A promising dehydrogenase-based bioanode for a glucose biosensor and glucose/O2 biofuel cell

A new type of dehydrogenase-based amperometric glucose biosensor was constructed using glucose dehydrogenase (GDH) which was immobilized on the edge-plane pyrolytic graphite (EPPG) electrode modified with poly(phenosafranin)-functionalized single-walled carbon nanotubes (PPS-SWCNTs). The PPS-SWCNT-modified EPPG electrode was prepared by electropolymerization of phenosafranin on the EPPG electrode which had been previously coated with SWCNTs. The performance of the GDH/PPS-SWCNT/EPPG bioanode was evaluated using cyclic voltammetry and amperometry in the presence of glucose. The GDH/PPS-SWCNT/EPPG electrode possesses promising characteristics as a glucose sensor: a wide linear dynamic range of 50 to 700 μM, low detection limit of 0.3 μM, fast response time (1-2 s), high sensitivity (96.5 μA cm(-2) mM(-1)), and anti-interference and anti-fouling abilities. Moreover, the performance of the GDH/PPS-SWCNT/EPPG bioanode was tested in a glucose/O(2) biofuel cell. The maximum power density delivered by the assembled glucose/O(2) biofuel cell could reach 64.0 μW cm(-2) at a cell voltage of 0.3 V with 40 mM glucose.     

Amperometric glucose biosensor based on electrodeposition of platinum nanoparticles onto covalently immobilized carbon nanotube electrode

A new amperometric biosensor for glucose was developed based on adsorption of glucose oxidase (GOx) at the gold and platinum nanoparticles-modified carbon nanotube (CNT) electrode. CNTs were covalently immobilized on gold electrode via carbodiimide chemistry by forming amide linkages between carboxylic acid groups on the CNTs and amine residues of cysteamine self-assembled monolayer (SAM). The fabricated GOx/Au_nano/Pt_nano/CNT electrode was covered with a thin layer of Nafion to avoid the loss of GOx in determination and to improve the anti-interferent ability. The immobilization of CNTs on the gold electrode was characterized by quartz crystal microbalance technique. The morphologies of the CNT/gold and Pt_nano/CNT/gold electrodes have been investigated by scanning electron microscopy (SEM), and the electrochemical performance of the gold, CNT/gold, Pt_nano/gold and Pt_nano/CNT/gold electrodes has also been studied by amperometric method. In addition, effects of electrodeposition time of Pt nanoparticles, pH value, applied potential and electroactive interferents on the amperometric response of the sensor were discussed.

The enzyme electrode exhibited excellent electrocatalytic activity and rapid response for glucose in the absence of a mediator. The linear range was from 0.5 to 17.5 mM with correction coefficient of 0.996. The biosensor had good reproducibility and stability for the determination of glucose.     

Amperometric glucose biosensor based on mediated electron transfer between immobilized glucose oxidase and plasma-polymerized thin film of dimethylaminomethylferrocene on sputtered gold electrode.

We propose an electron transfer-mediated amperometric enzyme biosensor based on plasma-polymerized thin film of dimethylaminomethylferrocene (DMAMF) on a sputtered gold electrode. The DMAMF plasma-polymerized film is deposited directly onto the surface of the electrode under dry conditions. The resulting thin film not only has redox sites but also is extremely thin (approximately 20 nm), adheres well onto the substrate (electrode), has a flat surface and a highly-crosslinked network structure, and is hydrophilic in nature. Glucose oxidase is densely immobilized onto the surface of DMAMF plasma-polymerized film on the gold electrode. From the electrochemical measurement, the biosensor can cover the wide range of glucose concentration (1.3 - 81 mM) at +350 mV of applied potential. The current response of the glucose biosensor was decreased by less than 5% in an aerobic solution as compared to that in an anaerobic solution. These show that the DMAMF plasma-polymerized films play a role as the electron transfer mediators between the reaction center of enzyme and the electrode.     

A low-cost,fast, disposable and sensitive biosensor study: flow injection analysis of glucose at poly-methylene blue-modified pencil graphite electrode

In this work, firstly methylene blue (MB) was electropolymerized onto pencil graphite electrode (PGE) surface for the electrocatalytic oxidation of NADH. Cyclic voltammograms show that oxidation potential of NADH at Poly-MB/PGE shifted to negative direction about 300 mV compared with bare PGE. These results indicate that Poly-MB/PGE exhibits a good electrocatalytic activity toward NADH oxidation. Then, a glucose biosensor study was performed based on the determination of enzymatically generated NADH by glucose dehydrogenase (GDH) which immobilized onto Poly-MB/PGE using glutaraldehyde cross-linking procedure. The biosensing of glucose in flow injection analysis (FIA) system was performed at GDH/Poly-MB/PGE for the first time. The electrocatalytic oxidation currents of enzymatically produced NADH obtained from FI amperometric current–time curves recorded at + 200 mV and in phosphate buffer solution at pH 7.0 containing 1.0 M KCl were linearly related to the concentration of glucose. Linear calibration plots are obtained in the concentration range from 0.01 to 1.0 mM. The limit of detection (LOD) was found to be 4.0 µM. A fast, sensitive, low-cost and disposable glucose biosensor was constructed in FIA system using GDH/Poly-MB/PGE; therefore, it might provide a new perspective for the fabrication of biosensor of other compounds such as glutamate, lactate and alcohol.     

Electrocatalytic Reduction of H2O2 and Oxygen on the Surface of Thionin Incorporated onto MWCNTs Modified Glassy Carbon Electrode: Application to Glucose Detection

A new H₂O₂ enzymeless sensor has been fabricated by incorporation of thionin onto multiwall carbon nanotubes (MWCNTs) modified glassy carbon electrode. First 50 μL of acetone solution containing dispersed MWCNTs was pipetted onto the surface of GC electrode, then, after solvent evaporations, the MWCNTs modified GC electrode was immersed into an aqueous solution of thionin (electroless deposition) for a short period of time <5–50 s. The adsorbed thin film of thionin was found to facilitate the reduction of hydrogen peroxide in the absence of peroxidase enzyme. Also the modified electrode shows excellent catalytic activity for oxygen reduction at reduced overpotential. The rotating modified electrode shows excellent analytical performance for amperometric determination of hydrogen peroxide, at reduced overpotentials. Typical calibration at ?0.3 V vs. reference electrode, Ag/AgCl/3 M KCl, shows a detection limit of 0.38 μM, a sensitivity of 11.5 nA/μM and a liner range from 20 μM to 3.0 mM of hydrogen peroxide. The glucose biosensor was fabricated by covering a thin film of sol–gel composite containing glucose oxides on the surface of thionin/MWCNTs modified GC electrode. The biosensor can be used successfully for selective detection of glucose based on the decreasing of cathodic peak current of oxygen. The detection limit, sensitivity and liner calibration rang were 1 μM, 18.3 μA/mM and 10 μM–6.0 mM, respectively. In addition biosensor can reach 90% of steady currents in about 3.0 s and interference effect of the electroactive existing species (ascorbic acid–uric acid and acetaminophen) is eliminated. The usefulness of biosensor for direct glucose quantification in human blood serum matrix is also discussed. This sensor can be used as an amperometric detector for monitoring oxidase based biosensors.     

An amperometric glucose biosensor based on glucose oxidase immobilized in electropolymerized poly(o-aminophenol) and carbon nanotubes composite film on a gold electrode.

An amperometric glucose biosensor is developed that is based on immobilization of glucose oxidase (GOD) in a composite film of poly(o-aminophenol) (POAP) and carbon nanotubes (CNT), which are electrochemically co-polymerized at a gold (Au) electrode. Because of the high surface per volume ratio and excellent electrical conductivity of CNT, the biosensor based on an Au/POAP/CNT/GOD electrode has lower detection limit (0.01 mM), larger maximum response current (0.24 mA cm(-2)) and higher sensitivity (11.4 mA M(-1) cm(-2)) than the values of the biosensor based on an Au/POAP/GOD electrode. Additionally, the biosensor shows fast response time, large response current, and good anti-interferent ability for ascorbic acid, uric acid and acetaminophen. Good reproducibility and stability of the biosensor are also observed.     

Highly Sensitive Amperometric Glucose Biosensor Based on Glassy Carbon Electrode with Copper/Palladium Coating

A highly sensitive amperometric glucose biosensor based on immobilizing glucose oxidase in electropolymerized poly(o‐phenylenediamine) film on glassy carbon electrode coated sequentially with copper and palladium layers has been developed. The steady‐state amperometric response to glucose was determined by means of the oxidation of hydrogen peroxide generated by the enzymatic reaction at a potential of either +0.70 or +0.40 V (vs. Ag|AgCl reference). The deposited copper/palladium layer showed great enhancement in the performance of the enzyme electrode, possibly due to its better electrocatalytic activity for hydrogen peroxide oxidation and large surface area. Effects of the relative loading of palladium, enzyme and polymer on the electrode performance were examined in detail. Sensitivity and detection limit for glucose determinations at +0.70 V were about 7.3 μA/mM and 0.1 μM, respectively. A wide linear range up to 6.0 mM glucose could be achieved. Electrode performance was superior to similar works reported in the literature. The response time was less than 2 s and its lifetime was longer than three months. The permeable polyphenylenediamine film also offered good anti‐interference ability to ascorbic acid, uric acid and acetaminophen, especially when a detection potential of +0.40 V was employed.     

Amperometric Glucose Biosensor on Layer by Layer Assembled Carbon Nanotube and Polypyrrole Multilayer Film

An amperometric glucose biosensor on layer by layer assembled carbon nanotube and polypyrrole multilayer film has been reported in the present investigation. Homogeneous and stable single wall carbon nanotubes (SWNTs) and polypyrrole (PPy) multilayer films were alternately assembled on platinum coated Polyvinylidene fluoride (PVDF) membrane. Since conducting polypyrrole has excellent anti‐interference ability, protection ability in favor of increasing the amount of the SWNTs on platinum coated PVDF membrane and superior transducing ability, a layer by layer approach of polypyrrole and carbon nanotubes has provided an excellent matrix for the immobilization of enzyme. The layer‐by‐layer assembled SWNTs and PPy‐modified platinum coated PVDF membrane is shown to be an excellent amperometric sensor over a wide range of concentrations of glucose. The glucose oxidase (GOx) was immobilized on layer by layer assembled film by a physical adsorption method by cross linking through Glutaraldehyde. The glucose biosensor exhibited a linear response range from 1 mM to 50 mM of glucose concentration with excellent sensitivity of 7.06 μA/mM.     

Amperometric urea biosensor based on urease and electropolymerized toluidine blue dye as a pH-sensitive redox probe

The electropolymerized toluidine blue film deposited on the glassy carbon electrode show amperometrically detectable pH sensitivity. This feature of polytoluidine blue (PTOB) film was used for a construction of an amperometric urea biosensor. We have observed a linear shift of the formal redox potential with increasing pH value between 4 and 8 giving the slope of 81 mV(Delta) pH(-1). Polytoluidine blue film has had a significantly increased stability and higher electrochemical activity compared to the adsorbed monomeric dye. The polytoluidine blue urea biosensor has been operating at a working potential of -200 mV vs. SCE. The sensitivity of the biosensor was 980 nA mM(-1) cm(-2). The biosensor showed linearity in concentration range up to 0.8 mM with the detection limit of 0.02 mM (S/N=3).     

Glucose Biosensor Based on Multi‐Walled Carbon Nanotube Modified Glassy Carbon Electrode

An amperometric glucose biosensor based on multi‐walled carbon nanotube (MWCNT) modified glassy carbon electrode has been developed. MWCNT‐modified glassy carbon electrode was obtained by casting the electrode surface with multi‐walled carbon nanotube materials. Glucose oxidase was co‐immobilized on the MWCNT‐modified glassy carbon surface by electrochemical deposition of poly(o‐phenylenediamine) film. Enhanced catalytic electroreduction behavior of oxygen at MWCNT‐modified electrode surface was observed at a potential of ?0.40 V (vs. Ag|AgCl) in neutral medium. The steady‐state amperometric response to glucose was determined at a selected potential of ?0.30 V by means of the reduction of dissolved oxygen consumed by the enzymatic reaction. Common interferents such as ascorbic acid, 4‐acetamidophenol, and uric acid did not interfere in the glucose determination. The linear range for glucose determination extended to 2.0 mM and the detection limit was estimated to be about 0.03 mM.     

Glucose Biosensor Based on Oxygen Electrode Part IV: In Vivo Evaluation of the Rechargeable Glucose Sensor

Abstract

A glucose monitoring system consisting of a pair of amperometric sensors: a glucose biosensor based on oxygen electrode and an oxygen sensor, two miniature potentiostats, an instrumentation amplifier and a data logger has been developed. The glucose sensor has linear response to the glucose concentration in vitro at 37°C up to 26 mM (480 mg/dL) in the phosphate buffer solution (pH 7.4), and linear range up to 21 mM (380 mg/dL) in undiluted bovine plasma. The system was evaluated in vivo with the sensors subcutaneously implanted in healthy mongrel dogs. During the implantation the system output was continuously recorded. The results of short-term subcutaneous implantation of the integrated system demonstrated good agreement between the glucose concentration measured by the biosensor and that obtained using standard glucose determination methods. The delay-time between the tissue glucose level (measured by the biosensor) and the blood glucose level (obtained by standard methodology) was 3 to 10 minutes. During the chronic implantation the biosensor was refilled in vivo. Rejuvenation of the sensor response after refilling was observed demonstrating the potential of such sensors for long-term implantation.     

Ceramic Carbon/Polypyrrole Materials for the Construction of Bienzymatic Amperometric Biosensor for Glucose

Amperometric enzymatic biosensors have high selectivity and simplicity in use. It has advantages over other analytical methods in biochemistry, pharmacology, so it evokes strong interests1,2. Generally, the detection mode involved in oxidase based biosensors is often based on the electrochemical detection of hydrogen peroxide directly3,4. However the direct oxidation of hydrogen peroxide requires a relative high working potential (exceeding ca. 0.6 V vs. SCE), at which many biological sub…     

Minimization of Interferences in Flow Injection Amperometric Glucose Biosensor Based on Oxidation of Enzymatically‐produced H2O2

One of the major problems in amperometric biosensors based on detection of H₂O₂ produced by enzymatic reaction between oxidase enzymes and substrate is the interference of redox active compounds such as ascorbic acid (AA), dopamine (DA) and uric acid (UA). To minimize these interferences, sodium bismuthate was used for the first time as an insoluble pre‐oxidant in the flow injection (FI) amperometric glucose biosensor at a Glucose oxidase (GOx) immobilized Pt/Pd bimetallic modified pre‐anodized pencil graphite electrode (p.PGE). In this context, these interfering compounds were injected into a flow injection analysis (FIA) system using an injector which was filled with NaBiO₃. Thus, these interferents were converted into their redox inactive oxidized forms before reaching the electrode in the flow cell. While glucose was not influenced by the pre‐oxidant in the injector, the huge oxidation peak currents of the interferents decreased significantly in the biosensor. FI amperometric current time curves showed that the AA, DA and UA were minimized by 96 %, 86 %, and 98 % respectively, in the presence of an equivalent concentration of interferences in a 1.0 mM glucose solution. The proposed FI amperometric glucose biosensor exhibits a wide linear range (0.01–10 mM, R²=0.9994) with a detection limit of 2.4×10^?3 mM. Glucose levels in the artificial serum and two real samples were successfully determined using the fabricated FI amperometric biosensor.     

An amperometric glucose biosensor based on the immobilization of glucose oxidase on the CuGeO3 nanowire modified electrode

A new amperometric biosensor based on adsorption of glucose oxidase (GOx) at the CuGeO₃ nanowires (NW) modified glassy carbon electrode (GCE) is reported in this article. The properties of the biosensor were characterized by Fourier transform infrared spectroscopy (FTIR), electrochemical impedance spectroscopy (EIS) and electrochemical techniques. The GCE modified with the CuGeO₃ NW/GOD showed excellent electrocatalytical response to the oxidation of glucose. Different parameters including GOx concentration, working potential and pH of supporting electrolyte that governed the analytical performance of the biosensor have been studied in detail and optimized. The biosensor was applied to detect glucose with a linear range of 0.5 to 7 mM. The biosensor exhibited excellent reproducibility and stability.     

Hydrogen peroxide and glucose biosensor based on silver nanowires synthesized by polyol process

Silver nanowires synthesized through a polyol process using polyvinylpyrrolidone as protection (PVP-AgNWs) were used as a new electrode material for constructing a sensor. Hydrogen peroxide (H(2)O(2)) and glucose were used as analytes to demonstrate the sensor performance of the PVP-AgNWs. It is found that the PVP-AgNWs-modified glassy carbon electrode (PVP-AgNWs/GCE) exhibits remarkable catalytic performance toward H(2)O(2) reduction. This sensor has a fast amperometric response time of less than 2 s and the catalytic current is linear over the concentration of H(2)O(2) ranging from 20 μM to 3.62 mM (R = 0.998) with a detection limit of 2.3 μM estimated on a signal-to-noise ratio of 3. A glucose biosensor was constructed by immobilizing glucose oxidase (GOD) onto the surface of the PVP-AgNWs/GCE. The resultant glucose biosensor can be used for glucose detection in human blood serum with a sensitivity of 15.86 μA mM(-1) cm(-2) and good selectivity and stability.     

Glucose Biosensor Based on Oxygen Electrode. Part I: Silastic Coated Polycarbonate Membranes for Biosensor Application

Abstract

An amperometric glucose biosensor based on the oxygen electrode principle has been developed. Polycarbonate membranes (pore size from 0.01 μm to 0.4 μm) were used as external glucose diffusion membranes in order to obtain direct proportionality of the amperometric signal to the substrate concentration in the entire physiological range. The commercially available membranes - standard (hydrophilic, treated with Polyvinylpyrrolidone/(PVP)) and PVP-free membranes were compared with membranes coated with a silicone elastomer (silastic). Spindrop coating technique was used to create stable, adhesive coatings over the polycarbonate membranes. These coated membranes achieved diffusion control of the glucose flux such that the amperometric signal of the biosensor was linearly proportional to the substrate concentration up to 16 mM glucose. The membrane parameters were optimized by varying the parameters of the coating process-spin rate of the membrane rotation and the silastic/water ratio in the coating emulsion.     

Automated determination of glucose in soluble coffee using Prussian Blue-glucose oxidase-Nafion((R)) modified electrode

A highly selective, fast and stable biosensor for determination of glucose in soluble coffee has been developed. The biosensor electrode consist of a thin film of ferric hexacyanoferrate (Prussian Blue or PB) electrodeposited on the glassy carbon electrode (GCE) (to provide a catalytic surface for the detection of hydrogen peroxide) glucose oxidase immobilized on top of the electrode and a Nafion^® polymer layer. The stability of the PB film and the biosensor was evaluated by injecting standard-solution (50 μM H₂O₂ and 0.5 mM glucose) during 4 h in a flow-injection system with the electrodes polarized at −50 mV versus Ag/AgCl. The system is able to handle about 60 samples per hour and is very stable and suitable for industrial control. Determination of glucose in the range 2.5 and 15% (w/v) in phosphate buffer with precision (r.s.d. < 1.5%) has been achieved and is in agreement with the conventional procedures. Linear calibration in the range of 0.15 and 2.50 mM with detection limits of ca. 0.03 mM has been obtained. The morphology of the enzyme glucose oxidase on the modified electrode has been analyzed by scanning electron microscopy (SEM) measurements.     

表面可更新的石墨陶瓷葡萄糖生物传感器研究

A surface-renewable glucose biosensor is reported. The glucose biosensor is developed using glucose oxidase (GOD) encapsulated in organically modified solgel glass (ORMOSIL) network in the composite matetial. The organic group in the ORMOSIL network controls the hydrophobicity of the electrode surface and thus limits the wettability of the electrode surface. The graphite powder provides the conductivity for the electrode.Ferrocenecarboxylic acid in phosphate buffer solution (pH 7.0) transfers electron between enzyme and electrode. Cyclic volammetry and amperometric measurements have been used to exmine the electrochemical behavior of glucose biosensor as shown in Fig. 1 and Fig.2. The electrode gives a linear response range of 1 -20mM glucose with a sensitivity of 3.26 μA· mM-1. The electrode can be renewed easily in reproducible manner by a simple polishing step.     

Pulsed Deposited Manganese and Vanadium Oxide Film Modified with Carbon Nanotube and Gold Nanoparticle: Chitosan and Ionic Liquid‐based Biosensor

Present study describes the synthesis of mixed oxide films of manganese and vanadium by electrochemical pulsed deposition technique on a glassy carbon electrode (GCE) modified with multiwall carbon nanotubes (MWCNT). The film was further decorated with gold nanoparticles to enhance the reduction signal of dissolved oxygen in pH 5.17 acetate buffer solution. All of the electrochemical synthesized modified electrodes have been characterized with Scanning electron microscopy(SEM), High‐resolution transmission electron microscopy (HRTEM), X‐Ray photoelectron spectroscopy (XPS), X‐Ray diffraction (XRD) techniques. The electrode obtained (AuNPs/MnOx?VOx/CNT/GCE) was utilized as a platform for glucose biosensor where the glucose oxidase enzyme was immobilized on the composite film with the aid of chitosan and an ionic liquid. The electrochemical performance of the biosensor was investigated by cyclic voltammetry and the relative parameters have been optimized by amperometric measurements in pH 5.17 acetate buffer solution. The developed biosensor exhibited a linear range for glucose between 0.1–1.0 mM and the limit of detection was calculated as 0.02 mM.     

Development of Amperometric Glucose Biosensor Based on Reconstitution of Glucose Oxidase on Polymeric 3‐Aminophenyl Boronic Acid Monolayer

An amperometric biosensor for determining glucose based on deflavination of the enzyme glucose oxidase and subsequent reconstitution of the apo‐protein with a complexed flavin adenine dinucleotide (FAD) monolayer is described. The GOx‐reconstituted electrode exhibited excellent electrocatalytic activities towards the reduction and oxidation of hydrogen peroxide as well. The prepared biosensor showed an excellent performance for glucose at +0.5 V with a high sensitivity (5.94 μA/mM) and relatively good response time (～12 s) in a wide concentration range of 1–17 mM (correlation coefficient of 0.9998). The applicability to blood analysis was also evaluated.