Peroxide Biosensor Research Articles

Nanobiocomposite Electrochemical Biosensor Utilizing Synergic Action of Neutral Red Functionalized Carbon Nanotubes

Abstract An amperometric hydrogen peroxide biosensor using a nanobiocomposite based on neutral red modified carbon nanotubes and co-immobilized glucose oxidase and horseradish peroxidase is reported. Modification of the nanobiocomposite electrode with neutral red resulted in a sensitive, low-cost and reliable H2O2 sensor. The use of carbon nanotubes, as the conductive part of the composite, facilitated fast electron transfer rates. The biosensor was characterized for the influence of pH, potential and temperature. A remarkable feature of the biosensor is the detection of H2O2 at low applied potentials where the noise level and interferences are minimal. The sensor has a fast steady-state measuring time of 10 s with a quick response (2 s). The biosensor showed a linear range from 15 nM to 45 mM of H2O2 and a detection limit of 5 nM. Nafion, which is used as a binder, makes the determination free from other electroactive substances. The repeatability, reproducibility, stability and analytical performance of the sensor are very good.

Reduced Graphene Oxide|Carbon Ceramic Electrode Modified with CdS‐Hemoglobin as a Sensitive Hydrogen Peroxide Biosensor

AbstractA sensitive hydrogen peroxide (H2O2) biosensor was developed based on a reduced graphene oxide|carbon ceramic electrode (RGO|CCE) modified with cadmium sulfide‐hemoglobin (CdS‐Hb). The electron transfer kinetics of Hb were promoted due to the synergetic function of RGO and CdS nanoparticles. The transfer coefficient (α) and the heterogeneous electron transfer rate constant (ks) were calculated to be 0.54 and 2.6 s−1, respectively, indicating a great facilitation achieved in the electron transfer between Hb and the electrode surface. The biosensor showed a good linear response to the reduction of H2O2 over the concentration range of 2–240 µM with a detection limit of 0.24 µM (S/N=3) and a sensitivity of 1.056 µA µM−1 cm−2. The high surface coverage of the CdS‐Hb modified RGO|CCE (1.04×10−8 mol cm−2) and a smaller value of the apparent MichaelisMenten constant (0.24 mM) confirmed excellent loading of Hb and high affinity of the biosensor for hydrogen peroxide.

An Electrochemical Hydrogen Peroxide Biosensor Based on Polydopamine-Entrapped G-Quadruplex-Hemin DNAzyme

The complex of G-quadruplex-hemin, named peroxidase-mimicking DNAzyme, shows a catalytic activity towards the electro-reduction of hydrogen peroxide. Although DNAzyme is popular for the preparation of colorimetric biosensors for hydrogen peroxide, it is rarely used as a catalyst in the assembly of electrochemical biosensors. Meanwhile, it is of significance to develop novel methods in immobilizing DNA for the purpose of fabrication of DNA-based biosensors. In this study, an electrochemical hydrogen peroxide biosensor was successfully constructed using polydopamine-entrapped G-quadruplex-hemin DNAzyme. G-quadruplex-hemin complex was achieved by mixing DNA and hemin. After the physical adsorption of DNAzyme on a glassy carbon electrode, a droplet of 10 μL containing 5 g L−1 dopamine in PBS (pH 8.0) was then cast onto the surface. Dopamine was oxidized by oxygen in air to form polydopamine, realizing the immobilization of DNAzyme. The influence of DNA sequence on the performance of the biosensor is investigated and different conformation-dependent response from electrochemical to optical sensors was observed. The proposed biosensor shows a linear range with the concentration from 10 μM to 1.5 mM and a detection limit of 2.2 μM for hydrogen peroxide. The results demonstrated the possibility of enzyme immobilization on an electrode through polydopamine and the substitution of G-quadruplex-hemin DNAzyme for natural enzyme.

Synthesis, characterisation and catalytic activity of gold and silver nanoparticles in the bio-sensor application

Most of the hydrogen peroxide (H2O2) bio-sensors developed till date are based on enzymes and proteins causing them to have a limited lifetime. Moreover, complex procedures are followed for sensor fabrication. Therefore, an inorganic material-based sensor, with a simple design and longer shelf life is highly desirable. In this work, surfactant-metal (gold and silver) nanoparticles are prepared in aqueous solutions containing cetyltrimethylammonium bromide. The particle sizes of the metal nanoparticles obtained are characterised by UV–Vis, HRTEM, X-ray diffraction and FTIR; the average sizes of gold and silver nanoparticles are 8 and 10 ± 0.2 nm, respectively. The nanoparticles are tested for H2O2 detection. The sensor is characterised and tested using samples from M to mM H2O2 range and a linear response is observed. Low-detection limits and high sensitivity are some of the advantages of this work. Same principle could be extended for the detection of other substrates as well.

A novel hydrogen peroxide sensor based on immobilization of haemoglobin on nano-TiO2/DTAB composite film

The direct electron transfer of immobilized haemoglobin (Hb) on nano-TiO2 and dodecyltrimethylammonium bromide (DTAB) film modified carbon paste electrode (CPE) and its application as a hydrogen peroxide (H2O2) biosensor were investigated. On nano-TiO2/DTAB/Hb/CPE, Hb displayed a rapid electron transfer process with participation of one proton and with an electron transfer rate constant which estimated as 0.29 s− 1. Thus, the proposed biosensor exhibited a high sensitivity and excellent electrocatalytic activity for the reduction of H2O2. The catalytic reduction current of H2O2 was proportional to H2O2 concentration in the range of 0.2–4.0 mM with a detection limit of 0.07 mM. The apparent Michaelis–Menten constant (Kmapp) of the biosensor was calculated to be 0.127 mM, exhibiting a high enzymatic activity and affinity. This sensor for H2O2 can potentially be applied in determination of other reactive oxygen species as well.

Highly sensitive and selective hydrogen peroxide biosensor based on hemoglobin immobilized at multiwalled carbon nanotubes–zinc oxide composite electrode

A highly sensitive and selective amperometric hydrogen peroxide (H2O2) biosensor based on immobilization of hemoglobin (Hb) at multiwalled carbon nanotubes–zinc oxide (MWCNT/ZnO) composite modified glassy carbon electrode (GCE) is reported. ZnO microsponges were electrochemically grown on MWCNT surface by the simple, cost-effective, green, electrochemical method at room temperature. The MWCNT/ZnO/Hb composite film showed a pair of well-defined, quasi-reversible redox peaks with a formal potential (E°′) of −0.336V, characteristic features of heme redox couple of Hb. The electron transfer rate constant (ks) of immobilized Hb was 1.26s−1. The developed biosensor showed a very fast response (>2s) toward H2O2 with good sensitivity, wide linear range, and low detection limit of 0.02μM. The fabricated biosensor showed interesting features, including high selectivity, acceptable stability, good reproducibility, and repeatability along with excellent conductivity, facile electron mobility of MWCNT, and good biocompatibility of ZnO. The fabrication method of this biosensor is simple and effective for determination of H2O2 in real samples with quick response, good sensitivity, high selectivity, and acceptable recovery.

A new electrochemical biosensor for hydrogen peroxide using HRP/AgNPs/cysteamine/p-ABSA/GCE self-assembly modified electrode

An electrochemical hydrogen peroxide biosensor was designed by immobilizing horseradish peroxidase (HRP) on Ag nanoparticles/cysteamine/p-aminobenzene sulfonic acid/glassy carbon (GC) electrode. Ag nanoparticles can act as tiny conduction centers on electrodes that adsorb redox enzymes, facilitating the transfer of electrons with no requiring any loss of biological activity. The forerunner film was first electropolymerized on the glassy carbon electrode with p-aminobenzene sulfonic acid (p-ABSA) by cyclic voltammetry. The cysteamine (CA) was bound on the surface of the film by electrostatic force, then Ag nanoparticles were immobilized on the cysteamine monolayer, and lastly HRP was adsorbed onto the surfaces of the Ag nanoparticles. A dramatic decrease in the overvoltage of H2O2 was observed with improved sensitivity, which makes the modified electrodes of great promise for oxidase-based amperometric biosensors. The biosensor responded to H2O2 in the linear range from 1.2×106 mol/L to 9.8×103 mol/L with a detection limit of 1.1×108 mol/L. Moreover, the obtained biosensor exhibited good accuracy and high sensitivity.

Hydrogen Peroxide Biosensor Based on Cellulose Diacetate Ionic Liquid Film Immobilizing Myoglobin

Hydrogen Peroxide Biosensor Based on Cellulose Diacetate Ionic Liquid Film Immobilizing Myoglobin

Fabrication of Hydrogen Peroxide Biosensors Based on Microelectrode Array of Silicon Dioxide Cavities

Fabrication of Hydrogen Peroxide Biosensors Based on Microelectrode Array of Silicon Dioxide Cavities

Electrochemistry and biosensing activity of cytochrome c immobilized on a mesoporous interface assembled from carbon nanospheres

We report on an amperometric biosensor for hydrogen peroxide. It is obtained via layer-by-layer assembly of ordered mesoporous carbon nanospheres and poly(diallyldimethylammonium) on the surface of an indium tin oxide (ITO) glass electrode and subsequent adsorption of cytochrome c. UV–vis absorption spectroscopy was applied to characterize the process of forming the assembled layers. Cyclic voltammetry revealed a direct and quasi-reversible electron transfer between cytochrome c and the surface of the modified ITO electrode. The surface-controlled electron transfer has an apparent heterogeneous electron-transfer rate constant (ks) of 5.9 ± 0.2 s−1 in case of the 5-layer electrode. The biosensor displays good electrocatalytic response to the reduction of H2O2, and the amperometric signal increase steadily with the concentration of H2O2 in the range from 5 μM to 1.5 mM. The detection limit is 1 μM at pH 7.4. The apparent Michaelis-Menten constant (Km) of the sensor is 0.53 mM. We assume that the observation of a direct electron transfer of cytochrome c on mesoporous carbon nanospheres may form the basis for a feasible approach for durable and reliable detection of H2O2.

Application of tubular tetrapod magnesium oxide in a biosensor for hydrogen peroxide

Tubular tetrapod magnesium oxide (tt-MgO) can be synthesized by thermal evaporation of Mg metal powder with a pre-grown tetrapod ZnO template. The morphology and structure of the tt-MgO were characterized by X-ray diffraction, scanning electron microscopy and transmission electron microscopy. A composite prepared from tt-MgO, nafion and horseradish peroxidase was employed to modify a gold electrode to result in an electrochemical biosensor for hydrogen peroxide that displays excellent sensitivity and rapid response in the presence of hydroquinone as a mediator. Its sensitivity is 335.4 μA mM-1 cm-2, its response is linear in the range from 1.0 to 450 μM, and the detection limit is 0.3 μM. These results demonstrate that tt-MgO provides a promising material for the designs of biosensors.

The electrochemiluminescent behavior of nickel phthalocyanine (NiTSPc)/H2O2 system on a heated ITO electrode

Abstract In this study, the electrochemiluminescent (ECL) behavior of Nickel(II) tetrasulfophthalocyanine (NiTSPc)/H2O2 on a heated indium tin oxide (ITO) electrode was investigated. The effect of pH value, electrochemical scan mode, concentration of NiTSPc and electrode surface temperature on the ECL intensities had been studied in detail. Based on the fact that the ECL of NiTSPc can be greatly enhanced by hydrogen peroxide at the ITO electrode, a new ECL biosensor for hydrogen peroxide has been developed. The possible mechanism for the ECL of NiTSPc has also been proposed.

Novel Amperometric Hydrogen Peroxide Biosensor Based on Horseradish Peroxidase Azide Covalently Immobilized on Ethynyl‐Modified Screen‐Printed Carbon Electrode via Click Chemistry

AbstractA novel, simple and versatile protocol for covalent immobilization of horseradish peroxidase (HRP) on screen‐printed carbon electrode (SPCE) based on the combination of diazonium salt electrografting and click chemistry has been successfully developed. The ethynyl‐terminated monolayers are obtained by diazonium salt electrografting, then, in the presence of copper (I) catalyst, the ethynyl modified surfaces reacted efficiently and rapidly with horseradish peroxidase bearing an azide function (azido‐HRP), thus forming a covalent 1,2,3‐triazole linkage by means of click chemistry. All the experimental results suggested that HRP was immobilized onto the electrode surface successfully without denaturation. Furthermore, the immobilized HRP showed a fast electrocatalytic reduction for H2O2. A linear range from 5.0 to 50.0 µM in a phosphate buffer (pH 5.5) with detection limit of 0.50 µM and sensitivity of 0.23 nA/µM were obtained. The heterogeneous electron transfer rate constant Kct was 1.52±0.22 s−1 and the apparent MichaelisMenten constant was calculated to be 0.028 mM. The HRP‐functionalized electrode demonstrated a good reproducibility and long‐term stability.

A new hydrogen peroxide biosensor based on synergy of Au@Au2S2O3 core–shell nanomaterials and multi-walled carbon nanotubes towards hemoglobin

A new H2O2 biosensor was constructed by immobilizing Hb in the Chit-MWNTs/Au@Au2S2O3 composite membrane. Owing to the superior performance of novel Au@Au2S2O3 core–shell nanomaterials and its synergy with multi-walled carbon nanotubes (MWNTs), the direct electron transfer of Hb in composite membrane was easily achieved. The Chit-MWNTs/Hb/Au@Au2S2O3/GCE displayed a pair of well-defined and reversible redox peaks in 0.1M pH 7.0 PBS. The electrocatalytic reduction to H2O2 of Chit-MWNTs/Hb/Au@Au2S2O3/GCE was also studied. Its apparent Michaelis–Menten constant for H2O2 was 0.014mM, showing a good affinity. The experimental results showed that the linear dependence of the biosensor was from 4.0×10−5 to 7.2×10−4M, and the detection limit of the sensor was 3.2×10−6M (S/N=3). Moreover, the biosensor showed a rapid response to H2O2, a good stability and reproducibility.

Rapid and sensitive amperometric determination of hydrogen peroxide with a biosensor based on a carboxyphenyl functionalised boron-doped diamond electrode

In the present work, a mediator-free biosensor for hydrogen peroxide (H2O2) based on a surface carboxyphenyl functionalised boron-doped diamond (BDD) electrode has been constructed via a simple electrochemical way in an aryl diazonium salt. The BDD surface was primarily interfacial, designed by grafting an aromatic ‘bridge’ of 4-carboxyphenyl (4-CP) groups by single-stepped electrochemical reduction of 4-CP diazonium cations. A locked-in carboxyl-terminated BDD interface was thus produced. Then the model biomolecule, cytochrome c (Cyt c) was attached to the interface through the ‘bridge’ by covalent combination between –NH2 terminal groups of Cyt c and –COOH groups of 4-CP, obtaining a Cyt c/4-CP/BDD biosensor. Well-defined peaks attributed to the FeIII/FeII couple of Cyt c were clearly observed on the biosensor which could not be obtained direct on BDD without carboxyphenyl functionalisation. The Cyt c/4-CP/BDD biosensor further exhibited remarkable biosensor features in the rapid and sensitive amperometric sensing of H2O2. The steady-state reduction current response to the addition of H2O2 could be achieved within 4 s. In the range from 4.0 × 10−6 to 3.6 × 10−5 M, the reduction current indicated good linear dependence with the concentration of H2O2. The detection limit was estimated to be as low as 6.1 × 10−7 M based on the signal to noise ratio of 3 and the apparent Michaelis constant was evaluated to be 1.02 ± 0.25 × 10−5 M according to the Lineweaver-Burke equation. All the results indicated that the covalent graft 4-carboxyphenyl group played an important role in BDD surface function to help the direct electron transfer of Cyt c with favourable biocompatibility and good bio-electrocatalytic affinity. A promising biosensor for hydrogen peroxide has thus been provided.

Synthesis and Ag-content-depended electrochemical properties of Ag/ZnO heterostructured nanomaterials

Ag/ZnO core/shell heterostructured nanomaterials were for the first time synthesized by Ag nanoparticle-mediated method, using which a novel hydrogen peroxide biosensor could be constructed. TEM and XRD measurements revealed that the Ag/ZnO heterojunctions were successfully acquired. Moreover, their sizes and shapes depended on Ag nanoparticles concentrations (i.e., Ag/ZnO-1, Ag/ZnO-2, and Ag/ZnO-3 heterojunctions were prepared using one-, two-, and three-fold Ag nanoparticles, respectively). Electrochemical measurements indicated that electrochemical properties of the Ag/ZnO-x-based biosensors also associated with the Ag nanoparticle content and the Hb–Nafion–Ag/ZnO-2 biosensor had the best electrochemical property, strongest redox peak currents and smallest ΔEp of about 40mV. Moreover, the Ag/ZnO-2-based biosensor displayed excellent electrocatalytic properties such as wide linear range (2–540μM), high sensitivity (83mAcm−2mol−1), low detection limit (0.18μM), and good stability and reproducibility.

Fabrication of hydrogen peroxide biosensor based on Ni doped SnO2 nanoparticles

Ni doped SnO2 nanoparticles (0–5wt%) have been prepared by a simple microwave irradiation (2.45GHz) method. Powder X-ray diffraction (XRD) and transmission electron microscopy (TEM) studies confirmed the formation of rutile structure with space group (P42/mnm) and nanocrystalline nature of the products with spherical morphology. Direct electrochemistry of horseradish peroxidase (HRP)/nano-SnO2 composite has been studied. The immobilized enzyme retained its bioactivity, exhibited a surface confined, reversible one-proton and one-electron transfer reaction, and had good stability, activity and a fast heterogeneous electron transfer rate. A significant enzyme loading (3.374×10−10molcm−2) has been obtained on nano-Ni doped SnO2 as compared to the bare glassy carbon (GC) and nano-SnO2 modified surfaces. This HRP/nano-Ni–SnO2 film has been used for sensitive detection of H2O2 by differential pulse voltammetry (DPV), which exhibited a wider linearity range from 1.0×10−7 to 3.0×10−4M (R=0.9897) with a detection limit of 43nM. The apparent Michaelis–Menten constant (KMapp) of HRP on the nano-Ni–SnO2 was estimated as 0.221mM. This excellent performance of the fabricated biosensor is attributed to large surface-to-volume ratio and Ni doping into SnO2 which facilitate the direct electron transfer between the redox enzyme and the surface of electrode.

Fabrication of a novel hydrogen peroxide biosensor based on Au-(PEO106PPO70PEO106) hairy nanospheres

Abstract In this case, the Au-F127 (triblock copolymer PEO 106 PPO 70 PEO 106 , PEO is short for polyethylene oxide and PPO is the abbreviation of polypropene oxide) hybrid hairy nanospheres were obtained by the redox reaction using pluronic F127 and HAuCl 4 as precursor. A novel hemoglobin (Hb) biosensor was fabricated by immobilizing the Hb onto the Au-F127 film on the surface of glassy carbon electrode (GCE). The Au-F127 nanocomposites and Hb were characterized by transmission electron microscopy (TEM), polycrystalline electron diffraction ring pattern and ultra-violet visible spectra (UV–vis). UV–vis spectra suggested that Hb in the film could keep its native secondary structure. The performances of Hb/(Au-F127) on GCE were characterized with cyclic voltammetry (CV) and typical amperometric response ( i – t ) measurements. The immobilized Hb maintained its bioactivity and displayed an excellent electrochemical behavior with a formal potential of −339 mV. The resulting electrode showed an electrocatalytic activity to hydrogen peroxide (H 2 O 2 ). The linear response range of the H 2 O 2 biosensor was from 3.0 × 10 −7 to 5.7 × 10 −4 M with a low detection limit of 4.0 × 10 −8 M.

Hydrogen Peroxide Biosensor Based on Enzymatic Modification of Electrode Using Deposited Silver Nano Layer

Hydrogen Peroxide Biosensor Based on Enzymatic Modification of Electrode Using Deposited Silver Nano Layer

A novel amperometric hydrogen peroxide biosensor based on pyrrole-PAMAM dendrimer modified gold electrode

Horseradish peroxidase (HRP) was immobilized into an electrochemically prepared copolymer of pyrrole–PAMAM (PAMAM; polyamidoamine) dendrimers for the construction of amperometric hydrogen peroxide biosensor. First, second, and third generation amidoamine–pyrrole dendrons having branched amine periphery and focal pyrrole functionality were synthesized via divergent pathway. Pyrrole dendrimers were covalently attached onto the electrode surface and polymerized by electrochemical copolymerization with pyrrole monomer. The synthesized dendrimers and copolymers have been characterized by FTIR-ATR and NMR. These copolymers have been utilized as conducting films for amperometric hydrogen peroxide sensing. The HRP retains its bioactivity after immobilization into the dendronized pyrrole-copolymers. Amperometric response was measured as a function of concentration of hydrogen peroxide, at fixed potential of +0.35V vs. Ag/AgCl in a phosphate buffered saline (pH 7.5). The effect of pH and temperature of the medium, storage, and reusability properties were investigated. The results indicate an efficient immobilization of enzyme onto the PAMAM type dendrimer modified surface containing pyrrole monomer, which leads to high enzyme loading, and increased lifetime stability of the electrode.