Electrochemiluminescence (ECL) biosensor based on tris(2,2'-bipyridyl)ruthenium(II) with glucose and lactate dehydrogenases encapsulated within alginate hydrogels.

Two dehydrogenase enzymes (glucose, GDH, and lactate, LDH, dehydrogenases) encapsulated within alginate hydrogels were deposited on glassy carbon electrodes. The as-prepared enzyme modified alginate hydrogels were utilized as electrochemiluminescence (ECL)-based biosensors for the indirect detection of glucose and lactic acid upon reaction between NADH and tris(2,2'-bipyridyl) ruthenium (II) [Ru(bpy)3]2+. The ECL response was obtained from the redox reaction between the substrate, the cofactor NAD+ and the encapsulated enzyme. The production of NADH resulting from the enzymatic reaction led to the ECL emission upon reaction with [Ru(bpy)3]2+. The biosensors showed good stability and repeatability, with linear range between 0.56 and 4.2 µM and limit of detection of 0.84 µM for glucose, and linear range between 5 and 30 µM with a limit of detection of 2.52 µM for lactic acid. These ECL-based biosensors showed good sensitivity when tested in the presence of common interfering species. These biosensors were utilized in artificial sweat and were characterized by good reproducibility and repeatability. The results herein presented suggest that the dehydrogenases encapsulated within alginate hydrogels have potential for the development of biocompatible sensors for detection of glucose and lactic acid in physiological fluids.

Enzymes Encapsulated within Alginate Hydrogels: Bioelectrocatalysis and Electrochemiluminescence Applications.

A simple procedure to incorporate enzymes (horseradish peroxidase, HRP, and lactate oxidase, LOx) within alginate hydrogels is reported with electrochemiluminescence (ECL) used to detect the enzymatic reactions with the corresponding substrates. First, HRP and LOx were successfully immobilized into CaCO3 microspheres, followed by the electrostatic layer-by-layer deposition of a nanoshell onto the microspheres, and finally by their dispersion into alginate solution. The as-prepared dispersion was drop cast onto the glassy carbon electrodes and cross-linked by the external and internal gelation methods using Ca2+ cations. The enzymes encapsulated within the alginate hydrogels were characterized using cyclic voltammetry and kinetic studies performed using ECL. The results showed that the enzymatic activity was significantly maintained as a result of the immobilization, with values of the apparent Michaelis-Menten constants estimated as 7.71 ± 0.62 and 8.41 ± 0.43 µM, for HRP and LOx, respectively. The proposed biosensors showed good stability and repeatability with an estimated limit of detection of 5.38 ± 0.05 and 0.50 ± 0.03 µM for hydrogen peroxide and lactic acid, respectively. The as-prepared enzymes encapsulated within the alginate hydrogels showed good stability up to 28 days from their preparation. The sensitivity and selectivity of the enzymes encapsulated within the alginate hydrogels were tested in real matrices (HRP, hydrogen peroxide, in contact lens solution; LOx, lactic acid in artificial sweat) showing the sensitivity of the ECL detection methods for the detection of hydrogen peroxide and lactic acid in real samples.

Application of Graphene Nanoplatelets in Supercapacitor Devices: A Review of Recent Developments.

As worldwide energy consumption continues to increase, so too does the demand for improved energy storage technologies. Supercapacitors are energy storage devices that are receiving considerable interest due to their appealing features such as high power densities and much longer cycle lives than batteries. As such, supercapacitors fill the gaps between conventional capacitors and batteries, which are characterised by high power density and high energy density, respectively. Carbon nanomaterials, such as graphene nanoplatelets, are being widely explored as supercapacitor electrode materials due to their high surface area, low toxicity, and ability to tune properties for the desired application. In this review, we first briefly introduce the theoretical background and basic working principles of supercapacitors and then discuss the effects of electrode material selection and structure of carbon nanomaterials on the performances of supercapacitors. Finally, we highlight the recent advances of graphene nanoplatelets and how chemical functionalisation can affect and improve their supercapacitor performance.

Photo- and Electrochemical Dual-Responsive Iridium Probe for Saccharide Detection.

Dual detection systems are of interest for rapid, accurate data collection in sensing systems and in vitro testing. We introduce an IrIII complex with a boronic acid receptor site attached to the 2-phenylpyridine ligand as an ideal probe with photo- and electrochemical signals that is sensitive to monosaccharide binding in aqueous solution. The complex displays orange luminescence at 618 nm, which is reduced by 70 and 40 % upon binding of fructose and glucose, respectively. The electro-chemiluminescent signal of the complex also shows a direct response to monosaccharide binding. The IrIII complex shows the same response upon incorporation into hydrogel matrices as in solution, thus demonstrating the potential of its integration into a device, as a nontoxic, simple-to-use tool to observe sugar binding over physiologically relevant pH ranges and saccharide concentrations. Moreover, the complex's luminescence is responsive to monosaccharide presence in cancer cells.

Voltammetry at Hexamethyl-P-Terphenyl Poly(Benzimidazolium) (HMT-PMBI)-Coated Glassy Carbon Electrodes: Charge Transport Properties and Detection of Uric and Ascorbic Acid.

We describe the voltammetric behavior of an anion-exchange membrane, hexamethyl-p-terphenyl poly(benzimidazolium) (HMT-PMBI). The anion-exchange properties of HMT-PMBI chemically modified electrodes were investigated using K4Fe(CN)6 and K2IrCl6 as redox probes. The permselectivity properties of HMT-PMBI chemically modified electrodes were ascertained using tris(2-2')bipyridyl-ruthenium(II) chloride Ru(bpy)32+. Cyclic voltammetry and chronoamperometry were utilized to extract parameters such as the concentration of the redox mediators inside the films and the apparent diffusion coefficients. We found the concentration of K4Fe(CN)6 and K2IrCl6 redox species within HMT-PMBI-coated films to be on the order of 0.04-0.1 mol·dm-3, and values of Dapp ca. 10-10-10-9 cm2·s-1. To evaluate the possibility of using such a polymer coating in electroanalysis, HMT-PMBI-modified electrodes were utilized for the voltammetric detection of uric acid in artificial urine, Surine® and ascorbic acid in Vitamin C samples. The results showed that HMT-PMBI-coated electrodes can detect uric acid in Surine® with a limit of detection (LoD) of 7.7 µM, sensitivity of 0.14 µA·µM-1·cm-2, and linear range between 5 µM and 200 µM, whereas for Vitamin C tablets, the LoD is 41.4 µM, the sensitivity is 0.08 µA·µM-1·cm-2, and the linear range is between 25 µM and 450 µM.

Voltammetric Detection of Caffeine in Beverages at Nafion/Graphite Nanoplatelets Layer-by-Layer Films.

We report for the first time a procedure in which Nafion/Graphite nanoplatelets (GNPs) thin films are fabricated using a modified layer-by-layer (LbL) method. The method consists of dipping a substrate (quartz and/or glassy carbon electrodes) into a composite solution made of Nafion and GNPs dissolved together in ethanol, followed by washing steps in water. This procedure allowed the fabrication of multilayer films of (Nafion/GNPs)n by means of hydrogen bonding and hydrophobic‒hydrophobic interactions between Nafion, GNPs, and the corresponding solid substrate. The average thickness of each layer evaluated using profilometer corresponds to ca. 50 nm. The as-prepared Nafion/GNPs LbL films were characterized using various spectroscopic techniques such as X-ray photoelectron spectroscopy (XPS), energy-dispersive X-ray spectroscopy (EDS), FTIR, and optical microscopy. This characterization highlights the presence of oxygen functionalities that support a mechanism of self-assembly via hydrogen bonding interactions, along with hydrophobic interactions between the carbon groups of GNPs and the Teflon-like (carbon‒fluorine backbone) of Nafion. We showed that Nafion/GNPs LbL films can be deposited onto glassy carbon electrodes and utilized for the voltammetric detection of caffeine in beverages. The results showed that Nafion/GNPs LbL films can achieve a limit of detection for caffeine (LoD) of 0.032 µM and linear range between 20‒250 µM using differential pulse voltammetry, whereas, using cyclic voltammetry LoD and linear range were found to be 24 µM and 50‒5000 µM, respectively. Voltammetric detection of caffeine in beverages showed good agreement between the values found experimentally and those reported by the beverage producers. The values found are also in agreement with those obtained using a standard spectrophotometric method. The proposed method is appealing because it allows the fabrication of Nafion/GNPs thin films in a simple fashion using a single-step procedure, rather than using composite solutions with opposite electrostatic charge, and also allows the detection of caffeine in beverages without any pre-treatment or dilution of the real samples. The proposed method is characterized by a fast response time without apparent interference, and the results were competitive with those obtained with other materials reported in the literature.

Analytical applications of nanomaterials in electrogenerated chemiluminescence.

This critical review covers the use of carbon nanomaterials (single-wall carbon nanotubes, multi-wall carbon nanotubes, graphene, and carbon quantum dots), semiconductor quantum dots, and composite materials based on the combination of the aforementioned materials, for analytical applications using electrogenerated chemiluminescence. The recent discovery of graphene and related materials, with their optical and electrochemical properties, has made possible new uses of such materials in electrogenerated chemiluminescence for biomedical diagnostic applications. In electrogenerated chemiluminescence, also known as electrochemiluminescence (ECL), electrochemically generated intermediates undergo highly exergonic reactions, producing electronically excited states that emit light. These electron-transfer reactions are sufficiently exergonic to enable the excited states of luminophores, including metal complexes, quantum dots and carbon nanocrystals, to be generated without photoexcitation. In particular, this review focuses on some of the most advanced and recent developments (especially during the last five years, 2010-2014) related to the use of these novel materials and their composites, with particular emphasis on their use in medical diagnostics as ECL immunosensors.

Nanomaterials for biosensing with electrochemiluminescence (ECL) detection.

Analytical applications of nanomaterials used in electrochemiluminescence (ECL)-based detection methods are reviewed. Among nanomaterials, carbon-based nanomaterials (carbon nanotubes, graphene), metal nanoparticles, quantum dots, inorganic metal complexes and conducting polymers are considered. The most common mechanisms of ECL detections are also described in this review. Finally, challenges and perspectives of the use of such materials in chemical analysis are discussed.

Enhanced electrochemiluminescence and charge transport through films of metallopolymer-gold nanoparticle composites.

Water-soluble 4-(dimethylamino) pyridine (DMAP) stabilized gold nanoparticles (DMAP-AuNP) were synthesized by ligand exchange and phase transfer (toluene/water). The DMAP-AuNPs are positively charged with the core diameter of 4 +/- 1 nm. Metallopolymer-gold nanocomposites were prepared by mixing gold nanoparticles and [Ru(bpy)(2)PVP(10)](ClO(4))(2), in water at different mole ratios; bpy is 2,2'-bipyridyl and PVP is poly (4-vinylpyridine). The photoluminescence emission intensity of the metallopolymer decreases with increasing AuNP loading and approximately 57% of the emission intensity is quenched when the Au NP:Ru mole ratio is 14.8 x 10(-2). The rate of homogeneous charge transfer through thin layers of the nanocomposite deposited on glassy carbon electrodes increases with increasing nanoparticle loading. The homogeneous charge transport diffusion coefficient, D(CT), for the composite (AuNP:Ru mole ratio 13.2 x 10(-2)) is (2.8 +/- 0.8) x 10(-11) cm(2) s(-1) and is approximately 3-fold higher than that found for the pure metallopolymer. Significantly, despite the ability of the metal nanoparticles to quench the ruthenium-based emission, the electrochemiluminescence of the nanocomposite with a AuNP:Ru mole ratio of 4.95 x 10(-2) is approximately three times more intense than the parent metallopolymer. This enhancement arises from the increased rate of charge transport that leads to a greater number of excited states per unit time while minimizing the quenching effects. The implications of these findings for the design of electrochemiluminescent sensors are discussed.

Nanostructured materials for electrochemiluminescence (ECL)-based detection methods: recent advances and future perspectives.

This review presents a general picture of the last advances and developments (2003-2008) related to novel nanostructured materials for electrochemiluminescence-based biosensors using. It briefly covers the basic mechanisms of ECL detection, and the recent developments in fabrication of solid-state ECL sensors using nanostructured materials such as carbon nanotubes, metal nanoparticles, quantum dots, thin films of metallopolymers and of inorganic metal complexes. Finally, challenges and perspectives of the use of such materials for biomedical diagnostics are discussed.

Electrogenerated chemiluminescence.

In electrogenerated chemiluminescence, also known as electrochemiluminescence (ECL), electrochemically generated intermediates undergo a highly exergonic reaction to produce an electronically excited state that then emits light. These electron-transfer reactions are sufficiently exergonic to allow the excited states of luminophores, including polycyclic aromatic hydrocarbons and metal complexes, to be created without photoexcitation. For example, oxidation of [Ru(bpy)(3)](2+) in the presence of tripropylamine results in light emission that is analogous to the emission produced by photoexcitation. This review highlights some of the most exciting recent developments in this field, including novel ECL-generating transition metal complexes, especially ruthenium and osmium polypyridine systems; ECL-generating monolayers and thin films; the use of nanomaterials; and analytical, especially clinical, applications.

Nafion-tris(2-2'-bipyridyl)ruthenium(II) Ultrathin Langmuir-Schaefer films: redox catalysis and electrochemiluminescent properties.

A simple procedure to incorporate tris(2-2'-bipyridyl)ruthenium(II), [Ru(bpy)3]2+, into Nafion Langmuir-Schaefer (LS) films is described. Nafion LS films (tens of nanometers thick) were formed on quartz glass and indium tin oxide (ITO) directly from Nafion-[Ru(bpy)3]2+ Langmuir films assembled at the water-air interface. This procedure allowed the direct incorporation of [Ru(bpy)3]2+ into Nafion films without the need for subsequent loading. UV-vis spectroscopy confirmed the successful incorporation of [Ru(bpy)3]2+ within the LS films and showed that the amount of [Ru(bpy)3]2+ immobilized in this way scaled with film thickness. Voltammetric studies on ITO-modified electrodes confirmed the successful incorporation of [Ru(bpy)3]2+ and demonstrated that [Ru(bpy)3]2+ was retained within the ultrathin films over a long time scale. These electrodes were tested for the electrocatalytic reduction of tripropylamine. Significant catalysis was observed due to the rapid turnover of [Ru(bpy)3]2+/3+ between the electrode surface and outer boundary of the film, as a direct consequence of the ultrathin film dimensions. Concomitant electrochemiluminescence (ECL) was demonstrated highlighting the potential of this material for sensing applications.

Formation and evaluation of electrochemically-active ultra-thin palladium-Nafion nanocomposite films.

A simple method for producing electrochemically-active palladium nanoparticles within ultra-thin Nafion films is described.

One-step formation of ultra-thin chemically functionalized redox-active Langmuir-Schaefer Nafion films.

A novel "one-step procedure" to incorporate different cationic redox-active species within ultra-thin Nafion films is reported. Ultra-thin films of Nafion containing trimethylammonioferrocene (FA), tris(2,2'-bipyridyl)ruthenium(), Ru(bpy), and hexaaminoruthenium(), [Ru(NH)], were prepared using the Langmuir-Schaefer (LS) technique. The morphology and thickness of the films were evaluated with atomic force microscopy (AFM). Film thicknesses were 1.9 ± 0.7, 1.8 ± 0.9, 1.5 ± 0.9 nm per layer for Nafion-FA, Nafion-Ru(bpy)and Nafion-[Ru(NH)], respectively. Electrochemical data confirmed the successful incorporation of the mediators into the films and proved that the redox activity was maintained during extensive voltammetric cycling. Key parameters such as surface coverage, concentration of the mediator within the films and apparent diffusion coefficients were extracted using cyclic voltammetry. To demonstrate the utility of the redox-active films for sensing applications, films with Ru(bpy) incorporated were utilized for the electrocatalysis of oxalate in aqueous solution.

Measurement of apparent diffusion coefficients within ultrathin nafion Langmuir-Schaefer films: comparison of a novel scanning electrochemical microscopy approach with cyclic voltammetry.

The use of scanning electrochemical microscopy (SECM) to evaluate the apparent diffusion coefficient, Dapp, of redox-active species in ultrathin Nafion films is described. In this technique, an ultramicroelectrode (UME) tip, positioned close to a film on a macroscopic electrode, is used to oxidize (or reduce) a species in bulk solution, causing the tip-generated oxidant (reductant) to diffuse to the film/solution interface. The oxidation (reduction) of film-confined species regenerates the reductant (oxidant) in solution, leading to feedback to the UME. A numerical model is developed that allows Dapp to be determined. For these studies, ultrathin films of Nafion were prepared using the Langmuir-Schaefer (LS) technique and loaded with an electroactive species, either the ferrocene derivative ferrocenyltrimethylammonium cation, FA+, or tris(2,2'-bipyridyl)ruthenium(II), Ru(bpy)32+. The morphology and the thickness of the Nafion LS films (1.5 +/- 0.2 nm per layer deposited) were evaluated using atomic force microscopy (AFM). For comparison with the SECM measurements, cyclic voltammetry (CV) was employed to evaluate the concentration of electroactive species within the Nafion LS films and to determine Dapp. The latter was found to be essentially invariant with film thickness, but the value for Ru(bpy)32+ was 1 order of magnitude larger than for FA+. CV and SECM measurements yield different values of Dapp, and the underlying reasons are discussed. In general, the Dapp values for these films are considerably smaller than for recast Nafion films, which can be attributed to the compactness of Nafion LS films. Nonetheless, the ultrathin nature of the films leads to fast response times, and we thus expect that these modified electrodes could find applications in sensing, electroanalysis, and electrocatalysis.

Adsorption dynamics and electrochemical and photophysical properties of thiolated ruthenium 2,2'-bipyridine monolayers.

A new complex [Ru(bpy)(2)(bpySH)](PF(6))(2), RuBpySH, has been prepared bearing two anchoring groups for surface attachment, where bpy is 2,2'-bipyridyl and bpySH is 5,5'-bis(mercaptomethyl)-2,2'-bipyridine. Monolayers of RuBpySH have been formed on micro and macro platinum electrodes by spontaneous adsorption from micromolar solutions of the complex in 50:50 v/v water/acetone. The monolayers can be reversibly switched between the Ru(2+) and the Ru(3+) forms. Cyclic voltammetry is well-defined with a peak-to-peak splitting of 30 +/- 5 mV and a full width at half-maximum of 110 +/- 10 mV being observed for scan rates up to 5 V s(-1) where the supporting electrolyte is 0.1 M tetrabutylammonium tetrafluoroborate in acetonitrile. Adsorption is irreversible in this system, and the saturation coverage obtained is 8.1 +/- 0.4 x 10(-11) mol cm(-2) when the complex concentration in the deposition solution is between 10 microM and 1.0 mM. The dynamics of adsorption depend markedly on the bulk concentration and are described in terms of irreversible adsorption. Dry monolayers display luminescence properties similar to those of powder samples of the complex, indicating that the monolayer has characteristics of the solid-state sample rather than the solution sample of the complex. Significantly, efficient electrochemiluminescence is generated using tripropylamine as the coreactant. The rate of electron transfer across the electrode/monolayer interface has been probed using high scan rate cyclic voltammetry. The standard heterogeneous electron-transfer rate constant, k degrees , is 0.9 +/- 0.1 x 10(4) s(-1), and there is weak adsorbate-electrode electronic communication.

Electrochemistry of cytochrome c incorporated in Langmuir-Blodgett films of Nafion and Eastman AQ 55.

Ultrathin films of Nafion and Eastman-AQ 55 loaded with cytochrome c (cyt c) were obtained and transferred on indium tin oxide (ITO) electrodes via the Langmuir-Blodgett (LB) technique. The pressure-area isotherms for mixed ionomer-protein films indicate that the miscibility of cyt c in the interfacial layer is better for Nafion than for AQ 55. Interestingly, these composite films maintain the electroactivity of cyt c without requiring the addition of promoters or mediators. Both for AQ 55-cyt c and Nafion-cyt c films, the half-wave potential for the reversible reduction of ferricytochrome c corresponds to the value expected for the weakly adsorbed protein. The modified electrodes show electrocatalytic reaction with ascorbate anion. Comparison with previous literature reports indicate that for Nafion the LB coating procedure is unique in keeping the electroactivity of cyt c.

Langmuir--Schaefer films of Nafion with incorporated TiO(2) nanoparticles.

An easy method of incorporating TiO(2) nanoparticles into Nafion perfluorinated ionomer is proposed. Ultrathin films of Nafion were prepared by employing the Langmuir-Schaefer (LS) technique. The pressure-area isotherm study of a Langmuir monolayer of Nafion at the air-water interface on different concentrations of NaCl as the subphase allowed us to find the best experimental conditions for the deposition of stable Langmuir-Schaefer films. Incorporation of TiO(2) nanoparticles was performed by dipping Nafion LS films in a solution of TiO(2) nanoparticles. The uniformity of the TiO(2) incorporation was detected by UV-visible spectroscopy. The morphology of the Nafion, Nafion/TiO(2) nanoparticles thin films, and the changes due to the annealing procedure were investigated by atomic force microscopy. Interestingly, the AFM investigation showed that Nafion and Nafion/TiO(2) LS films have thermal stability up to 600 degrees C.

Bacteriorhodopsin-based Langmuir-Schaefer films for solar energy capture.

The photovoltaic (PV) solar cell, converting incident solar radiation directly into electrical energy, today represents the most common power source for the earth-orbiting spacecraft, and the utilization of organic materials in this context is here explored in comparison with the present state of the art placing emphasis in organic nanotechnology. Poly[3-3'(vinylcarbazole)] (PVK) was synthesized by oxidative polymerization with ferric chloride of N-vinylcarbazole. The resulting polymer was then deposited on solid support by using the Langmuir-Schaefer (LS) technique. The pressure-area isotherm of PVK revealed the possibility of compact monolayer formation at the air-water interface. Different layers of PVK were doped with iodine vapors. The cyclic voltammetry investigation of PVK-doped I2 showed a distinctive electrochemical behavior. The photoinduced charge transfer across a donor/acceptor (D/A) hybrid interface provided an effective method to study the PV properties of the composite LS films. The results are compared with other approaches within the biological framework, such as bacteriorhodopsin (BR), and organic nanostructured materials.