Electrochemical oxidation of chlorophenols at a boron-doped diamond electrode and their determination by high-performance liquid chromatography with amperometric detection

Anodically pretreated diamond electrodes have been used for the detection of chlorophenols (CPs) in environmental water samples after high-performance liquid chromatographic (HPLC) separation. The anodization of as-deposited boron-doped polycrystalline diamond thin-film electrodes has enabled the stable determination of phenols over a wide concentration range. Prior to the HPLC analysis, a comparative study with ordinary glassy carbon, as-deposited diamond, and anodized diamond was made to examine the oxidative behavior of phenols by cyclic voltammety and flow injection analysis with amperometric detection. At anodized diamond electrodes, reproducible, well-defined cyclic voltammograms were obtained even at high CP concentration (5 mM), due to a low proclivity for adsorption of the oxidation products on the surface. In addition, after prolonged use, the partially deactivated diamond could be reactivated on line by applying a highly anodic potential (2.64 Vvs SCE) for 4 min, which enabled the destruction of the electrodeposited polymer deposits. Hydroxyl radicals produced by the high applied potential, in which oxygen evolution occurs, are believed to be responsible for the oxidation of the passivating layer on the surface. When coupled with flow injection analysis (FIA), anodized diamond exhibited excellent stability, with a response variability of 2.3% (n = 100), for the oxidation of a high concentration (5 mM) of chlorophenol. In contrast, glassy carbon exhibited a response variability of 39.1%. After 100 injections, the relative peak intensity, for diamond decreased by 10%, while a drastic decrease of 70% was observed for glassy carbon. The detection limit obtained in the FIA mode for 2,4-dichlorophenol was found to be 20 nM (S/N = 3), with a linear dynamic range up to 100 microM. By coupling with the column-switching technique, which enabled on-line preconcentration (50 times), the detection limit was lowered to 0.4 nM (S/N = 3). By use of this technique, anodized diamond electrodes were demonstrated for the analysis of CPs in drainwater that was condensed from the flue gas of waste incinerators.     

Electrochemical Oxidation of NADH at Highly Boron-Doped Diamond Electrodes

Conductive boron-doped chemical vapor-deposited diamond thin films, already known to have superior properties for general electroanalysis, including low background current and a wide potential window, are here shown to have additional advantages with respect to electrochemical oxidation of nicotinamide adenine dinucleotide (NADH), including high resistance to deactivation and insensitivity to dissolved oxygen. Cyclic voltammetry, amperometry, and the rotating disk electrode technique were used to study the reaction in neutral pH solution. Highly reproducible cyclic voltammograms for NADH oxidation were obtained at as-deposited diamond electrodes. The response was stable over several months of storage in ambient air, in contrast to glassy carbon electrodes, which deactivated within 1 h. The diamond electrode exhibited very high sensitivity for NADH, with an amperometric detection limit of 10 nM (S/N = 7). The response remained stable, even in the very low concentration range, for several months. In addition, interference effects due to ascorbic acid were minimal when the concentrations of NADH and ascorbic acid were comparable. An NADH-mediated dehydrogenese-based ethanol biosensor incorporating an unmodified diamond electrode is demonstrated. The present results indicate that diamond is a useful electrode material for the analytical detection of NADH, making it attractive for use in sensors based on enzyme-catalyzed reactions involving NADH as a cofactor.     

Integrated all-diamond ultramicroelectrode arrays: optimization of faradaic and capacitive currents

Integrated all-diamond ultramicroelectrode arrays (UMEAs) were fabricated using standard photolithography processes. The array consisted of typically 45 ultramicroelectrodes with a diameter of 10 μm and with a center-to-center spacing of 60 μm. The quasi-reference and counter electrodes were made from conductive diamond and were integrated on a 5 × 5 mm(2) chip. On the UMEA, a high ratio of faradaic current to capacitive current was achieved on heavily boron-doped and hydrogen-terminated diamond surfaces at slow scan rates and in high concentration of supporting electrolyte. A sensitive and reproducible detection of dopamine was achieved on hydrogen-terminated diamond UMEA at slow scan rates. The detection limit of dopamine in the presence of ascorbic acid was 1.0 nM, which is 50-100 times lower than that obtained on the macrosized boron-doped diamond electrodes. This array is promising for sensitive and reproducible detection of analytes in solutions with low detection limits.     

Electrochemical oxidation of oxalic acid at highly boron-doped diamond electrodes

Electrochemical oxidation of oxalic acid has been investigated at bare, highly boron-doped diamond electrodes. Cyclic voltammetry and flow injection analysis with amperometric detection were used to study the electrochemical reaction. Hydrogen-terminated diamonds exhibited well-defined peaks of oxalic acid oxidation in a wide pH range. A good linear response was observed for a concentration range from 50 nM to 10 microM, with an estimated detection limit of approximately 0.5 nM (S/N = 3). In contrast, oxygen-terminated diamonds showed no response for oxalic acid oxidation inside the potential window, indicating that surface termination contributed highly to the control of the oxidation reaction. An investigation with glassy carbon electrodes was conducted to confirm the surface termination effect on oxalic acid oxidation. Although a hydrogen-terminated glassy carbon electrode showed an enhancement of signal-to-background ratio in comparison with untreated glassy carbon, less stability of the current responses was observed than that at hydrogen-terminated diamond.     

Characterization of electrode fouling and surface regeneration for a platinum electrode on an electrophoresis microchip

A method of electrochemically cleaning noble metal electrodes is presented and characterized for electrophoresis microchips with electrochemical detection. First, the loss of sensitivity due to electrode fouling by serotonin is characterized as a function of injection number and analyte concentration. Signal attenuation is observed to be greater at high concentrations (100 microM) and negligible at very low concentrations (approximately 1 microM). Next, an electrochemical treatment procedure is optimized to yield sensitive and reproducible amperometric detection of the highly adsorptive compounds, serotonin and histamine. Thus, the performance of the electrode is reproducibly regenerated following as much as a approximately 99% reduction in surface activity. Utilizing the optimized three-level waveform, derived from that used for pulsed amperometric detection, detection limits as low as 78 nM and 17 microM have been obtained for serotonin and histamine, respectively. In the case of serotonin, this represents the lowest detection limit for a neurotransmitter by microchip electrophoresis with amperometric detection and the first report of amperometric detection of histamine detection at an unmodified platinum electrode. Repeated use of the electrode and application of electrochemical treatment did not appear to measurably affect the noise, longevity, metal adhesion, or physical appearance of the electrode.     

Electrochemical detection of arsenic(III) using iridium-implanted boron-doped diamond electrodes

Iridium-modified, boron-doped diamond electrodes fabricated by an ion implantation method have been developed for electrochemical detection of arsenite (As(III)). Ir+ ions were implanted with an energy of 800 keV and a dose of 10(15) ion cm(-2). An annealing treatment at 850 degrees C for 45 min in H2 plasma (80 Torr) was required to rearrange metastable diamond produced by an implantation process. Characterization was investigated by SEM, AFM, Raman, and X-ray photoelectron spectroscopy. Cyclic voltammetry and flow injection analysis with amperometric detection were used to study the electrochemical reaction. The electrodes exhibited high catalytic activity toward As(III) oxidation with the detection limit (S/N = 3), sensitivity, and linearity of 20 nM (1.5 ppb), 93 nA microM(-1) cm(-2), and 0.999, respectively. The precision for 10 replicate determinations of 50 microM As(III) was 4.56% relative standard deviation. The advantageous properties of the electrodes were its inherent stability with a very low background current. The electrode was applicable for analysis of spiked arsenic in tap water containing a significant amount of various ion elements. The results indicate that the metal-implanted method could be promising for controlling the electrochemical properties of diamond electrodes.     

Electrochemical determination of pharmaceuticals in spiked water samples

The electrochemical behaviour of acidic and neutral pharmaceutical active compounds (PhACs) was studied by cyclic voltammetry and pulse voltammetric techniques on mercury, carbon nanotube paste, carbon paste and gold electrodes. The best results, in terms of sensitivity, linearity range and detection limits, were obtained by differential pulse voltammetry (DPV) for ofloxacin (LOD 5.2 microM), differential pulse polarography (DPP) for clofibric acid (LOD 4.7 microM) and normal pulse voltammetry (NPV) for diclofenac (LOD 0.8 microM) and propranolol (LOD 0.5 microM). An enrichment step of approximately two orders of magnitude was performed by a solid-phase extraction procedure (SPE) in order to concentrate the samples. The developed method was optimized and tested on spiked river water samples.     

Monocrystalline diamond paste-based electrodes and their applications for the determination of Fe(II) in vitamins

A new class of electrochemical sensors, namely, electrodes based on diamond paste, was designed using monocrystalline diamond (natural diamond 1 microm and synthetic diamond, 50 microm (synthetic-1) and 1 microm (synthetic-2)) powder and paraffin oil. The characterization of the electrodes was performed using cyclic voltammetry and differential pulse voltammetry. Fe(II) was determined by differential pulse voltammetry (DPV) at 75 mV (vs Ag/AgCl) using all diamond paste-based electrodes. The linear concentration range was between 10(-8) and 10(-4) mol/L for both the natural diamond and synthetic-2 with detection limits of 10(-10) and 10(-9) mol/L, respectively, whereas the linear concentration range for synthetic-1 was between 10(-7) and 10(-3) mol/L with a detection limit of 10(-8) mol/L Fe(II) was determined successfully from four types of pharmaceutical products. The recovery values of Fe(II) in the pharmaceutical products were higher than 98.00% with relative standard deviation values < 5%.     

Application of diamond microelectrodes for end-column electrochemical detection in capillary electrophoresis

Highly boron-doped diamond microelectrodes were employed in an end-column electrochemical detector for capillary electrophoresis (CE). The diamond microline electrodes were fabricated from conducting diamond thin films (exposed surface area, 300 x 50 microm), and their analytical performance as CE detectors was evaluated in a laboratory-made CE installation. The CE-ED system exhibited high separation efficiency for the detection of several catecholamines, including dopamine (DA), norepinephrine (NE), and epinephrine (E), with excellent analytical performance, for example, 155,000 theoretical plates for DA. The diamond-based electrochemical detection system also displayed low detection limits (approximately 20 nM for E at S/N = 3) and a highly reproducible current response with 10 repetitive injections of mixed analytes containing DA, NE, and E (each 50 microM), with relative standard deviations (RSD) of approximately 5%. The performance of the diamond detector in CE was also evaluated in the detection of chlorinated phenols (CP). When compared to the carbon fiber microelectrode, the diamond electrode exhibited lower detection limits in an end-column CE detection resulting from very low noise levels and highly reproducible analyses without electrode polishing due to analyte fouling, which makes it possible to perform easier and more stable CE analysis.     

Direct electrochemical oxidation of disulfides at anodically pretreated boron-doped diamond electrodes

Anodically oxidized diamond electrodes have been used to oxidize disulfides, thiols, and methionine in aqueous acidic media and tested for amperometric detection of these compounds after chromatographic separation. Cyclic voltammetric signals for 1 mM glutathione disulfide (GSSG) were observed at 1.39 and 1.84 V vs SCE, the values being less positive than those of its as-deposited counterpart as well as glassy carbon electrode. The voltammetric and chronocoulometric results have indicated the high stability of the electrode with negligible adsorption. A positive shift in the peak potential with increasing pH indicated the attractive electrostatic interaction between the anodically oxidized diamond surface and the positively charged GSSG in acidic media that promoted its analytical performance. The results of the electrolysis experiments of disulfides and thiols showed that the oxidation reaction mechanism of glutathione (GSH) and GSSG involves oxygen transfer. Following separation by liquid chromatography (LC), the determination of both GSH and GSSG in rat whole blood was achieved at a constant potential (1.50 V vs Ag/AgCl), and the limits of detection for GSH and GSSG were found to be 1.4 nM (0.028 pmol) and 1.9 nM (0.037 pmol) with a linear calibration range up to 0.25 mM. These detection limits were much lower than those reported for the amperometry using Bi-PbO2 electrodes and LC-mass spectrometry, and the LC method using diamond electrodes were comparable with enzymatic assay in real sample analysis. The high response stability and reproducibility together with the possibility of regeneration of the electrode surface by on-line anodic treatment at 3 V for 30 min further support the applicability of anodically pretreated diamond for amperometric detection of disulfides.     

Simultaneous measurement of dopamine and ascorbate at their physiological levels using voltammetric microprobe based on overoxidized poly(1,2-phenylenediamine)-coated carbon fiber

Overoxidized poly-(1,2-phenylenediamine) (OPPD)-coated carbon fiber microelectrodes (CFMEs) exhibit, in combination with square-wave voltammetry (SWV) detection mode, the attractive ability to simultaneously measure low nM dopamine (DA) and mM ascorbate (AA) in a pH 7.4 medium. The PPD polymer film is electrodeposited onto a carbon fiber at a constant potential of 0.8 V versus Ag/AgCl using a solution containing sodium dodecylsulfate as the dopant. After overoxidation using cyclic voltammetry (CV) in the potential range from 0 to 2.2 V at a scan rate of 10 V/s, the resulting OPPD-CFME displays a high SWV current response to cationic DA at approximately 0.2 V and has a favorably low response to anionic AA at approximately 0.0 V vs Ag/AgCl. The preparation of the new OPPD-sensing film has been carefully studied and optimized. The OPPD properties and behavior were characterized using CV and SWV under various conditions and are discussed with respect to DA and AA detection. The linear calibration range for DA in the presence of 0.3 mM AA is 50 nM to 10 microM, with a correlation coefficient of 0.998 and a detection limit of 10 nM using 45-s accumulation. The detection limit for DA in the absence of AA was estimated to be 2 nM (S/N = 3). The linear range for AA in the presence of 100 nM DA is 0.2-2 mM, with a correlation coefficient of 0.999 and a detection limit of 80 microM. The reproducibilities of SWV measurements at OPPD-CFCMEs are 1.6% and 2.5% for 100 nM DA and 0.3 mM AA, respectively. Potential interfering agents, such as 3,4-dihydroxyphenylacetic acid, uric acid, oxalate, human serum proteins, and glucose, at their physiologically relevant or higher concentrations did not have any effect. These favorable features offer great promise for in vitro and in vivo application of the proposed OPPD-coated microprobe.     

Insulin oxidation and determination at carbon electrodes

The redox chemistry of insulin was investigated at glassy carbon (GC) electrodes that were coated with films of chitosan (CHIT) and multiwalled carbon nanotubes (CNT). While bare electrodes deactivated quickly during insulin oxidation, the GC electrodes coated with CHIT and CHIT-CNT films generated stable insulin currents. The GC/CHIT-CNT electrodes were used for investigating the electrooxidation process of insulin and amperometric determination of insulin. The mass spectrometric, electron paramagnetic resonance, and separation studies of electrolyzed insulin solutions suggested that the loss of 4 mass units upon insulin oxidation at CNT could be accounted for by the formation of two dityrosine cross-links intramolecularly. At a potential of 0.700 V and physiological pH 7.40, the GC/CHIT-CNT electrodes displayed a detection limit of approximately 30 nM insulin (S/N = 3), sensitivity of 135 mA M(-1) cm(-2), linear dynamic range from 0.10 to 3.0 microM (R2 = 0.995), and superior operational and long-term stability. The CNT-based electrodes are promising new insulin detectors for diabetes-related studies such as fast chromatographic analysis of therapeutic insulin formulations or evaluation of quality of pancreatic islets prior to their transplantation.     

Programmed potential sweep voltammetry for lower detection limits

We report a novel programmed potential sweep voltammetry for a much lower detection limit than those achieved by any other known electroanalyitcal techniques. In this technique, an input waveform is programmed such that the background current would become flat or any other predefined form in the potential region of interest where the peak current arising from the analyte is observed, followed by the amplification of the background subtracted peak current. The current thus obtained showed a much better signal integrity at very low analyte concentrations than those obtained by the traditional linear sweep voltammetric and other related voltammetric techniques. The technique was applied to the analysis of dopamine at a carbon ultramicroelectrode (10-microm diameter). The background-compensated currents showed excellent dynamic linearity for dopamine concentrations of more than 3 orders of magnitudes between 500 pM and 100 nM with an estimated detection limit of 127 pM. This method can provide a convenient way for determining biogenic amines in real time with a much higher sensitivity.     

Aptamer-based immunosensor on the ZnO nanorods networks

This paper presents the fabrication and characteristics of a new aptamer-based electrochemical immunosensor on the patterned zinc oxide nanorod networks (ZNNs) for detecting thrombin. Aptamers are single-stranded RNA or DNA sequence that binds to target materials with high specificity and affinity. An antibody-antigen-aptamer sandwich structure was employed to this immunosensor for detecting thrombin. First, hydrothermally grown ZNNs were patterned on the patterned 0.02 cm2 Au/Ti electrodes on a glass substrate by lift-off process. The high isoelectric point (IEP, approximately 9.5) of nanostructured ZnO makes it suitable for immobilizing proteins with low IEP. Then 5 microL of the 500 nM antibody was immobilized on the ZNNs electrode. 5 micro/L of the mixture of 1 microM aptamer labeled by ferrocene (Fc) and thrombin was dropped on the electrode for antibody-antigen binding. The peak oxidation currents of the immunosensors at various thrombin concentrations were measured by using cyclic voltammetry. The peak oxidation current was observed at 340 mV versus Ag/AgCl electrode, and the peak oxidation current increased linearly from 62.26 nA to 354.13 nA with the logarithmic concentration of thrombin in the range from 100 pM to 250 nM. Fabrication of an aptamer-based immunosensor for thrombin detection is a new attempt and the characteristics of the fabricated immunosensors showed that the fabricated aptamer-baded immunosensor worked electrochemically well and had a low detection limit (approximately 91.04 pM) and good selectivity.     

Selective voltammetric and amperometric detection of uric acid with oxidized diamond film electrodes

Electrochemically anodized diamond film electrodes were used for selective detection of uric acid (UA) in the presence of high concentrations of ascorbic acid (AA) by differential pulse voltammetry and chronoamperometry. Because the oxidation peak potential for AA is approximately 450 mV more positive than that for UA at anodized diamond electrodes, UA can be determined with very good selectivity. By use of chronoamperometry, linear calibration curves were obtained for UA over the concentration range up to 1 x 10(-6) M in 0.1 M HClO4 solution, with the lowest experimental value measured being 5 x 10(-8) M. This is consistent with the fact that a statistical analysis of the calibration curve yielded a detection limit of 1.5 x 10(-8) M (S/N = 3). AA in less than 20-fold excess does not interfere. The practical analytical utility of the method is demonstrated by the measurement of UA in human urine and serum without any preliminary treatment.     

Strategies for low detection limit measurements with cyclic voltammetry.

Cyclic voltammetry of Nafion-coated, carbon-fiber electrodes is used to detect trace concentrations of dopamine, both in a flow injection apparatus and in the brain of an anaesthetized rat. To improve signal-to-noise ratios, the sources of noise during cyclic voltammetry have been determined and strategies have been developed to decrease the noise. With the potentiostat employed, the measured noise is comparable to that expected for Johnson noise from the feedback resistor of the current transducer. Additional noise arises from the waveform generator employed and, in some cases, line noise. Line noise is discriminated against by starting each cyclic voltammogram either in phase or 180 degrees out of phase with the line frequency. When used in vivo, additional noise also arises from the physiological activity of the animal. Detection limits are found to closely correspond to those predicted on the basis of simulation of the voltammetric shape and the measured noise. Detection limits are improved by the use of appropriate analog and digital filtering, ensemble averaging, and appropriate timing of repetitive cyclic voltammograms. The combined use of these techniques enables the in vivo detection of approximately 100 nM of dopamine with a signal-to-noise ratio of 25.     

Fabrication and electrocatalysis of self-assembly directed gold nanoparticles anchored carbon nanotubes modified electrode

A self-assembly directed approach was adopted to modify glassy carbon electrode (GC) with gold nanoparticles incorporation and the electrocatalytic performance of self-assembly modified electrode, GC/SA-Au-ME was critically evaluated for the oxidation of ascorbic acid (AA). The modification involves the dispersion of multi-wall carbon nanotube (MWNT) and an inclusion complex, beta-cyclodextrin-4-aminothiophenol on the surface of GC electrode in the presence of cetyltrimethylammonium bromide (CTAB). Gold nanoparticles were deposited into the self-assembled sites to fabricate the modified electrode, GC/SA-Au-ME. Another electrode (GC-Au-ME) was fabricated under similar conditions in the absence of CTAB. The electrocatalytic activity of the modified electrodes (GC/SA-Au-ME and GC-Au-ME) towards the oxidation of AA was critically compared. Cyclic voltammetry, chronoamperometry, and double potential chronoamperometry were used to evaluate the characteristics of the modified electrodes. The self-assembled electrode (GC/SA-Au-ME) shows excellent electrocatalytic activity over the other electrode, GC-Au-ME. Augmented current response, faster electron transfer kinetics (with a rate constant for electron transfer process as 3.25 x 10(4) cm3 mol(-1) s(-1)), linear range of response for the analyte (1-50 mM with an extended detection limit to 1 microM), better sensitivity, and selectivity were witnessed for the self-assembly directed modified electrode.     

Reproducible fabrication of robust, renewable vertically aligned multiwalled carbon nanotube/epoxy composite electrodes

We describe the reproducible fabrication of robust, vertically aligned multiwalled carbon nanotube (VACNT)/epoxy composite electrodes. The electrodes are characterized by cyclic voltammetry, impedance spectroscopy, and scanning electron and atomic force microscopies. Low background currents are obtained at the electrodes, and common redox probe molecules and NADH show excellent voltammetric behavior. When electrode performance deteriorates due to fouling, the electrode surfaces can be reproducibly renewed by mechanical polishing followed by O(2) plasma treatment. The electrochemical performance of the electrodes is maintained after more than 100 cycles of use and renewal.     

Voltammetric determination of L-cysteine at conductive diamond electrodes

Boron-doped diamond (BDD) electrodes were used to examine L-cysteine (CySH) oxidation in alkaline media. The results of the voltammetric and polarization measurements showed that at BDD electrodes the overall CySH oxidation reaction is controlled by the initial electrochemical step, i.e., the oxidation of the CyS- electroactive species. The same conclusion was supported by the results of a study of pH effects. Conversely, at glassy carbon (GC) electrodes, the same reaction is controlled by the desorption of the reaction products. These results account for the poor response for CySH determination at GC compared to BDD. It was found that BDD exhibits excellent behavior for CySH determination, clearly outperforming GC. The results demonstrate that measurement of the peak current for CySH oxidation can be used as a basis for simple method for determining CySH in the micromolar concentration range by the use of BDD electrodes.     

Simultaneous determination of serotonin and dopamine at the PEDOP/MWCNTs-Pd nanoparticle modified glassy carbon electrode

Electrochemical determination of dopamine (DA) and serotonin (5-HT) have been studied at a modified glassy carbon electrode (GCE) in 0.1 M phosphate buffer solution (PBS) using cyclic voltammetry (CV) and differential pulse voltammetry (DPV) at pH 7.4, all over the interfering biomolecule ascorbic acid (AA). The GCE was modified by palladium-functionalized, multi-walled carbon nanotubes (MWCNTs-Pd) with electrochemical deposition of poly 3,4-ethylenedioxy pyrrole (PEDOP), denoted as PEDOP/MWCNTs-Pd/GCE, and investigated by SEM and EIS experiments. The highly electrocatalytic activity of the modified electrode toward 5-HT and DA was demonstrated from the sensitive and well-separated voltammetric experiment. The oxidation peaks found were 0.165 and 0.355 mV for DA and 5-HT, respectively. The composite film shows a significant accumulation effects on two species, as well as the mutual interference among the analytes. This biosensor was best in response compared to other modified electrodes made in the same lab. The lowest detection limits were found to be 5.0 x 10(-9) and 1.0 x 10(-8) for 5-HT and DA, respectively. The respective linear ranges were determined as 1.0 x 10(-7) to 2.0 x 10(-4) and 1.0 x 10(-7) to 2.0 x 10(-4) for 5-HT and DA.