Effect of Poly(amine)s on the Redox Properties of Carboxymethylcellulose and Ferrocene‐modified Poly(amine) Layer‐by‐Layer Films

Multilayer films consisting of carboxymethylcellulose (CMC) and ferrocene‐modified poly(ethyleneimine) (Fc‐PEI) or poly(allylamine hydrochloride) (Fc‐PAH) were successfully prepared on a gold electrode to examine their redox properties. The redox current of (Fc‐PEI/CMC)_n film‐coated electrodes increased with the number of layers, while the (Fc‐PAH/CMC)_n film‐coated electrodes exhibited increased response only for the first eight bilayers. The (Fc‐PEI/CMC)_n and (Fc‐PAH/CMC)_n films deposited on the surface of Fc‐free multilayer film‐coated electrodes also showed a redox response. The (PEI/CMC)₅ film‐coated electrode showed redox responses in Fc‐PEI and Fc‐PAH solutions, confirming the uptake of the Fc‐polymers in the inner film. In contrast, the uptake of the Fc‐polymers in the (PAH/CMC)₅ film was severely suppressed, suggesting that different permeability of (PEI/CMC)₅ and (PAH/CMC)₅ films.     

Facile Synthesis of Poly (N‐isopropylacrylamide) Coated SiO2 Core‐shell Microspheres via Surface‐initiated Atom Transfer Radical Polymerization for H2O2 Biosensor Applications

In this study, inorganic/organic composites containing poly (N‐isopropylacrylamide) coated core‐shell SiO₂ microspheres were prepared via surface‐initiated atom transfer radical polymerization (ATRP). The thermal responsive polymer, N‐isopropylacrylamide was treated with methanol, water and CuBr/CuBr₂/1,1,4,7,7‐pentamethyldiethylenetriamine (PMDETA) at room temperature to form PNIPAM@SiO₂ microspheres. The as‐prepared PNIPAM@SiO₂ microspheres were characterized by FT‐IR, TGA, XPS, SEM, TEM analyses. Hemoglobin (Hb) was immobilized onto the surfaces of PNIPAM@SiO₂ microspheres via hydrophobic and π‐π stacking interactions. The as‐prepared Hb/PNIPAM@SiO₂ electrode exhibits well‐defined redox peak at a formal potential of −0.38 V, validating the direct electrochemistry of Hb. The Hb immobilized composite film retained its bioelectroactivity without any significant loss of catalytic activity. The modified electrode detects H₂O₂ over a wide linear concentration range (0.1 μM to 333 μM) with a detection limit of 0.07 μM. This modified electrode also successfully detects H₂O₂ from food and disinfectant samples with appreciable recovery values, validating its practicality. We believe that PNIPAM@SiO₂ composite has great potential to be used in the detection of H₂O₂ and development of other enzyme based biosensors.     

Functionalization of Single‐Walled Carbon Nanotubes with Cubic Prussian Blue and Its Application for Amperometric Sensing

In this work, we synthesized electroactive cubic Prussian blue (PB) modified single‐walled carbon nanotubes (SWNTs) nanocomposites using the mixture solution of ferric‐(III) chloride and potassium ferricyanide under ambient conditions. The successful fabrication of the PB‐SWNTs nanocomposites was confirmed by scanning electron microscopy (SEM), transmission electron microscopy (TEM), UV‐vis absorption spectroscopy, Fourier transform infrared (FTIR) spectroscopy, and cyclic voltammetry (CV). PB nanocrystallites are observed to be finely attached on the SWNTs sidewalls in which the SWNTs not only act as a carrier of PB nanocrystallites but also as Fe(III)‐reducer. The electrochemical properties of PB‐SWNTs nanocomposites were also investigated. Using the electrodeposition technique, a thin film of PB‐SWNTs/chitosan nanocomposites was prepared onto glassy carbon electrode (GCE) for the construction of a H₂O₂ sensor. PB‐SWNTs/chitosan nanocomposites film shows enhanced electrocatalytic activity towards the reduction of H₂O₂ and the amperometric responses show a linear dependence on the concentration of H₂O₂ in a range of 0.5–27.5 mM and a low detection limit of 10 nM at the signal‐to‐noise ratio of 3. The time required to reach the 95% steady state response was less than 2 s. CV studies demonstrate that the modified electrode has outstanding stability. In addition, a glucose biosensor is further developed through the simple one‐step electrodeposition method. The observed wide concentration range, high stability and high reproducibility of the PB‐SWNTs/chitosan nanocomposites film make them promising for the reliable and durable detection of H₂O₂ and glucose.     

Redox Properties of Ferricyanide Ion on Layer‐by‐Layer Thin Film‐Coated Gold Electrodes; Effects of Type of Polycationic Components in the Film

The surface of a gold (Au) electrode was coated with layer‐by‐layer (LbL) thin films composed of poly(vinyl sulfate) (PVS) and different type of poly(amine)s including poly(allylamine) (PAH), poly(ethyleneimine) (PEI) and poly(diallyldimethylammonium chloride) (PDDA) and redox properties of ferricyanide ion ([Fe(CN)₆]^3?) on the LbL film‐coated Au electrodes were studied. The LbL film‐coated electrodes exhibited redox response to [Fe(CN)₆]^3? ion when the outermost surface of the LbL film was covered with the cationic poly(amine)s while virtually no response was observed on the LbL film‐coated electrodes whose outermost surface was covered with PVS due to an electrostatic repulsion between [Fe(CN)₆]^3? ion and the negatively‐charged PVS layer. The redox properties of [Fe(CN)₆]^3? ion on the LbL film‐coated electrodes significantly depended on the type of polycationic materials in the LbL film. The LbL film‐coated electrodes which had been immersed in the [Fe(CN)₆]^3? solution for 15 min exhibited redox response even in a [Fe(CN)₆]^3? ion‐free buffer solution, suggesting that [Fe(CN)₆]^3? ion is confined in the films. In the buffer solution, redox peaks were observed between +0.1 and 0.4 V depending on the type of polycations in the film. Thus, [Fe(CN)₆]^3? ion can be confined in the film and the redox potential is polycation‐dependent.     

Extremely Stable Copper–Polymelamine Composite Material for Amperometric Hydrogen Peroxide Sensing

For the first time, electroactive poly(melamine) film has been prepared and evaluated to function as a matrix to immobilize “spherical” copper nanoclusters and also as a ligand to form a copper complex with enhanced electrocatalytic activity. This is a simple two‐step process that involves first polymerization of melamine on a screen‐printed carbon electrode followed by electrodeposition of copper without using harmful and environmentally toxic chemicals. It is remarkable that this electrode can electrocatalytically reduce H₂O₂ at approximately ?0.2 V versus Ag/AgCl in pH 7 PBS and possesses superior stability in amperometric analysis of H₂O₂ under the continuous wall‐jet flow of 1 mM of H₂O₂ for more than 5 h. Most importantly, we have demonstrated that the as‐prepared copper‐poly(melamine)‐modified electrode can solve the setbacks of copper‐based H₂O₂ sensors reported so far. © 2013 Wiley Periodicals, Inc. J. Polym. Sci., Part B: Polym. Phys. 2013 , 51, 1639–1646     

Self‐Assembled Prussian Blue Nanoparticles Based Electrochemical Sensor for High Sensitive Determination of H2O2 in Acidic Media

In this paper, self‐assembled Prussian blue nanoparticles (PBNPs) on carbon ceramic electrode (CCE) were developed as a high sensitive hydrogen peroxide (H₂O₂) electrochemical sensor. The PBNPs film was prepared by a simple dipping method. The morphology of the PBNPs‐modified CCE was characterized by scanning electron microscopy (SEM). The self‐assembled PB film exhibited sufficient mechanical, electrochemical stability and high sensitivity in compare with other PB based H₂O₂ sensors. The sensor showed a good linear response for H₂O₂ over the concentration range 1 μM–0.26 mM with a detection limit of ca. 0.7 μM (S/N=3), and sensitivity of 754.6 mA M^?1 cm^?2. This work demonstrates the feasibility of self‐assembled PBNPs‐modified CCE for practical sensing applications.     

Carbon Nanotubes‐Based Amperometric Cholesterol Biosensor Fabricated Through Layer‐by‐Layer Technique

A carbon nanotubes‐based amperometric cholesterol biosensor has been fabricated through layer‐by‐layer (LBL) deposition of a cationic polyelectrolyte (PDDA, poly(diallyldimethylammonium chloride)) and cholesterol oxidase (ChOx) on multi‐walled carbon nanotubes (MWNTs)‐modified gold electrode, followed by electrochemical generation of a nonconducting poly(o‐phenylenediamine) (PPD) film as the protective coating. Electrochemical impedance measurements have shown that PDDA/ChOx multilayer film could be formed uniformly on MWNTs‐modified gold electrode. Due to the strong electrocatalytic properties of MWNTs toward H₂O₂ and the low permeability of PPD film for electroacitve species, such as ascorbic acid, uric acid and acetaminophen, the biosensor has shown high sensitivity and good anti‐interferent ability in the detection of cholesterol. The effect of the pH value of the detection solution on the response of the biosensor was also investigated. A linear range up to 6.0 mM has been observed for the biosensor with a detection limit of 0.2 mM. The apparent Michaelis‐Menten constant and the maximum response current density were calculated to be 7.17 mM and 7.32 μA cm^?2, respectively.     

Biosensor Based on Self‐Assembling Glucose Oxidase and Dendrimer‐Encapsulated Pt Nanoparticles on Carbon Nanotubes for Glucose Detection

A novel amperometric glucose biosensor based on layer‐by‐layer (LbL) electrostatic adsorption of glucose oxidase (GOx) and dendrimer‐encapsulated Pt nanoparticles (Pt‐DENs) on multiwalled carbon nanotubes (CNTs) was described. Anionic GOx was immobilized on the negatively charged CNTs surface by alternatively assembling a cationic Pt‐DENs layer and an anionic GOx layer. Transmission electron microscopy images and ζ‐potentials proved the formation of layer‐by‐layer nanostructures on carboxyl‐functionalized CNTs. LbL technique provided a favorable microenvironment to keep the bioactivity of GOx and prevent enzyme molecule leakage. The excellent electrocatalytic activity of CNTs and Pt‐DENs toward H₂O₂ and special three‐dimensional structure of the enzyme electrode resulted in good characteristics such as a low detection limit of 2.5 μM, a wide linear range of 5 μM–0.65 mM, a short response time (within 5 s), and high sensitivity (30.64 μA mM^?1 cm^?2) and stability (80% remains after 30 days).     

Sensitive and Selective Sensing of Hydrogen Peroxide with Iron‐Tetrasulfophthalocyanine‐Graphene‐Nafion Modified Screen‐Printed Electrode

We developed a novel iron‐tetrasulfophthalocyanine‐graphene‐Nafion (FeTSPc‐GR‐Nafion) modified screen‐printed electrode to determine hydrogen peroxide (H₂O₂) with high sensitivity and selectivity. The nanocomposite film (FeTSPc‐GR‐Nafion) exhibits an excellent electrocatalytic activity towards oxidation of H₂O₂ at a potential of +0.35 V in the absence of enzyme. A comparative study reveals that the FeTSPc‐GR complexes play a dual amplification role. Amperometric experiment indicates that the sensors possess good sensitivity and selectivity, with a linear range from 2.0×10^?7 M to 5.0×10^?3 M and a detection limit of 8.0×10^?8 M. This sensor has been successfully used to develop the glucose biosensor and has also been applied to determine H₂O₂ in sterile water.     

Hemin Modified SnO2 Films on ITO‐PET with Enhanced Activity for Electrochemical Sensing

We present a simple and efficient method for the preparation of hemin‐modified SnO₂ films on low cost, flexible, conducting ITO‐PET substrates to enable the development of a sensitive electrochemical sensor for the determination of H₂O₂. Using a hydrothermal processing method meant that the SnO₂ films can be prepared at low temperatures, compatible with the PET substrate. The properties of the electrodes enable a high hemin loading to be achieved in a stable and functional way, allowing the direct reduction and oxidation of the immobilized hemin and maintaining its high electrocatalytic activity in the reduction of H₂O₂ on the surface. The results showed a sensitive response linearly proportional to the concentration of H₂O₂ in the range 1.5 to 90 μM.     

Electrochemical Fabrication of Prussian Blue Nanocube‐decorated Electroreduced Graphene Oxide for Amperometric Sensing of NADH

In this study, Prussian blue (PB) film on the electroreduced graphene oxide (ERGO)‐modified Au electrode surface (ERGO/PB) is easily prepared by means of cyclic voltammetric technique in the mixture of K₃Fe(CN)₆ and FeCl₃. Its electrochemical behaviors for NADH biosensor are studied. The structural and morphological characters of modified electrode material are analyzed with using of XPS, XRD, Raman, EDS, and SEM techniques. ERGO/PB hybrid nanocomposite for NADH biosensor is exhibited to the higher catalytic effect (linear range from 1.0 to 100 μM, detection limit of 0.23 μM at S/N=3) compared to naked Au, ERGO‐modified Au, and PB‐modified Au electrodes. In addition to, ERGO/PB electrode was used to voltammetric and amperometric detection of H₂O₂. ERGO/PB electrodes also showed the same behavior as the NADH sensor. This ERGO/PB‐modified electrode supplied a simple, new, and low‐cost route for amperometric sensing of both NADH and H₂O₂.     

A Highly Selective Amperometric Hydrogen Peroxide Sensor Based on Silicomolybdate‐Doped‐Glutaraldehyde‐Cross‐Linked Poly‐L‐Lysine Film Modified Glassy Carbon Electrode

Silicomolybdate‐doped‐glutaraldehyde‐cross‐linked poly‐L ‐lysine (PLL‐GA‐SiMo) film modified glassy carbon electrode was prepared by means of electrostatically trapping the silicomolybdate anion in the cationic film. The PLL‐GA‐SiMo film was stable and the charge transport through the film was fast. The modified electrode shows excellent electrocatalytic activity towards hydrogen peroxide reduction with significant reduction of overpotential, however, not responded to potential interferrents such as dopamine, ascorbic acid and uric acid. This unique feature of PLL‐GA‐SiMo modified electrode allowed for the development of a highly selective method for the determination of H₂O₂ in the presence of interferents.     

Urchin‐like Ag Nanowires as Non‐enzymatic Hydrogen Peroxide Sensor

Urchin‐like Ag nanowires were prepared by reacting AgNO₃(aq) with Cu metal in the presence of cetyltrimethylammonium chloride and HNO₃(aq) on a screen printed carbon electrode at room temperature. The diameters of the nanowires were about 100 nm, while the lengths were up to 10 μm. Cyclic voltammetric experiments using the Ag nanowires as the working electrode showed electrocatalytic H₂O₂ reduction. The electrode exhibited a high sensitivity of 4705 μA mM^‐1 mg^‐1 cm^‐2 from 50 μM to 10.35 mM and a measurable detection limit of 10 μM in amperometric detection. This is the first report on Ag NWs for non‐enzymatic H₂O₂ sensing.     

Silicomolybdate‐Doped PEDOT Modified Electrode: Electrocatalytic Reduction of Bromate and Oxidation of Ascorbic Acid

Electrically conducting poly(3,4‐ethylenedioxythiophene) (PEDOT) film doped with silicomolybdate (SiMo₁₂O₄₀^4? or SiMo₁₂) was synthesized by electrochemical polymerization. The synthesized film is capable of fast charge propagation during redox reactions in strong acid medium 0.2 M H₂SO₄ solution. The modified electrode was used towards reduction of bromate and successfully employed as an amperometric sensor for bromate and also above modified electrode was investigated for ascorbic acid oxidation.     

Electron‐Transfer Mediator Microbiosensor Fabrication Based on Immobilizing HRP‐Labeled Au Colloids on Gold Electrode Surface by 11‐Mercaptoundecanoic Acid Monolayer

In this paper, a new strategy for constructing a mediator‐type amperometric hydrogen peroxide (H₂O₂) microbiosensor was described. An electropolymerized thionine film (PTH) was deposited directly onto a gold electrode surface. The resulting redox film was extremely thin, adhered well onto a substrate (electrode), and had a highly cross‐linked network structure. Consequently, horseradish peroxidase (HRP) was successfully immobilized on nanometer‐sized Au colloids, which were supported by thiol‐tailed groups of 11‐mercaptoundecanoic acid (11‐MUA) monolayer covalently bound onto PTH film. With the aid of the PTH mediator, HRP‐labeled Au colloids microbiosensor displayed excellent electrocatalytical response to the reduction of H₂O₂. This matrix showed a biocompatible microenvironment for retaining the native activity of the covalent HRP and a very low mass transport barrier to the substrate, which provided a fast amperometric response to H₂O₂. The proposed H₂O₂ microbiosensor exhibited linear range of 3.5 μM–0.7 mM with a detection limit of 0.05 μM (S/N=3). The response showed a Michaelis‐Menten behavior at larger H₂O₂ concentrations. The K_M^app value for the biosensors based on 24 nm Au colloids was found to be 47 μM, which demonstrated that HRP immobilized on Au colloids exhibited a high affinity to H₂O₂ with no loss of enzymatic activity. This microbiosensor possessed good analytical performance and storage stability.     

Nonenzymatic H2O2 Electrochemical Sensor Based on SnO2‐NPs Coated Polyethylenimine Functionalized Graphene

This paper demonstrated using polyethylenimine (PEI)‐functionalized graphene (Gr) incorporating tin oxide (SnO₂) hybrid nanocomposite as a platform for nonenzymatic H₂O₂ electrochemical sensor. The results of UV‐vis spectroscopy and X‐ray diffraction (XRD) confirmed the simultaneous formation of tin oxide (SnO₂) nanocomposite and reduction of graphene oxide (GO). Transmission electron microscopy (TEM) images showed a uniform distribution of nanometer‐sized tin oxide nanoparticles on the grapheme sheets, which could be achieved using stannous chloride (SnCl₂) complex instead of tin oxide as precursor. The electrochemical measurements, including cyclic voltammetry (CV) and amperometric performance (I‐t), showed that the PEI‐functionalized Gr supported SnO₂ (SnO₂‐PEI‐Gr) exhibited an excellent electrocatalytic activity toward the H₂O₂. The corresponding calibration curve of the current response showed a linear detection range of 9×10⁻⁶∼1.64×10⁻³ mol L⁻¹, while the limit of detection was estimated to be 1×10⁻⁶ mol L⁻¹. Electrochemical studies indicated that SnO₂ and functionalized Gr worked synergistically for the detection of H₂O₂.     

Silver Nanograins Incorporated PEDOT Modified Electrode for Electrocatalytic Sensing of Hydrogen Peroxide

A silver nanograins (Ag_NGs) incorporated poly[3,4‐ethylenedioxythiophene] (PEDOT) modified electrode was prepared by a simple electrochemical method without using any stabilizer or reducing agent. The surface morphology and thickness of the resulting modified electrode was characterized by using AFM. It was found that the size of the silver particles in the PEDOT modified electrode was smaller than that in the bare electrode. AFM studies also revealed that Ag_NGs were uniformly distributed in the PEDOT modified electrode and the thickness of the film was found to be 35 nm. The Ag_NGs incorporated PEDOT modified electrode exhibited good electrocatalytic activity towards the reduction of hydrogen peroxide without an enzyme or mediator immobilized in the electrode. It has shown good amperometric response to hydrogen peroxide (H₂O₂) with a detection limit of 7 μM and a response time of 5 s.     

A Third‐Generation Hydrogen Peroxide Biosensor Based on Horseradish Peroxidase Covalently Immobilized on Electrografted Organic Film on Screen‐Printed Carbon Electrode

A new convenient strategy to fabricate a third‐generation hydrogen peroxide biosensor was described. The screen‐printed carbon electrode (SPCE) was first modified with a layer of 4‐nitrophenyl assembled from the 4‐nitroaniline diazonium salt synthesized in situ in acidic aqueous solution. Next, the nitro groups were converted to amines followed by crosslinking to the horseradish peroxidase (HRP) by glutaraldehyde. The redox chemistry of the active center of the HRP was observed and the HRP‐modified electrode displayed electrocatalytic activity towards the reduction of hydrogen peroxide (H₂O₂) without any mediators. H₂O₂ was determined in a linear range from 5.0 μM to 50.0 μM, with a detection limit of 1.0 μM. Furthermore, the biosensor exhibited fast amperometric response, good reproducibility and long‐term stability.     

Direct Electrochemistry and Bioelectrocatalysis of Microperoxidase‐11 Immobilized on Chitosan‐Graphene Nanocomposite

A bioelectrochemical platform has been constructed for the direct electron transfer and biosensing purposes of microperoxidase‐11 (MP‐11) immobilized on the chitosan dispersed multilayer graphene nanocomposite. The immobilized MP‐11 at the modified gold electrode displays a well‐defined and quasireversible redox peaks, with a formal potential of ?0.38 V/SCE in a buffer solution (pH 7.0). MP‐11 absorbed on the electrode surface exhibits high electrocatalytic activity toward the reduction of both oxygen and hydrogen peroxide and also shows good analytical performance for the amperometric detection of H₂O₂ with a linear range from 2.5 to 135 μM. These results indicate the graphene modified electrode might be used as a third generation biosensor for H₂O₂ detection.     

3,4‐Dihydroxy‐L‐phenylalanine for Preparation of Gold Nanoparticles and as Electron Transfer Promoter in H2O2 Biosensor

3,4‐Dihydroxy‐L ‐phenylalanine (dopa) and 2‐(3,4‐dihydroxyphenyl)ethylamine (dopamine) were investigated as reducing agent and stabilizer for synthesis of gold nanoparticles (AuNPs) by one‐pot heating of a solution of HAuCl₄/dopa or dopamine. AuNPs with different sizes were obtained by controlling the mass ratios of HAuCl₄/dopa or dopamine. The formation mechanism for AuNPs was also proposed. Immobilization of horseradish peroxidase (HRP) and promotion of its electron transfer by polydopa film were investigated for preparation of H₂O₂ biosensor. Alkaline dopa solution was dropped onto a gold electrode for the formation of polydopa film. HRP was immobilized on the polydopa film through interactions between heme centre of HRP and the amine and carboxyl groups in polydopa. The AuNPs embedded in the polydopa film improved the electron transfer efficiency. These two factors allowed successful development of a H₂O₂ sensor with HRP@polydopa‐AuNPs electrode. Due to its biocompatibility, the polydopa‐AuNPs film provided good retention of enzyme activity and long‐term stability of the sensor. A rapid catalytic response (3 s) and a linear range from 0.006 to 5.0 mmol L^?1 were obtained for H₂O₂. This facile preparation strategy can be extended to other enzyme‐based biosensors.