Analytical Characteristics of Electrochemical Biosensor Using Pt‐Dispersed Graphene on Boron Doped Diamond Electrode

An electrochemical biosensor was developed using Pt‐nanoparticles (Pt‐NPs) dispersed graphene based on a boron‐doped diamond thin film electrode. To compare its performances with those of other biosensors, glucose was used as a target analyte. This biosensor exhibited a wide linear range, a low detection limit and a higher sensitivity compared to other amperometric biosensors using graphene‐based electrodes. In addition, the biosensor promotes a direct electron transfer between the redox enzymes and the electrode surface and detects low concentration analytes. The excellent performance of the biosensor is attributed to the synergistic effect of the Pt‐NPs, graphene sheet and the BDD thin film. Therefore, it can be a promising application for electrochemical detection of analytes.     

Interaction between co-deposited metals during stripping voltammetry at boron-doped diamond electrodes

Electrochemical processes, which underlie the use of conductive diamond electrodes for the simultaneous detection of two or more metal ions in solution by anodic stripping voltammetry (ASV), have been investigated. The model analyte system studied contains the two metal species, Ag+(aq) and Pb2+(aq), and the experimental techniques employed include cyclic and square wave voltammetries, along with X-ray photoelectron spectroscopy and secondary electron microscopy. Although the bulk metallic forms of Ag and Pb are immiscible, several interactions in the system between the two metal species present are observed, which significantly influence the electrodeposition and electrodissolution processes which underlie ASV. The subsequent nucleation and growth of a given metal on the electrode surface is enhanced by the presence of the second metal on the surface. The encapsulation of one metal by the other, within the metal particulates that form on the electrode surface, significantly reduces the stripping yield at the potentials characteristic of the individual metals. The stripping potentials are also influenced by bonding interactions between deposited Ag and Pb, which broaden the characteristic stripping peaks in cyclic voltammetry, as well as producing underpotential deposition and stripping. Given these interactions, the extent to which ASV at diamond electrodes can be used to determine the solution concentrations of Ag+(aq) and Pb2+(aq) is considered.     

Multi-walled Carbon Nanotubes and Gold Nanorod Decorated Biosensor for Detection of microRNA-126

In this article, we introduced a novel electrochemical biosensor for the detection of microRNA-126. The biosensor utilizes a hybridization assay combined with multi-walled carbon nanotubes and gold nanorod-decorated screen-printed carbon electrodes. For electrode preparation, gold nanorods were first immobilized onto the surface of bare and multi-walled carbon nanotube-modified screen-printed carbon electrodes, and the thiol tagged-capture probe was immobilized on the electrode surface through gold and thiol group interaction. After the immobilization, thiol tagged-capture probe hybridized with the target sequence. Under optimum conditions, we determined limit of detection (LOD) and limit of quantification (LOQ) as high as 11 nM and 36 nM, respectively.     

Pt-polyaniline nanocomposite on boron-doped diamond electrode for amperometic biosensor with low detection limit

Boron-doped diamond electrodes covered with a nanostructured Pt nanoparticle-polyaniline composite have been fabricated and employed as sensitive amperometric sensors with low detection limit. A highly conductive boron-doped diamond thin film (BDD) was prepared by chemical vapor deposition, and its morphology was characterized by scanning electron microscopy and transmission electron microscopy. The nanostructured composite layer was grown on the BDD electrode by electrochemical deposition of polyaniline and Pt nanoparticles. Glucose oxidase (GOx) was then adsorptively immobilized on the modified BDD electrode. The biosensor displays a large surface area, high catalytic activity of the Pt nanoparticles, efficient electron mediation through the conducting polymer, and low background current of the electrode. The biosensor exhibits an excellent response to glucose, with a broad linear range from 5.9 μM to 0.51 mM, a sensitivity of 5.5 μA·mM^?1, a correlation coefficient (R) of 0.9947, and a detection limit of 0.10 μM. The apparent Michaelis-Menten constant (K _M ^app ) and the maximum current density of the electrode are 4.1 mM and 0.021 mA, respectively. This suggests that the immobilized GOx possesses a higher affinity for glucose at the lower K _M ^app , and that the enzymatic reaction rate constitutes the rate-limiting step of the response.     

Monitoring metal ion contamination onset in hydrofluoric acid using silicon-diamond and dual silicon sensing electrode assembly.

Potentiometric detection of trace levels of metallic contamination onset in hydrofluoric acid using a silicon-based sensor in conjunction with two non-contaminating reference electrode systems is presented in this paper. In the first case, conductive diamond was used as a non-contaminating reference electrode. Cyclic voltammetry and open-circuit potential experiments demonstrated the feasibility of using a conductive diamond film electrode as a quasi-reference electrode in the HF solution. In the second case, a dual silicon electrode system was used with one of the silicon-based electrodes protected with an anion permeable membrane behaving as the quasi-reference electrode. The dual silicon sensing electrode system possessed an additional operational advantage of being unaffected by the solution acidity. Though both sensing configurations were able to detect the metal ion contamination onsets at the parts-per-trillion to parts-per-billion levels, the dual silicon electrode design showed a greater compatibility for the on-line detection of metallic impurities in HF etching baths commonly used in semiconductor processing.     

Diamond Electrodes for High Sensitivity Mercury Detection in the Aquatic Environment: Influence of Surface Preparation and Gold Nanoparticle Activity

This work compares polished and unpolished boron doped diamond (BDD) electrodes decorated with two sizes of gold nanoparticles (AuNPs) for use as robust mercury sensors in aquatic environments. The size of the catalytically active AuNPs on the electrode surfaces was demonstrated to have a less significant effect on the sensitivity for mercury detection than the surface preparation of the BDD. The lowest limits of detection were achieved with the polished BDD electrodes, which both detected mercury at a concentration of 1 pM, six orders of magnitude greater sensitivity than the lowest detection limit of 5 μM achieved with an unpolished BDD electrode, and high in comparison with other reported electrode systems.     

Edge Plane Pyrolytic Graphite Electrodes for Stripping Voltammetry: a Comparison with Other Carbon Based Electrodes

The first examples of using edge plane pyrolytic graphite electrodes for anodic and cathodic stripping voltammetry (ASV and CSV) are presented, notably the ASV of silver and the CSV of manganese. In the former example, detection limits for silver (based on 3σ) of 8.1 nM and 0.185 nM for 120 s and 300 s accumulation time, respectively, were achievable using the edge plane electrode, which were superior to those observed on glassy carbon, basal plane pyrolytic graphite and boron‐doped diamond electrodes. In the second example, a detection limit for manganese of 0.3 μM was possible which was comparable with that achievable with a boron‐doped diamond electrode but with an increased sensitivity. Comparison of the edge plane pyrolytic graphite electrode with boron‐doped diamond electrodes reveals that the edge plane electrode has comparable detection limits and sensitivities whilst exhibiting a lower signal‐to‐noise ratio and large potential window for use in trace analysis suggesting boron‐doped diamond can be conveniently replaced by edge plane pyrolytic graphite as an electrode material in many applications.     

Fabrication of Pt nanoparticles-decorated CVD diamond electrode for biosensor applications

An electrochemical biosensor was developed using boron-doped diamond (BDD) as an electrode material. To enhance the electrical performance of the electrode, the BDD electrode was decorated with Pt-nanoparticles (Pt-NPs) by electrochemical deposition. Their morphology according to the applied potentials for the synthesis of Pt-NPs was characterized by SEM. To identify the performance of the electrode modified with Pt-NPs, glucose detection was used as a sample sensing process, and the results were compared with those of a gold electrode and a bare BDD electrode. The electrochemical characteristics of the modified electrode were examined by cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The BDD electrode with the Pt-NPs showed higher sensitivity and a lower detection limit than the Au electrode and BDD electrode. The proposed biosensor based on the Pt-NPs decorated BDD electrode showed high sensitivity, a low detection limit, fast direct electron transfer and good stability.     

Mediator-free amperometric determination of glucose based on direct electron transfer between glucose oxidase and an oxidized boron-doped diamond electrode

A mediator-free glucose biosensor, termed a “third-generation biosensor,” was fabricated by immobilizing glucose oxidase (GOD) directly onto an oxidized boron-doped diamond (BDD) electrode. The surface of the oxidized BDD electrode possesses carboxyl groups (as shown by Raman spectra) which covalently cross-link with GOD through glutaraldehyde. Glucose was determined in the absence of a mediator used to transfer electrons between the electrode and enzyme. O₂ has no effect on the electron transfer. The effects of experimental variables (applied potential, pH and cross-link time) were investigated in order to optimize the analytical performance of the amperometric detection method. The resulting biosensor exhibited fast amperometric response (less than 5 s) to glucose. The biosensor provided a linear response to glucose over the range 6.67×10⁻⁵ to 2×10⁻³ mol/L, with a detection limit of 2.31×10⁻⁵ mol/L. The lifetime, reproducibility and measurement repeatability were evaluated and satisfactory results were obtained.     

Microfluidic based impedance biosensor for pathogens detection in food products

A MEMS‐based impedance biosensor was designed, fabricated, and tested to effectively detect the presence of bacterial cells including E. coli O157:H7 and Salmonella typhimurium in raw chicken products using detection region made of multiple interdigitated electrode arrays. A positive dielectrophoresis based focusing electrode was used in order to focus and concentrate the bacterial cells at the centerline of the fluidic microchannel and direct them toward the detection microchannel. The biosensor was fabricated using surface micromachining technology on a glass substrate. The results demonstrate that the device can detect Salmonella with concentrations as low as 10 cells/mL in less than 1 h. The device sensitivity was improved by the addition of the focusing electrodes, which increased the signal response by a factor between 6 and 18 times higher than without the use of the focusing electrodes. The biosensor is selective and can detect other types of pathogen by changing the type of the antibody immobilized on the detection electrodes. The device was able to differentiate live from dead bacteria.     

Synthetic boron-doped carbonado,a new electrode material: synthesis in C–metal–B growth systems aimed at the lowering of the synthesis temperature without loss of electrochemical activity

New electrode material—boron-doped synthetic carbonado (bulk polycrystalline diamond)—was synthesized at high pressures and high temperatures in the C–metal (Co, Ni, or Fe) –B growth systems. The metal borides were used as the growth medium-forming substances for graphite-to-diamond transformation at a temperature of ~?1300 °C and pressure of 8 GPa. For comparison, etalon carbonado-type electrode with nearly limiting concentration of boron in diamond was synthesized by subjecting the mixture of amorphous boron with graphite to much higher temperatures (2200–2500 °C) under the same pressure. Despite the lower content of boron in diamond synthesized in the presence of metal borides, these new boron-doped carbonado electrodes are not inferior to the etalon compact in their electrochemical activity, as judging by the onset potential of anodic chlorine evolution from KCl solution. The presence of metal-containing structural defects in boron-doped diamond matrix is supposed to be responsible for the somewhat enhanced catalytic activity of the electrodes. High-pressure synthesis of bulk metal-modified boron-doped diamond opens a new avenue in the development of superior functional electrode materials.     

Recent electroanalytical applications of boron-doped diamond electrodes

This review overviews recent reports on the electroanalytical applications of boron-doped diamond (BDD) electrodes. Because BDD electrodes have excellent features for electroanalysis, such as wide potential window, low background current, electrochemical stability, and fouling resistance, they can be useful for sensitive and stable detection of various substances, including drugs, bio-related substances, metal ions, and organic pollutants. Many articles have reported high-sensitivity detection of real samples, demonstrating that this electrode material is practically applicable. Surface modification of the BDD electrodes using metal nanoparticles, nanocarbons, and polymers can increase the sensitivity of the electrochemical detection. Furthermore, research on the electroanalytical device equipped with BDD electrodes will be expanded by combining peripheral technologies related to the device fabrication.     

检测痕量Hg~(2+)的DNA电化学生物传感器

通过自组装方法将修饰有二茂铁基团的富T序列DNA分子(DNA-Fc)固定在金电极表面,得到了一种基于DNA修饰电极的电化学汞离子(Hg2+)传感器.当溶液中有Hg2+存在时,Hg2+可与修饰电极上DNA的T碱基发生较强的特异结合,形成T-Hg2+-T发卡结构,使DNA分子构象发生改变,其末端具有电化学活性的二茂铁基团远离电极表面,电化学响应随之发生变化.示差脉冲伏安法(DPV)结果显示:DNA末端二茂铁基团的还原峰在0.26V(vs饱和甘汞电极(SCE))附近,峰电流随溶液中Hg2+浓度的增加而降低;Hg2+浓度范围在0.1nmol·L-1-1μmol·L-1时,电流相对变化率与Hg2+浓度的对数呈现良好的线性关系.该修饰电极对Hg2+的检测限为0.1nmol·L-1,可作为痕量Hg2+检测的电化学生物传感器.干扰实验也表明,该传感器对Hg2+具有良好的特异性与灵敏度.     

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.     

Detection of zeptomolar concentrations of alkaline phosphatase based on a tyrosinase and horse-radish peroxidase bienzyme biosensor

A bienzyme biosensor based on tyrosinase and horse-radish peroxidase is described in a flow injection analysis and cyclic voltammetry for measurement of phenol. Tyrosinase and horse-radish peroxidase were immobilized on the surface of a glassy carbon electrode by bovine serum albumin and glutaric dialdehyde. Phenol was oxidized by tyrosinase and horse-radish peroxidase via catechol to o-quinone in the presence of oxygen and hydrogen peroxide. The o-quinone was reduced to produce catechol (the substrate recycling) on the electrode surface. The enhanced sensitivity of the bienzyme electrode to phenol was observed in the flow injection system comparing with tyrosinase and horse-radish peroxidase monoenzyme electrodes. The mechanisms for enhanced amperometric response to phenol of bienzyme electrode were discussed. The biosensor was used to detect alkaline phosphatase (ALP). A detection limit of 1.4×10⁻¹⁵ M ALP (140 zmol/100 μl) was obtained after 1 h incubation with phenyl phosphate.     

Carbon nanotube-enhanced electrochemical DNA biosensor for DNA hybridization detection

A novel and sensitive electrochemical DNA biosensor based on multi-walled carbon nanotubes functionalized with a carboxylic acid group (MWNTs-COOH) for covalent DNA immobilization and enhanced hybridization detection is described. The MWNTs-COOH-modified glassy carbon electrode (GCE) was fabricated and oligonucleotides with the 5'-amino group were covalently bonded to the carboxyl group of carbon nanotubes. The hybridization reaction on the electrode was monitored by differential pulse voltammetry (DPV) analysis using an electroactive intercalator daunomycin as an indicator. Compared with previous DNA sensors with oligonucleotides directly incorporated on carbon electrodes, this carbon nanotube-based assay with its large surface area and good charge-transport characteristics dramatically increased DNA attachment quantity and complementary DNA detection sensitivity. This is the first application of carbon nanotubes to the fabrication of an electrochemical DNA biosensor with a favorable performance for the rapid detection of specific hybridization.     

Supramolecular immobilization of glucose oxidase on gold coated with cyclodextrin-modified cysteamine core PAMAM G-4 dendron/Pt nanoparticles for mediatorless biosensor design

Cysteamine core polyamidoamine G-4 dendron branched with β-cyclodextrins was chemisorbed on the surface of Au electrodes and further coated with Pt nanoparticles. Adamantane-modified glucose oxidase was subsequently immobilized on the nanostructured electrode surface by supramolecular association. This enzyme electrode was used to construct a reagentless amperometric biosensor for glucose, making use of the electrochemical oxidation of H₂O₂ generated in the enzyme reaction. The amperometric response of the biosensor was rapid (6 s) and a linear function of glucose concentration between 5 and 705 μmol?L^?1. The biosensor had a low detection limit of 2.0 μmol?L^?1, sensitivity of 197 mA?mol^?1?L?cm^?2, and retained 94 % of its initial response after storage for nine days at 4 °C.     

Dielectrophoretic cell trapping for improved surface plasmon resonance imaging sensing

The performance of conventional surface plasmon resonance (SPR) biosensors can be limited by the diffusion of the target analyte to the sensor surface. This work presents an SPR biosensor that incorporates an active mass‐transport mechanism based on dielectrophoresis and electroosmotic flow to enhance analyte transport to the sensor surface and reduce the time required for detection. Both these phenomena rely on the generation of AC electric fields that can be tailored by shaping the electrodes that also serve as the SPR sensing areas. Numerical simulations of electric field distribution and microparticle trajectories were performed to choose an optimal electrode design. The proposed design improves on previous work combining SPR with DEP by using face‐to‐face electrodes, rather than a planar interdigitated design. Two different top‐bottom electrode designs were experimentally tested to concentrate firstly latex beads and secondly biological cells onto the SPR sensing area. SPR measurements were then performed by varying the target concentrations. The electrohydrodynamic flow enabled efficient concentration of small objects (3 μm beads, yeasts) onto the SPR sensing area, which resulted in an order of magnitude increased SPR response. Negative dielectrophoresis was also used to concentrate HEK293 cells onto the metal electrodes surrounded by insulating areas, where the SPR response was improved by one order of magnitude.     

Boron-doped carbon nanotubes modified electrode for electroanalysis of NADH

Boron-doped carbon nanotubes (BCNTs) as a novel carbon nanomaterial have higher catalytic activity. Electroanalysis of dihydronicotinamide adenine dinucleotide (NADH) based on the BCNTs modified electrode has been investigated. Comparing with the bare glassy carbon (GC) and carbon nanotubes (CNTs)/GC electrodes, the BCNTs/GC electrode allowed highly sensitive amperometric detection of NADH at the lower applied potential, and minimization of surface contamination. Therefore, BCNTs are useful and promising material for the detection of NADH and are attractive for dehydrogenase-based amperometric biosensor or other analytical applications.     

基于多壁碳纳米管的三电极血乙醇生物传感器的研究

采用丝网印刷技术在PVC基板上印制三电极,将多壁碳纳米管、麦尔多拉蓝、乙醇脱氢酶(ADH)以及氧化型辅酶Ⅰ(NAD+)依次修饰在工作电极表面,然后在该三电极表面贴一层亲水膜,形成一个5μL反应池,制成血乙醇检测新型生物传感器检测条。结果表明,此生物传感器具有良好的准确性和稳定性;检测线性范围为0.5～20mmol/L,r=0.99493;检出限为0.22mmol/L;电流达到95%稳态时间小于15s。考察了pH值、温度及干扰物对生物传感器的影响。用此生物传感器和顶空气相色谱法对10份全血标本乙醇浓度平行测试,两者相关性良好r=0.97583。利用虹吸现象吸取微量全血直接定量测定乙醇浓度是此生物传感器的特点。