Electrochemical recognition and quantification of cytochrome c expression in Bacillus subtilis and aerobe/anaerobe Escherichia coli using N,N,N',N'-tetramethyl-para-phenylene-diamine (TMPD).

The colorimetric identification of pathogenic and non-pathogenic bacteria in cell culture is commonly performed using the redox mediator N,N,N',N'-tetramethyl-para-phenylene-diamine (TMPD) in the so-called oxidase test, which indicates the presence of bacterial cytochrome c oxidases. The presented study demonstrates the ability of electrochemistry to employ TMPD to detect bacteria and quantify the activity of bacterial cytochrome c oxidases. Cyclic voltammetry studies and chronoamperometry measurements performed on the model organism Bacillus subtilis result in a turnover number, calculated for single bacteria. Furthermore, trace amounts of cytochrome c oxidases were revealed in aerobically cultured Escherichia coli, which to our knowledge no other technique is currently able to quantify in molecular biology. The reported technique could be applied to a variety of pathogenic bacteria and has the potential to be employed in future biosensing technology.

Nanoelectrode array formation by electrolytic nanoparticle impacts.

We report the fabrication of functional nanoelectrode arrays by the electrolysis of AgBr nanoparticles (NPs) impacting on a glassy carbon electrode from suspension in aqueous solution. The impacted NPs result in Ag NP deposits of similar size to the originating NP, with the coverage of these arrays easily controlled by the time of the deposition step. The NPs constituting the array are deposited randomly across the surface with little aggregation or agglomeration. The fabricated arrays are themselves electrochemically active, mediating the reduction of hydrogen peroxide, H2O2.

Transient electrochemistry: beyond simply temporal resolution.

Some physicochemical intrigues for which transient electrochemistry was necessary to solve the problem are summarized in this feature article. First, we highlight the main constraints to be aware of to access to low time scales, and particularly focus on the effects of stray capacitances. Then, the electron transfer rate constant measured for redox molecules in a self-assembled monolayer configuration is compared to the conductance measured through the same systems, but at the single molecule level. This evidences strong conformational changes when molecules are trapped in the nanogap created between both electrodes. We also report about dendrimers, for which a short electrochemical perturbation induces creation of a diffusion layer within the molecule, allowing the electron hopping rate to be measured and analyzed in terms of molecular motions of the redox centers. Finally, we show that transient electrochemistry provides also useful information when coupled to other methodologies. For example, when an ultrasonic field drives very fast movements of a bubble situated above the electrode surface, the motion can be detected indirectly through a modification of the diffusion flux. Another field concerns pulse radiolysis, and we describe how the reactivity (at the electrode or within the solution) of radicals created by a radiolytic pulse can be quantified, widening the possibilities of electrochemistry to operate in biological media.

Exploring the mineral-water interface: reduction and reaction kinetics of single hematite (α-Fe2O3) nanoparticles.

In spite of their natural and technological importance, the intrinsic electrochemical properties of hematite (α-Fe2O3) nanoparticles are not well understood. In particular, particle agglomeration, the presence of surface impurities, and/or inadequate proton concentrations are major obstacles to uncover the fundamental redox activities of minerals in solution. These are particularly problematic when samples are characterized in common electrochemical analyses such as cyclic voltammetry in which nanoparticles are immobilized on a stationary electrode. In this work, the intrinsic reaction kinetics and thermodynamics of individual hematite nanoparticles are investigated by particle impact chronoamperometry. The particle radius derived from the integrated area of spikes recorded in a chronoamperogram is in excellent agreement with electron microscopy results, indicating that the method provides a quantitative analysis of the reduction of the nanoparticles to the ferrous ion. A key finding is that the suspended individual nanoparticles undergo electrochemical reduction at potentials much more positive than those immobilized on a stationary electrode. The critical importance of the solid/water interface on nanoparticle activity is further illustrated by a kinetic model. It is found that the first electron transfer process is the rate determining step of the reductive dissolution of hematite nanoparticles, while the overall process is strongly affected by the interfacial proton concentration. This article highlights the effects of the interfacial proton and ferrous ion concentrations on the reductive dissolution of hematite nanoparticles and provides a highly effective method that can be readily applied to study a wide range of other mineral nanoparticles.

In situ detection of salicylate in Ocimum basilicum plant leaves via reverse iontophoresis.

The quantitative analysis of salicylate provides useful information for the evaluation of metabolic processes in plants. We report a simple, noninvasive method to measure salicylate in situ in Ocimum basilicum leaves using reverse iontophoresis in combination with cyclic voltammetry at disposable screen-printed electrodes and the concentration of salicylate in basil leaves was found to be 3 mM.

Ion insertion into individual 7,7,8,8-tetracyanoquinodimethane nanoparticles.

We report the quantification of partial ion insertion into individual 7,7,8,8-tetracyanoquinodimethane nanoparticles. It is shown that both potassium and sodium ions can be inserted into single TCNQ nanoparticles from aqueous solution. The extent of both potassium and sodium insertion into individual nanoparticles is quantitatively measured and shown to be partial and sodium ion shows a higher extent of insertion. The insertion process is inferred to be limited and controlled by the formation of a thin shell of salt, Na(+)/K(+) TCNQ˙(-) formed at the surface of the nanoparticle.

Electrochemical detection of single micelles through 'nano-impacts'.

A new class of 'soft' particles, micelles, is detected electrochemically via 'nano-impacts' for the first time. Short, sharp bursts of current are used to indicate the electrical contact of a single CTAB (cetyltrimethylammonium bromide) micelle with an electrode via the oxidation of the bromide content. The variation in CTAB concentration for such 'nano-impact' experiments shows that a significant number of 'spikes' are observed above the CMC (critical micelle concentration) and this is attributed to the formation of micelles. A comparison with dynamic light scattering is also reported.

The selective electrochemical detection of homocysteine in the presence of glutathione, cysteine, and ascorbic acid using carbon electrodes.

The detection of homocysteine, HCys, was achieved with the use of catechol via 1,4-Michael addition reaction using carbon electrodes: a glassy carbon electrode and a carbon nanotube modified glassy carbon electrode. The selective detection of homocysteine was investigated and achieved in the absence and presence of glutathione, cysteine and ascorbic acid using cyclic voltammetry and square wave voltammetry. A calibration curve of homocysteine detection was determined and the sensitivity is (0.20 ± 0.02) µA µM(-1) and the limit of detection is 660 nM within the linear range. Lastly, commercially available multi walled carbon nanotube screen printed electrodes were applied to the system for selective homocysteine detection. This work presents a potential practical application towards medical applications as it can be highly beneficial towards quality healthcare management.

A disposable sticky electrode for the detection of commercial silver NPs in seawater.

The ability to perform efficient and affordable field detection and quantification of nanoparticles in aquatic environmental systems remains a significant technical challenge. Recently we reported a proof of concept of using 'sticky' electrodes for the detection of silver nanoparticles (Tschulik et al 2013 Nanotechnology 29 295502). Now a disposable electrode for detection and quantification of commercial Ag nanoparticles in natural seawater is presented. A disposable screen printed electrode is modified with cysteine and characterized by sticking and stripping experiments, with silver nanoparticle immobilization on the electrode surface and subsequent oxidative stripping, yielding a quantitative determination of the amount of Ag nanoparticles adhering to the electrode surface. The modified electrode was applied to natural seawater to mimic field-based environmental monitoring of Ag NPs present in seawater. The results demonstrated that commercial Ag NPs in natural seawater can be immobilized, enriched and quantified within short time period using the disposable electrodes without any need for elaborate experiments.

Electrochemical detection of commercial silver nanoparticles: identification, sizing and detection in environmental media.

The electrochemistry of silver nanoparticles contained in a consumer product has been studied. The redox properties of silver particles in a commercially available disinfectant cleaning spray were investigated via cyclic voltammetry before particle-impact voltammetry was used to detect single particles in both a typical aqueous electrolyte and authentic seawater media. We show that particle-impact voltammetry is a promising method for the detection of nanoparticles that have leached into the environment from consumer products, which is an important development for the determination of risks associated with the incorporation of nanotechnology into everyday products.

Effects of convergent diffusion and charge transfer kinetics on the diffusion layer thickness of spherical micro- and nanoelectrodes.

Nuances of the linear diffusion layer approximation are examined for slow charge transfer reactions at (hemi)spherical micro- and nanoelectrodes. This approximation is widely employed in Electrochemistry to evaluate the extent of electrolyte solution perturbed by the electrode process, which is essential to the understanding of the effects arising from thin-layer diffusion, convergent diffusion, convection, coupled chemical reactions and the double layer. The concept was well established for fast charge transfer processes at macroelectrodes, but remains unclear under other conditions such that a thorough assessment of its meaning was necessary. In a previous publication [A. Molina, J. González, E. Laborda and R. G. Compton, Phys. Chem. Chem. Phys., 2013, 15, 2381-2388] we shed some light on the influence of the reversibility degree. In the present work, the meaning of the diffusion layer thickness is investigated when very small electrodes are employed and so the contribution of convergent diffusion to the mass transport is very important. An analytical expression is given to calculate the linear diffusion layer thickness at (hemi)spherical electrodes and its behaviour is studied for a wide range of conditions of reversibility (from reversible to fully-irreversible processes) and electrode size (from macro- to nano-electrodes). Rigorous analytical solutions are deduced for true concentration profiles, surface concentrations, linear diffusion layer thickness and current densities when a potential pulse is applied at (hemi)spherical electrodes. The expressions for the magnitudes mentioned above are valid for electrodes of any size (including (hemi)spherical nanoelectrodes) and for any degree of reversibility, provided that mass transport occurs exclusively via diffusion. The variation of the above with the electrode size, applied potential and charge transfer kinetics is studied.

On the meaning of the diffusion layer thickness for slow electrode reactions.

A key concept underpinning electrochemical science is that of the diffusion layer - the zone of depletion around an electrode accompanying electrolysis. The size of this zone can be found either from the simulated or measured concentration profiles (yielding the 'true' diffusion layer thickness) or, in the case of the Nernst ('linear') diffusion layer by extrapolating the concentration gradient at the electrode surface to the distance at which the concentration takes its bulk value. The latter concept is very well developed in the case of fast (so-called reversible) electrode processes, however the study of the linear diffusion layer has received scant attention in the case of slow charge transfer processes, despite its study being of great interest in the analysis of the influence of different experimental variables which determine the electrochemical response. Analytical explicit solutions for the concentration profiles, surface concentrations and real and linear diffusion layers corresponding to the application of a potential step to a slow charge transfer process are presented. From these expressions the dependence of the diffusion layer thickness on the potential, pulse time, heterogeneous rate constant and ratio of bulk concentrations of electroactive species and of diffusion coefficients is quantified. A profound influence of the reversibility degree of the charge transfer on the diffusion layer thickness is clear, showing that for non-reversible processes the real and linear diffusion layers reveal a minimum thickness which coincides with the equilibrium potential of the redox couple in the former case and with the reversible half-wave potential in the latter one.

Direct electrochemical detection and sizing of silver nanoparticles in seawater media.

We report proof-of-concept measurements relating to the impact of nanoparticles with an electrode potentiostatted at a value corresponding to the diffusion controlled oxidation of silver nanoparticles in authentic seawater media. The charge associated with the oxidation reveals the number of atoms in the nanoparticle and thus its size and state of aggregation.

Square wave voltammetry at disc microelectrodes for characterization of two electron redox processes.

Analytical explicit solutions are presented for the use of square wave voltammetry (SWV) at disc microelectrodes to study two-electron reversible redox processes. This combines the advantages of SWV (minimization of capacitative effects, peak-shaped response and quick experiments) with those of microelectrodes (reduction of capacitative and ohmic drop effects, enhanced mass transport and measurements of small volumes). Further, the analytical expressions are very easy to implement in comparison with the numerical methods usually employed for simulation of electrochemical experiments at microdisc electrodes. From the theory, the effects of the technique parameters (frequency, pulse amplitude) are examined and procedures are given for the characterization of the redox system from the values of the peak current, peak potential and half-peak width. Finally, the theory is applied to the experimental study of the two-electron reduction of anthraquinone-2-sulfonate in aqueous media. For this system, the formal potentials of the redox centres in aqueous solutions can be tuned by means of the electrolyte cation.

Electrochemical formation and investigation of a self-assembled [60]fullerene monolayer.

The formation and characterisation of a C(60) monolayer at the electrode|electrolyte interface has been studied by cyclic voltammetry, potential step chronoamperometry and ac voltammetry. The presence of the monolayer is evidenced by the presence of a very sharp peak P in the voltammogram, attributed to the faradaic phase formation of an ordered monolayer, and of a reduction post peak Q associated with the reduction of adsorbed species. The chronoamperograms exhibit a well-defined maximum, characteristic of a nucleation and growth mechanism. By comparison with existing models of phase transitions, a progressive polynucleation and growth mechanism is demonstrated. The monolayer is proposed to consist of a 2D fulleride salt. It is suggested that the formation of the monolayer can take place for a broad range of solution compositions, but requires an atomically smooth substrate such as mercury.

Voltammetry in room temperature ionic liquids: comparisons and contrasts with conventional electrochemical solvents.

The recent literature is surveyed to explore the nature of voltammetry in room temperature ionic liquids. The extent of similarities with conventional electrochemical solvents is reported and some surprising differences are noted.

Effect of deposited bismuth on the potential of maximum entropy of Pt(111) single-crystal electrodes.

The effect of bismuth adsorption on the entropy of formation of the double layer on Pt(111) electrodes has been studied with the laser-induced temperature jump method. The coulostatic response to the temperature change induced by pulsed laser illumination allows the estimation of the sign and magnitude of the thermal coefficient of the potential drop at the interphase. This is related to the entropy of formation of the double layer, and the particular potential where this thermal coefficient becomes zero can be identified with the potential of maximum entropy of double-layer formation (pme). The effect of bismuth adsorption on the pme depends on the adatom coverage. At high coverages, a marked decrease of the pme is observed. This trend follows the change of the potential of zero charge expected from work function measurements, and it is likely due to the change in the orientation of solvent molecules induced by surface dipoles originated between the adatom and the substrate. At low coverage, the pme increases with the bismuth coverage. The disruption of the water structure due to the presence of the bismuth adatoms is tentatively proposed as the most likely explanation for this behavior.

Silver nanoparticle assemblies supported on glassy-carbon electrodes for the electro-analytical detection of hydrogen peroxide.

Electrochemical detection of hydrogen peroxide using an edge-plane pyrolytic-graphite electrode (EPPG), a glassy carbon (GC) electrode, and a silver nanoparticle-modified GC electrode is reported. It is shown, in phosphate buffer (0.05 mol L(-1), pH 7.4), that hydrogen peroxide cannot be detected directly on either the EPPG or GC electrodes. However, reduction can be facilitated by modification of the glassy-carbon surface with nanosized silver assemblies. The optimum conditions for modification of the GC electrode with silver nanoparticles were found to be deposition for 1 min at -0.5 V vs. Ag from 5 mmol L(-1) AgNO3/0.1 mol L(-1) TBAP/MeCN, followed by stripping for 2 min at +0.5 V vs. Ag in the same solution. A wave, due to the reduction of hydrogen peroxide on the silver nanoparticles is observed at -0.68 V vs. SCE. The limit of detection for this modified nanosilver electrode was 2.0 x 10(-6) mol L(-1) for hydrogen peroxide in phosphate buffer (0.05 mol L(-1), pH 7.4) with a sensitivity which is five times higher than that observed at a silver macro-electrode. Also observed is a shoulder on the voltammetric wave corresponding to the reduction of oxygen, which is produced by silver-catalysed chemical decomposition of hydrogen peroxide to water and oxygen then oxygen reduction at the surface of the glassy-carbon electrode.

Bioanalytical utility of sonovoltammetry.

Dopamine dissolved within egg homogenate was used as a model system to study the effects of electrode contamination and its subsequent reactivation through ultrasonically mediated in situ cleaning effects. The merits in conducting electroanalytical investigations under the influence of the ultrasonic field were also appraised. Maintaining the ultrasound field during oxidative measurements was found to yield hydrodynamic profiles that were linear over the concentration range 2-20 microM dopamine. The resulting sonolinear sweep voltammograms were compared with conventional rotating disk measurements, with the former found to provide significantly increased limiting currents that were attenuable through the manipulation of the field intensity. The problem of retaining selectivity in the presence of high concentrations of ascorbate was also assessed with the addition of cupric ion prior to commencing the measurements found to efficiently negate an otherwise substantive interference.