲基咪唑四氟硼酸盐为离子液体,通过添加乙腈溶剂配置了具有不同摩尔分数ρIL的电解液(从0.25到1.0)。结果表明,溶剂效应对超级电容性能的影响与电压扫描速率或电流密度密切相关。低扫描速率下,溶剂对储能基本没有影响,而高扫描速率下,添加溶剂可显著提升比电容(在ρIL=0.25时,增加~2倍)。这是由于溶剂削弱了离子-离子间交互作用,从而降低了电解液粘度(~29倍),内阻(~5.5倍)和介电弛豫时间(~6.3倍)。在ρIL=0.25时,超级电容最大能量和功率密度分别为65.2 Wh·kg~(-1)和18066.6 W·kg~(-1),显著优于近期文献报道结果。特别地,当工作温度提升到50°C时,其能量密度将达到85.5 Wh·kg~(-1),显著高于传统水系、有机电解液超级电容和铅酸电池,与镍金属氢化物和锂离子电池性能相当。