Peroxide Biosensor Research Articles

A non-enzymatic hydrogen peroxide biosensor based on cerium metal-organic frameworks, hemin, and graphene oxide composite

This study presents the development of a novel non-enzymatic electrochemical biosensor for the real-time detection of hydrogen peroxide (H2O2) based on a composite of cerium metal–organic frameworks (Ce-MOFs), hemin, and graphene oxide (GO). The Ce-MOFs served as an efficient matrix for hemin encapsulation, while GO enhanced the conductivity of the composite. Characterization techniques including scanning electron microscopy (SEM), X-ray diffraction (XRD), Fourier-transform infrared (FTIR) spectroscopy, UV–vis spectroscopy, and thermogravimetric analysis (TGA) confirmed the successful integration of hemin into the Ce-MOFs. The Ce-MOFs@hemin/GO-modified sensor demonstrated sensitive H2O2 detection due to the exceptional electrocatalytic activity of Ce-MOFs@hemin and the high conductivity of GO. This biosensor exhibited a linear response to H2O2 concentrations from 0.05 to 10 mM with a limit of detection (LOD) of 9.3 μM. The capability of the biosensor to detect H2O2 released from human prostate carcinoma cells was demonstrated, highlighting its potential for real-time monitoring of cellular oxidative stress in complex biological environments. To further assess its practical applicability, the sensor was tested in human serum samples, yielding promising results with recovery values ranging from 94.50 % to 103.29 %. In addition, the sensor showed excellent selectivity against common interfering compounds due to the outstanding peroxidase-like activity of the composite.

A Non-Disposable Electrochemical Sensor Based on Laser-Synthesized Pd/LIG Nanocomposite-Modified Screen-Printed Electrodes for the Detection of H2O2.

There have been many studies on the significant correlation between the hydrogen peroxide content of different tissues or cells in the human body and the risk of disease, so the preparation of biosensors for detecting hydrogen peroxide concentration has been a hot topic for researchers. In this paper, palladium nanoparticles (PdNPs) and laser-induced graphene (LIG) were prepared by liquid-phase pulsed laser ablation and laser-induced technology, respectively. The complexes were prepared by stirring and used for the modification of screen-printed electrodes to develop a non-enzymatic hydrogen peroxide biosensor that is low cost and mass preparable. The PdNPs prepared with anhydrous ethanol as a solvent have a uniform particle size distribution. The LIG prepared by laser direct writing has good electrical conductivity, and its loose porous structure provides more adsorption sites. The electrochemical properties of the modified electrode were characterized by cyclic voltammetry, chronoamperometry, and electrochemical impedance spectroscopy. Compared with bare screen-printed electrodes, the modified electrodes are more sensitive for the detection of hydrogen peroxide. The sensor has a linear response range of 5 µM-0.9 mM and 0.9 mM-5 mM. The limit of detection is 0.37 µM. The above conclusions indicate that the hydrogen peroxide electrochemical biosensor prepared in this paper has great advantages and potential in electrochemical catalysis.

Ni/WS2/WC Composite Nanosheets as an Efficient Catalyst for Photoelectrochemical Hydrogen Peroxide Sensing and Hydrogen Evolution.

It is highly attractive to develop a photoelectrochemical (PEC) sensing platform based on a non-noble-metal nano array architecture. In this paper, a PEC hydrogen peroxide (H2O2) biosensor based on Ni/WS2/WC heterostructures was synthesized by a facile hydrothermal synthesis method and melamine carbonization process. The morphology, structural and composition and light absorption properties of the Ni/WS2/WC catalyst were investigated by scanning electron microscopy (SEM), X-ray diffraction (XRD), X-ray photoelectron spectroscopy (XPS) and UV-visible spectrophotometer. The average size of the Ni/WS2/WC nanosheets was about 200 nm. Additionally, the electrochemical properties toward H2O2 were studied using an electrochemical workstation. Benefiting from the Ni and C atoms, the optimized Ni/WS2/WC catalyst showed superior H2O2 sensing performance and a large photocurrent response. It was found that the detection sensitivity of the Ni/WS2/WC catalyst was 25.7 μA/cm2/mM, and the detection limit was 0.3 mmol/L in the linear range of 1-10 mM. Simultaneously, the synthesized Ni/WS2/WC electrode displayed excellent electrocatalytic properties in hydrogen evolution reaction (HER), with a relatively small overpotential of 126 mV at 10 mA/cm2 in 0.5 M H2SO4. This novel Ni/WS2/WC electrode may provide new insights into preparing other efficient hybrid photoelectrodes for PEC applications.

A new strategy to build electrochemical enzymatic biosensors using a nanohybrid material based on carbon nanotubes and a rationally designed schiff base containing boronic acid

We report a nanohybrid material obtained by non-covalent functionalization of multi-walled carbon nanotubes (MWCNTs) with the new ligand (((1E,1′E)-(naphthalene-2,3-diylbis(azaneylylidene))bis(methaneylylidenedene)) bis(4-hydroxy-3,1-phenylene))diboronic acid (SB-dBA), rationally designed to mimic some recognition properties of biomolecules like concanavalin A, for the development of electrochemical biosensors based on the use of glycobiomolecules as biorecognition element. We present, as a proof-of-concept, a hydrogen peroxide biosensor obtained by anchoring horseradish peroxidase (HRP) at a glassy carbon electrode (GCE) modified with the nanohybrid prepared by sonication of 2.0 mg mL−1 MWCNTs and 0.50 mg mL−1 SB-dBA in N,N-dimethyl formamide (DMF) for 30 min. The hydrogen peroxide biosensing was performed at −0.050 V in the presence of 5.0 × 10−4 M hydroquinone. The analytical characteristics of the resulting biosensor are the following: linear range between 0.175 μM and 6.12 μM, detection limit of 58 nM, and reproducibility of 2.0 % using the same nanohybrid (6 biosensors), and 9.0 % using three different nanohybrids. The sensor was successfully used to quantify hydrogen peroxide in enriched milk and human blood serum samples and in a commercial disinfector.

A catalase enzyme biosensor for hydrogen peroxide at a poly(safranine T)-ternary deep eutectic solvent and carbon nanotube modified electrode

A new electrochemical enzyme biosensor has been developed to monitor hydrogen peroxide. The sensor uses a multiwalled carbon nanotube modified glassy carbon electrode covered by a poly(safranine T) polymer film prepared by potential cycling electropolymerization in ternary deep eutectic solvent (DES), prepared from choline chloride (ChCl), ethylene glycol (EG) plus malonic acid (MA) or lactic acid (LA) with composition optimization. It is shown that this leads to greater polymer growth and improved film properties compared to binary DES; the process was monitored by an electrochemical quartz crystal microbalance. The best ternary DES composition was found to be 16 % ChCl:MA / 84 % ChCl:EG. Electrochemical characterisation was done by cyclic voltammetry and electrochemical impedance spectroscopy, and the morphology of the synthesized polymer films was examined by scanning electron microscopy. Catalase enzyme was immobilized on the modified electrode and the enzyme biosensor used for measurement of hydrogen peroxide, with a very low detection limit of 34 nM. The selectivity was found to be excellent in relation to common interferents and the biosensor showed very good recovery in peroxide measurements in commercial samples.

Interfacing Silver Nanoparticles with Hematene Nanosheets for the Electrochemical Sensing of Hydrogen Peroxide

Background: Hydrogen peroxide (H2O2) is an important metabolite that plays a crucial role in enzymatic reactions in living organisms. However, it acts as a reactive oxygen species (ROS) that causes various chronic diseases. The main challenging aspects in detecting H2O2 in body cells are the ultra-lowlevel concentrations and its reactivity. Hence, it is highly essential to develop a platform for H2O2 with high sensitivity and selectivity. Objective: In this work, we report an electrochemical biosensor for hydrogen peroxide (H2O2) by interfacing 3-dimensional silver nanoparticles (Ag-NPs) with 2-dimensional hematene (HMT) nanosheets. Methods: The two-dimensional nanomaterial, HMT, was exfoliated from natural iron ore hematite (α- Fe2O3) and characterized by Raman spectroscopy. The morphology of the Ag nanoparticles and HMT was imaged by scanning electron microscope. Electrochemical characterization of Ag/HMT modified glassy carbon electrode (GCE) was performed by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). Results: The fabricated sensor showed a wide linearity range of H2O2 concentrations from 0.99 μM to 1110 μM and a low detection limit of 0.16 μM using CV. Further, the sensor was successfully applied for the electrochemical sensing of hydrogen peroxide using chronoamperometry (CA) from 20 μM to 1110 μM (LOD 5.5 μM). Conclusion: The proposed electrochemical sensor for H2O2 is fast responding with a high sensitivity, and shows selectivity in the presence of biologically important molecules. These consequences suggested that the formation of heterostructures between 2D and 3D nanomaterials unveils the possibility of stable and selective electrochemical sensors for bioanalytics.

Hydrogen Peroxide (H2O2) Biosensor Based on the Conducting Polymer Using Self-Assembly Technique

Biosensors are in principle fabricated by immobilized biomaterials on a detector membrane and combining them with electrochemical equipment. The applications of enzyme-based biosensors can be explored such as in the process of gas detection, medicine, pathogen detection and detection of toxic levels of substances before and after bioremediation. In this study, H2O2 detection was performed using HRP/PANI, HRP/PPY, HRP/PT and HRP/PT/PPY/PANI layers. The HRP/PANI layer from Variable Pressure Scanning Electron Microscopy (VPSEM) image exhibited a dry surface. The HRP/PPY layer exhibited a surface with agglomerate molecules. The HRP/PT layer, on the hand, exhibited a layer surface with almost the same molecular size. This is confirmed by the higher surface roughness value for HRP/PPY compared to other layers obtained via characterization with Atomic Force Microscopy (AFM). The increasing current response for all three layers was arranged in HRP/PANI> HRP/PT> HRP/PPY. VPSEM and AFM images exhibited surfaces with molecules being in an agglomeration state after the H2O2 detection process. In terms of current response, the response rate of H2O2on the surface of the HRP/PT/PPY/PANI electrode caused the current response obtained to be fast. The roughness value increased with time due to the reaction that took place between the surface of the HRP/PT/PPY/PANI layer with H2O2. The day-based current response showed that day 1 to day 14 exhibited a uniform graph pattern but from day 21 to day 30 there was a change in the graph pattern due to the HRP/PT/PPY/PANI layer undergoing degradation. The activity of the HRP enzyme was studied by looking at its absorption effect for 30 days. From day 1 to day 14, there was a difference in the overall rate of absorption. However, from day 21 to day 30, the rate of absorption remained constant which explains the slowing down of HRP activity.

Hydrogen peroxide biosensors with novel photophysical properties

Hydrogen peroxide biosensors with novel photophysical properties

Evaluation of soybean peroxidase - Copper phosphate mediated organic-inorganic hybrid for hydrogen peroxide biosensor application

An electrochemical biosensor for H2O2 detection was developed by using soybean peroxidase-copper phosphate mediated organic inorganic hybrid (OIH). The characterization of OIH was carried out by FESEM and FTIR. FESEM analysis showed a flower-like porous morphology of the OIH and FTIR confirmed the presence of soybean peroxidase and copper phosphate in the OIH. For sensor development, the paste of OIH was formulated in the presence of phosphate buffered saline (PBS) and was screen printed on indium tin oxide (ITO) coated glass substrate. Cyclic voltammetry analysis showed that the developed biosensor could detect H2O2 in the linear range of 20–100 ​μM with R2 value of 0.963. The limit of detection (LOD) and sensitivity values calculated for H2O2 are 0.19 ​μM and 27.44 μA/(μM.cm2) respectively. Along with cyclic voltammetry experiments, electrochemical impedance spectroscopy (EIS) analysis was also carried out to study the sensing mechanism. The developed biosensor showed better selectivity towards H2O2 when tested against d-glucose, l-tyrosine, l-lysine, and l-ascorbic acid.

A whole-cell hydrogen peroxide biosensor and its application in visual food analysis

<p>Hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>) is broadly used in the food industry for bleaching, sterilization, and deodorization. Detection of H<sub>2</sub>O<sub>2</sub> in food/drinks is important for food safety. In this study, the H<sub>2</sub>O<sub>2</sub>-inducible whole-cell biosensor KT2440 [p<i>PahpC</i>] was constructed based on the bacterial strain <i>Pseudomonas putida</i> KT2440. The H<sub>2</sub>O<sub>2</sub>-inducible promoter <i>PahpC</i> was fused with the reporter gene cluster <i>luxCDABE</i> to produce an H<sub>2</sub>O<sub>2</sub>-inducible bioluminescent biosensor. KT2440 [p<i>PahpC</i>] semi-quantitatively detected H<sub>2</sub>O<sub>2</sub> in the range of 10 - 2000 ?M. This H<sub>2</sub>O<sub>2</sub> biosensor exhibited high specificity and no response to other commonly used redox agents, such as KMnO<sub>4</sub>, Ca(ClO)<sub>2</sub>, and thiourea. This KT2440 [p<i>PahpC</i>] biosensor was used to detect H<sub>2</sub>O<sub>2</sub> in food samples, demonstrating its robust performance. The whole-cell biosensor provides a new approach to the detection of H<sub>2</sub>O<sub>2</sub> in the food industry.</p>

Upconversion luminescent sensor for endogenous H2O2 detection in cells based on the inner filter effect of coated silver layer

We design and construct an upconversion luminescence sensor for hydrogen peroxide detection based on lanthanide-doped nanoparticles (LDN-OA). After modification with an electron-donating ligand, the silver was coated on the surface of LDN-OA in-situ by photo-induced free-electron reduction under the excitation of ultraviolet light. Then, the core-shell structure of upconversion nanoparticles growing with the silver metal surface is formed and modified with polyethylene-glycol, leading to the final hydrogen peroxide biosensor (denoted as LDN@Ag-PEG). The sensor is constructed based on the luminescence inner filter effect, using the LDN-OA as an energy donor and the coated silver layer as an energy acceptor. The confocal images were analyzed in vitro as well, presenting that the LDN@Ag-PEG sensor could achieve the sensing of endogenous hydrogen peroxide under 980 nm excitation. Thus the upconversion luminescence sensor provides a powerful idea for the development of intracellular biomolecule sensors upon near-infrared light irradiation as well.

A sensitive hydrogen peroxide biosensor based on a new electron mediator 1-aminoethoxy-5-ethylphenazine dioctyl sulfosuccinate

Hydrogen peroxide (H2O2) plays a vital role in the fields of pharmaceutical production and biomedicine. The level of H2O2 in cells directly affects the growth and apoptosis of cells, so the development of H2O2 detection technology is very important. In this paper, a new type of electron mediator, 1-aminoethoxy-5-ethylphenazine dioctyl sulfosuccinate (aePEDS) was designed and synthesized and a biosensor based on a glass carbon (GC)/aePEDS/ horseradish peroxidase (HRP) modified electrode for the detection of H2O2 was developed. H2O2 was detected by the biosensor at a low working potential of 0.11 V (vs. Ag/AgCl), avoiding interference from other electroactive substances. Under optimal conditions, the limit of detection was 0.2 μM and the sensitivity was 0.001 μA/mM. The developed sensor was used to evaluate the activity of antitumor drugs by detecting the level of hydrogen peroxide in the cell culture medium after drug stimulation of lung cancer cells. Compared with the hydrogen peroxide kit method, the constructed electrochemical biosensor can more sensitively and accurately reflect the impact of the change of hydrogen peroxide content on cells after drug stimulation and is a more promising method for cell detection and screening of drug activity.

Photolithographic 3D microarray electrode-based high-performance non-enzymatic H2O2 sensor

In recent years, electrochemical biosensors have been widely used in health care, and, due to the increasing demand for their use, new sensing performance requirements have been proposed. Widening detection range and speeding up response rate while improving the sensitivity of these sensors is a major current challenge. In this study, a high-performance three-dimensional (3D) non-enzymatic hydrogen peroxide biosensor was fabricated based on a simple photolithographic process. Neatly arranged cylindrical microarrays (MA) with clear edges were fabricated using SU-8 photoresist. Subsequently, a gold thin film was sputtered onto the MA to form a 3D Au/microarray electrode (Au/MAE). Additionally, we investigated the optimal size of the MA (aspect ratio = 2, diameter = 30 µm) required to obtain an Au/MAE with the best electron transfer properties. Based on these results, a 3D CuNPs/Au/MAE sensor was constructed via the electrochemical in-situ reduction of copper nanoparticles (CuNPs). Due to its increased specific surface area and contact sites, this 3D sensor with high stability and good selectivity for detecting hydrogen peroxide exhibited higher sensitivity and wider range of detection than 2D CuNPs/AuE sensor. Consequently, the design of 3D microarray structure provides a new strategy for the fabrication of high-performance sensors that can be used in clinical applications.

Bioelectrochemistry in Undergraduate Curriculum and Research

Electrochemistry has a wide range of uses and applications, but undergraduate students do not appreciate this when the topic is first introduced in General Chemistry. Introducing more electrochemical methods in later chemistry courses can encourage interest in the field. In the chemistry department at Lebanon Valley College, the students first encounter electrochemistry through the typical introduction of redox reactions, the Nernst equation, and galvanic cells. The topic is expanded in Analytical Chemistry for third years during which the students learn about and perform electroanalytical methods, including potentiometric analysis, coulometry, amperometry, and voltammetry. Students who choose the advanced elective Chemical Sensors and Biosensors learn about electrochemical sensors as one unit of the course. This unit also includes a section on bioelectrochemical sensors. Students develop a hydrogen peroxide biosensor and use amperometry to evaluate its performance, giving them a better understanding of concepts like limit of detection, sensitivity, and dynamic linear range.In addition to the chemistry curriculum, students can choose to do electrochemical research in the department. Recent projects include fabricating and evaluating enzyme electrodes for a wide range of biologically relevant molecules, designing self-powered biosensors, and designing a biophotovoltaic device for solar energy conversion with plants. We also currently have a research project with the focus of developing an electrosynthesis experiment to be added to the organic lab sequence. Throughout these research projects, students gain experience with several electrochemical methods, analysis of electrochemical data, and general lab skills. They are also required to present their work, either in posters or oral presentations, a skill that will prepare them for their careers after college. Overall this combination of curriculum and research leads to more students being interested in electrochemistry. They learn that these methods can be used for every area of chemistry, not just analytical. It has lead to an increase in the number of graduating students who continue working in the field, whether in industry or academia.

Durable Hydrogen Peroxide Biosensors Based on Polypyrrole-Decorated Platinum/Palladium Bimetallic Nanoparticles

Durable Hydrogen Peroxide Biosensors Based on Polypyrrole-Decorated Platinum/Palladium Bimetallic Nanoparticles

Highly Sensitive Hydrogen Peroxide Biosensor Based on Tobacco Peroxidase Immobilized on p‐Phenylenediamine Diazonium Cation Grafted Carbon Nanotubes: Preventing Fenton‐like Inactivation at Negative Potential

AbstractHerein, we present a novel electrode platform for H2O2 detection based on the immobilization of recombinant Tobacco Peroxidase (r‐TOP) onto graphite electrodes (G) modified with p‐phenylenediamine (p‐PD) diazonium cation grafted multi‐walled carbon nanotubes (MWCNTs). The employment of both p‐phenylenediamine moieties and covalent cross‐linking by using glutaraldehyde allowed us to enhance the sensitivity, stability, and selectivity toward H2O2 detection, as well as preventing enzyme inactivation due to the electro‐Fenton reaction. This reaction continuously produces hydroxyl radicals, whose high and unselective reactivity is likely to reduce drastically the operating life of the biosensor. The protection against the electro‐Fenton reaction is mainly ascribed to a beneficial enzyme immobilization leading to a correct orientation achieved through cross‐linking the enzyme in combination with interaction between the uncoupled ‐NH2 groups (mainly uncharged at pH 7, considering a pKa of 4.6) available on the electrode surface and the enzyme. In particular, the electrode based on the r‐TOP/p‐PD/MWCNTs/G platform showed a lower limit of detection of 1.8 μM H2O2, an extended linear range between 6 and 900 μM H2O2, as well as a significant increase in sensitivity (63.1±0.1 μA mM−1 cm−2) compared with previous work based on TOP. Finally, the r‐TOP/p‐PD/MWCNTs/G electrode was tested in several H2O2 spiked food samples as a screening analytical method for the detection of H2O2.

Validation and Verification of Nanopolymer Based Hydrogen Peroxide Biosensor

Validation and Verification of Nanopolymer Based Hydrogen Peroxide Biosensor

Fully Inkjet-Printed Biosensors Fabricated with a Highly Stable Ink Based on Carbon Nanotubes and Enzyme-Functionalized Nanoparticles.

Enzyme inks can be inkjet printed to fabricate enzymatic biosensors. However, inks containing enzymes present a low shelf life because enzymes in suspension rapidly lose their catalytic activity. Other major problems of printing these inks are the non-specific adsorption of enzymes onto the chamber walls and stability loss during printing as a result of thermal and/or mechanical stress. It is well known that the catalytic activity can be preserved for significantly longer periods of time and to harsher operational conditions when enzymes are immobilized onto adequate surfaces. Therefore, in this work, horseradish peroxidase was covalently immobilized onto silica nanoparticles. Then, the nanoparticles were mixed into an aqueous ink containing single walled carbon nanotubes. Electrodes printed with this specially formulated ink were characterized, and enzyme electrodes were printed. To test the performance of the enzyme electrodes, a complete amperometric hydrogen peroxide biosensor was fabricated by inkjet printing. The electrochemical response of the printed electrodes was evaluated by cyclic voltammetry in solutions containing redox species, such as hexacyanoferrate (III/II) ions or hydroquinone. The response of the enzyme electrodes was studied for the amperometric determination of hydrogen peroxide. Three months after the ink preparation, the printed enzyme electrodes were found to still exhibit similar sensitivity, demonstrating that catalytic activity is preserved in the proposed ink. Thus, enzyme electrodes can be successfully printed employing highly stable formulation using nanoparticles as carriers.

Franz cells for facile biosensor evaluation: A case of HRP/SWCNT-based hydrogen peroxide detection via amperometric and wireless modes

Reducing animal use in biosensor research requires broader use of in vitro methods. In this work, we present a novel application of Franz cells suitable for biosensor development and evaluation in vitro. The work describes how Franz cell can be equipped with electrodes enabling characterization of biosensors in close proximity to skin. As an example of a sensor, hydrogen peroxide biosensor was prepared based on horseradish peroxidase (HRP)/single-walled carbon nanotube (SWCNT)-modified textile. The electrode exhibited lower detection limit of 0.3 μM and sensitivity of 184 μA mM−1 cm−2. The ability of this biosensor to monitor H2O2 penetration through skin and dialysis membranes was evaluated in Franz cell setup in amperometric and wireless modes. The results also show that catalase activity present in skin is a considerable problem for epidermal sensing of H2O2. This work highlights opportunities and obstacles that can be addressed by assessment of biosensors in Franz cell setup before progressing to their testing in animals and humans.

Non-enzymatic colorimetric biosensor for hydrogen peroxide using lignin-based silver nanoparticles tuned with ionic liquid as a peroxidase mimic

Hydrogen peroxide (H2O2) is a byproduct of oxidative metabolism, acts as a signaling molecule, and can induce protein and DNA damage by oxidative stress. Ionic liquid-coated lignin stabilized silver nanoparticles (LAgNPs) have been used for the colorimetric detection of H2O2. Both ionic liquid and lignin moieties have a synergistic effect on the conductivity of silver nanoparticles to display intrinsic peroxidase-like activity with the enhanced potential to catalyze the oxidation of substrate 3,3′,5,5′-tetramethylbenzidine (TMB). LAgNPs have been synthesized by the green synthesis method and the prepared nanoparticles have been characterized by different techniques such as FTIR, EDX, TGA, SEM, and XRD. 1-H-3-methylimidazolium acetate (ionic liquid) was prepared and used for tuning of AgNPs. By using ionic liquid capped LAgNPs the colorimetric detection of H2O2 was done with the assistance of TMB solution and results were analyzed by UV–Vis spectrophotometer. Different parameters such as (amount of capping agent and TMB, pH, H2O2 concentration, and time) were optimized to get the best results of the proposed sensor. The H2O2 biosensor exhibited a linear response in the range of 1 × 10−9–3.6 × 10−7 M, with a detection limit of 1.379 × 10−8 M and a quantification limit of 4.59 × 10−8 M, and R2 of 0.999. The sensor gave a short response time of 5 min for colorimetric detection of H2O2 at pH 7.5 and room temperature. For the detection of hydrogen peroxide, the proposed sensor showed good sensitivity and selectivity and was successfully employed for the detection of H2O2 in the blood serum samples of hypertension patients.