Júlia M. C. S. Magalhães

Simultaneous electrochemical determination of arsenic, copper, lead and mercury in unpolluted fresh waters using a vibrating gold microwire electrode

In this work, a simple, rapid, reliable and low cost method for simultaneous electrochemical determination of As, Cu, Hg and Pb ions, on a vibrating gold microwire electrode combined with stripping voltammetry, is described for the first time. The multi-element detection was performed in the presence of oxygen by differential pulse anodic stripping voltammetry (DPASV) in HCl 0.1 M with NaCl 0.5 M. This media was found optimum in terms of peak resolution, peak shape and sensitivities, and has a composition similar to seawater to which the method could potentially be applied. The gold microwire electrode presented well defined, undistorted, sharp and reproducible peaks for trace concentrations of Cu, Hg and Pb and As presented a reproducible peak with a small shoulder. Using a gold vibrating microwire electrode of 25 μm diameter and 30s deposition time, the detection limits of As, Cu, Hg and Pb were 0.07, 0.4, 0.07 and 0.2 μgL(-1), respectively. Possible effects of Al, Cd, Cr, Fe, Mn, Ni, Sb and Zn were investigated but did not cause any significant interferences. Finally, the method was applied for the simultaneous determination of these four metals in unpolluted river water samples and the results were validated by Atomic Absorption Spectroscopy with Electrothermal Atomization (AAS-EA) or by Inductively Coupled Plasma Mass Spectrometry (ICP-MS).

UREA POTENTIOMETRIC BIOSENSOR BASED ON UREASE IMMOBILIZED ON CHITOSAN MEMBRANES

Potentiometric biosensors based on urease (E.C. 3.5.1.5.) immobilized on chitosan membranes coupled to all-solid-state nonactin ammonium ion selective electrodes are described. The enzyme was immobilized on the chitosan membranes by four procedures: (A) adsorption; (B) adsorption followed by reticulation with dilute aqueous glutaraldehyde solution; (C) activation with glutaraldehyde followed by contact with the enzyme solution; and (D) activation with glutaraldehyde, contact with the enzyme solution and reduction of the Schiff base with sodium borohydride. The response characteristics of the biosensors obtained with these enzymatic membranes were determined and compared. The biosensor with best response characteristics, obtained by procedure (B), showed the following characteristics of response to urea: (i) linearity in the 10(-4) to 10(-2) M range; (ii) slope of up to 56 mV per decade; (iii) response time between 30 s and 2 min; and (iv) lifetime of 2 months. This biosensor was tested in the determination of urea in blood serum samples.

MIP-graphene-modified glassy carbon electrode for the determination of trimethoprim

A novel sensitive electrochemical sensor was developed by electropolymerization of pyrrole (PY) and molecularly imprinted polymer (MIP) which was synthesized onto a glassy carbon electrode (GCE) in aqueous solution using cyclic voltammetry in the presence of Trimethoprim (TMP) as template molecules. Furthermore, a previous electrode modification was performed by deposition of a suspension of graphene on the electrodes surface. The performance of the imprinted and non-imprinted (NIP) films was evaluated by impedance spectroscopy (EIS) and cyclic voltammetry (CV) of a ferric solution. The molecularly imprinted film exhibited a high selectivity and sensitivity toward TMP. The sensor presented a linear range, between peak current intensity and logarithm of TMP concentration between 1.0 × 10(-6) and 1.0 × 10(-4)M. The results were accurate (with recoveries higher than 94%), precise (with standard deviations less than 5%) and the detection limit was 1.3 × 10(-7)M. The new sensor is selective, simple to construct and easy to operate. The MIP sensor was successfully applied to quantify TMP in urine samples.

Array of potentiometric sensors for the analysis of creatinine in urine samples

The development of potentiometric biosensors for creatinine based on creatinine iminohydrolase (E.C. 3.5.4.21) immobilized on chitosan membranes coupled to a nonactin based ammonium ion selective electrode is described. The response characteristics of three types of biosensors with the enzyme immobilized by three different procedures were evaluated. The biosensors with better response characteristics were obtained by coupling the ammonium ion selective electrodes to chitosan membranes with the enzyme immobilized by adsorption. The linear response range of these biosensors to creatinine was 10(-4) to 10(-2) M, the response time was between 30 and 60 s, they showed an operational lifetime of 44 days and the slope of the response to creatinine in the first day varied between 50 and 52 mV decade-1. An array of six potentiometric sensors, constituted by two creatinine biosensors and four ion selective electrodes for potassium, sodium, ammonium and calcium was calibrated and a multivariate model based on PLS1 for the response to creatinine was obtained and validated. The array was used for the analysis of creatinine in urine samples and the results were compared with the results of a clinical analysis laboratory, based on the Jaffé reaction.

Identification of enzyme produced firefly oxyluciferin by reverse phase HPLC

Firefly oxyluciferin (2-(6′-hydroxybenzothiazolyl)-4-hydroxythiazole) was chemically synthesized and characterized by means of 13C and 1H NMR, UV–vis spectrometry and RP-HPLC using different pH elution conditions. One of the chromatographic peaks observed in luciferase-catalyzed reaction mixtures was identified as corresponding to oxyluciferin.

Array of potentiometric sensors for simultaneous analysis of urea and potassium.

Urea biosensors based on urease immobilized by crosslinking with BSA and glutaraldehyde coupled to ammonium ion-selective electrodes were included in arrays together with potassium, sodium and ammonium PVC membrane ion-selective electrodes. Multivariate calibration models based on PCR and PLS2 were built and tested for the simultaneous determination of urea and potassium. The results show that it is possible to obtain PCR and PLS2 calibration models for simultaneous determination of these two species, based on a very small set of calibration samples (nine samples). Coupling of biosensors with ion-selective electrodes in arrays of sensors raises a few problems related to the limited stability of response and unidirectional cross-talk of the biosensors, and this matter was also subjected to investigation in this work. Up to three identical urea biosensors were included in the arrays, and the data analysis procedure allowed the assessment of the relative performance of the sensors. The results show that at least two urea biosensors should be included in the array to improve urea determination. The prediction errors of the concentration of urea and potassium in the blood serum samples analyzed with this array and a PLS2 calibration model, based on nine calibration samples, were lower than 10 and 5%, respectively.

Electrochemical biosensors for Salmonella : State of the art and challenges in food safety assessment

According to the recent statistics, Salmonella is still an important public health issue in the whole world. Legislated reference methods, based on counting plate methods, are sensitive enough but are inadequate as an effective emergency response tool, and are far from a rapid device, simple to use out of lab. An overview of the commercially available rapid methods for Salmonella detection is provided along with a critical discussion of their limitations, benefits and potential use in a real context. The distinguished potentialities of electrochemical biosensors for the development of rapid devices are highlighted. The state-of-art and the newest technologic approaches in electrochemical biosensors for Salmonella detection are presented and a critical analysis of the literature is made in an attempt to identify the current challenges towards a complete solution for Salmonella detection in microbial food control based on electrochemical biosensors.

A potentiometric magnetic immunoassay for rapid detection of Salmonella typhimurium

Potentiometric detection with homemade polymeric membrane microelectrodes was coupled to a magnetic sandwich immunoassay for Salmonella typhimurium determination. Cadmium and sodium ion selective electrodes were used respectively as indicator and pseudo-reference electrodes and were prepared in pipette tips to allow potentiometric measurements in microliter sample volumes. In the proposed method, the concentration of S. typhimurium was proportional to the amount of cadmium released upon dissolution of a CdS nanoparticle labeled to the secondary detection antibody. The limit of detection was 2 cells per 100 μL. The immunomagnetic assay with potentiometric detection is suitable for sensitive and rapid (average total time per assay of 75 minutes) detection of S. typhimurium in milk samples. The proposed method is easy to perform, safe, sensitive, and low cost and has potential for in situ analysis.

Study of the temperature behaviour of all-solid-state nonactin ammonium electrodes with PVC membrane applied to graphite–epoxy supports of varied composition

Abstract A study of the effect of the temperature on the response of all-solid-state ammonium selective electrodes prepared by casting PVC membranes with nonactin on supports of graphite-epoxy composites containing a variable amount of graphite up to 60% is presented. It is shown that while supports made of composites with 30% or less of graphite provide no adequate response characteristics, the variation of the percentage of graphite between 40 and 60% has no influence neither on the slope and standard potential temperature coefficients of the electrodes, nor on their isopotential point concentrations. The temperature hysteresis of the electrodes was found to be limited and also independent of the amount of graphite in the composite.

Gingival hyperplasia as a first manifestation of Crohn's disease

The mouth can be affected in patients with Crohn’s disease (CD), underlining the possibility of involvement of any segment of the gastrointestinal (GI) tract. The most common oral manifestation of CD is aphtosis, but it is also present in a multitude of other diseases (e.g., Behçet’s disease, sarcoidosis) and in healthy individuals. Therefore it is a very unspecific sign. Other, less common manifestations include gingival hyperplasia, diffuse gingival swelling, mucosal hyperplasia, fissuring cheilitis, and pyostomatitis vegetans. We report the case of a young man who presented with gingival hyperplasia followed by lip edema and oral aphtosis. These findings, together with the presence of granuloma in a biopsy of the gums, led to the diagnosis of CD, although the patient did not complain of any GI symptoms.