Adriana N. Correia

Biosensor based on multi-walled carbon nanotubes paste electrode modified with laccase for pirimicarb pesticide quantification

This study focused on the development of a sensitive enzymatic biosensor for the determination of pirimicarb pesticide based on the immobilization of laccase on composite carbon paste electrodes. Multi-walled carbon nanotubes (MWCNTs) paste electrode modified by dispersion of laccase (3%, w/w) within the optimum composite matrix (60:40%, w/w, MWCNTs and paraffin binder) showed the best performance, with excellent electron transfer kinetic and catalytic effects related to the redox process of the substrate 4-aminophenol. No metal or anti-interference membrane was added. Based on the inhibition of laccase activity, pirimicarb can be determined in the range 9.90 × 10(-7) to 1.15 × 10(-5) mol L(-1) using 4-aminophenol as substrate at the optimum pH of 5.0, with acceptable repeatability and reproducibility (relative standard deviations lower than 5%). The limit of detection obtained was 1.8 × 10(-7) mol L(-1) (0.04 mg kg(-1) on a fresh weight vegetable basis). The high activity and catalytic properties of the laccase-based biosensor are retained during ca. one month. The optimized electroanalytical protocol coupled to the QuEChERS methodology were applied to tomato and lettuce samples spiked at three levels; recoveries ranging from 91.0 ± 0.1% to 101.0 ± 0.3% were attained. No significant effects in the pirimicarb electroanalysis were observed by the presence of pro-vitamin A, vitamins B1 and C, and glucose in the vegetable extracts. The proposed biosensor-based pesticide residue methodology fulfills all requisites to be used in implementation of food safety programs.

Studies of the hydrogen evolution reaction on smooth Co and electrodeposited Ni–Co ultramicroelectrodes

Abstract The hydrogen evolution reaction (HER) was studied on smooth Co and on electrodeposited Ni–Co ultramicroelectrodes (UMEs) in alkaline solutions at several temperatures by steady-state polarisation curves. The real electrochemical area was previously estimated by cyclic voltammetry to account for the large difference in roughness factor of the two surfaces. The values obtained for the Tafel slopes were very close to 2.303 RT / βnF while the ‘apparent’ energies of activation were 59 and 41 kJ mol −1 for Co and Ni–Co, respectively. A common Volmer–Heyrovsky mechanism with Heyrovsky as the rate-determining step (RDS) was initially proposed. This was confirmed when the experimental results were mathematically treated by a non-linear fitting procedure using the kinetic equations derived for that mechanism. The calculations revealed that Ni–Co is a more efficient catalyst for the HER then pure Co, with a rate constant value of 0.16×10 −10 mol s −1 cm −2 at 25°C for the slow step. Although this value is more than one order of magnitude smaller than that already reported for deposited Ni, it is considerably larger than the one measured here (0.02×10 −10 mol s −1 cm −2 ) for pure Co at 25°C.

Sensitive bi-enzymatic biosensor based on polyphenoloxidases–gold nanoparticles–chitosan hybrid film–graphene doped carbon paste electrode for carbamates detection

A bi-enzymatic biosensor (LACC-TYR-AuNPs-CS/GPE) for carbamates was prepared in a single step by electrodeposition of a hybrid film onto a graphene doped carbon paste electrode (GPE). Graphene and the gold nanoparticles (AuNPs) were morphologically characterized by transmission electron microscopy, X-ray photoelectron spectroscopy, dynamic light scattering and laser Doppler velocimetry. The electrodeposited hybrid film was composed of laccase (LACC), tyrosinase (TYR) and AuNPs entrapped in a chitosan (CS) polymeric matrix. Experimental parameters, namely graphene redox state, AuNPs:CS ratio, enzymes concentration, pH and inhibition time were evaluated. LACC-TYR-AuNPs-CS/GPE exhibited an improved Michaelis-Menten kinetic constant (26.9±0.5M) when compared with LACC-AuNPs-CS/GPE (37.8±0.2M) and TYR-AuNPs-CS/GPE (52.3±0.4M). Using 4-aminophenol as substrate at pH5.5, the device presented wide linear ranges, low detection limits (1.68×10(-9)±1.18×10(-10)-2.15×10(-7)±3.41×10(-9)M), high accuracy, sensitivity (1.13×10(6)±8.11×10(4)-2.19×10(8)±2.51×10(7)%inhibitionM(-1)), repeatability (1.2-5.8% RSD), reproducibility (3.2-6.5% RSD) and stability (ca. twenty days) to determine carbaryl, formetanate hydrochloride, propoxur and ziram in citrus fruits based on their inhibitory capacity on the polyphenoloxidases activity. Recoveries at two fortified levels ranged from 93.8±0.3% (lemon) to 97.8±0.3% (orange). Glucose, citric acid and ascorbic acid do not interfere significantly in the electroanalysis. The proposed electroanalytical procedure can be a promising tool for food safety control.

Laccase-Prussian blue film-graphene doped carbon paste modified electrode for carbamate pesticides quantification.

A novel enzymatic biosensor for carbamate pesticides detection was developed through the direct immobilization of Trametes versicolor laccase on graphene doped carbon paste electrode functionalized with Prussian blue films (LACC/PB/GPE). Graphene was prepared by graphite sonication-assisted exfoliation and characterized by transmission electron microscopy and X-ray photoelectron spectroscopy. The Prussian blue film electrodeposited onto graphene doped carbon paste electrode allowed considerable reduction of the charge transfer resistance and of the capacitance of the device. The combined effects of pH, enzyme concentration and incubation time on biosensor response were optimized using a 2(3) full-factorial statistical design and response surface methodology. Based on the inhibition of laccase activity and using 4-aminophenol as redox mediator at pH 5.0, LACC/PB/GPE exhibited suitable characteristics in terms of sensitivity, intra- and inter-day repeatability (1.8-3.8% RSD), reproducibility (4.1 and 6.3% RSD), selectivity (13.2% bias at the higher interference:substrate ratios tested), accuracy and stability (ca. twenty days) for quantification of five carbamates widely applied on tomato and potato crops. The attained detection limits ranged between 5.2×10(-9)molL(-1) (0.002mgkg(-1) w/w for ziram) and 1.0×10(-7)molL(-1) (0.022mgkg(-1) w/w for carbofuran). Recovery values for the two tested spiking levels ranged from 90.2±0.1 (carbofuran) to 101.1±0.3% (ziram) for tomato and from 91.0±0.1% (formetanate) to 100.8±0.1% (ziram) for potato samples. The proposed methodology is appropriate to enable testing pesticide levels in food samples to fit with regulations and food inspections.

Corrosion study of electrodeposited Zn and Zn-Co coatings in chloride medium

The corrosion behavior of electrodeposited Zn and Zn-Co coatings in 0.1 mol dm-3 NaCl aqueous solutions was investigated using scanning electron microscopy (SEM), energy dispersive X-ray (EDAX), galvanostatic electrochemical stripping (GES), electrochemical impedance spectroscopy (EIS) techniques and open circuit measurements (Eoc). All coatings were electrodeposited on mild steel. Using GES it was possible to identify Zn-Co intermetallic phases in the coatings with Co content higher than 1 at.%. The GES, SEM and EDX results revealed that the corrosion mechanism of the Zn-Co layers with high Co content is related to the appearance of cracks due to the dissolution of the g Zn-rich phases, while the corrosion of the Zn and Zn-1Co layers was characterized by dissolution of the coatings. The Zn and Zn-Co coatings were found to contain the same insoluble corrosion products, which act as a corrosion barrier that delays but does not prevent corrosion. The better corrosion resistance of the Zn-Co coatings obtained in the bath without additive, in comparison with Zn-1Co obtained in the bath containing additive is due to the high Co content which leads to an ennoblement of the coating. All Zn-Co electrodeposits showed greater corrosion resistance than the Zn electrodeposits. The Zn-18Co layer showed the best corrosion resistance, with a lifetime about three times longer than that of the Zn layer.

Square wave voltammetric determination of nitrofurantoin in pharmaceutical formulations on highly boron-doped diamond electrodes at different boron-doping contents.

The influence of the boron-doping levels in boron-doped diamond film electrodes on the electrochemical response of nitrofurantoin (NFT) and the development of an electroanalytical procedure for NFT determination were investigated. The investigations were carried out using the techniques of cyclic voltammetry and square wave voltammetry on diamond film electrodes with different boron-doping levels (i.e., 5000, 10,000 and 20,000 mg L(-1)). The level of boron-doping in the diamond film electrodes influenced the electrochemical reduction of NFT. The appropriate cyclic voltammetric response of NFT was obtained with Britton-Robinson buffer at pH 4 and for diamond films doped with 10,000 and 20,000 mg L(-1) of boron. These two films were selected for the development of the electroanalytical procedure. The use of square wave voltammetry with the optimized parameters demonstrated a good linear relationship between the peak current and the NFT concentration for a wide range of concentration. The lower limit of detection for the electrodes doped with 10,000 and 20,000 mg L(-1) of boron were 2.69 x 10(-8) mol L(-1) (6.40 microg L(-1)) and 8.15 x 10(-9) mol L(-1) (1.94 microg L(-1)), respectively, while the lower limits of quantification were 8.96 x 10(-8) mol L(-1) (21.33 microg L(-1)) and 2.72 x 10(-8) mol L(-1) (6.47 microg L(-1)), respectively. The applicability of the proposed procedure was tested using a commercial pharmaceutical formulation of NFT, and the results were compared with the procedure recommended by the British Pharmacopeia. The proposed procedure was sensitive, accurate and precise for analysis of NFT and did not require complex preparations or renovations of the electrode surface. This presents the advantage of eliminating mercury waste and minimizing the adsorptive problems related to the use of other electrodic solid surfaces.

Simple laccase-based biosensor for formetanate hydrochloride quantification in fruits

This work describes the development of an electrochemical enzymatic biosensor for quantification of the pesticide formetanate hydrochloride (FMT). It is based on a gold electrode modified with electrodeposited gold nanoparticles and laccase. The principle behind its development relies on FMTs capacity to inhibit the laccase catalytic reaction that occurs in the presence of phenolic substrates. The optimum values for the relevant experimental variables such as gold nanoparticles electrochemical deposition (at -0.2V for 100s), laccase immobilization (via glutaraldehyde cross-linking), laccase concentration (12.4mg/mL), substrate selection and concentration (5.83×10(-5)M of aminophenol), pH (5.0), buffer (Britton-Robinson), and square-wave voltammetric parameters were determined. The developed biosensor was successfully applied to FMT determination in mango and grapes. The attained limit of detection was 9.5×10(-8)±9.5×10(-10)M (0.02±2.6×10(-4)mg/kg on a fresh fruit weight basis). Recoveries for the five tested spiking levels ranged from 95.5±2.9 (grapes) to 108.6±2.5% (mango). The results indicated that the proposed device presents suitable characteristics in terms of sensitivity (20.58±0.49A/μM), linearity (9.43×10(-7) to 1.13×10(-5)M), accuracy, repeatability (RSD of 1.4%), reproducibility (RSD of 1.8%) and stability (19days) for testing of compliance with established maximum residue limits of FMT in fruits and vegetables.

Characterisation of electrodeposited and heat-treated Ni−Mo−P coatings

The electrodeposition, hardness and corrosion resistance properties of Ni−Mo−P coatings were investigated. Characterisations of the electrodeposited coatings were carried out using scanning electron microscopy, X-ray diffraction and energy dispersive X-ray analysis techniques. Corrosion tests were performed at room temperature in 10-1 mol dm-3 NaCl solutions and by potentiodynamic linear polarisation. Amorphous Ni−Mo−P coatings were successfully obtained by electrodeposition using direct current. The coating composition showed to be dependent on the bath composition, current density and bath temperature. Both P and Mo contents contribute for the hardness properties of the Ni−Mo−P coatings and the absence of cracks is a requirement to produce electrodeposited Ni−Mo−P coatings with good hardness properties. The hardness values increase with heat-treatment temperature due to the precipitation of Ni, Ni3P and NiMo phases during the heat treatment. The corrosion resistance of the electrodeposited Ni−Mo−P amorphous coatings increases with P content in the layer. Among the electrodeposited Ni−Mo−P amorphous coatings, Ni78Mo10P12 presented the best hardness and corrosion-resistance properties. The results showed that the addition of P is beneficial for the hardness and corrosion resistance properties of the Ni−Mo-based coatings.

Square-wave adsorptive voltammetry of dexamethasone: Redox mechanism, kinetic properties, and electroanalytical determinations in multicomponent formulations

The electrochemical reduction behavior of dexamethasone at a hanging mercury drop electrode was investigated by cyclic and square-wave adsorptive voltammetries in a Britton-Robinson buffer at pH 2.0. The optimized experimental conditions consisted of a pulse potential frequency of 100 s(-1), a pulse amplitude of 15 mV, and a potential step height of 2 mV, with E(acc)=-0.60V and t(acc)=15s. From these parameters, it was also possible to develop a detailed study about the kinetic and mechanistic events involved in the reduction process. Two well-defined peaks were observed in the cathodic scan, and peak 2 was used to obtain analytical curves. A linear range between 4.98×10(-8) and 6.10×10(-7)mol L(-1), with a detection limit of 2.54×10(-9)mol L(-1) and a quantification limit of 8.47×10(-9)mol L(-1), was observed. Moreover, it was possible to achieve a simple, selective, and versatile methodology adaptable to the quantification of dexamethasone because common excipients used in multicomponent commercial formulations caused no interference. The satisfactory recoveries and the low relative standard deviation data reflected the high accuracy and precision of the proposed method for the determination of dexamethasone in injectable eye drops and elixir samples.

Morphological, structural, microhardness and corrosion characterisations of electrodeposited Ni-Mo and Cr coatings

The corrosion resistance of electrodeposited Cr and Ni-Mo coatings and the influence of heat treatment on the crystallographic structure, morphology and microhardness properties were investigated here. The characterisations were carried out using scanning electron microscopy (SEM), X-ray diffraction (XRD) and energy dispersive X-ray analysis (EDX) techniques. Corrosion tests were performed at room temperature in 10-1 mol dm-3 NaCl solutions and by potentiodynamic linear polarization technique.TheMo content in the layer and current efficiency increased with the molybdate ion concentration in the plating solution, while the surface morphology evolved from rough and homogeneous to cracked surface with the increase of the amount of Mo in the layer. The electrochemical corrosion tests showed that the Cr coatings have better corrosion resistance than the Ni-Mo coatings in chloride medium and that all the studied coatings corrode in chloride medium. Ni-13Mo coating has the nobler corrosion behavior among the studied Ni-Mo coatings. The microhardness of the Ni-13Mo coatings increased as the annealing temperature increased which is related with the precipitation of Ni, Ni4Mo and NiMo phases during the heat treatment of this coating. Ni-13Mo coating is a potential substitute for chromium coating in industrial applications when operating at temperatures higher than 100 oC and good microhardness properties are required.