Novel poly (neutral red) nanowires as a sensitive electrochemical biosensing platform for hydrogen peroxide determination

Poly (neutral red) nanowires (PNRNWs) have been synthesized for the first time by the method of cyclic voltammetric electrodeposition using porous anodic aluminum oxide (AAO) template and were examined by scanning electron microscopy (SEM) and transmission electron microscope (TEM). Moreover, horseradish peroxidase (HRP) was encapsulated in situ in PNRNWs (denoted as PNRNWs–HRP) by electrochemical copolymerization for potential biosensor applications. The PNRNWs showed excellent efficiency of electron transfer between the HRP and the glassy carbon (GC) electrode for the reduction of H₂O₂ and the PNRNWs–HRP modified GC electrode showed to be excellent amperometric sensors for H₂O₂ at −0.1 V with a linear response range of 1 μM to 8 mM with a correlation coefficient of 0.996. The detection limit (S/N = 3) and the response time were determined to be 1 μM and <5 s and the high sensitivity is up to 318 μA mM⁻¹ cm⁻².     

A sensitivity-controlled hydrogen peroxide sensor based on self-assembled Prussian Blue modified electrode

In this communication, a hydrogen peroxide (H₂O₂) sensor based on self-assembled Prussian Blue (PB) modified electrode was reported. Thin film of PB was deposited on the electrode by self-assembly process including multiple sequential adsorption of ferric ions and hexacyanoferrate ions. The as-prepared PB modified electrode displayed sufficient stability for practical sensing application. At an applied potential of ?0.05 V vs. Ag/AgCl (sat. KCl), PB modified electrode with 30 layers exhibited a linear dependence on H₂O₂ concentration in the range of 1 × 10^?6–4 × 10^?4 M (r = 0.9998) with a sensitivity of 625 mA M^?1 cm^?2. It was found that the sensitivity of H₂O₂ sensors could be well controlled by adjusting the number of deposition cycles for PB preparation. This work demonstrates the feasibility of self-assembled PB modified electrode in sensing application, and provides an effective approach to control the sensitivity of PB-based amperometric biosensors.     

Non-enzymatic detection of hydrogen peroxide using a functionalized three-dimensional graphene electrode

A novel three-dimensional (3D) electrochemical sensor was developed for highly sensitive detection of hydrogen peroxide (H₂O₂). Monolithic and macroporous graphene foam grown by chemical vapor deposition (CVD) served as the electrode scaffold. Using in-situ polymerized polydopamine as the linker, the 3D electrode was functionalized with thionine molecules which can efficiently mediate the reduction of H₂O₂ at close proximity to the electrode surface. Such stable non-enzymatic sensor is able to detect H₂O₂ with a wide linear range (0.4 to 660 μM), high sensitivity (169.7 μA mM^− 1), low detection limit (80 nM), and fast response (reaching 95% of the steady current within 3 s). Furthermore, this sensor was used for real-time detection of dynamic release of H₂O₂ from live cancer cells in response to a pro-inflammatory stimulant.     

Activity and stability of horseradish peroxidase in hydrophilic room temperature ionic liquid and its application in non-aqueous biosensing

The activity and stability of horseradish peroxidase (HRP) were investigated in a hydrophilic room temperature ionic liquid 1-butyl-3-methylimidazolium tetrafluroborate ([bmim][BF₄]) by electrochemical methods. Although no detectable activity exhibited in anhydrous [bmim][BF₄], HRP was active in the presence of a small amount of water (4.53%, v/v). And its activity can be improved by immobilization in agarose hydrogel. The immobilized HRP possesses excellent activity at 65 °C. It remained 80.2% of its initial activity after being immersed for 10.5 h in an aqueous mixture of [bmim][BF₄] with some hydrogen peroxide (H₂O₂) under room temperature, implying extremely high stability. Moreover, the immobilized HRP was found to be very sensitive and stable in H₂O-containing [bmim][BF₄] for the detection of H₂O₂, with a wide linear range of 6.10 × 10⁻⁷ to 1.32 × 10⁻⁴ mol l⁻¹ and low detection limit of 1.0 × 10⁻⁷ mol l⁻¹.     

Improved direct electron transfer and electrocatalytic activity of horseradish peroxidase immobilized on gemini surfactant–polyvinyl alcohol composite film

The voltammetric and electrocatalytic behavior of horseradish peroxidase (HRP) immobilized on a cationic gemini surfactant (i.e. C₁₂H₂₅N(CH₃)₂–C₁₂H₂₄–N(CH₃)₂C₁₂H₂₅Br₂, C₁₂–C₁₂–C₁₂)–polyvinyl alcohol (PVA) composite film-coated glassy carbon electrode (GCE) has been studied. It is found that on the novel composite film HRP presents excellent electroactivity and can exhibit a pair of well-defined voltammetric peaks in 0.10 M pH 7.0 phosphate buffer solution (PBS). The immobilized HRP also presents good bioelectrocatalytic activity, and it can catalyze the reduction of oxygen (O₂), hydrogen peroxide (H₂O₂), nitrite ion (NO₂^?) and trichloroacetic acid (TCA). For H₂O₂ the catalytic current is linear to its concentration in the range of 0.195–97.5 μM, and the detection limit is down to 6.5 × 10^?8 M. The response shows Michaelis–Menten feature and the apparent Michaelis–Menten constant is estimated to be 110.5 μM. Similarly, the electrode can sense NO₂^? and TCA. In addition, it is observed that the spacer group of gemini surfactant affects the electroactivity of HRP significantly. A spacer group with higher flexibility and hydrophility is favorable to the electron transfer of HRP. UV–vis spectrum indicates that the structure of HRP in the PVA–C₁₂–C₁₂–C₁₂ film is similar to that of native HRP. Thus the C₁₂–C₁₂–C₁₂–PVA composite possesses good biocompatibility and has promising application in fabricating biosensor and bioelectronics.     

Iron (III) nanocomposites for enzyme-less biomimetic cathode: A promising material for use in biofuel cells

In this paper, we discuss the synthesis and electrochemical properties of a new material based on iron oxide nanoparticles stabilized with poly(diallyldimethylammonium chloride) (PDAC); this material can be used as a biomimetic cathode material for the reduction of H₂O₂ in biofuel cells. A metastable phase of iron oxide and iron hydroxide nanoparticles (PDAC–FeOOH/Fe₂O₃-NPs) was synthesized through a single procedure. On the basis of the Stokes–Einstein equation, colloidal particles (diameter: 20 nm) diffused at a considerably slow rate (D = 0.9 × 10^? 11 m s^? 1) as compared to conventional molecular redox systems. The quasi-reversible electrochemical process was attributed to the oxidation and reduction of Fe³⁺/Fe²⁺ from PDAC–FeOOH/Fe₂O₃-NPs; in a manner similar to redox enzymes, it acted as a pseudo-prosthetic group. Further, PDAC–FeOOH/Fe₂O₃-NPs was observed to have high electrocatalytic activity for H₂O₂ reduction along with a significant overpotential shift, ΔE = 0.68 V from ? 0.29 to 0.39 V, in the presence and absence of PDAC–FeOOH/Fe₂O₃-NPs. The abovementioned iron oxide nanoparticles are very promising as candidates for further research on biomimetic biofuel cells, suggesting two applications: the preparation of modified electrodes for direct use as cathodes and use as a supporting electrolyte together with H₂O₂.     

Gold nanoparticles decorated carbon fiber mat as a novel sensing platform for sensitive detection of Hg(II)

The three-dimensional fibril-like carbon fiber mat electrode (CFME) decorated with Au nanoparticles (AuNPs) was employed to construct Hg(II) sensing platform for the first time. The highly porous feature of CFME combining the high affinity of AuNPs for mercury endowed the sensing platform with high sensitivity and good reproducibility. Under optimal conditions, the prepared AuNPs/CFME was capable of sensing Hg(II) with a detection limit of 0.1 μg L^− 1 (S/N = 3) using differential pulse anodic stripping voltammetry (DPASV). Finally, the AuNPs/CFME was successfully demonstrated for the determination of Hg(II) in real water samples with satisfactory results.     

One-step fabrication of chitosan–hematite nanotubes composite film and its biosensing for hydrogen peroxide

This work reports on a novel chitosan–hematite nanotubes composite film on a gold foil by a simple one-step electrodeposition method. The hybrid chitosan–hematite nanotubes (Chi–HeNTs) film exhibits strong electrocatalytic reduction activity for H₂O₂. Interestingly, two electrocatalytic reduction peaks are observed at −0.24 and −0.56 V (vs SCE), respectively, one controlled by surface wave and the other controlled by diffusion process. The Chi–HeNTs/Au electrode shows a linear response to H₂O₂ concentration ranging from 1 × 10⁻⁶ to 1.6 × 10⁻⁵ mol L⁻¹ with a detection limit of 5 × 10⁻⁸ mol L⁻¹ and a sensitivity as high as 1859 μA μM⁻¹ cm⁻².     

Sensing characteristics of aged zirconia-based hydrogen sensor utilizing Zn–Ta-based oxide sensing-electrode

We report here the enhanced sensing characteristics to H₂ for a potentiometric sensor using an yttria-stabilized zirconia (YSZ) solid electrolyte and a ZnO(+ 84 wt.% Ta₂O₅) sensing electrode (SE) after aging at 500 °C. The emf response toward 400 ppm H₂ was found to gradually increase up to − 800 mV after 40 days operation (aging) and was stabilized at this value until the 90th day. The aged and stabilized sensor exhibited highly sensitive response to H₂, with minor responses toward other examined gases such as NOx and HCs. The 90% response time toward 100 ppm H₂ was approximately 70 s. The H₂ sensitivity of the stabilized sensor was hardly affected by changes in water vapor as well as O₂ concentration, with repeatable and reproducible responses to H₂.     

Amperometric enzyme biosensor for hydrogen peroxide via Ugi multicomponent reaction

A novel strategy based on the Ugi multicomponent reaction was employed for immobilizing horseradish peroxidase on sodium alginate-coated gold electrode. The electrode was employed for constructing an amperometric biosensor device using 1 mM hydroquinone as electrochemical mediator. The electrode showed linear response (poised at −300 mV vs Ag/AgCl) toward H₂O₂ concentration between 70 μM and 8.8 mM at pH 7.0. The biosensor reached 95% of steady-state current in about 12 s and its sensitivity was 33.8 mA/M cm². The electrode retained full initial activity after 30 days of storage at 4 °C in 50 mM sodium phosphate buffer, pH 7.0.     

Studies on Fe–Co spinel electrodes

FeCo₂O₄ spinel oxide pelleted electrodes were prepared from the respective powders, obtained by low-temperature coprecipitation method. X-ray diffraction studies suggest the coexistence of two spinel phases, with different a-cell parameters. The samples show semiconductor-type behaviour, in the range 530–340 K. The estimated activation energy for conduction is about 0.7 eV. These phases are stable, after being used as electrode materials, as the XRD and SEM/EDS results show. Cyclic voltammetry has been used to investigate the electrochemical behaviour of the FeCo₂O₄ electrodes in 1 mol dm⁻³ KOH aqueous solutions. The voltammetric data allowed finding out the redox reactions occurring at the electrode surface, namely Fe₃O₄·4H₂O/Fe(OH)₂ or Fe₃O₄/Fe₂O₃ and CoO₂/CoOOH by comparing the experimental results with those referred in the literature.     

A disposable amperometric enzyme immunosensor for rapid detection of Vibrio parahaemolyticus in food based on agarose/Nano-Au membrane and screen-printed electrode

Recent outbreaks of foodborne illnesses continue to support the need for rapid and sensitive methods for detection of foodborne pathogens. A disposable electrochemical immunosensor for detection of Vibrio parahaemolyticus (VP) based on the screen-printed electrode (SPE) coated with agarose/Nano-Au membrane and horseradish peroxidase (HRP) labeled VP antibody (HRP-anti-VP) has been developed in this paper. Then, the immunosensor was characterized by cyclic voltammetry (CV) and laser scanning confocal microscope (LSCM). The immunosensor was incubated with the one-step immunoassay format involving VP for 30 min at room temperature (25 ± 0.5 °C). The access of the active center of HRP catalyzing the oxidation reaction of thionine by H₂O₂ was partly inhibited by VP, which connected on the surface of the immunosensor by immunoreaction. VP could be quantificationally detected according to the shift of reduction current while CV was used as electrochemical means to detect the products of the enzymatic reaction. Under the optimum conditions of immunoreaction and electrochemical detection, VP was rapidly detectable by sigmoidal curve method and form a linear calibration between 10⁵ and 10⁹ cfu/ml with an associated detection limit of 7.374 × 10⁴ cfu/ml (S/N = 3). The immunosensor had acceptable specificity, reproducibility, stability and accuracy, indicating that the immunosensor could satisfy the need of practical sample detection.     

Direct electron transfer of cytochrome c and its biosensor based on gold nanoparticles/room temperature ionic liquid/carbon nanotubes composite film

A robust and effective composite film based on gold nanoparticles (GNPs)/room temperature ionic liquid (RTIL)/multi-wall carbon nanotubes (MWNTs) modified glassy carbon (GC) electrode was prepared by a layer-by-layer self-assembly technique. Cytochrome c (Cyt c) was successfully immobilized on the RTIL-nanohybrid film modified GC electrode by electrostatic adsorption. Direct electrochemistry and electrocatalysis of Cyt c were investigated. The results suggested that Cyt c could be tightly adsorbed on the modified electrode. A pair of well-defined quasi-reversible redox peaks of Cyt c was obtained in 0.10 M, pH 7.0 phosphate buffer solution (PBS). RTIL-nanohybrid film showed an obvious promotion for the direct electron transfer between Cyt c and the underlying electrode. The immobilized Cyt c exhibited an excellent electrocatalytic activity towards the reduction of H₂O₂. The catalysis currents increased linearly to the H₂O₂ concentration in a wide range of 5.0 × 10⁻⁵– 1.15 × 10⁻³ M. Based on the multilayer film, the third-generation biosensor could be constructed for the determination of H₂O₂.     

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.     

A comparative study for treatment of white liquor by different applications of Fenton process

In this paper, the treatability of white liquor by conventional (CFP), modified (MFP) and electro-Fenton oxidation processes (EFP) was investigated depending on the COD parameter. Based on the experimental results, up to 62.4%, 58.4% and 54.9% COD removals by the CFP, MFP and EFP were achieved, respectively. It was observed that adjustment of initial pH to acidic values is not required in the CFP. The optimal operational conditions were found to be [Fe²⁺] = 500 mg/L, [H₂O₂] = 1000 mg/L at pH 7.3 (original pH) in the CFP, [Fe⁰] = 1250 mg/L, [H₂O₂] = 1000 mg/L at pH 3 in the MFP, and I = 1.0 A, [H₂O₂] = 1500 mg/L at pH 3 in the EFP, respectively. As a result, the CFP has been determined as a more efficient alternative treatment method.     

Potentiometric coulometry based on charge accumulation with a peroxidase/osmium polymer-immobilized electrode for sensitive determination of hydrogen peroxide

The charge accumulation due to peroxidase (POD)-catalyzed reduction of H₂O₂ in a test solution (4 μL) by Os(II) in a POD/PVI[Os(dmebpy)₂Cl]-immobilized layer on an electrode (PVI = poly(1-vinylimidazole), dmebpy = 4,4′-dimethyl-2,2′-bipyridine) was monitored potentiometrically for the detection of H₂O₂. Before potentiometry, the Os(II)/Os(III) ratio of the modified electrode was controlled by pre-electrolysis at a given potential in a separated electrolysis cell. The redox potential of the Os polymer film in the test solution shifted to the positive side on the addition of H₂O₂ and reached a constant value due to the accumulation of Os(III) in the film. The total amount of the accumulated charge was determined from the area of the portion corresponding to the redox potential shift on a reversible cyclic voltammogram recorded separately. The low detection limit (5 pmol H₂O₂) was realized with 82–90% of the recovery percentage.     

Quasi-reversible two-electron reduction of oxygen at gold electrodes modified with a self-assembled submonolayer of cysteine

The electrochemical reduction of molecular oxygen (O₂) has been performed at gold electrodes modified with a submonolayer of a self-assembly (sub-SAM/Au) of a thiol compound (typically cysteine (CYST)) in O₂-saturated 0.5 M KOH. At bare gold electrode O₂ reduction reaction proceeds irreversibly, while this reaction is totally hindered at gold electrodes with a compact structure of CYST over its surface. The partial reductive desorption of the compact CYST monolayer was achieved by controlling the potential of the CYST/Au electrode, leading to the formation of a submonolayer coverage of the thiol compound over the Au electrode surface (sub-SAM/Au), at which the CYST molecules selectively block the Au(1 0 0) and Au(1 1 0) fractions (the so-called rough domains) of the polycrystalline Au while the Au(1 1 1) component (the so-called smooth domains) remains bare (i.e., uncovered with CYST). This sub-SAM/Au electrode extraordinarily exhibits a quasi-reversible two-electron reduction of molecular oxygen (O₂) in alkaline medium with a peak separation (ΔE_p) between the cathodic and anodic peak potentials (E_p^c,E_p^a) of about 60 mV. The ratio of the anodic current to the cathodic one is close to unity. The formal potential (E^o′) of this reaction is found to equal −150 mV vs. Ag/AgCl/KCl(sat.).     

Porous cuprous oxide microcubes for non-enzymatic amperometric hydrogen peroxide and glucose sensing

Using porous cuprous oxide (Cu₂O) microcubes, a simple non-enzymatic amperometric sensor for the detection of H₂O₂ and glucose has been fabricated. Cyclic voltammetry (CV) revealed that porous Cu₂O microcubes exhibited a direct electrocatalytic activity for the reduction of H₂O₂ in phosphate buffer solution and the oxidation of glucose in an alkaline medium. The non-enzymatic amperometric sensor used in the detection of H₂O₂ with detection limit of 1.5 × 10^?6 M over wide linear detection ranges up to 1.5 mM and with a high sensitivity of 50.6 μA/mM. This non-enzymatic voltammetric sensor was further utilized in detection of glucose with a detection limit of 8.0 × 10^?7 M, a linear detection range up to 500 μM and with a sensitivity of ?70.8 μA/mM.     

Application of carboxyl functionalized graphene oxide as mimetic peroxidase for sensitive voltammetric detection of H2O2 with 3,3′,5,5′-tetramethylbenzidine

A sensitive electrochemical method for H₂O₂ determination was proposed with carboxyl functionalized graphene oxide (GO-COOH) as mimetic peroxidase and 3,3′,5,5′-tetramethylbenzidine (TMB) as substrate. GO-COOH exhibited intrinsic peroxidase-like activity that could catalyze the oxidation of TMB with H₂O₂. The generated product exhibited a sensitive second order derivative linear sweep voltammetric reduction peak at − 0.93 V (vs. Ag/AgCl) in Britton–Robinson buffer. Under the optimal conditions the reduction peak current was proportional to H₂O₂ concentration in the linear range from 0.006 to 0.8 μmol L^− 1 with the detection limit of 1.0 nmol L^− 1 (3σ). This proposed method was further applied to determine H₂O₂ content in fresh milk samples with satisfactory results.     

Direct electrochemistry of hemoglobin and glucose oxidase in electrodeposited sol–gel silica thin films on glassy carbon

This work points out that electrogeneration of silica gel (SG) films on glassy carbon electrodes (GCEs) can be applied to immobilize biomolecules – hemoglobin (Hb) or glucose oxidase (GOD) or both of them in mixture – without preventing their activity. These proteins were physically entrapped in the sol–gel material in the course of the electro-assisted deposition process applied to form the thin films onto the electrode surface. SG films were prepared from a precursor solution by applying a suitable cathodic potential likely to induce a local pH increase at the electrode/solution interface, accelerating thereby polycondensation of the silica precursors with concomitant film formation. Successful immobilization of proteins was checked by various physico-chemical techniques. Both Hb and GOD were found to undergo direct electron transfer, as demonstrated by cyclic voltammetry. GCE–SG–Hb gave rise to well-defined peaks at potentials E_c = −0.29 V and E_a = −0.17 V in acetate buffer, corresponding to the Fe^III/Fe^II redox system of heme group of the protein, while GCE–SG–GOD was characterized by the typical signals of FAD group at E_c = −0.41 V and E_a = −0.33 V in phosphate buffer. These two redox processes were also evidenced on a single voltammogram when both Hb and GOD were present together in the same SG film. Hb entrapped in the silica thin film displayed an electrocatalytic behavior towards O₂ and H₂O₂ in solution, respectively in the mM and μM concentration ranges. Immobilized GOD kept its biocatalytic properties towards glucose. Combined use of these two proteins in mixture has proven to be promising for detection of glucose in solution via the electrochemical monitoring of oxygen consumption (decrease of the oxygen electrocatalytic signal).