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.     

Polyaniline nanotubes for impedimetric triglyceride detection

Polyaniline nanotubes (PANI-NT) based film electrophoretically deposited onto indium–tin–oxide (ITO) coated glass plate has been utilized for covalent immobilization of lipase (LIP), via glutaraldehyde (Glu), for triglyceride detection using impedimetric technique. It is shown that fatty acid molecules produced during triglyceride hydrolysis result in change in charge transfer resistance (R_CT) of PANI-NT film with varying triglyceride concentration. LIP/Glu/PANI-NT/ITO bioelectrode has linearity as 25–300 mg dL^?1, sensitivity as 2.59 × 10^?3 KΩ^?1 mg^?1 dL, response time as 20 s and regression coefficient as 0.99. A low value of apparent Michaelis–Menten constant (～0.62 mM) indicates high enzyme affinity to tributyrin. The LIP/Glu/PANI-NT/ITO bioelectrode has been utilized to estimate triglyceride in serum samples.     

Glucose biosensor based on new carbon nanotube-gold-titania nano-composites modified glassy carbon electrode

In this paper, a novel biosensor was prepared by immobilizing glucose oxidase （GOx） on carbon nanotube-gold-titania nanocomposites （CNT/Au/TiO2） modified glassy carbon electrode （GCE）. SEM was initially used to investigate the surface morphology of CNT/Au/TiO2 nanocomposites modified GCE, indicating the formation of the nano-porous structure which could readily facilitate the attachment of GOx on the electrode surface. Cyclic voltammogram （CV） and electrochemical impedance spectrum （EIS） were further utilized to explore relevant electrochemical activity on CNT]Au/TiO2 nanocomposites modified GCE. The observations demonstrated that the immobilized GOx could efficiently execute its bioelectrocatalytic activity for the oxidation of glucose. The biosensor exhibited a wider linearity range from 0.1 mmol L-1 to 8 mmol L^-1 glucose with a detection limit of 0.077 mmol L^- 1.     

Electrodeposition of monodispersed metal nanoparticles in a nafion film: Towards highly active nanocatalysts

We have explored a new and facile method for the fabrication of metal nanoparticles on the electrode surface. The approach for fabricating metal nanoparticles was carried out by two steps consisting of ion-exchange in nafion film coated on the electrode and subsequent reduction of metal ions to metallic nanoparticles by electrochemical method. The results of characterization by TEM show that metal nanoparticles were nearly monodispersed in the whole nafion film. The average diameters of Cu, Co and Ni nanoparticles were statistically measured to be 5.1 nm ± 0.2 nm, 4.6 nm ± 0.2 nm and 4.7 nm ± 0.2 nm, respectively. The amount of metal nanoparticles can be readily controlled by the amount of nafion coated on the electrode. By performing the H₂O₂ reduction at the obtained Cu nanoparticles, the high electrocatalytic activity of metal nanoparticles fabricated has been confirmed.     

A molecular wire modified glassy carbon electrode for achieving direct electron transfer to native glucose oxidase

Here we describe a strategy for achieving direct electron transfer to native glucose oxidase (GOx), an enzyme in which the redox active centre is buried deep within the glycoprotein. To achieve this a glassy carbon electrode is modified with a mixed monolayer of 4-carboxyphenyl and a 20 Å long oligo(phenylethynyl) molecular wire (MW), assembled from the respective aryl diazonium salts. Subsequently GOx is adsorbed to the interface, followed by covalent attachment. The redox chemistry of the active centre of glucose oxidase, flavin adenine dinucleotide, was observed at an E_1/2 of –443 mV (vs. Ag|AgCl). The enzyme was shown to retain its activity. Most importantly, in the absence of oxygen the electrode was still able to biocatalytically turn over glucose at −400 mV, thereby demonstrating that the enzyme was being recycled back to its catalytically active oxidized form from its inactive reduced form. The rate of enzyme turnover was 1.1 s⁻¹.     

Multilayer structured carbon nanotubes/poly-l-lysine/laccase composite cathode for glucose/O2 biofuel cell

Multilayer film of laccase, poly-l-lysine (PLL) and multi-walled carbon nanotubes (MWNTs) were prepared by a layer-by-layer self-assembly technique. The results of the UV–vis spectroscopy and scanning electron microscopy studies demonstrated a uniform growth of the multilayer. The catalytic behavior of the modified electrode was investigated. The (MWNTs/PLL/laccase)_n multilayer modified electrode catalyzed four-electron reduction of O₂ to water, without any mediator. The possible application of the laccase-catalyzed O₂ reduction at the (MWNTs/PLL/laccase)_n multilayer modified ITO electrode was illustrated by constructing a glucose/O₂ biofuel cell with the (MWNTs/thionine/AuNPs)₈ GDH film modified ITO electrode as a bioanode and the (MWNTs/PLL/laccase)₁₅ film modified ITO electrode as a biocathode. The open-circuit voltage reached to 700 mV, and the maximum power density achieved 329 μW cm⁻² at 470 mV of the cell voltage.     

Synergistic contributions of multiwall carbon nanotubes and gold nanoparticles in a chitosan–ionic liquid matrix towards improved performance for a glucose sensor

We report an ingenious approach for the fabrication of a promising glucose sensor, GOx/Au/CS–IL–MWNT(SH), that exploits the synergistic beneficial characteristics of multiwalled-carbon nanotubes (MWNTs), gold nanoparticles (AuNPs), chitosan (CS) and room temperature ionic liquid (RTIL). Direct electron transfer between glucose oxidase (GOx) and electrode was achieved. Scanning electron microscopy and atomic force microscopy images of GOx/Au/CS–IL–MWNT(SH) reveal that MWNTs and AuNPs are dispersed in CS–IL matrix. Cyclic voltammetry, impedance spectroscopy and chronoamperometry were used to evaluate the performance of biosensor. The GOx/Au/CS–IL–MWNT(SH) biosensor exhibits a linear current response to glucose concentration (1–10 mM) at a low potential of 0.10 V and precludes interferences from uric acid and ascorbic acid. The GOx/Au/CS–IL–MWNT(SH) biosensor has superior performances over GOx/CS–IL–MWNT(SH).     

Membrane-less and mediator-free enzymatic biofuel cell using carbon nanotube/porous silicon electrodes

Membrane-less and mediator-free direct electron transfer enzymatic biofuel cells (BFCs) with bioelectrodes comprised of single wall carbon nanotubes (SWNTs) deposited by two methods on porous silicon (pSi) substrates, are reported. In one method the SWNTs were grown by chemical vapor deposition (CVD) and then functionalized with carboxylic groups, and in the second method, pre-synthesized carboxylated SWNTs (c-SWNTs) were electrophoretically deposited on gold-coated pSi. Anodic glucose oxidase (GOx) and cathodic laccase (Lac) were immobilized on the pSi/SWNT substrates to form BFCs in pH 7 phosphate buffer solution. A peak power density of 1.38 μW/cm² (with a lifetime of 24 h) down to 0.3 μW/cm² was obtained for a BFC comprised of c-SWNT/enzyme electrodes in 4 mM glucose solution as fuel, corresponding to normal blood sugar concentration, and air as oxidant. BFCs of this relatively simple architecture have the potential for further optimization of power output and lifetime.     

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.     

Direct-growth of polyaniline nanowires for enzyme-immobilization and glucose detection

Amperometric enzyme biosensor based on the glucose oxidase (GOx) incorporated polyaniline nanowires (PANI-NWs) on carbon cloth (CC) electrode was demonstrated. The simple, direct-growth of PANI-NWs on CC, via electrochemical polymerization, provides free-standing, template-independent, hence almost (interfacial) defects-free nanostructures. The defect-free interfaces, along with the excellently sensitive organic nanostructured-surface, as evident from its significantly large effective surface area (24 times the geometric area) for redox-sensing, allows efficient entrapment/immobilization and sensing of biomolecules, via rapid electron-transfer at NWs-CC. The GOx-immobilized PANI-NWs/CC, even in initial unoptimized stage, exhibited an excellent sensitivity, ～2.5 mA mM^?1 cm^?2, to glucose, over detection range 0–8 mM, adequate for clinical monitoring of human glucose levels. The report clearly reveals a cost-effective simple system possessing enormous potentiality for biosensors, bioenergy and bioelectronics applications.     

Direct electrochemistry of glucose oxidase on a graphite nanosheet–Nafion composite film modified electrode

The direct electrochemistry of glucose oxidase (GOD) immobilized in a modified electrode based on a composite film of exfoliated graphite nanosheets (GNSs) and Nafion has been investigated for the first time. Direct electron communication between GOD and the electrode was achieved with a fast electron transfer rate (12.6 s^?1). In addition, the bioactivity of GOD was retained after immobilization in the composite film and glucose could be determined based on the decrease of the electrocatalytic response of the reduced form of GOD to dissolved oxygen. The resulting biosensor exhibited higher sensitivity (3.4 μA mM^?1). Considering much lower cost of GNSs and ready preparation from graphite, the GNSs-based modified electrode described here is superior to the carbon nanotubes (CNTs)-based modified electrodes and should have wide applications in third-generation biosensors, bioelectronics and electrocatalysis.     

Electrooxidation of insulin at silicon carbide nanoparticles modified glassy carbon electrode

For the first time silicon carbide nanoparticles (SiC) was used for electrode modification and electrocatalytic oxidation of insulin. In comparison to bare glassy carbon (GC) electrode, the oxidation of insulin at GC electrode modified with SiC nanoparticles occurred at reduced overpotentials. The modified electrode was applied for insulin detection using cyclic voltammetry, differential pulse voltammetry (DPV) and flow injection analysis (FIA). Flow injection amperometric determination of insulin at this modified electrode yielded a calibration curve with the following characteristics; linear dynamic range up to 600 pM, sensitivity of 710 pA pM^?1 cm^?2 and detection limit of 3.3 pM. In addition interference effect of the electroactive existing species (uric acid, glucose, lactic acid, l-cysteine and cholesterol) was diminished and for ascorbic acid eliminated by covering the surface of modified electrode with nafion film. This electrode shows many advantages as an insulin sensor such as simple preparation method without using any specific electron transfer mediator or specific reagent, high sensitivity, excellent catalytic activity, short response time, long term stability and remarkable antifouling property toward insulin and its oxidation product. Sensitivity, detection limit and antifouling properties of this insulin sensor are better than all of the reports in the literature for insulin detection at physiological pH solutions.     

An ionophore–Nafion modified bismuth electrode for the analysis of cadmium(II)

A novel ionophore–Nafion modified bismuth electrode is described for sensitive and selective anodic stripping analysis of cadmium(II). The electrode is prepared by coating the glassy carbon electrode with the cadmium ionophore N,N,N′,N′-tetrabutyl-3,6-dioxaoctanedi(thioamide) and Nafion composite. Bismuth is deposited in situ on the electrode surface by plating simultaneously with cadmium in sample solution. Numerous key variables affecting the current response of cadmium have been optimized. The electrode has a linear concentration range of 0.5–10 nM with a deposition time of 180 s. The detection limit is 1.3 × 10^?10 M and the relative standard deviations for 0.5 and 7 nM cadmium are 6.5% and 4.5%, respectively. The proposed electrode shows excellent selectivity over other heavy metals, such as copper, lead and indium. The attractive performance of such electrode offers a feasible way to monitor trace cadmium(II) rapidly and precisely in complex matrixes.     

Electrodeposition of gold nanoparticles at a solid|ionic liquid|aqueous electrolyte three-phase junction

Gold nanoparticles have been electrodeposited on an electrode through electrogeneration at an ITO|AuCl₄^? solution in an ionic liquid|aqueous electrolyte three-phase junction. The electrodeposition was carried out by inverted double-pulse potential chronoamperometry. The direct reduction of AuCl₄^? ions at the electrode is followed by a counterion transfer through the liquid|liquid interface. Contrary to the electrodeposition from a single ionic liquid phase, scanning electron microscopy reveals that the shape of the resulting nanoparticles is highly angular and well-developed with a diameter of 110 ± 30 nm. Catalytic oxidation of glucose on the modified electrode is demonstrated.     

Functionalization of multi-walled carbon nanotubes with poly(amidoamine) dendrimer for mediator-free glucose biosensor

By grafting with poly(amidoamine) (PAMAM) dendrimer, novel carbon nanotube (CNT) nano-composites have been successfully prepared. The novel functionalized matrix with plenty amino groups circumvents the troublesome solubility problem of CNTs in solvents, especially in water, greatly expanding the scope of the application of carbon nanotubes. The GO_x and HRP immobilized CNT-PAMAM based on the functional CNTs was synthesized. The bi-enzymatic CNT-PAMAM nano-composites are highly dispersible in water and show very promising applications in the fabrication of mediator-free bi-enzymatic biosensors for sensitive detection of glucose. The cooperation of nano-composite between CNT and high dense GO_x and HRP results in very high sensitivity to glucose with a current response of 2200 nA mM⁻¹ and fast response (∼1 s). The modified electrode exhibits a wide linear response range for glucose from 4.0 μM to 1.2 mM (R = 0.9971, N = 15), with a detection limit of 2.5 μM. The negative electrode potential of −0.34 V is favorable for glucose detection in real samples without interference caused by other biomolecules.     

Far-infrared studies on Nafion and perfluoroimide acid (PFIA) and their alkali salts

The terahertz/far-infrared spectra (<300 cm⁻¹) of perfluorinated sulfonic acid (Nafion NR211 polymer) and perfluoroimide acid (PFIA polymer) and their alkali (M⁺) salts have been analyzed and the results are presented. Pronounced features in the spectra of these ionomers that correlate systematically with the corresponding cation mass are reported and from their spectral position the force constants are derived. The average vibrational force constants for Nafion/M⁺ and PFIA/M⁺ are found to be 54 ± 7 and 39 ± 4 N/m, respectively. Such terahertz/far-infrared signatures probe the detailed structure of the Nafion/M⁺ and PFIA/M⁺ ionic clusters and, in turn, provide benchmarks for elucidating the ionomer “water channels” or water molecules located in the ionomer–water interface upon hydration. Qualitative trends in the vibrational energies of Nafion and PFIA can be explained by consideration of electronic and/or structural (ionic domain-size) effects.     

A Novel Enzymatic Glucose Biosensor and Nonenzymatic Hydrogen Peroxide Sensor Based on (3‐Aminopropyl) Triethoxysilane Functionalized Reduced Graphene Oxide

In the present study, a novel enzymatic glucose biosensor using glucose oxidase (GOx) immobilized into (3‐aminopropyl) triethoxysilane (APTES) functionalized reduced graphene oxide (rGO‐APTES) and hydrogen peroxide sensor based on rGO‐APTES modified glassy carbon (GC) electrode were fabricated. Nafion (Nf) was used as a protective membrane. For the characterization of the composites, Fourier transform infrared spectroscopy (FTIR), X‐ray powder diffractometer (XRD), and transmission electron microscopy (TEM) were used. The electrochemical properties of the modified electrodes were investigated using electrochemical impedance spectroscopy, cyclic voltammetry, and amperometry. The resulting Nf/rGO‐APTES/GOx/GC and Nf/rGO‐APTES/GC composites showed good electrocatalytical activity toward glucose and H₂O₂, respectively. The Nf/rGO‐APTES/GC electrode exhibited a linear range of H₂O₂ concentration from 0.05 to 15.25 mM with a detection limit (LOD) of 0.017 mM and sensitivity of 124.87 μA mM⁻¹ cm⁻². The Nf/rGO‐APTES/GOx/GC electrode showed a linear range of glucose from 0.02 to 4.340 mM with a LOD of 9 μM and sensitivity of 75.26 μA mM⁻¹ cm⁻². Also, the sensor and biosensor had notable selectivity, repeatability, reproducibility, and storage stability.     

Electrodeposition of PtCo alloy nanoparticles on inclusion complex film of functionalized cyclodextrin–ionic liquid and their application in glucose sensing

Platinum–cobalt (PtCo) alloy nanoparticles (NPs) are successfully fabricated by ultrasonic-electrodeposition method, using an inclusion complex (IC) film of functionalized cyclodextrin (CD)–ionic liquid (IL) as support. The morphology and composition of the PtCo alloy NPs are characterized by scanning electron microscopy, energy dispersive X-ray spectroscopy and X-ray diffraction, respectively. It is found that they are well-dispersed on the CD–IL surface and exhibit many unique features. The resulting modified glassy carbon electrode shows excellent catalytic activity for glucose oxidation. Under the physiological condition, the oxidation current of glucose is linear to its concentration up to 20 mM with sensitivity of 13.7 μA mM^?1 cm^?2. In addition, the interference from the oxidation of ascorbic acid and uric acid could be effectively avoided. Therefore, it is promising as a nonenzymatic glucose sensor.     

Highly efficient enzyme immobilization by nanocomposites of metal organic coordination polymers and carbon nanotubes for electrochemical biosensing

New nanocomposites with multi-walled carbon nanotubes (MWCNTs) embedded in metal-organic coordination polymers (MOCPs) were successfully prepared as highly efficient matrices of enzyme immobilization for sensitive electrochemical biosensing. NaAuCl₄ was pre-adsorbed on the MWCNTs to act as anchor sites to further coordinate with ligand benzenedithiol and form MOCPs. The formation of MWCNTs-MOCPs one-pot entrapped glucose oxidase (GOx) with a ratio close to 100% and exhibited enhanced mass-transfer over MOCPs. Thus MWCNTs-MOCPs-modified electrodes present superior enzymatic catalysis performance of greatly enhanced sensitivity (136 μA cm^− 2 mM^− 1) and magnitudes-lower detection limit (48 nM), being superior to most analogues.     

The role of diazonium interface chemistry in the design of high performance polypyrrole-coated flexible ITO sensing electrodes

Benzene sulfonic acid-doped polypyrrole (PPyBSA) films were electrochemically prepared on aminobenzenediazonium-modified flexible ITO in aqueous media in a simple and efficient way. Diazonium electroreduction resulted in aminophenyl-modified ITO (ITO-NH₂), which served as a platform for the electrodeposition of a PPyBSA layer. Without diazonium pretreatment, the PPyBSA coating delaminates readily upon sonication or during ultrahigh vacuum outgassing prior to XPS analysis. In contrast, a pretreatment involving electrografting for 45 s at − 800 mV resulted in excellent adhesion of PPyBSA, which withstood 75 min of ultrasonication. The robust ITO-NH₂-PPy electrode was employed for the fast, simple and accurate detection of lead (II) by differential pulse voltammetry. The potentiometric response was linear in the Pb(II) concentration range 10^− 8 to 2.5 × 10^− 7 mol·L^− 1 and the detection limit was 9.94 × 10^− 10 mol·L^− 1 at S/N = 3.This work demonstrates the central role of diazonium chemistry in producing an adhesive aryl monolayer on low-cost, flexible ITO electrodes, allowing the fabrication of robust, high-performance electrochemical-sensing conductive-polymer films.