Binary and Ternary Deep Eutectic Solvents for Methylene Green Electropolymerization on Multiwalled Carbon Nanotubes: Optimization, Characterization and Application.

Choline chloride (ChCl) based binary and ternary deep eutectic solvents (DES) were evaluated for methylene green electropolymerization with oxalic acid (OA) and ethylene glycol (EG) as hydrogen bond donors. Binary DES ChCl:OA in molar ratios 1:1 and 2:1 and ChCl:EG 1:2 and ternary DES (tDES) in different molar ratios and percentages of water were evaluated. The highest polymer growth was in ChCl:OA:EG-tDES with added water, that had a lower viscosity and higher ionic conductivity when associated with HCl as dopant. This enhanced the formation of more cation radicals and, consequently, more polymer formation. The PMG/MWCNT/GCE-tDES sensor was successfully applied to the simultaneous determination of 5-aminosalicylic acid (5-ASA) and acetaminophen (APAP) by differential pulse voltammetry in the concentration range 2 µM - 200 µM, with detection limits of 0.37 µM and 0.49 µM for 5-ASA and APAP, respectively. The sensor demonstrated good repeatability, reproducibility and stability, and was successfully applied in pharmaceutical formulations.

Fluorinated Mn(III)/(II)-Porphyrin with Redox-Responsive 1 H and 19 F Relaxation Properties.

A new fluorinated manganese porphyrin, (Mn-TPP-p-CF3 ) is reported capable of providing, based on the Mn(III)/Mn(II) equilibrium, dual 1 H relaxivity and 19 F NMR response to redox changes. The physical-chemical characterization of both redox states in DMSO-d6 /H2 O evidenced that the 1 H relaxometric and 19 F NMR properties are appropriate for differential redox MRI detection. The Mn(III)-F distance (dMn-F =9.7-10 Å), as assessed by DFT calculations, is well tailored to allow for adequate paramagnetic effect of Mn(III) on 19 F T1 and T2 relaxation times. Mn-TPP-p-CF3 has a reversible Mn(II)/Mn(III) redox potential of 0.574 V vs. NHE in deoxygenated aqueous HEPES/ THF solution. The reduction of Mn(III)-TPP-p-CF3 in the presence of ascorbic acid is slowly, but fully reversed in the presence of air oxygen, as monitored by UV-Vis spectrometry and 19 F NMR. The broad 1 H and 19 F NMR signals of Mn(III)-TPP-p-CF3 disappear in the presence of 1 equivalent ascorbate replaced by a shifted and broadened 19 F NMR signal from Mn(II)-TPP-p-CF3 . Phantom 19 F MR images in DMSO show a MRI signal intensity decrease upon reduction of Mn(III)-TPP-p-CF3 , retrieved upon complete reoxidation in air within ~24 h. 1 H NMRD curves of the Mn(III)/(II)-TPP-p-CF3 chelates in mixed DMSO/water solvent have the typical shape of Mn(II)/Mn(III) porphyrins.

Novel Amperometric Mercury-Selective Sensor Based on Organic Chelator Ionophore.

A novel amperometric sensor for the direct determination of toxic mercury ions, Hg2+, based on the organic chelator ionophore N, N di (2-hydroxy-5-[(4-nitrophenyl)diazenyl]benzaldehyde) benzene-1,2-diamine (NDBD), and multiwalled carbon nanotubes (MWCNT) immobilized on a glassy carbon electrode surface was developed. The parameters influencing sensor performance including the ionophore concentration, the applied potential, and electrolyte pH were optimized. The sensor response to Hg2+ was linear between 1-25 µM with a limit of detection of 60 nM. Interferences from other heavy metal ions were evaluated and the sensor showed excellent selectivity towards Hg2+. The method was successfully applied to the determination of mercury ions in milk and water samples.

Electrochemical Impedance Spectroscopy in the Characterisation and Application of Modified Electrodes for Electrochemical Sensors and Biosensors.

Electrochemical impedance spectroscopy is finding increasing use in electrochemical sensors and biosensors, both in their characterisation, including during successive phases of sensor construction, and in application as a quantitative determination technique. Much of the published work continues to make little use of all the information that can be furnished by full physical modelling and analysis of the impedance spectra, and thus does not throw more than a superficial light on the processes occurring. Analysis is often restricted to estimating values of charge transfer resistances without interpretation and ignoring other electrical equivalent circuit components. In this article, the important basics of electrochemical impedance for electrochemical sensors and biosensors are presented, focussing on the necessary electrical circuit elements. This is followed by examples of its use in characterisation and in electroanalytical applications, at the same time demonstrating how fuller use can be made of the information obtained from complete modelling and analysis of the data in the spectra, the values of the circuit components and their physical meaning. The future outlook for electrochemical impedance in the sensing field is discussed.

Highly sensitive and selective nanostructured microbiosensors for glucose and lactate simultaneous measurements in blood serum and in vivo in brain tissue.

Highly sensitive and selective nanostructured lactate and glucose microbiosensors for their in vivo simultaneous determination in rat brain were developed based on carbon fiber microelectrodes (CFM) modified with nanoporous gold (NPG) using the Dynamic Hydrogen Bubble Template (DHBT) method. Electrodeposition of platinum nanoparticles (PtNP) onto the NPG film enhances the sensitivity and the electrocatalytic properties towards H2O2 detection. The nanostructured microelectrode platform was modified by glucose oxidase (GOx) and lactate oxidase (LOx) enzyme immobilization. High selective measurements were achieved by covering with a perm-selective layer of electropolymerized m-phenylenediamine, deposition of a Nafion® film and by using a null sensor. The morphological characteristics and electroanalytical performance of the microbiosensors were assessed, by scanning electron microscopy and electrochemical techniques, respectively. The PtNP/NPG/CFM shows a high sensitivity to H2O2 (5.96 A M-1 cm-2) at 0.36 V vs. Ag/AgCl, with a linear range from 0.2 to 200 µM, and an LOD of 10 nM. The microbiosensors were applied to the simultaneous determination of lactate and glucose in blood serum samples. Moreover, the basal extracellular concentrations of lactate and glucose were measured in vivo in four different rat brain structures. These results support the potential of the microbiosensor to be used as a valuable tool to investigate brain neurochemicals in vivo.

Magnetic-field-assisted deposition of self-assembling crystallite layers of Co2+-containing layered double hydroxides.

Precipitation of nanocrystallites of cobalt-aluminium layered double hydroxides in a magnetic field has been studied. In a magnetic field perpendicular to the substrate, dense and homogeneous films have been obtained. Magnetic anisotropy of the crystallites is explained by deviation from the statistical cation distribution in favour of honeycomb-like coordination of cobalt.

Polyphenazine and polytriphenylmethane redox polymer/nanomaterial-based electrochemical sensors and biosensors: a review.

In recent years, an increasing number of studies has demonstrated that redox polymers can be used in simple and effective electrochemical sensing platforms due to their fast electron transfer and electrocatalytic ability. To develop more sensitive and selective electrochemical (bio)sensors, the electrocatalytic properties of redox polymers and the electrical, mechanical, and catalytic properties of various nanomaterials are combined. This review aims to summarize and contribute to the development of (bio)sensors based on polyphenazine or polytriphenylmethane redox polymers combined with nanomaterials, including carbon-based nanomaterials, metal/metal oxide, and semiconductor nanoparticles. The synthesis, preparation, and modification of these nanocomposites is presented and the contribution of each material to the performance of (bio)sensor has been be examined. It is explained how the combined use of these redox polymers and nanomaterials as a sensing platform leads to improved analytical performance of the (bio)sensors. Finally, the analytical performance characteristics and practical applications of polyphenazine and polytriphenylmethane redox polymer/nanomaterial-based electrochemical (bio)sensors are compared and discussed.

Hybrid Nanocomposite Platform, Based on Carbon Nanotubes and Poly(Methylene Blue) Redox Polymer Synthesized in Ethaline Deep Eutectic Solvent for Electrochemical Determination of 5-Aminosalicylic Acid.

A novel hybrid composite of conductive poly(methylene blue) (PMB) and carbon nanotubes (CNT) was prepared for the detection of 5-aminosalicylic acid (5-ASA). Electrosynthesis of PMB with glassy carbon electrode (GCE) or with carbon nanotube modified GCE was done in ethaline deep eutectic solvent of choline chloride mixed with ethylene glycol and a 10% v/v aqueous solution. Different sensor architectures were evaluated in a broad range of pH values in a Britton-Robinson (BR) buffer using electrochemical techniques, chronoamperometry (CA), and differential pulse voltammetry (DPV), to determine the optimum sensor configuration for 5-ASA sensing. Under optimal conditions, the best analytical performance was obtained with CNT/PMBDES/GCE in 0.04 M BR buffer pH 7.0 in the range 5-100 µM 5-ASA using the DPV method, with an excellent sensitivity of 9.84 µA cm-2 µM-1 (4.9 % RSD, n = 5) and a detection limit (LOD) (3σ/slope) of 7.7 nM, outclassing most similar sensors found in the literature. The sensitivity of the same sensor obtained in CA (1.33 µA cm-2 µM-1) under optimal conditions (pH 7.0, Eapp = +0.40 V) was lower than that obtained by DPV. Simultaneous detection of 5-ASA and its analogue, acetaminophen (APAP), was successfully realized, showing a catalytic effect towards the electro-oxidation of both analytes, lowering their oxidation overpotential, and enhancing the oxidation peak currents and peak-to-peak separation as compared with the unmodified electrode. The proposed method is simple, sensitive, easy to apply, and economical for routine analysis.

A novel nanostructured poly(thionine)-deep eutectic solvent/CuO nanoparticle film-modified disposable pencil graphite electrode for determination of acetaminophen in the presence of ascorbic acid.

A new electrochemical sensor based on thionine (TH), an electroactive polymer, and CuO nanoparticle (CuONP)-modified pencil graphite electrode (PGE) has been developed. Poly(thionine) (PTH) was formed on the CuO/PGE surface by electropolymerisation in ethaline deep eutectic solvent (DES) containing acetic acid dopant to form PTHEthaline/CuO/PGE. Cyclic voltammetry, electrochemical impedance spectroscopy, and differential pulse voltammetry were utilized to evaluate the fabrication process, electrochemical properties, and performance parameters of the modified electrodes. The analytical performance of the PTHEthaline/CuO/PGE was evaluated with respect to linear range, limit of detection, repeatability, and reproducibility for the detection of acetaminophen (APAP) by electrooxidation in the presence of ascorbic acid (AA). Analytical parameters such as pH were optimized. The combined use of PTH and CuONP led to enhanced performance towards APAP due to the large electroactive surface area and synergistic catalytic effect, with a wide linear working range and low detection limit. The reliability of the proposed sensor for the detection of APAP was successfully tested in pharmaceutical samples containing APAP and AA, with very good recoveries. Graphical abstract.

Electrochemical synthesis and characterization of poly(thionine)-deep eutectic solvent/carbon nanotube-modified electrodes and application to electrochemical sensing.

Electropolymerization of thionine (TH) on multiwalled carbon nanotube (MWCNT)-modified glassy carbon electrodes (GCE) in ethaline deep eutectic solvent (DES) was carried out for the first time, to prepare poly(thionine) (PTH) films with different nanostructured morphologies. PTH films were formed on MWCNT/GCE by potential cycling electropolymerization in ethaline with the addition of different acid dopants CH3COOH, HClO4, HNO3, H2SO4 and HCl, acetic acid being the best. The electropolymerization process was monitored with an electrochemical quartz crystal microbalance. The polymerization scan rate was a key factor affecting the electrochemical and morphological properties of the PTHEthaline-CH3COOH/MWCNT/GCE; electrodeposition at 200 mV s-1 showing the best performance. The PTH/MWCNT/GCE platform was characterized using cyclic and differential pulse voltammetry, electrochemical impedance spectroscopy and scanning electron microscopy. The analytical characteristics of the PTH films were evaluated for sensing of ascorbic acid and biosensing of uric acid. The developed sensor exhibited a low detection limit (1.1 µM), wide linear range (2.8-3010 µM) and high sensitivity (1134 µA cm-2 mM-1) for ascorbic acid. After immobilization of uricase, UOx, on PTH/MWCNT/GCE, the biosensor was successfully applied to the determination of uric acid, with fast response (˂ 7 s), good sensitivity (450 µA cm-2 mM-1, wide linear range (0.48-279 µM) and low detection limit (58.9 nM), better than in the literature and than with PTH prepared in aqueous solution. The determination of uric acid in synthetic urine samples was successfully tested and the mean analytical recovery was 100.8 ± 1.4%. This is a promising approach for the determination of uric acid in real samples. Graphical abstract.

Biotoxic trace metal ion detection by enzymatic inhibition of a glucose biosensor based on a poly(brilliant green)-deep eutectic solvent/carbon nanotube modified electrode.

A highly sensitive glucose oxidase (GOx) electrochemical biosensor for the determination of the biotoxic trace metal ions Hg2+, Cd2+, Pb2+ and CrVI by enzyme inhibition has been developed. GOx was immobilized on a novel sensing platform consisting of poly(brilliant green) films formed by potential cycling electropolymerization in sulfuric acid doped ethaline deep eutectic solvent on multiwalled carbon nanotube modified glassy carbon electrodes. Polymer films produced in this medium presented more uniform morphology and better electrochemical sensing properties than those prepared in aqueous solution. The inhibitor concentration necessary to give 50% inhibition, I50, was used for the determination of the type of reversible inhibition, and the relationship between I50 and the inhibition constant Ki is discussed. The new biosensor was successfully used for the determination of biotoxic trace metal ions with a nanomolar limit of detection, lower than in the literature, very good repeatability, stability and selectivity, and was applied successfully to detection of the toxic trace metal species in milk samples.

A biocompatible redox MRI probe based on a Mn(ii)/Mn(iii) porphyrin.

For the development of redox responsive MRI probes based on the MnIII/MnII couple, stable complexation of both reduced and oxidized forms of the metal ion and appropriate tuning of the redox potential in the biologically relevant range are key elements. The water soluble fluorinated Mn-porphyrin derivative Mn-3 satisfies both requirements. In aqueous solutions, it can reversibly switch between MnIII/MnII oxidation states. In the presence of ascorbic acid or ß-mercaptoethanol, the MnIII form undergoes reduction, which is slowly but fully reversed in the presence of air oxygen. A UV-Vis kinetic study of MnIII/MnII reduction under oxygen-free conditions yielded second-order rate constants, k2, of 46.1 M-1 s-1 and 13.8 M-1 s-1 for the reaction with ascorbic acid and ß-mercaptoethanol, respectively. This could correspond, in the absence of oxygen, to a half-life of a few minutes in blood plasma and a few seconds in circulating immune cells where ascorbic acid reaches 20-40 µM and a few mM concentrations, respectively. In contrast to expectations based on the redox potential, reduction with glutathione or cysteine does not occur. It is prevented by the coordination of the glutathione carboxylate group(s) to MnIII in the axial position, as was evidenced by NMR data. Therefore, MnIII-3 acts as an ascorbate specific turn-on MRI probe, which in turn can be re-oxidized by oxygen. The relaxivity increase from the oxidized to the reduced form is considerably improved at medium frequencies (up to 80 MHz) with respect to the previously studied Mn-TPPS4 analogues; at 20 MHz, it amounts to 150%. No in vitro cytotoxicity is detectable for Mn-3 in the typical MRI concentration range. Finally, 19F NMR resonances of MnIII-3 are relatively sharp which could open further opportunities to exploit such complexes as paramagnetic 19F NMR probes.

Tyrosinase based amperometric biosensor for determination of tyramine in fermented food and beverages with gold nanoparticle doped poly(8-anilino-1-naphthalene sulphonic acid) modified electrode.

The aim of the present work was to develop an amperometric biosensor for tyramine (Tyr) measurement in food and beverages. The biosensor architecture is based on tyrosinase (Tyrase) immobilization on glassy carbon electrode modified by a nanocomposite consisting of gold nanoparticles (AuNP) synthesized by a green method and poly(8-anilino-1-naphthalene sulphonic acid) modified glassy carbon electrode. Under optimized experimental conditions for fixed potential amperometric detection, the biosensor exhibited a linear response to tyramine in the range 10-120 µM and the limit of detection was estimated to be 0.71 µM. The novel platform showed good selectivity, long-term stability, and reproducibility. The strong interaction between tyrosinase and the nanocomposite was revealed by the high value of the Michaelis-Menten constant (79.3 µM). The fabricated biosensor was successfully applied to the determination of Tyr in dairy products and fermented drinks with good recoveries, which makes it a promising biosensor for quantification of tyramine.

Impedimetric sensor for tyramine based on gold nanoparticle doped-poly(8-anilino-1-naphthalene sulphonic acid) modified gold electrodes.

A novel impedimetric sensor for the determination of tyramine (Tyr), a biogenic amine, on the surface of gold nanoparticle-poly-(8-anilino-1-napthalene sulphonic acid), AuNP-PANSA modified gold electrode (AuE) is presented for the first time. The AuNP were successfully synthesized by a green synthesis method. Their characterization and optimization were conducted using X-ray diffraction, scanning electron microscopy, transmission electron microscopy, electrochemical impedance spectroscopy and cyclic voltammetry. Under optimal conditions, the impedimetric sensor revealed a relatively broad linear range from 0.8 to 80 µM similar to more complex architectures found in the literature and the limit of detection of 0.04 µM was the lowest achieved until now. In order to test the reliability of the proposed method, real sample application studies were conducted using dairy products and fermented drinks. It was found that the sensor presented a good selectivity and recovery. Furthermore, the impedimetric sensor shows good reproducibility, stability, selectivity and very small interferences which augur well for its application in food safety control processes.

A novel amperometric enzyme inhibition biosensor based on xanthine oxidase immobilised onto glassy carbon electrodes for bisphenol A determination.

A novel and simple biosensor for the determination of bisphenol A (BPA) based on xanthine oxidase (XOD) enzymatic inhibition has been developed. The biosensor was prepared from xanthine oxidase immobilised by crosslinking with glutaraldehyde, with hypoxanthine as enzyme substrate, and was successfully applied to the determination of BPA using fixed potential amperometry. Biosensor performance was optimised with respect to the applied potential, influence of pH of the electrolyte solution, XOD loading and the substrate concentration. The enzyme inhibition mechanism was evaluated from Cornish-Bowden plus Dixon plots and was found to be reversible and competitive with an apparent inhibition constant of 8.15 nM. Under optimised conditions, the determination of BPA can be achieved in the linear range up to 41 nM with a detection limit of 1.0 nM, which is equal to the lowest reported in the literature, with very good repeatability and reproducibility. The selectivity of the biosensor was evaluated by performing an interference study and found to be excellent; and stability was investigated. It was successfully applied to the detection of BPA in mineral water and in river water.

Catalase based hydrogen peroxide biosensor for mercury determination by inhibition measurements.

A new amperometric hydrogen peroxide enzyme inhibition biosensor for the indirect determination of toxic mercury ions, Hg2+, based on catalase immobilized on a glassy carbon electrode surface by cross-linking with glutaraldehyde and bovine serum albumin, is reported. The parameters influencing biosensor performance were optimized, including enzyme loading, the amount of hydrogen peroxide, the applied potential and electrolyte pH. It was shown that the inhibition of catalase by Hg2+ species is irreversible, with a linear inhibition response between 5×10-11 and 5×10-10M. The limit of detection calculated as 10% inhibition was 1.8×10-11M and is the lowest reported until now. Electrochemical impedance spectroscopy was successfully used as a diagnostic of inhibition. Interferences from other heavy metal ions and organic pesticides were evaluated and the inhibition showed very good selectivity towards Hg2+. The method was successfully applied to the determination of mercury ions in different types of water sample.

A novel sensitive amperometric choline biosensor based on multiwalled carbon nanotubes and gold nanoparticles.

A novel amperometric biosensor for choline determination has been developed, exploiting the electrocatalytic properties of multiwalled carbon nanotubes (MWCNT) and gold nanoparticles (GNP). Chitosan (Chit), a natural biocompatible polymer, was used to disperse CNT, then Chit-MWCNT was dropped on the surface of a glassy carbon electrode (GCE), followed by GNP; finally, choline oxidase (ChOx) was immobilized by glutaraldehyde crosslinking. The ChOx/(GNP)4/MWCNT/GCE exhibited linear response to choline from 3 to 120µM, the sensitivity was 204µAcm-2mM-1 and the detection limit was 0.6µM. The biosensor exhibited good intra and inter-electrode precision, and excellent selectivity and stability. Electrochemical impedance spectroscopy (EIS) was also used to measure choline at 0.0V and this is the first report on choline determination by EIS. Successful measurement in milk samples was performed.

Ceramic-Based Multisite Platinum Microelectrode Arrays: Morphological Characteristics and Electrochemical Performance for Extracellular Oxygen Measurements in Brain Tissue.

Ceramic-based multisite Pt microelectrode arrays (MEAs) were characterized for their basic electrochemical characteristics and used for in vivo measurements of oxygen with high resolution in the brain extracellular space. The microelectrode array sites showed a very smooth surface mainly composed of thin-film polycrystalline Pt, with some apparent nanoscale roughness that was not translated into an increased electrochemical active surface area. The electrochemical cyclic voltammetric behavior was characteristic of bulk Pt in both acidic and neutral media. In addition, complex plane impedance spectra showed the required low impedance (0.22 MΩ; 10.8 Ω cm2) at 1 kHz and very smooth electrode surfaces. The oxygen reduction reaction on the Pt surface proceeds as a single 4-electron reduction pathway at -0.6 V vs Ag/AgCl reference electrode. Cyclic voltammetry and amperometry demonstrate excellent electrocatalytic activity toward oxygen reduction in addition to a high sensitivity (-0.16 ± 0.02 nA µM-1) and a low limit of detection (0.33 ± 0.20 µM). Thus, these Pt MEAs provide an excellent microelectrode platform for multisite O2 recording in vivo in the extracellular space of the brain, demonstrated in anaesthetized rats, and hold promise for future in vivo studies in animal models of CNS disease and dysfunction.

ß-Cyclodextrin carbon nanotube-enhanced sensor for ciprofloxacin detection.

A simple and expedite electrochemical methodology was developed for the determination of ciprofloxacin, based on a glassy carbon (GC) electrode modified by a combination of multi-walled carbon nanotubes (MWCNT) with ß-cyclodextrin (ß-CD) incorporated in a polyaniline film. The combined use of ß-CD and MWCNT in the electrochemical sensor leads to a significant signal improvement. The ß-CD/MWCNT modified GC electrode exhibited efficient electrocatalytic behavior in the oxidation of ciprofloxacin with relatively high sensitivity, stability and lifetime. Molecular modeling studies showed that ciprofloxacin binds preferably to ß-CD rather than to CNT edges, leading to an improved sensitivity of the sensor. Under optimized conditions, a linear calibration curve was obtained for ciprofloxacin in the concentration range 10-80 µM with a detection limit of 50 nM. The analytical performance of this sensor was evaluated for the detection of ciprofloxacin in a wastewater treatment plant effluent.

New CNT/poly(brilliant green) and CNT/poly(3,4-ethylenedioxythiophene) based electrochemical enzyme biosensors.

A combination of the electroactive polymer poly(brilliant green) (PBG) or conducting polymer poly(3,4-ethylenedioxythiophene) (PEDOT) with carbon nanotubes to obtain CNT/PBG and CNT/PEDOT modified carbon film electrodes (CFE) has been investigated as a new biosensor platform, incorporating the enzymes glucose oxidase (GOx) as test enzyme, alcohol oxidase (AlcOx) or alcohol dehydrogenase (AlcDH). The sensing parameters were optimized for all biosensors based on CNT/PBG/CFE, CNT/PEDOT/CFE platforms. Under optimized conditions, both GOx biosensors exhibited very similar sensitivities, while in the case of AlcOx and AlcDH biosensors, AlcOx/CNT/PBG/CFE was found to give a higher sensitivity and lower detection limit. The influence of dissolved O2 on oxidase-biosensor performance was investigated and was shown to be different for each enzyme. Comparisons were made with similar reported biosensors, showing the advantages of the new biosensors, and excellent selectivity against potential interferents was successfully demonstrated. Finally, alcohol biosensors were successfully used for the determination of ethanol in alcoholic beverages.