Disposable Injection‐Moulded Cell‐on‐a‐Chip Microfluidic Devices with Integrated Conducting Polymer Electrodes for On‐Line Voltammetric and Electrochemiluminescence Detection

This work reports the construction and characterization of plastic electrochemical micro‐flow‐cells with integrated injection‐moulded polymer electrodes. The three electrodes (working, auxiliary, and reference) were fabricated by injection‐moulding from a conducting grade of polystyrene loaded with carbon fibers. On‐chip reference electrodes were prepared by coating one of the conducting polymer electrodes with a Ag/AgCl layer (implemented either by e‐beam evaporation of Ag followed by electrochemical formation of AgCl or by applying a Ag/AgCl paste). Working electrodes were either polymer electrodes coated with Au by e‐beam evaporation or bare conducting polymer electrodes. The electrodes were integrated into the micro‐flow‐cells by an over‐moulding process followed by ultrasonic welding. The devices were characterized by optical and electrochemical techniques. Studies by cyclic voltammetry (CV), anodic stripping voltammetry (ASV) and electrochemiluminescence (ECL) demonstrate ‘proof–of‐principle’ of the micro‐flow‐cells as electrochemical sensors.     

集成化金膜阵列电极的制作研究

以聚乙烯不干胶掩膜版法结合金属溅射沉积技术在FR-4玻璃纤维版上制作了由6个金膜工作电极(1 mm×2 mm)、1个大面积金膜对电极(2 mm×13 mm)和1个厚膜Ag/AgCl参比电极构成的集成化金膜阵列电极系统,并利用电化学手段对阵列电极系统进行了考察。研究结果表明,K3Fe(CN)6在厚膜Ag/AgCl/1.0 mol/L NaCl参比电极上的式电位与商业Ag/AgCl/3.0 mol/L NaCl参比电极相差0.067 V;参比电极放置1个月后,测量电位未发生明显变化。利用扫描电化学显微镜对工作电极表面平整度进行考察,结果表明工作电极表面具有较好的平整度。通过测量H2SO4还原峰面积评价了工作电极电化学面积的批内、批间一致性;通过K3Fe(CN)6在电极上的Ipa/Ipc比值评价了工作电极电化学特性的批内、批间一致性。结果表明,阵列电极面积和电化学特性具有良好的批内和批间一致性。对集成化金膜阵列电极系统的研究结果表明,聚乙烯不干胶掩膜版法结合金属溅射沉积技术制作的阵列电极能够满足电化学电极的要求,可作为电化学生物传感器的基础电极。     

Investigation of a Thin‐film Quasi‐reference Electrode Fabricated by Combined Sputtering‐evaporation Approach

This paper presents the development of a thin‐film quasi‐reference electrode (tQRE), which was fabricated by sputtering silver (Ag) on a conducting gold layer. The Ag layer was subsequently covered by silver chloride (AgCl) with the aid of e‐beam evaporation. The functionality of the tQREs as reliable reference electrodes was confirmed by observing the potential response in solutions with various chloride ion concentrations. The influence of solution pH on the potential change of the tQREs was evaluated. In the solution with controlled ionic strength, the tQREs were able to provide stable and consistent potential outputs. Experimental investigation of the electrochemical sensors with integrated tQREs demonstrated potential applicability of the tQREs to be incorporated into miniaturized and disposable lab‐on‐a‐chip sensors for point‐of‐care/in‐situ measurements.     

Fabrication of a Simple and Cheap Screen-printed Silver/Silver Chloride (Ag/AgCl) Quasi-reference Electrode

In this work, a silver/silver chloride ink is fabricated using two steps. First the silver ink is prepare using silver, nail polish and acetone. Then the silver ink is painted in a paper substrate and a silver chloride layer is deposited using a bleach solution. The result is the silver/silver chloride conductive ink. The silver ink is cheap ($2.49/g), well-dispersive and very easy to fabricate. The materials were characterized by SEM and XRD. The Ag ink showed the formation of a continuous network throughout the silver ink film with fewer agglomeration. The effective chlorination process was also observed in the Ag/AgCl characterization. Since the Ag/AgCl substrate will be used as a quasi-reference electrode, it is important to investigate the electrical properties. The Ag ink showed an average ohmic resistance of 2.27 Ω. The addition of the AgCl layer decreases the conductivity, as expected. In summary, the Ag/Ag/Cl ink developed is simple, well-dispersed, cheap and with good conductivity. Therefore, it can be used as a conductive ink in the fabrication of quasi-reference electrodes.     

Properties of Ag/AgCl electrodes fabricated with IC-compatible technologies

The purpose of this work is to fabricate and characterize Ag/AgCl electrodes made on a silicon chip at the wafer level with integrated circuit-compatible fabrication techniques. Such electrodes are useful as reference electrodes in several kinds of chemical sensors. Two types of electrode were investigated. The first type uses an evaporated AgCl layer that is patterned with lift-off photolithography. The second type is formed by exposing a selected part of the silver substrate to a KCrO₃Cl solution. Both types of electrode give the thermodynamically expected potential response to variations of Cl⁻ ion concentration. The potential generated by the KCrO₃Cl-formed electrodes was more stable, however. Auger electron spectroscopy depth profiles indicate that immersion in a KCrO₃Cl solution produces a thin layer of AgCl on top of a layer of AgO. The low electronic resistance of AgO then reduces the measured series resistance of the KCrO₃Cl-formed electrodes. Impedance plane plots and the impedance as a function of frequency were measured for both types of electrode, and the impedance of the evaporated AgCl electrodes was indeed considerably higher. The impedance measurements could be successfully modelled by assuming a Randles equivalent circuit for the AgCl/electrolyte interface. For the KCrO₃Cl-formed electrodes, the impedance was modified by the porosity these electrodes manifested.     

The microporous structure of silver/silver chloride electrodes and the implications for Harned cell operation

The stability of silver/silver chloride reference electrodes when moved between different electrolyte solutions is limited by the small amount of solution transferred. The time required for the Ag/AgCl electrode potential to stabilize has been shown to increase with electrode size in an empirical fashion. The results presented suggest the presence of a microporous structure in the electrodes that limits the rate at which traces of any previous solutions are diluted by any new solution environment. Operational implications for the stability and accuracy of the Harned Cell used as the primary standard for pH measurements and the certification of primary reference buffers are discussed. Presented to the CCQM Electrochemical Analysis Working Group, Paris, April 2004.     

Amperometric determination of ascorbic acid on screen-printing ruthenium dioxide electrode

Ruthenium oxide (RuO₂) is commonly used in resistive pastes for screen printing. The electrochemical properties of a screen-printing planar RuO₂ electrode have hardly been studied. In this communication, planar electrochemical sensors with a RuO₂ working electrode, an Ag|AgCl reference electrode and a RuO₂ counter electrode are fabricated by screen printing. The electro-oxidation of ascorbic acid, uric acid, and hydrogen peroxide on these RuO₂ electrodes is investigated by means of cyclic voltammetry. Compared to uric acid and hydrogen peroxide, ascorbic acid can be easily oxidized at the low operating potential (<150 mV versus Ag|AgCl). The amperometric measurement of ascorbic acid at 100 mV on a RuO₂ electrode shows fast response and good linearity in the 0–4 mM range. Meanwhile, the electrochemical interference from uric acid and hydrogen peroxide at this potential is very small.     

Analytical quality solid-state composite reference electrode manufactured by injection moulding

An injection moulding method was used for fabricating solid-state reference electrodes (Ag/AgCl type) based on a polymer/inorganic salt composite. In this method, a silver/silver chloride wire was placed inside a mould into which the mixture of polymer and inorganic salt was injected. The obtained solid-state composite reference electrodes were extensively tested to study the influence of different parameters such as solution composition, the concentrations and mobility of ions and pH on the potential stability of the electrodes. These experiments revealed that the composite reference electrodes are insensitive to the matrix effect, have excellent potential readings stability and considerably reduced leakage of inorganic salt. The composite reference electrodes were compared favourably to high-quality commercial reference electrodes. It was concluded that the composite reference electrodes obtained by injection moulding are of analytical quality allowing for continuous, prolonged and intensive usage.     

非标记凝血酶阵列电化学适体传感器的研究

基于自制的集成化三阵列金膜电极,构建了一个简单、灵敏、非标记的凝血酶阵列电化学适体传感器。以聚乙烯不干胶掩膜版法结合金属溅射沉积技术,在FR-4玻璃纤维板上制作了由3个金膜工作电极、1个大面积金膜对电极和1个厚膜Ag/AgCl参比电极构成的集成化金膜阵列电极系统。以集成化金膜阵列电极作为基础电极,采用巯基自组装技术将带巯基的凝血酶适体固定在3个金工作电极表面,巯基己醇封闭后获得三阵列凝血酶适体传感器,以电活性物质铁氰化钾作为电化学探针,基于凝血酶适体和凝血酶结合前后铁氰化钾在电极表面传质的不同导致电流变化进行凝血酶的测定。采用方波脉冲伏安法,铁氰化钾氧化峰电流的变化值与凝血酶浓度在 1.52~63 nmol/L范围内呈良好的线性关系,检出限(S/N=3)为0.92 nmol/L。     

Voltammetric characterization of a fully integrated, patterned single walled carbon nanotube three-electrode system on a glass substrate

A single walled carbon nanotube (SWCNT)-based three-electrode system was fully integrated on glass substrates using a standard microfabrication process and electrochemically characterized using cyclic voltammetry. O(2) plasma functionalization of the SWCNT film working electrode for achieving high sensitivity was voltammetrically optimized with respect to the plasma power and treatment time. Chlorination of a Ag thin-film was done in an acidic solution for different dip times to form a thin-film Ag/AgCl reference electrode. The Nernstian behavior of as-prepared and seven-day-aged Ag/AgCl thin-film electrodes was investigated for seeking the optimum reference electrode with long-term stability and was compared to a commercial reference electrode. A quality control evaluation and a performance assessment of the fully integrated SWCNT-transferred sensing systems were performed using cyclic voltammetry. The proposed SWCNT-based three electrode device exhibited clear electrochemistry under voltammetric conditions, and is therefore a candidate for use in all electrochemical biosensors.     

Copper(II) ion sensor based on electropolymerized undoped conducting polymers

A solid state copper(II) ion sensor is reported based on the application of electropolymerized undoped (neutral) polycarbazole (PCz) and polyindole (PIn) modified electrodes. The new sensor shows high selectivity to Cu²⁺ ions with a detection limit of 10^–5 M. PCz and PIn are formed respectively by the anodic oxidation of 50 mM carbazole and 5 mM indole monomers in dichloromethane containing 0.1 M tetrabutylammonium perchlorate on a platinum electrode using a single compartment cell. Potentiostatic polymerization of both the monomers are carried out at 1.3 V and 1.0 V vs. Ag/AgCl, respectively. Perchlorate ions were electrochemically removed from the polymer films by applying – 0.2 V vs. Ag/AgCl. Polymer-coated electrodes are incubated in 1 M KCl solution for 8 h followed by incubation in distilled water for 2 h before using as a metal ion sensor. The undoped PCz and PIn electrodes were found to be highly selective and sensitive for Cu²⁺ ions with little selectivity for Pb²⁺ and negligible response towards Ag⁺, Hg²⁺, Cu⁺, Ni²⁺, Co²⁺, Fe²⁺, Fe³⁺ or Zn²⁺. Potentiometric responses for Cu²⁺ ions are recorded for both the sensor electrodes together with a double-junction Ag/AgCl reference electrode. Calibration curves for Cu²⁺ are reported for both ion sensors. The polymer-modified electrodes were found to be stable for several weeks. Electronic Publication     

Performance characteristics of a planar ‘clark-type’ oxygen sensor

A planar two-electrode oxygen sensor is fabricated using standard IC technology. It consists of an Ag cathode and an Ag/AgCl/Cl⁻ reference anode. The electrochemical characterization of the cathode and the long-term behaviour, i.e., stability, of the reference electrode are determined in order to optimize the fabrication procedure. The device is then completed with a hydrogel layer (poly 2-hydroxyethyl methacrylate, pHEMA) and a silicone rubber membrane. Voltammetric and chronoamperometric tests of the complete sensor are performed and its O₂ response determined.     

On-chip multi-electrochemical sensor array platform for simultaneous screening of nitric oxide and peroxynitrite

In this work we report on the design, microfabrication and analytical performances of a new electrochemical sensor array (ESA) which allows for the first time the simultaneous amperometric detection of nitric oxide (NO) and peroxynitrite (ONOO(-)), two biologically relevant molecules. The on-chip device includes individually addressable sets of gold ultramicroelectrodes (UMEs) of 50 μm diameter, Ag/AgCl reference electrode and gold counter electrode. The electrodes are separated into two groups; each has one reference electrode, one counter electrode and 110 UMEs specifically tailored to detect a specific analyte. The ESA is incorporated on a custom interface with a cell culture well and spring contact pins that can be easily interconnected to an external multichannel potentiostat. Each UME of the network dedicated to the detection of NO is electrochemically modified by electrodepositing thin layers of poly(eugenol) and poly(phenol). The detection of NO is performed amperometrically at 0.8 V vs. Ag/AgCl in phosphate buffer solution (PBS, pH = 7.4) and other buffers adapted to biological cell culture, using a NO-donor. The network of UMEs dedicated to the detection of ONOO(-) is used without further chemical modification of the surface and the uncoated gold electrodes operate at -0.1 V vs. Ag/AgCl to detect the reduction of ONOOH in PBS. The selectivity issue of both sensors against major biologically relevant interfering analytes is examined. Simultaneous detection of NO and ONOO(-) in PBS is also achieved.     

An Arrayed Micro‐glutamate Sensor Probe Integrated with On‐probe Ag/AgCl Reference and Counter Electrodes

Complete all‐in‐one multi‐arrayed glutamate (Glut) sensors have been constructed on a silicon‐based micromachined probe composed of micro‐platinum (Pt) working electrodes, a micro‐silver/silver chloride (Ag/AgCl) reference electrode (RE), and a micro‐Pt counter electrode (CE). The OCP shift of the electrodeposited Ag/AgCl on‐probe micro‐reference electrode compared with a Ag/AgCl wire is <0.1 mV/h. The composition ratio of Ag, Cl, and Pt on the electrodeposited on‐probe micro‐reference electrode is observed to be 1.00 : 0.48 : 0.02 analyzed by EDS. The miniaturized amperometric Glut biosensors were constructed on working electrode sites (electrode area: ∼8.5×10⁻⁵ cm²) of the microprobe modified with glutamate oxidase (GlutOx) enzyme layers for the selective, fast, and continuous detection of L‐glutamate. The sensor selectivity towards common electroactive interferents has been improved significantly by coating the electrode surface with perm‐selective polymer layers, overoxidized polypyrrole (PPY) and Nafion®. The sensitivity, detection range, and response time of the proposed all‐in‐one Glut biosensors are 204.7±5.8 nA μM⁻¹ cm⁻² (N=5), 4.99–109 μM, and 2.7±0.3 sec, respectively and no interferent signals of AA and DA were observed. The sensor can be reused over 19 times of continuous repetitive operation (total measurement time: ∼4 hours) and the sensor sensitivity can retain up to four weeks of storage.     

Electrochemical Detection of MS2 Phage Using a Bead‐based Immunoassay and a NanoIDA

A microbead based sandwich immunoassay for MS2 bacteriophage was developed using an interdigitated array (IDA) electrode with nanoscale dimensions (220 nm electrode width, 620 nm gap). The IDA was fabricated using an electron beam lithographic lift‐off technique. After an antibody‐assisted capture of MS2 using paramagnetic microbeads, a β‐galactosidase labeled secondary antibody was used to convert p‐aminophenyl‐β‐D ‐galactopyranoside (PAPG) into the redox active p‐aminophenol (PAP). Amperometric detection of PAP with IDA electrodes at +300 and ?200 mV vs. a Ag/AgCl reference electrode was used to measure the result, detecting MS2 concentrations as low as 10 ng/mL.     

Improvement of the silver/silver chloride reference electrode and its application to pH measurement

When a silver/silver chloride (Ag AgCl ) reference electrode was used continuously in a low conductivity solution or reductive solution, it was often observed that stability of the liquid junction potential was lost. This phenomenon was remarkable with a Ag AgCl reference electrode compared to a calomel reference electrode. We found that 340 mg l(-1) of silver was dissolved in 3 M potassium chloride (KCl) internal solution as silver complex ions (AgCl(-(x-1))(x)) for x = 2 or 3. However, only 1.93 mg l(-1) of silver chloride (AgCl) can theoretically be dissolved in water. The complex ion that effused into the sample solution through the liquid junction clogged the liquid junction (e.g. porous ceramic) as AgCl, or as metallic silver (Ag) in reducing solution. Therefore, the constant effusion of KCl internal solution was inhibited, and the liquid junction potential became unstable or fluctuating. A new reference electrode was developed, which can eliminate AgCl(-(x-1))(x) in 3 M KCl internal solution by the use of chelating resins. A combination of this reference electrode with a pH electrode made long-term stable pH measurements possible.     

Ag/ZnO Catalysts with Different ZnO Nanostructures for Non‐enzymatic Detection of Urea

Silver coated ZnO nanorods and nanoflakes with different crystallographic orientations were synthesized by a combination of sputter deposition and solution growth process. Catalytic properties of morphology‐dependent Ag/ZnO nanostructures were then investigated for urea sensors without enzyme. Ag/ZnO nanorods on carbon electrodes exhibit a higher catalytic activity and an improved efficiency than Ag/ZnO nanoflakes on carbon electrodes. Ag/ZnO nanorod catalysts with more electrochemically surface area (169 cm² mg^?1) on carbon electrode facilitate urea electrooxidation due to easier electron transfer, which further promotes the urea electrolysis. The Ag/ZnO nanorod catalysts also show a significant reduction in the onset voltage (0.410 V vs. Ag/AgCl) and an increase in the current density (12.0 mA cm^?2 mg^?1) at 0.55 V vs Ag/AgCl. The results on urea electrooxidation show that Ag/ZnO nanostructures can be a potential catalyst for non‐enzymatic biosensors and fuel cells.     

Environmentally benign chitosan as precursor and reductant for synthesis of Ag/AgCl/N-doped carbon composite photocatalysts and their photocatalytic degradation performance

In this study, we reported a novel Ag/AgCl loaded N-doped carbon composite photocatalyst (Ag/AgCl/NC) which was fabricated by a facile and green method. The composite was prepared only by two simple steps. Firstly, the Ag/N-doped carbon (Ag/NC) was prepared by one-step hydrothermal treatment; during this progress the environmentally benign and renewable natural chitosan was used as not only reducer and stabilizer, but also as a nitrogen source and carbon source. Secondly, Ag/AgCl/NC composite was synthesized via in situ oxidation reaction by adding FeCl₃. The Ag/AgCl/NC composite was characterized using X-ray diffraction, transmission electronic microscopy, energy dispersive X-ray spectra, UV-visible diffused reflectance spectra, X-ray photoelectron spectroscopy and nitrogen adsorption-desorption measurements, respectively. The obtained Ag/AgCl/NC composite exhibited a superior photocatalytic activity and stability for the degradation of rhodamine B (RhB) under visible light irradiation.     

The fabrication and performance of an Ag/AgCl reference electrode in human serum

A simple set of electric circuits was used to assemble a pulse generator. With pulse potentials and under galvanostatical control, a clean silver wire was anodized electrochemically for 0.2–0.5 min in 1.0 mol l⁻¹ HCl with a pulse current density of 20 mA cm⁻², and the pulse wave parameters of t_a/t_c = 1 and a cycle of 4 s forming an Ag/AgCl reference electrode. Even though the AgCl layer was consumed during the working period when the Ag/AgCl electrode was used as a cathode, the AgCl layer could be in situ recovered electrochemically in serum used when a reversed potential was applied to the electrode system immediately after the measuring program was finished. The current response curve of the anode indicated that an AgCl layer in high density was basically accomplished during the first 6 pulse cycles in human serum. In order to keep a stable and uniform AgCl layer on the reference electrode after each measuring cycle, the ratio of the recovery time (t_r) to the working time (t_w) was measured and the smallest value was obtained at 0.03. The open-circuit potential of the Ag/AgCl electrode with respect to a SCE in 0.1 mol l⁻¹ KCl was monitored over a period of 14 days and the mean value was 40.09 mV vs SCE with a standard deviation of 2.55 mV. The potential of the Ag/AgCl reference electrode did remain constant when the measurements were repeated more than 600 times in undiluted human serum with a standard deviation of 1.89 mV. This study indicated that the Ag/AgCl reference electrode could been rapidly fabricated with a pulse potential and could be used as a reference electrode with long-term stable properties in human serum samples.     

Bifunctional AgCl/Ag composites for SERS monitoring and low temperature visible light photocatalysis degradation of pollutant

With the assistance of Polyvinylpyrrolidone (PVP), AgCl/Ag composites were fabricated in N, N-Dimethylformamide (DMF) solvent via a photoactivated route. The size of AgCl particles was in the range of 500 nm to 1 μm and the Ag particle's diameter was about 10–20 nm. Different from those core–shell structures reported before, the Ag nanoparticles were dispersed uniformly both on the surface and in the body of AgCl particles. The generation of such kind of composites was resulted from the reducing ability of DMF and light irradiation during the formation of AgCl particles. The as-obtained AgCl/Ag composites presented great activity for both surface-enhanced Raman scattering (SERS) detection and visible light photocatalytic degradation of organic dyes. Additionally, the AgCl/Ag composites could maintain high photocatalytic activity even though the ambient temperature was as low as 15 °C and recycle photocatalysis experiments indicated that the photocatalyst exhibited higher stability. Such kind of AgCl/Ag composites holds great potential for environmental monitoring devices and pollutant treatments.