Functional groups modulate the sensitivity and electron transfer kinetics of neurochemicals at carbon nanotube modified microelectrodes

The surface properties of carbon-based electrodes are critically important for the detection of biomolecules and can modulate electrostatic interactions, adsorption and electrocatalysis. Carbon nanotube (CNT) modified electrodes have previously been shown to have increased oxidative sensitivity and reduced overpotential for catecholamine neurotransmitters, but the effect of surface functionalities on these properties has not been characterized. In this study, we modified carbon-fiber microelectrodes (CFMEs) with three differently functionalized single-wall carbon nanotubes and measured their response to serotonin, dopamine, and ascorbic acid using fast-scan cyclic voltammetry. Both carboxylic acid functionalized and amide functionalized CNTs increased the oxidative current of CFMEs by approximately 2-6 fold for the cationic neurotransmitters serotonin and dopamine, but octadecylamine functionalized CNTs resulted in no significant signal change. Similarly, electron transfer was faster for both amide and carboxylic acid functionalized CNT modified electrodes but slower for octadecylamine CNT modified electrodes. Oxidation of ascorbic acid was only increased with carboxylic acid functionalized CNTs although all CNT-modified electrodes showed a trend towards increased reversibility for ascorbic acid. Carboxylic acid-CNT modified disk electrodes were then tested for detection of serotonin in the ventral nerve cord of a Drosophila melanogaster larva, and the increase in sensitivity was maintained in biological tissue. The functional groups of CNTs therefore modulate the electrochemical properties, and the increase in sensitivity from CNT modification facilitates measurements in biological samples.     

Microelectrode Sensing of Adenosine/Adenosine‐5′‐triphosphate with Fast‐Scan Cyclic Voltammetry

Adenosine and adenosine‐5′‐triphosphate (ATP) are important extracellular signaling molecules. Here, we studied adenosine and ATP using fast‐scan cyclic voltammetry at carbon‐fiber microelectrodes. Although ATP and adenosine have similar oxidation potentials, ATP oxidation current was highly dependent on buffer pH and divalent cation concentrations but adenosine current was not. Therefore, they can be distinguished by adding a divalent cation chelator or calibrating electrodes at different pH values. The enzymatic degradation of adenosine by adenosine deaminase was monitored in a mixture of adenosine and ATP in presence of EDTA (ethylenediaminetetraacetate). This sensing method is promising for enzyme kinetics or in vitro studies.     

Carbon-fiber microelectrodes modified with 4-sulfobenzene have increased sensitivity and selectivity for catecholamines

Elliptical and cylindrical geometries of carbon-fiber microelectrodes were modified by covalent attachment of 4-sulfobenzenediazonium tetrafluoroborate following its electroreduction. Elliptical electrodes fabricated from Thornel P-55 carbon fibers show the highest amount of 4-sulfobenzene attached to the electrode. Fast-scan cyclic voltammetry was used to compare the response to dopamine and other neurochemicals at these modified carbon-fiber microelectrodes. The grafted layer causes an increased sensitivity to dopamine and other positively charged analytes that is due to increased adsorption of analyte in the grafted layer. However, this layer remains permeable to negatively charged compounds. Modified electrodes retain the increased sensitivity for dopamine during measurements in mouse brain tissue.     

Carbon nanotube-modified microelectrodes for simultaneous detection of dopamine and serotonin in vivo

Dopamine and serotonin are important neurotransmitters that interact in the brain. While dopamine is easily detected with electrochemical sensors, the detection of serotonin is more difficult because reactive species formed after oxidation can adsorb to the electrode, reducing sensitivity. Carbon nanotube treatments of electrodes have been used to increase the sensitivity, promote electron transfer, and reduce fouling. Most methods have focused on nanotube coatings of large electrodes and slower electrochemical techniques that are not conducive to measurements in vivo. In this study, we investigated carbon-fiber microelectrodes modified with single-walled carbon nanotubes for the co-detection of dopamine and serotonin in vivo. Using fast-scan cyclic voltammetry, S/N ratios for the neurotransmitters increased after nanotube coating. Electrocatalytic effects of nanotubes were not apparent at fast scan rates but faster kinetics were observed with slower scanning. Nanotube-modified microelectrodes showed significantly less fouling after exposure to serotonin than bare electrodes. The nanotube-modified electrodes were used to monitor stimulated dopamine and serotonin changes simultaneously in the striatum of anesthetized rat after administration of a serotonin synthetic precursor. These studies show that nanotube-coated microelectrodes can be used with fast scanning techniques and are advantageous for in vivo measurements of neurotransmitters because of their greater sensitivity and resistance to fouling.     

Chitosan coated carbon fiber microelectrode for selective in vivo detection of neurotransmitters in live zebrafish embryos

We report the development of a chitosan modified carbon fiber microelectrode for in vivo detection of serotonin. We find that chitosan has the ability to reject physiological levels of ascorbic acid interferences and facilitate selective and sensitive detection of in vivo levels of serotonin, a common catecholamine neurotransmitter. Presence of chitosan on the microelectrode surface was investigated using scanning electron microscopy (SEM) and cyclic voltammetry (CV). The electrode was characterized using differential pulse voltammetry (DPV). A detection limit of 1.6 nM serotonin with a sensitivity of 5.12 nA/μM, a linear range from 2 to 100 nM and a reproducibility of 6.5% for n=6 electrodes were obtained. Chitosan modified microelectrodes selectively measure serotonin in presence of physiological levels of ascorbic acid. In vivo measurements were performed to measure concentration of serotonin in the live embryonic zebrafish intestine. The sensor quantifies in vivo intestinal levels of serotonin while successfully rejecting ascorbic acid interferences. We demonstrate that chitosan can be used as an effective coating to reject ascorbic acid interferences at carbon fiber microelectrodes, as an alternative to Nafion, and that chitosan modified microelectrodes are reliable tools for in vivo monitoring of changes in neurotransmitter levels.     

Electrochemical Preparation of Brilliant‐Blue‐Modified Poly(diallyldimethylammonium Chloride) and Nafion‐Coated Glassy Carbon Electrodes and Their Electrocatalytic Behavior Towards Oxygen and L‐Cysteine

Brilliant blue FCF‐modified glassy carbon electrodes have been prepared by cycling the Nafion (or poly(diallyldimethylammonium chloride) (PDDAC)) coated electrodes repeatedly 15 cycles in brilliant blue FCF (BB FCF) dye solution. The BB FCF molecules are incorporated into Nafion coating by cycling the film‐covered electrode between +0.3 to 1.2 V (vs. Ag/AgCl) in pH 1.5 BB FCF solution while PDDAC‐coated electrode cycled between 0 to ?1.0 V (vs. Ag/AgCl) in pH 6.5 BB FCF solution to immobilize the dye. Electrostatic interaction between dye molecule and PDDAC was predominant in PDDAC coating whereas immobilization of dye in Nafion film attributed to the combined effect of electrostatic and hydrophobic interactions. The voltammetric features of BB FCF‐modified electrodes resemble that of surface‐confined redox couples. The peak potentials of BB FCF‐incorporated PDDAC‐coated electrode were shifted to more positive potential region with decreasing pH of contacting solution. BB FCF‐modified electrodes showed electrocatalytic activity towards reduction of oxygen and oxidation of L ‐cysteine with significant decease of overvoltage compared to unmodified electrode. The BB FCF‐modified Nafion‐coated electrode was tested for its analytical applications toward determination of L ‐cysteine. The linear range of calibration plot at BB FCF‐modified Nafion‐coated electrode is 10 to 100 μM, which coincides with L ‐cysteine levels in biological fluids. Sensitivity and detection limit of the electrode are 111 nA μM^?1 and 0.5 μM, respectively.     

Comparison of electrode materials for the detection of rapid hydrogen peroxide fluctuations using background-subtracted fast scan cyclic voltammetry

Hydrogen peroxide (H(2)O(2)) is a critically important signaling molecule. Endogenous H(2)O(2) mediates diverse physiological processes both intra- and intercellularly; and enzymatically generated H(2)O(2) is a widely used reporter molecule at biosensors that rely on enzymes to detect non-electroactive species. However, the development and application of electroanalytical methods for the direct detection of this molecule has been challenging because the electron transfer kinetics for the irreversible oxidation of H(2)O(2) are slow. We comparatively characterize the electrochemical oxidation of H(2)O(2) on bare and Nafion(?)-coated platinum and carbon-fiber microdisc electrodes using fast-scan cyclic voltammetry (FSCV). Using a waveform ranging from +0.2 to +1.3 V at 400 V s(-1), the electrocatalytic properties of the platinum surface were not readily apparent, and the carbon-fiber microelectrode demonstrated greater sensitivity and selectivity toward H(2)O(2). Nafion(?)-coating further enhanced detection on carbon electrodes. These results confirm that platinum electrodes, with or without Nafion(?), will not work acceptably with this approach, and confirm the value of carbon-fiber microelectrodes relative to more traditionally used platinum electrodes in the direct detection of rapid H(2)O(2) fluctuations using FSCV.     

Electrooxidation of catecholamines at carbon nanotube-modified indium tin oxide electrodes

In this study, we prepared carbon nanotube (CNT)/Nafion-modified ITO electrodes and investigated their electrochemical behavior. The CNTs were dissolved in a solution of the ionic polymer Nafion and then CNT/Nafion composite films were deposited onto ITO electrodes through spin-coating of this homogeneous solution. We studied the effects of chemical pretreatment of the CNTs and the pH of the buffer on the electroanalytical behavior of the CNT/Nafion-modified ITO electrodes toward catecholamines. The modified electrodes enhanced the peak current and lowered the overpotentials. We observed high electrooxidative performance for the modified ITO electrodes: the oxidative currents of the catecholamines were up to 125-fold higher than those obtained using bare ITO electrodes.     

Influence of Nafion Coatings and Surfactant on the Stripping Voltammetry of Heavy Metals at Bismuth‐Film Modified Carbon Film Electrodes

Nafion polymer coated bismuth‐film‐modified carbon film electrodes have been investigated for reducing the influence of contaminants such as surfactants in the anodic stripping voltammetry of trace metal ions. The influence of the coating on electrode response has been tested with both ex situ and in situ bismuth film deposition, with and without the polymer coating. The electrode assemblies and interfacial characteristics in the presence of the non‐ionic surfactant Triton‐X‐100 have been probed with electrochemical impedance spectroscopy. The Nafion coating successfully decreases the adsorption of Triton on the bismuth film surface, and demonstrates that this strategy allows measurement of these trace metals in environmental samples containing surfactants.     

Outer-sphere oxidation of ascorbate with os(bpy)33+ incorporated in nafion coatings on graphite electrodes

The electro-oxidation of ascorbate proceeds very slowly at graphite electrodes coated with Nafion. Incorporation of Os(bpy)₃²⁺ (bpy = 2,2′-bipyridine) in the coating produces a catalyzed oxidation of ascorbate at potentials where Os(bpy)₃²⁺ is oxidized to Os(bpy)₃³⁺. Analysis of the kinetic data demonstrates that the reaction between catalyst (Os(bpy)₃³⁺) and substrate (ascorbate) proceeds only within the outermost layer of the coating at the coating/solution interface. As the substrate concentration is increased, the limiting oxidation currents at coated rotating disk electrodes do not increase as rapidly as expected on the basis of current models of the behavior of polymer coated electrodes. Some possible reasons for this deviant behavior of cast Nafion coatings are suggested. Some implications of the results on the general utility of Nafion-coated electrodes in electro-catalytic applications are presented.     

Single‐Walled Carbon Nanotubes on Tungsten Wires: A New Class of Microelectrochemical Sensors

This paper reports the fabrication and the outstanding performance characteristics of novel microelectrodes consisting of tungsten (W) wires coated with homogeneous layers of single‐walled C nanotubes (SWNT). A series of studies using cyclic voltammetry indicate that the SWNT‐modified W electrodes possess interesting electrochemical features. In fact, they are able to catalyse electron transfer reactions involving a series of inorganic and biological molecules. These electrodes are characterized by a fast electron transfer, a wide working potential window, and a low background current. Moreover they demonstrate excellent reproducibility, good stability in various chemical media, and very high sensitivity towards a series of inorganic and organic compounds. The SWNT modified microelectrodes have been tested for the capacity to electrochemically detect ferrocene monocarboxylic acid and potassium hexacyanoferrate as well of a series of interesting biological molecules which include catechol, caffeic acid, DOPAC, ascorbic acid, L ‐tyrosine, acetaminophen, guanine, uric acid, and the neurotransmitters dopamine, epinephrine, and serotonin (5‐HT) hydrochloride. The advantages of the SWNT‐modified W electrodes are illustrated by comparing their analytical performance with that of conventional electrodes.     

A Microchip‐Based System for Immobilizing PC 12 Cells and Amperometrically Detecting Catecholamines Released After Stimulation with Calcium

In this paper, we describe a microchip‐based system for amperometrically monitoring the amount of catecholamines released from rat pheochromocytoma (PC 12) cells. Key to this system is a novel, yet simple method for the immobilization of PC 12 cells in poly(dimethylsiloxane) (PDMS)‐based microchannels. The procedure involves selectively coating microchannels with collagen followed by introduction of PC 12 cells over the PDMS structure, with the cells being immobilized only on the coated portion of the channels. The cell‐coated microchannels can then be reversibly sealed to a glass plate containing electrodes for amperometric detection, resulting in an immobilized cell reactor with integrated microelectrodes. Nafion‐coated microelectrodes made by micromolding of carbon inks were used to measure calcium‐induced catecholamine release from the cells. Varying concentrations of PC 12 cells immobilized in the microchannels led to a catecholamine release ranging from 20 to 160 μM when the cells were stimulated with a calcium solution. This microchip approach leads to a three‐dimensional culture that can be used with this or other cells lines to study the effect of external stimuli on neurotransmitter release.     

Electrochemical behavior of methyl viologen at graphite electrodes modified with Nafion sol–gel composite

Electrodes were prepared by spin-coating spectroscopic graphite rods with a Nafion doped sol. Coating solutions consisting of Nafion:TEOS (tetraethoxysilane) ratios of 3:1 and 4:1 gave smooth films on the electrode surface. These modified electrodes were evaluated and compared with Nafion modified and bare spectroscopic graphite electrodes using methyl viologen (MV²⁺) as a representative cationic electroactive probe. Substantial partitioning of MV²⁺ into the Nafion:sol–gel matrix to the electrode surface was observed by cyclic voltammetry and square wave voltammetry. Cyclic voltammograms of MV²⁺ in 0.1 M NaCl at Nafion:sol–gel 4:1 modified electrodes showed a reversible reduction to MV⁺ with E^0′=−0.695 V vs. Ag/AgCl. Results of scan rate variation showed the wave to be characterized by semi-infinite diffusion for scan rates in the range 50–500 mV/s. Slowing the scan rate below 50 mV/s resulted in a transition to thin-layer behavior. MV²⁺ partitioned much more quickly into the sol–gel-Nafion modified electrodes compared to pure Nafion modified electrodes. Reversibility of the MV²⁺-loaded modified Nafion-doped sol–gel coatings on electrodes was obtained by soaking in 1 M NaCl solution. Concentration calibration plots for MV²⁺ at the sol–gel-Nafion modified electrodes were nonlinear. Substantial enhancement of current signal at low concentrations was observed by square wave voltammetry.     

Effect of the Dispersing Agent on the Electrochemical Response of Glassy Carbon Electrodes Modified with Dispersions of Carbon Nanotubes

The electrochemical response of a glassy carbon electrode modified with carbon nanotubes (CNT) dispersed in two solvents, water and DMF, and two polymers, chitosan and Nafion is reported. The films were homogeneous when the dispersing agent was water or DMF. In the case of polymers, the surfaces present areas with different density of CNTs. A more sensitive electrochemical response was obtained when CNTs are dispersed in the solvents. In the case of CNT dispersed with polymers, the nature of the polymer demonstrated to be a critical parameter not only for dispersing the nanotubes but also for the electrochemical activity of the resulting electrodes.     

Ring-modified carbon fiber microelectrodes and multi-microelectrode devices

Two new types of modified microelectrodes were used alone and in multi-microelectrode devices. Carbon fibers with diameters of 7.2 μm were modified either by electropolymerization, or by thermal polymerization of a mixture of monomers to give a thick coating of modifier around the cylindrical fiber. The modified fibers were then coated with an insulating layer. The tips of the electrodes were polished perpendicular to the axis of the fiber to give ring-modified disk electrodes. Copolymers of poly(vinylferrocene) and poly(vinylpyridine) with crosslinked polystyrene were used, and demonstrated behavior similar to that for surface modified electrodes, except that the electrodes could be polished to renew the surface. Multi-microelectrode devices were prepared. For example, a ring-modified working electrode, a silver/silver chloride coated practical reference electrode, and a platinum auxiliary electrode were used in a molded epoxy probe in electrolyte solution to give cyclic voltammograms that were similar to those expected for a ring-modified working electrode using traditional reference and auxiliary electrodes.     

Critical evaluation of electrode design and matrix effects on monitoring organophosphate pesticides using composite carbon nanotube-modified electrodes

Carbon nanotubes (CNT)/Nafion-modified glassy carbon (GC) electrodes were used to immobilize the enzyme acetylcholinesterase (AChE) by crosslinking with glutaraldehyde. The CNT-modified electrodes exhibited a sensitive and stable electrocatalytic behavior towards thiocholine (TCh). Compared to ordinary GC electrodes modified with Nafion, a substantial (500-mV) decrease in the overvoltage of the TCh oxidation reaction is observed, along with a tenfold enhancement in the amperometric response. The CNT/Nafion/AChE electrode has very good stability of at least a month compared to surfaces made without crosslinking in the absence and presence of Nafion. Under optimal loadings of CNT, Nafion, AChE, and glutaraldehyde, a solution of CNT/Nafion in N,N-dimethylformamide (DMF) containing 4 mg/mL CNT and 0.01% Nafion was used to construct the electrodes in order to maximize the sensitivity of the biosensor for inhibition studies. An optimal enzyme loading of 0.137 U and crosslinking in 0.01% glutaraldehyde for 1 h was also needed to achieve this goal. The prepared electrodes had very good reproducibility to 1.0 mM acetylthiocholine (ATCh) (relative standard deviation [RSD] <5% for eight electrodes). Using paraoxon as a model pesticide, the biosensor was able to detect as low as 1.0 nM after 30 min of incubation at 30 °C. Using a log scale, the biosensor had good linearity in the concentration range 50?C800 nM, with a correlation coefficient of 0.99. The prepared biosensor was used to test real water samples spiked with paraoxon and showed good correlation with a calibration curve using phosphate buffer.     

Silica‐modified Electrodes for Electrochemical Detection of Malachite Green

New silica‐modified glassy carbon electrodes prepared with three different sorts of ordered mesoporous silica (OMS) were characterized and tested for the electrochemical detection of Malachite Green (MG). The electrodes were prepared by drop casting using silica suspensions and, for stability sake, a Nafion coating was deposited on the electrode top by the same technique. Square wave anodic stripping voltammetry was used to investigate the effect of various experimental parameters (deposition time, solution pH, silica type and concentration) on the performance of the modified electrodes. The best electrode (GC/MCM‐41‐NH₂/Nafion) with detection limit 0.36 μM, sensitivity 0.164±0.003 A/M; linear domain 1–6 μM was applied to detect MG in a commercial product commonly used as biocide in aquaria for ornamental fish.     

Decreased ascorbate sensitivity with nafion-coated carbon fibre electrodes in combination with copper(II) ions for the electrochemical determination of electroactive substances in vivo

A Nafion coating is applied on voltammetric carbon fibre electrodes. This treatment lowered the sensitivity to ascorbate and 3,4-dihydroxyphenylacetic acid (DOPAC), and increased that for dopamine. Electrochemical oxidation with a 70-Hz, 3-V, anodic triangular-wave voltage was used for pretreatment before the Nafion coating. The inclusion of copper ions in the Nafion film further decreased the sensitivity to ascorbic acid, but did not significantly affect to dopamine or DOPAC. In-vitro equilibration with a buffer of appropriate copper concentration is recommended for pretreatment of carbon fibre electrodes before their use in in-vivo voltammetry.     

基于BCNTs/GC电极的鸟嘌呤与腺嘌呤电化学行为及其同时检测

利用掺硼碳纳米管（BCNTs）/GC电极研究了鸟嘌呤（G）和腺嘌呤（A）的电化学氧化行为. 与GC和CNTs/GC电极相比,BCNTs/GC电极具有更强的电催化活性,且响应电流明显增加. 两混合样品在BCNTs/GC电极上的氧化峰间隔较大,可实现对A和G的同时检测.     

Enhanced Hydrogen Peroxide Sensing Based on Tetraruthenated Porphyrins/Nafion/Glassy Carbon‐modified Electrodes via Incorporating of Carbon Nanotubes

The incorporation of carbon nanotubes to a Nafion/tetraruthenated cobalt porphyrin/ glassy carbon electrode (GC/Nf/CoTRP vs GC/Nf/CNTCoTRP) enhanced the amperometric determination of hydrogen peroxide. Both electrodes produced a decrease in the overpotential required for the hydrogen peroxide oxidation in about 100 mV compared to glassy carbon under the same experimental conditions. Nevertheless, for GC/Nf/CNT/CoTRP, the increase in the current is remarkable. The GC/Nf/CoTRP modified electrode gave no significant analitycal signal for hydrogen peroxide reduction. Moreover, a great increase in current is observed with GC/Nf/CNT/CoTRP at ?150mV which suggests a significant increase in the sensitivity of the modified electrode. Scanning electrochemical microscopy (SECM) revealed an enhancement in the electroactivity of the GC/Nf/CNT/CoTRP modified electrode. This fact has been explained in terms of enhanced homogeneity of the electrodic surface as a consecuence of better dispersibility of CNT‐CoTRP produced by a Nafion polyelectrolyte.