Microchip capillary electrophoresis with electrochemical detection

A novel microchip capillary electrophoresis system with electrochemical detection, using the replaceable microelectrode, is first reported. This kind of electrode can be fabricated in general laboratories and can be replaced quickly with electrodes of different materials according to the requirements of experiments. The end-column electrochemical detection on microchip CE was achieved by fixing the working electrode (such as carbon fiber, Pt, or Au, etc.) through a guide tube on the end of the separation channel. The experiment results indicate that the alignment of the electrode with the channel outlet can be carried out accurately and reproducibly, and therefore, the detection device has low noise and good reproducibility. The detection limit of dopamine is 2.4 x 10(-7) M, which is the lowest result reported so far. The separation and detection of dopamine, 5-hydroxytryptamine and epinephrine using carbon fiber and Pt microdisk electrodes within 50 s was successfully performed.     

Following the biocatalytic activities of glucose oxidase by electrochemically cross-linked enzyme-Pt nanoparticles composite electrodes

An integrated platinum nanoparticles (NPs)/glucose oxidase (GOx) composite film associated with a Au electrode is used to follow the biocatalytic activities of the enzyme. The film is assembled on a Au electrode by the electropolymerization of thioaniline-functionalized Pt NPs and thioaniline-modified GOx. The resulting enzyme/Pt NPs-functionalized electrode stimulates the O 2 oxidation of glucose to gluconic acid and H 2O 2. The modified electrode is then implemented to follow the activity of the enzyme by the electrochemical monitoring of the generated H 2O 2. The effect of the composition of the Pt NPs/GOx cross-linked nanostructures and the optimal conditions for the preparation of the electrodes are discussed.     

Capillary electrophoresis with an integrated on-capillary tubular detector based on a carbon sol-gel-derived platform

An integrated on-capillary tubular electrochemical detector for capillary electrophoresis systems has been fabricated based on sol-gel technique. It consists of a sol-gel carbon composite tubular electrode attached permanently onto the outlet of the separation capillary. The device greatly eases the setting up of capillary electrophoresis with electrochemical detection (CEEC) as it makes possible electrode/capillary alignment without the aid of a micromanipulator since this integrated unit can be simply immersed in the CE separation buffer in an ordinary three-electrode stationary cell. To improve analytical performance of the integrated unit, the external wall of the exit capillary was etched with HF after the polyimide coating of the capillary had been removed. Influences of the working electrode length and the wall thickness at the outlet of capillary on the separation efficiency and amperometric sensitivity were assessed and optimized. The practical applicability of this configuration is demonstrated with the detection of both catecholamines and carbohydrates. The advantages, namely, versatility, convenience, ease of operation, and low-cost, of the new design combined with an excellent performance lead to high stability and low detection limits.     

Integrated electrophoresis chips/amperometric detection with sputtered gold working electrodes

An on-chip electrochemical detector for micromachined capillary electrophoresis (CE) systems, based on sputtering a gold working electrode directly onto the capillary outlet, is described. The new on-chip detector preparation requires no microfabrication or alignment procedures nor a decoupling mechanism. The attractive performance of the integrated electrophoresis chips/amperometric detection was demonstrated for the anodic detection of neurotransmitters. The response for dopamine was linear from 20 to 200 μM, with a LOD of 1.0 μM and a sensitivity of 52 pA/μM. Such intimate coupling of capillary electrophoresis chips and electrochemical detection facilitates the realization of complete integrated microanalytical devices.     

Nanofabrication of bio-self assembled monolayer and its electrochemical property for toxicant detection

Cyclic voltammetry (CV) has been used to investigate the electrochemical behavior of a glutathione (GSH) self assembled monolayer on modified gold electrodes (Bio-SAM). The GSH monolayer exhibits an influence on electrode surface activity. Electrochemically immobilized dsDNA onto a Cyt c/GSH-SAM/Au electrode, which is useful for the fabrication of a nanobiosensing device. The immobilized Cyt c followed by dsDNA immobilized films maintained its surface activity and finally dsDNA/Cyt c/GSH-SAM/Au electrode, targeted for the detection of toxicants. The films were characterized by CV, DPV, and AFM. The differential pulse voltammetry (DPV) technique was applied to detect three kinds of common toxins, 2-aminoanthracene (2-AA), 3-bromobenzanthrone (3-BBA) and bisphenol A (BPhA). The electrochemical signals showed good inverse relationship with the increase of concentrations of toxicants. Our proposed system based on electrochemical method with nanoscale film technology can be applied at highly sensitive biosensor for detecting various toxic chemicals.     

Three-dimensional mesoporous gold film to enhance the sensitivity of electrochemical detection

Cell-cell and cell-extracellular matrix (ECM) adhesion are fundamental and important in the development of a cell-based chip. In this study, a novel, simple, rapid, and one-step technique was developed for the fabrication of a uniform three-dimensional mesoporous gold thin film (MPGF) onto a gold (Au) coated glass plate based on an electrochemical deposition method. Scanning electron microscopy images demonstrated that the resulting MPGF electrode had uniformly distributed pores with diameters of about 20 nm. The cyclic voltammetric behavior of [Fe(CN)(6)](4-/3-) coupled onto MPGF and Au electrodes demonstrated that the MPGF electrode had a higher electrocatalytic sensitivity and reversibility than the bare Au electrode. The Arg-Gly-Asp (RGD) sequence containing the peptide was immobilized on the MPGF and bare Au substrates. HeLa cancer cells were then cultured on the RGD peptide layer. The successful immobilization of the peptide and cells was confirmed by atomic force microscopy. The cell proliferation and viability were evaluated by cyclic voltammetry and Trypan blue dyeing assay. These results indicated that the RGD/MPGF modified electrodes showed an electrochemical sensitivity in the detection of cancer cells which is approximately three times higher, especially at low cell density, than RGD/Au electrodes. This much improved sensitivity of the MPGF modified electrode demonstrates the potential for the fabrication of a highly sensitive and low-cost cell-based chip for rapid cancer detection.     

Electrochemical properties of carbon nanotube (CNT) film electrodes prepared by controllable adsorption of CNTs onto an alkanethiol monolayer self-assembled on gold electrodes

This paper describes electrochemical properties, such as electrode reactivity, electrode dimensions, and interfacial capacitance, of multiwalled carbon nanotube (MWNT) film electrodes prepared by controllable adsorption of the MWNTs onto the self-assembled monolayer (SAM) of n-octadecyl mercaptan (C18H37SH) deposited onto Au electrodes. The adsorption of the MWNTs onto the SAM-modified Au electrode substantially restores heterogeneous electron transfer between bare Au electrode and redox species in solution phase that is almost totally blocked by the SAM of C18H37SH, and as a result, the prepared MWNT/SAM-modified electrode possesses good electrode reactivity without a remarkable barrier to heterogeneous electron transfer. In addition, the surface coverage of the MWNTs is readily controlled by adjusting the immersion time for the adsorption of the MWNTs onto the SAM of C18H37SH, which essentially endows the prepared MWNT/SAM-modified electrodes with tunable electrode dimensions ranging from a nanoelectrode array to a macro-sized conventional electrode. On the other hand, the MWNT/SAM-modified electrode is found to possess a largely reduced interfacial capacitance, as compared with the MWNT film electrodes prepared with existing methods by directly confining the MWNTs onto electrode surface. This demonstration offers a new approach to fabrication of stable MWNT film electrodes with excellent electrochemical properties that are believed to be very attractive for electrochemical studies and electroanalytical applications.     

Simultaneous determination of tryptophan and glutathione in individual rat hepatocytes by capillary zone electrophoresis with electrochemical detection at a carbon fiber bundle--Au/Hg dual electrode

A method for single-cell analysis was developed by combining capillary zone electrophoresis (CZE) with electrochemical detection (ECD) using a dual electrode consisting of two different types of electrode material (carbon fiber and Au/Hg). In this method, the parallel mode was used. Different potentials were applied to both electrodes of the dual electrode. Tryptophan and glutathione, which could not be simultaneously detected by normal CZE-ECD, could be simultaneously and selectively detected by CZE-ECD at the dual electrode in one run, respectively. The CZE-ECD system with the dual electrode was applied to determine them in individual rat hepatocytes.     

Fabrication and characterization of platinum black and mesoporous platinum electrodes for in-vivo and continuously monitoring electrochemical sensor applications

Most electrochemical biosensors are disposable due to enzymes that are living creatures. Thus, these are limited to use in in-vivo and continuously monitoring biosensor system applications. The mesoporous (pores with a size of 2-50 nm) platinum (Pt) structure formed on a rod-shaped Pt microelectrode was reported for developments glucose sensors without any enzymes. In this paper, plane Pt electrode (non-treated), Pt black electrode, and mesoporous Pt electrode are fabricated and characterized on a silicon substrate in order to check their usability as enzymeless sensing electrodes for in-vivo and continuously monitoring electrochemical biosensors integrated with silicon CMOS read-out circuitry. The Pt black electrode with rough surface was fabricated by using an electrodeposition technique with hexachloroplatinic acid hydrate (HCPA) solutions. The proposed mesoporous Pt electrode with approximately 3 nm in pore diameter was fabricated by using an electrodeposition technique with nonionic surfactant octaethylene glycol monohexadecyl ether (C₁₆EO₈) and HCPA. The measured current responses at 40 mM glucose solution of the fabricated plane Pt, Pt black, and mesoporous Pt electrodes are approximately 12.4 nA/mm², 2.1 μA/mm², and 2.8 μA/mm², respectively. These data indicate that the mesoporous Pt electrode is much more sensitive than the other Pt electrodes and has strong potential for enzymeless electrochemical sensor applications.     

Templated synthesis, characterization, and sensing application of macroscopic platinum nanowire network electrodes

Novel platinum nanowire network electrodes have been fabricated through electrodeposition using mesoporous silica thin films as templates. These electrodes were characterized by X-ray diffraction, transmission electron microscope, and scanning electron microscope. The electrochemical properties of the electrodes, such as electrochemical active area and methanol oxidation, have also been studied. Compared with conventional polycrystalline Pt electrodes, these novel nanowire network electrodes possess high electrochemical active areas and demonstrate higher current densities and a lower onset potential for methanol electro-oxidation. Enzymatic Pt nanowire-network-based sensors show higher sensitivity for glucose detection than that using conventional polycrystalline Pt electrode. Such macroscopic nanowire network electrodes provide ideal platforms for sensing and other device applications.     

Detection of hydrogen peroxide produced during electrochemical oxygen reduction using scanning electrochemical microscopy

The substrate-generation/tip-collection mode of scanning electrochemical microscopy was used to detect hydrogen peroxide formed as an intermediate during oxygen reduction at various electrodes. The experiment is conceptually similar to rotating ring-disk experiments but does not require the production of a ring-disk assembly for the specific electrode material in question. In order to limit the extension of the diffusion layer above the sample, the sample electrode potential is pulsed while the Pt ultramicroelectrode probe (UME) is held at a constant potential for oxidative amperometric detection of hydrogen peroxide. The signal at UME is influenced by the sample region within the diffusion length of hydrogen peroxide during the pulse of 2.5 s. The method is tested with three model electrodes showing different behavior with respect to the oxygen reduction reaction (ORR) in acidic solution. Simple analytical models were used to extract effective rate constants for the most important reaction paths of ORR at gold and palladium-cobalt samples from the chronoamperometric response of the UME to a reduction pulse at the sample electrode.     

Determination of alcohols using a Ni-Pt alloy amperometric sensor

An amperometric sensor for monitoring ethanol concentration using a Ni-Pt alloy electrode has been developed. The films of Ni-Pt alloy were electrodeposited under various potentials on an Au/Al₂O₃ substrate. To examine the effects of Ni-Pt alloy films on the sensing performance, the electrodes with various Ni:Pt atomic proportions of 100:0, 25:75, 70:30, 82:18 and 0:100 were prepared and tested. X-ray diffraction and scanning electron microscope analyses indicate that Ni atoms of Ni-Pt alloy electrodes are inserted in the Pt lattice and a pyramid-like structure is formed with an increase of Pt content in the film. All the prepared electrodes have a linear relationship between response current and ethanol concentration for the detection range of 50 to 300 ppm ethanol in alkaline solutions. With an increase of Pt content in the film, the response time of the Ni-Pt alloy electrodes was reduced whereas the sensitivity was decreased. The sensor with 70 at.% Pt in the film was most stable with the duration over a 63-day-period. The sensitivity of the Ni-Pt alloy electrodes for detecting glucose, various alcohols and acids have also been studied.     

Electrogenerated chemiluminescence. 66. The role of direct coreactant oxidation in the ruthenium tris(2,2')bipyridyl/tripropylamine system and the effect of halide ions on the emission intensity

We describe the electrogenerated chemiluminescence (ECL) processes of the Ru(bpy)3(2+) (bpy = 2,2'-bipyridyl)/ tripropylamine (TPrA) system at glassy carbon, platinum, and gold electrodes. The electrochemical behavior of TPrA on different electrode materials and its influence on the ECL process are demonstrated. At glassy carbon electrodes, the direct oxidation of TPrA began at approximately 0.6 V vs SCE and exhibited a broad irreversible anodic peak. Two ECL waves were observed, one in the potential region more negative than 1.0 V vs SCE and one at more positive potentials. The first ECL process apparently occurs without the electrogeneration of Ru(bpy)3(3+), in contrast to that of the second ECL wave. At Pt and Au electrodes, however, the formation of surface oxides significantly blocked the direct oxidation of TPrA. An ECL wave below 1.0 V did not appear at Pt and was very weak at gold. The ECL peaks at potentials of 1.1-1.2 V were also much weaker than those observed at the glassy carbon electrode. These results showed that the direct oxidation of TPrA played an important role in the ECL processes. Therefore, the enhancement of the TPrA oxidation current might lead to an increase in the ECL intensity. Small amounts of halide species were found to inhibit the growth of surface oxides on Pt and gold electrodes and led to an obvious increase of TPrA oxidation current. The anodic dissolution of gold in halide-containing solution was also important in activating the gold electrode surface. The electrochemical catalytic effect of bromide further promoted the oxidation of TPrA. A halide effect on ECL at Pt and Au electrodes was also evident. The most effective enhancement of ECL was observed at Au electrode in a bromide-containing solution. This effect was also found in an commercial flow-through instrument (IGEN) and provided a simple way to improve the detection sensitivity at low concentrations of Ru(bpy)3(2+).     

Capillary electrophoresis coupled with electrochemiluminescence detection using porous etched joint

A new setup to couple capillary electrophoresis (CE) with electrochemiluminescence (ECL) detection is described in which the electrical connection of CE is achieved through a porous section at a distance of 7 mm from the CE capillary outlet. Because the porous capillary wall allowed the CE current to pass through and there was no electric field gradient beyond that section, the influence of CE high-voltage field on the ECL procedure was eliminated. The porous section formed by etching the capillary with hydrofluoric acid after only one side of the circumference of 2-3 mm of polyimide coating of the capillary was removed, while keeping the polyimide coating on the other part to protect the capillary from HF etching makes the capillary joint much more robust since only a part of the circumference of it is etched. A standard three-electrode configuration was used in experiments with Pt wire as a counter electrode, Ag/AgCl as a reference electrode, and a 300-microm diameter Pt disk as a working electrode. Compared with CE-ECL conventional decoupler designs, the present setup with a porous joint has no added dead volume created. Moreover, the dead volume can be increasingly decreased by shortening the distance ( approximately 100 microm) between the working electrode and the end of the separation capillary. The versatility in choice of capillaries and separation buffers within this design is the main advantage over the use of small i.d. capillary and low conductivity buffer in some CE-ECL systems. The performance of this setup is illustrated by the analyses of tripropylamine and proline.     

Electrochemical detectors prepared by electroless deposition for microfabricated electrophoresis chips

Microfabricated capillary electrophoresis (CE) chips with integrated electrochemical detection have been developed on glass substrates. An electroless deposition procedure was used to deposit a gold film directly onto the capillary outlet to provide high-sensitivity electrochemical detection for catechol and several nitroaromatic explosives. Scanning electron microscopy revealed that the electroless gold film contains nanoscopic gold aggregates (100-150 nm) with an average thickness of 79 nm. The electroless deposition procedure can be easily and routinely performed in any wet-chemistry laboratory, and electroless gold can be deposited onto complex and internal surfaces. Intimate coupling of electrochemical detection and CE chips obviates the need for a coupling mechanism or tedious alignment procedures. With nitroaromatic compounds as a working model, microchip capillary electrophoresis equipped with electroless gold has proven to provide high sensitivity and fast response times for sensor applications. The CE microchip system was capable of separation and determination of explosive compounds including TNT in less than 130 s with detection limits ranging from 24 to 36 microg/L, i.e., 4-fold enhancements in detection efficiency in comparison to thick-film technology.     

Fully integrated on-chip electrochemical detection for capillary electrophoresis in a microfabricated device

Microfabricated lab-on-a-chip devices employing a fully integrated electrochemical (EC) detection system have been developed and evaluated. Both capillary electrophoresis (CE) channels and all CE/EC electrodes were incorporated directly onto glass substrates via traditional microfabrication techniques, including photolithographic patterning, wet chemical etching, DC sputtering, and thermal wafer bonding. Unlike analogous CE/EC devices previously reported, no external electrodes were required, and critical electrode characteristics, including size, shape, and placement on the microchip, were established absolutely by the photolithography process. For the model analytes dopamine and catechol, detection limits in the 4-5 microM range (approximately 200 amol injected) were obtained with the Pt EC electrodes employed here, and devices gave stable analytical performance over months of usage.     

用单壁纳米碳管制备葡萄糖生物传感器的研究

以单壁纳米碳管为代表材料,对利用纳米碳管制备葡萄糖生物传感器中纳米碳管的作用和纳米碳管修饰电极的方法、酶的固定化方法及电极种类等因素对传感器性能的影响进行了研究.研究结果表明,纳米碳管的加入能有效地改善传感器的电化学性能,利用二茂铁和单壁纳米碳管共同修饰电极所制得的传感器的性能要好于仅用单壁纳米碳管修饰电极制得的传感器.在酶的固定化方法中,戊二醛交联法要略好于明胶包埋法;而利用铂电极制备出的生物传感器对葡萄糖的响应电流要明显高于利用金电极和玻碳电极制备出的生物传感器.这些结论对于开发纳米碳管在生物传感领域及生命科学相关领域的应用有参考价值.     

Comparison of Micro- and Nano-Pore Platinum Working Electrodes for CMOS Integrated Nondisposable Biosensor Applications

In this paper, micro-pore and nano-pore platinum (Pt) electrodes are fabricated, characterized, and compared in order to check the effects of the pore sizes of the working electrode in the electrochemical analysis. These electrodes are also fabricated on silicon substrate for developing nondisposable and continuous monitoring biosensors fully integrated with a CMOS read out circuit. The micro-pore Pt electrode was fabricated by using micromachining technique. The nano-pore Pt electrode was fabricated by using a nonionic surfactant C ₁₆EO₈ (octaethylene glycol monohexadecyl ether) and an electrodeposition technique. The micro-pore array Pt electrode has approximately 10 mum in pore diameter and 80 mum in height, while the nano-pore array Pt electrode has approximately 2.5 nm in diameter. The measured current responses of the fabricated nano-pore, micro-pore, and plane Pt electrodes are approximately 4600 nA/mm² , 92.4 nA/mm², and 9.9 nA/mm² in 0.1 M phosphate buffered saline solution mixed with 10 mM glucose solution, respectively     

Multiwalled carbon nanotube-based bi-enzyme electrode for total cholesterol estimation in human serum

This study aims at fabricating multiwalled carbon nanotubes (MWCNTs) based enzymatic bioelectrode for total cholesterol estimation in human serum. For this purpose, a gold (Au) electrode was modified with MWCNTs uniformly dispersed in nafion (Nf) matrix. Cholesterol oxidase (ChOx) and cholesterol esterase (ChEt) were immobilised onto this Nf–MWCNTs film-modified Au electrode using layer-by-layer technique to fabricate the final bioelectrodes. The immobilisation of ChOx and ChEt onto the electrodes was demonstrated by scanning electron microscopy. The fabricated bioelectrodes were electrochemically characterised using cyclic voltammetry. The bioelectrodes offer reliable response characteristics towards cholesterol and stable electrochemical properties in terms of extended linear response range of 0.080–0.950?mM, detection limit up to 0.01?mM and optimum storage stability up to three weeks. Experimental results reveal that the fabricated bioelectrode offer optimum repeatability and reproducibility towards the cholesterol estimation and can also efficiently exclude interference by the commonly coexisting ascorbic acid, uric acid, lactic acid, glucose and urea, which is favourable for its efficient use in the highly selective analysis of total cholesterol in serum samples.     

Highly selective and sensitive electrochemical detection of dopamine using a nafion coated hybrid macroporous gold modified electrode with platinum nanoparticles

A coral-like macroporous Au electrode with electroplated Pt nanoparticles (hybrid macroporous Au-/nPts) coated with Nafion has been fabricated for the first time and used for highly selective and sensitive determination of dopamine (DA). The physically characterized results indicated that the electroplated Pt nanoparticles were dispersed uniformly on the macroporous Au electrode. The porosity and window pore size of the fabricated macroporous Au electrode were 50% and 100-300 nm, respectively. Also the electroplated Pt nanoparticles size was approximately 10-20 nm. The cyclic voltammograms results showed that the hybrid macroporous Au-/nPts exhibited a much larger surface activation area, a roughness factor (RF) of 2024.7, much higher than that of the macroporous Au electrode, which is 46.07. The electrochemical experimental results showed that the hybrid macroporous Au-/nPts coated with Nafion exhibited a dramatic electrocatalytic effect on the oxidation of DA. At 0.1 V, it responded linearly to DA concentrations ranging from 20 μ M to 160 μ M with a detection sensitivity of 90.9 μA mM (-1) cm (-2). Furthermore, it showed wide detection ranging from 20 nM to 900 μ M. At the same time, the interference of ascorbic acid (AA) was effectively avoided because of the Nafion film coated on the surface of the hybrid electrode.