壳聚糖-纳米金-葡萄糖氧化酶生物传感器的制备及其应用于葡萄糖的测定

采用了一种简便快捷的电沉积方法制备了壳聚糖-纳米金复合膜并应用于葡萄糖生物传感器的构建.氯金酸和壳聚糖的混合液在玻碳电极表面电化学还原为金纳米粒子,再将葡萄糖氧化酶通过戊二醛交联的方式固定在纳米金复合膜修饰的玻碳电极表面,制成一种新型的葡萄糖氧化酶生物传感器.该传感器对葡萄糖的响应十分快速,在5 S内即达到平衡.测定葡萄糖的线性范围为20μmol·L-1～5 mmol·L-1,检出限(3S/N)为12μmol·L-1.     

葡萄糖氧化酶在石墨烯-纳米氧化锌修饰玻碳电极上的直接电化学及对葡萄糖的生物传感

采用滴涂法和电沉积法制备了石墨烯/纳米氧化锌复合膜修饰玻碳电极,再将葡萄糖氧化酶固定在修饰电极表面制成了电化学生物传感器,用于葡萄糖的灵敏测定。用循环伏安法在-0.7～-0.1 V范围内研究了葡萄糖氧化酶在修饰电极上的直接电化学行为。结果表明,石墨烯/纳米氧化锌复合膜能很好地保持葡萄糖氧化酶的生物活性,并显著促进了其电化学过程。在0.1 mol/L磷酸盐缓冲溶液(pH 7.0)中,固定在修饰电极上的葡萄糖氧化酶呈现出一对近乎可逆的氧化还原峰,并且对葡萄糖的氧化具有良好的催化作用。葡萄糖氧化酶在修饰电极上的电子转移常数ks为1.42 s-1,修饰电极对葡萄糖催化的米氏常数Kampp为14.2μmol/L。线性范围为2.5×10-6～1.5×10-3mol/L,检出限为2.4×10-7mol/L(S/N=3)。此修饰电极具有良好的导电性能、稳定性和重现性,可用于实际样品的分析测定。     

葡萄糖氧化酶在羟基磷灰石／Nation复合膜修饰电极上的直接电化学及其对葡萄糖的生物传感

将葡萄糖氧化酶固定于羟基磷灰石（HAp）-Nation复合膜,构建了高灵敏、高选择性的葡萄糖传感器．羟基磷灰石和Nation良好的协同作用,可以有效地提高传感器的稳定性与灵敏度．实验结果表明：固定在复合膜修饰电极上的葡萄糖氧化酶呈现出一对较好的近乎可逆的氧化还原峰,并且对葡萄糖的氧化有良好的催化作用,同时消耗溶解氧,从而导致溶解氧还原峰的降低．在-0.8V处,随葡萄糖浓度的增加,葡萄糖氧化酶催化葡萄糖氧化时消耗溶解氧的量增加,溶解氧还原电流逐渐降低,因此该修饰电极可以作为葡萄糖传感器实现对葡萄糖的高灵敏检测．在0．12～2．16mmol·L^-1浓度范围内,溶解氧还原电流的降低与葡萄糖的浓度成正比,据此可以测定出溶液中葡萄糖的浓度,该传感器的检出限和灵敏度分别为0．02mmol·L^-1（SIN=3）和6．75mA·mol·L^-1．因此,HAp-Nation复合膜为酶的固定和直接电化学研究提供了一个新的有效平台,在构建新型无试剂葡萄糖传感器方面具有较大的应用前景．     

氧化锌纳米簇-金纳米颗粒-壳聚糖复合膜修饰电极用于示差脉冲伏安法测定吗啡

将制备的氧化锌纳米簇和金纳米颗粒分散在壳聚糖中并滴涂在玻碳电极表面,制备了氧化锌纳米簇-金纳米颗粒-壳聚糖复合膜修饰电极(Au-ZnO-CHIT/GCE)。采用循环伏安法研究了吗啡在修饰电极上的电化学行为。结果表明:吗啡在该修饰电极上出现了一个氧化峰,提出了用示差脉冲伏安法测定吗啡的方法。吗啡浓度在5.3×10-6~6.5×10-4mol.L-1范围内与氧化峰电流呈线性关系,检出限(3S/N)为1.8×10-6mol.L-1。修饰电极用于尿液中吗啡的测定,回收率在80.0%~99.6%之间。     

基于壳聚糖-纳米金/纳米普鲁士蓝/L-半胱氨酸修饰的 葡萄糖传感器的研究

在金电极表面修饰一层L-半胱氨酸,再利用静电吸附作用固定纳米普鲁士蓝(nano-PB),然后利用壳聚糖-纳米金复合膜将葡萄糖氧化酶(GOD)固定于修饰电极表面,制成新型的葡萄糖传感器.通过交流阻抗技术,循环伏安法和计时电流法考察了电极的电化学特性.在优化的实验条件下,该传感器在葡萄糖浓度为3.0×10-6～1.0×10-3 mol/L范围内有线性响应,检测下限为1.6×10-6 mol/L.此外该传感器具有响应快、稳定性好和选择性良好的特点,能有效排除常见干扰物质如抗坏血酸、尿酸等对测定的影响.     

基于金纳米棒-壳聚糖复合膜的葡萄糖生物传感器

本文采用金纳米棒-壳聚糖复合膜固定葡萄糖氧化酶构建电流型葡萄糖生物传感器.通过电化学交流阻抗法和循环伏安法对酶膜状态进行了表征,得到了相应的等效电路和动力学参数.实验结果表明,金纳米棒-壳聚糖复合膜可以辅助电子传递,提高电极的电流响应,并使生物传感器的使用温度范围有很大的扩展.此传感器表现出对葡萄糖溶液浓度的优良响应,线性范围在2.78×10^-5mol/L—2.22×10^-3mol/L,响应灵敏度约为7.819μA·cm^-2(mmol/L)^-1,表观米氏常数为10mmol/L.本工作还研究了温度和溶液pH值对电极电流响应的影响.     

纳米金胶增强葡萄糖生物传感器的构建

将纳米金胶(AuNPs)和羟基磷灰石(HAp)按一定比例混合制备了新型复合膜用于葡萄糖氧化酶(GOD)的固定,构建了高灵敏的葡萄糖传感器。由于纳米金胶的存在,葡萄糖氧化酶的直接电化学性质得以增强,在去除氧气的PBS(pH 7.0)介质中,固定在复合膜内的GOD表现出一对良好的氧化还原峰。在饱和氧气条件下,当加入一定量的葡萄糖时,由于GOD催化葡萄糖氧化消耗溶液中的溶解氧,-0.8 V处溶解氧的还原峰电流降低,且峰电流降低的量与葡萄糖浓度在0.02～1.62 mmol/L范围内呈线性相关,检出限为5.0μmol/L,检测灵敏度达9.91 mA.mol-1.L,可实现对葡萄糖的快速检测。     

肌红蛋白-壳聚糖-金胶纳米复合修饰电极的电催化

在pH 5.4的HAc-NaAc缓冲溶液中,肌红蛋白-壳聚糖-金胶薄膜修饰电极(Mb-Ch itosan-Au colloid/GCE)于-0.20 V(vs.Ag/AgC l)处有一对准可逆的氧化还原峰,即Mb血红素辅基Fe(Ⅲ)/Fe(Ⅱ)电对的特征峰.本实验条件下,肌红蛋白与玻碳电极之间的电子传递明显加快,并考察了扫速、溶液pH及支持电解质浓度等因素对肌红蛋白电子传递的影响.紫外光谱图表明:肌红蛋白在壳聚糖-金胶溶液中依然保持其原始构象.该肌红蛋白-壳聚糖-金胶纳米修饰电极还能电催化溶解氧的还原.     

基于纳米复合材料/壳聚糖膜的葡萄糖生物传感器研究

采用纳米普鲁士蓝/金纳米粒子/壳聚糖(nano-PB/AuNPs/Chit)复合膜固定葡萄糖氧化酶(GOD)构建新型葡萄糖生物传感器。通过电化学阻抗谱以及电流-时间曲线法(I-t)研究了传感器的电化学特性。结果表明,传感器在葡萄糖浓度为0.01～1.0 mmol/L范围内呈线性,响应灵敏度为68.15μA.(mmol/L)-1.cm-2,表观米氏常数为5.1 mmol/L。该传感器可用于糖尿病人血糖的测定。     

以天青Ⅰ为介体的纳米金颗粒增强的葡萄糖传感器

采用层层自组装的方法和异种电荷互相吸引的原理,将Nafion修饰在金电极上固载带正电荷的天青Ⅰ,并利用天青Ⅰ中的氨基固载纳米金,再通过纳米金将酶固定在金电极表面,制成了葡萄糖传感器.采用循环伏安法和交流阻抗法,研究了金电极表面组装各层之后的电化学特征,以及电极对葡萄糖的电化学催化作用. 结果表明,天青Ⅰ不仅可以固定酶和纳米金,而且还可以在酶和电极之间有效地传递电子.在优化的实验条件下,该传感器对葡萄糖响应的线性范围为5.1×10-6 ~4.0×10-3 mol/L,检出限(S/N=3)为1.0 μmol/L.该生物传感器显示出较好的稳定性和抗干扰能力,将其用于人体血清中葡萄糖的测定,结果令人满意.     

Direct electrochemistry of glucose oxidase immobilized on NdPO4 nanoparticles/chitosan composite film on glassy carbon electrodes and its biosensing application

The direct electrochemistry of glucose oxidase (GOx) immobilized on a composite matrix based on chitosan (CHIT) and NdPO(4) nanoparticles (NPs) underlying on glassy carbon electrode (GCE) was achieved. The cyclic voltammetry and electrochemical impedance spectroscopy were used to characterize the modified electrode. In deaerated buffer solutions, the cyclic voltammetry of the composite films of GOx/NdPO(4) NPs/CHIT showed a pair of well-behaved redox peaks that are assigned to the redox reaction of GOx, confirming the effective immobilization of GOx on the composite film. The electron transfer rate constant was estimated to be 5.0 s(-1). The linear dynamic range for the detection of glucose was 0.15-10 mM with a correlation coefficient of 0.999 and the detection limit was estimated at about 0.08 mM (S/N=3). The calculated apparent Michaelis-Menten constant was 2.5 mM, which suggested a high affinity of the enzyme-substrate. The immobilized GOx in the NdPO(4) NPs/CHIT composite film retained its bioactivity. Furthermore, the method presented here can be easily extended to immobilize and obtain the direct electrochemistry of other redox enzymes or proteins.     

Direct electrochemistry of glucose oxidase immobilized on porous carbon nanofiber/room temperature ionic liquid/chitosan composite film and its biosensing application

The direct electron transfer of glucose oxidase (GOD) immobilized on a composite matrix based on porous carbon nanofibers (PCNFs), room-temperature ionic liquid (RTIL), and chitosan (CHIT) underlying on a glassy carbon electrode was achieved. The combination of the PCNFs, RTIL, and CHIT provided a suitable microenvironment for GOD to transfer electron directly. In deaerated buffer solutions (pH 7.0), the cyclic voltammetry of the GOD/PCNFs/RTIL/CHIT composite films showed a pair of well-defined redox peaks with the formal potential of −0.45 V (vs. SCE). The synergistic effort of the PCNFs, RTIL, and CHIT also promoted the stability of GOD in the composite film and retained its bioactivity.     

Tannic Acid Modified Electrochemical Biosensor for Glucose Sensing Based on Direct Electrochemistry

A novel glucose biosensor was constructed through the immobilization of glucose oxidase (GOx) on gold nanoparticles (Au NPs) deposited, and chemically reduced graphene oxide (rGO) nanocomposite. In the synthesis, tannic acid (TA) was used for the reduction of both graphene oxide, and Au³⁺ to rGO, and Au NPs, respectively. Also, by harnessing the π‐π interaction between graphene oxide and TA, and protein‐TA interaction, a novel nanocomposite for the fabrication of a third generation biosensor was successfully constructed. Upon the oxidation of TA to quinone, which is easily reducible at the negative potential range, enhanced electron transfer was obtained. The cyclic voltammetry (CV) results demonstrated a pair of well‐defined and quasi‐reversible redox peaks of active site molecule of GOx. The biosensor exhibited a linear response to glucose concentrations varying from 2 to 10 mM with a sensitivity of 18.73 mA mM⁻¹ cm⁻². The fabricated biosensor was used for the determination of glucose in beverages.     

Dendrimer-encapsulated Pt nanoparticles on mesoporous silica for glucose detection

A hybrid system of mesoporous silica (MS) particle incorporated with poly(amidoamine) dendrimer-encapsulated platinum nanoparticles (Pt-DENs) was constructed in a neutral aqueous solution through electrostatic interaction. The MS/Pt-DENs composite particles immobilized with glucose oxidase (GOx) were used to modify a glassy carbon electrode for detecting the electrocatalytic response to the reduction of glucose. Pt-DENs can improve the conductivity of MS and enhance the electron transfer between redox centers in enzymes and electrode surfaces. The structure of composite particles and the performance of MS/Pt-DEN-modified electrodes were characterized by transmission electron microscopy, N₂ sorption characterization method, electrochemical impedance spectroscopy, cyclic voltammetry and amperometric measurements. The MS/Pt-DENs/GOx-modified electrodes, which had a fast response of GOx less than 3?s, could be used for the determination of glucose ranging from 0.02 to 10?mM. The detection limits were 4???M at signal-to-noise ratio of 3.     

An electrochemically preanodized screen-printed carbon electrode for achieving direct electron transfer to glucose oxidase

Here we report the unique property of a preanodized screen-printed carbon electrode (SPCE1) that can allow direct electron transfer (DET) reaction of glucose oxidase (GOx). The GOx can be immobilized in the composite of oxygen functionalities and edge plane sites generated during preanodization without additional cross-linking agents. The electron transfer rate of GOx is greatly enhanced to 4.38 s⁻¹ as a result of the conformational change of GOx in the microenvironment enabling the accessibility of active site for GOx to the electrode. The analytical versatility is further improved with the aid of Nafion film. As a consequence, the as-prepared electrode can be used as a glucose biosensor and the number of potential foreign species is then restricted by molecular size, permeation and/or (bio)chemical reaction. Most importantly, the disposable nature of the proposed electrode is expected to promote the DET-related researches.     

Direct Electrochemistry of Glucose Oxidase on Nail‐like Carbon and Its Biosensing for Glucose

Nail‐like carbon (NLC) was synthesized by a simple hydrothermal method. It was the first time that a novel electrochemical biosensing of glucose was explored based on the glucose oxidase (GOx)‐NLC‐chitosan (CHIT) glassy carbon electrode. Morphology and structure of NLC were characterized by scanning electron microscope; meanwhile the chemical composition was determined by X‐ray diffraction and energy dispersive X‐ray spectroscopy. The cyclic voltammetry of immobilized GOx showed a pair of quasireversible redox peaks with the formal potential (E°′) of ?0.458 V and the peak‐to‐peak potential separation was 47 mV at a scan rate of 100 mV s^?1. The present biosensor has a linear range of glucose from 0.02 to 1.84 mM (correlation coefficient of 0.9991) and detection limit of 0.01 mM (S/N=3). Compared with the previous reports based on the carbon material biosensor, it has a high sensitivity of 165.5 μA mM^?1 cm^?2 and low apparent Michaelis–Menten constant of 0.506 mM. Thus, the NLC may have potential applications in the field of bioelectrochemistry, bioelectronics and biofuels.     

Direct electrochemistry of glucose oxidase and sensing glucose using a screen-printed carbon electrode modified with graphite nanosheets and zinc oxide nanoparticles

We have studied the direct electrochemistry of glucose oxidase (GOx) immobilized on electrochemically fabricated graphite nanosheets (GNs) and zinc oxide nanoparticles (ZnO) that were deposited on a screen printed carbon electrode (SPCE). The GNs/ZnO composite was characterized by using scanning electron microscopy and elemental analysis. The GOx immobilized on the modified electrode shows a well-defined redox couple at a formal potential of −0.4 V. The enhanced direct electrochemistry of GOx (compared to electrodes without ZnO or without GNs) indicates a fast electron transfer at this kind of electrode, with a heterogeneous electron transfer rate constant (K_s) of 3.75 s⁻¹. The fast electron transfer is attributed to the high conductivity and large edge plane defects of GNs and good conductivity of ZnO-NPs. The modified electrode displays a linear response to glucose in concentrations from 0.3 to 4.5 mM, and the sensitivity is 30.07 μA mM⁻¹ cm⁻². The sensor exhibits a high selectivity, good repeatability and reproducibility, and long term stability.

Graphical representation for the fabrication of GNs/ZnO composite modified SPCE and the immobilization of GOx

     

Construction of glucose biosensor based on sorption of glucose oxidase onto multilayers of polyelectrolyte/nanoparticles

A new approach to constructing an enzyme-containing film on the surface of a gold electrode for use as a biosensor is described. A basic multilayer film (BMF) of (PDDA/GNPs) _n /PDDA was first constructed on the gold electrode by electrostatic layer-by-layer self-assembly of poly(diallyldimethylammonium chloride) (PDDA) and gold nanoparticles (GNPs). Glucose oxidase (GOx) was then sorbed into this BMF by dipping the BMF-modified electrode into a GOx solution. The assembly of the BMF was monitored and tested via UV-vis spectroscopy and cyclic voltammetry (CV). The ferrocenemethanol-mediated cyclic voltammograms obtained from the gold electrode modified with the (PDDA/GNPs) _n /PDDA/GOx indicated that the assembled GOx remained electrocatalytically active for the oxidation of glucose. Analysis of the voltammetric signals showed that the surface coverage of active enzyme was a linear function of the number of PDDA/GNPs bilayers. This result confirmed the penetration of GOx into the BMF and suggests that the BMF-based enzyme film forms in a uniform manner. Electrochemical impedance measurements revealed that the biosensor had a lower electron transfer resistance (R _et) than that of a sensor prepared by layer-by-layer assembly of PDDA and GOx, due to the presence of gold nanoparticles. The sensitivity of the biosensor for the determination of glucose, which could be controlled by adjusting the number of PDDA/GNPs bilayers, was investigated.     

Amperometric glucose sensor based on glucose oxidase immobilized on gelatin-multiwalled carbon nanotube modified glassy carbon electrode

We investigated the direct electrochemistry of glucose oxidase (GOx) at gelatin-multiwalled carbon nanotube (GCNT) modified glassy carbon electrode (GCE). GOx was covalently immobilized onto GCNT modified GCE through the well known glutaraldehyde (GAD) chemistry. The immobilized GOx showed a pair of well-defined reversible redox peaks with a formal potential (E⁰′) of ? 0.40 V and a peak to peak separation (ΔE_p) of 47 mV. The surface coverage concentration (Г) of GOx in GCNT/GOx/GAD composite film modified GCE was 3.88 × 10^? 9 mol cm^? 2 which indicates the high enzyme loading. The electron transfer rate constant (k_s) of GOx immobilized onto GCNT was 1.08 s^? 1 which validates a rapid electron transfer processes. The composite film shows linear response towards 6.30 to 20.09 mM glucose. We observed a good sensitivity of 2.47 μA mM^?1 cm^? 2 for glucose at the composite film. The fabricated biosensor displayed two weeks stability. Moreover, it shows no response to 0.5 mM of ascorbic acid (AA), uric acid (UA), acetaminophen (AP), pyruvate (PA) and lactate (LA) which shows its potential application in the determination of glucose from human serum samples. The composite film exhibits excellent recovery for glucose in human serum at physiological pH with good practical applicability.     

A Glucose Biosensor Based on Immobilization of Glucose Oxidase into 3D Macroporous TiO2

3D macroporous TiO₂ inverse opals have been derived from a sol‐gel procedure using polystyrene colloidal crystals as templates. EDS and SEM showed a face‐centered cubic (FCC) structure TiO₂ inverse opal was obtained. Glucose oxidase (GOx) was successfully immobilized on the surface of indium‐tin oxide (ITO) electrode modified by TiO₂ inverse opal (TiO_2(IO)). Electrochemical properties of GOx/TiO_2(IO)/ITO electrode were characterized by using the three electrodes system. The result of cyclic voltammetry showed that a couple of stable and well‐defined redox peaks for the direct electron transfer of GOx in absence of glucose, and the redox peak height enhanced in presence of 0.1 μM glucose. Compare with the ordinary structured GOx/TiO₂/ITO electrode, inverse opal structured GOx/TiO_2(IO)/ITO electrode has a better respond to the glucose concentration change. Under optimized experimental conditions of solution pH 6.8 and detection potential at 0.30 V versus saturated calomel electrode (SCE), amperometric measurements were performed. The sensitivity and the detection limit of glucose detection was 151 μA cm^?2 mM^?1 and 0.02 μM at a signal‐to‐noise ratio of 3, respectively. The good response was due to the good biocompatibility of TiO₂ and the large effective surface of the three‐dimensionally ordered macroporous structure.