Development of a novel microextraction by packed sorbent-based approach followed by ultrahigh pressure liquid chromatography as a powerful technique for quantification phenolic constituents of biological interest in wines

A novel analytical approach, based on a miniaturized extraction technique, the microextraction by packed sorbent (MEPS), followed by ultrahigh pressure liquid chromatography (UHPLC) separation combined with a photodiode array (PDA) detection, has been developed and validated for the quantitative determination of sixteen biologically active phenolic constituents of wine. In addition to performing routine experiments to establish the validity of the assay to internationally accepted criteria (linearity, sensitivity, selectivity, precision, accuracy), experiments are included to assess the effect of the important experimental parameters on the MEPS performance such as the type of sorbent material (C2, C8, C18, SIL, and M1), number of extraction cycles (extract-discard), elution volume, sample volume, and ethanol content, were studied. The optimal conditions of MEPS extraction were obtained using C8 sorbent and small sample volumes (250 μL) in five extraction cycle and in a short time period (about 5 min for the entire sample preparation step). The wine bioactive phenolics were eluted by 250 μL of the mixture containing 95% methanol and 5% water, and the separation was carried out on a HSS T3 analytical column (100 mm × 2.1 mm, 1.8 μm particle size) using a binary mobile phase composed of aqueous 0.1% formic acid (eluent A) and methanol (eluent B) in the gradient elution mode (10 min of total analysis). The method gave satisfactory results in terms of linearity with r2-values > 0.9986 within the established concentration range. The LOD varied from 85 ng mL−1 (ferulic acid) to 0.32 μg mL−1 ((+)-catechin), whereas the LOQ values from 0.028 μg mL−1 (ferulic acid) to 1.08 μg mL−1 ((+)-catechin). Typical recoveries ranged between 81.1 and 99.6% for red wines and between 77.1 and 99.3% for white wines, with relative standard deviations (RSD) no larger than 10%. The extraction yields of the MEPSC8/UHPLC–PDA methodology were found between 78.1 (syringic acid) and 99.6% (o-coumaric acid) for red wines and between 76.2 and 99.1% for white wines. The inter-day precision, expressed as the relative standard deviation (RSD%), varied between 0.2% (p-coumaric and o-coumaric acids) and 7.5% (gentisic acid) while the intra-day precision between 0.2% (o-coumaric and cinnamic acids) and 4.7% (gallic acid and (−)-epicatechin). On the basis of analytical validation, it is shown that the MEPSC8/UHPLC–PDA methodology proves to be an improved, reliable, and ultra-fast approach for wine bioactive phenolics analysis, because of its capability for determining simultaneously in a single chromatographic run several bioactive metabolites with high sensitivity, selectivity and resolving power within only 10 min. Preliminary studies have been carried out on 34 real whole wine samples, in order to assess the performance of the described procedure. The new approach offers decreased sample preparation and analysis time, and moreover is cheaper, more environmentally friendly and easier to perform as compared to traditional methodologies.

Development of a new analytical approach based on cellulose membrane and chelator for differentiation of labile and inert metal species in aquatic systems

A new procedure was developed in this study, based on a system equipped with a cellulose membrane and a tetraethylenepentamine hexaacetate chelator (MD-TEPHA) for in situ characterization of the lability of metal species in aquatic systems. To this end, the DM-TEPHA system was prepared by adding TEPHA chelator to cellulose bags pre-purified with 1.0 mol L-1 of HCl and NaOH solutions. After the MD-TEPHA system was sealed, it was examined in the laboratory to evaluate the influence of complexation time (0-24 h), pH (3.0, 4.0, 5.0, 6.0 and 7.0), metal ions (Cu, Cd, Fe, Mn and Ni) and concentration of organic matter (15, 30 and 60 mg L-1) on the relative lability of metal species by TEPHA chelator. The results showed that Fe and Cu metals were complexed more slowly by TEPHA chelator in the MD-TEPHA system than were Cd, Ni and Mn in all pH used. It was also found that the pH strongly influences the process of metal complexation by the MD-TEPHA system. At all the pH levels, Cd, Mn and Ni showed greater complexation with TEPHA chelator (recovery of about 95-75%) than did Cu and Fe metals. Time also affects the lability of metal species complexed by aquatic humic substances (AHS); while Cd, Ni and Mn showed a faster kinetics, reaching equilibrium after about 100 min, and Cu and Fe approached equilibrium after 400 min. Increasing the AHS concentration decreases the lability of metal species by shifting the equilibrium to AHS-metal complexes. Our results indicate that the system under study offers an interesting alternative that can be applied to in situ experiments for differentiation of labile and inert metal species in aquatic systems. (c) 2006 Elsevier B.V. All rights reserved.

Development and application of a Saccharomyces cerevisiae-expressed nucleocapsid protein-based enzyme-linked immunosorbent assay for detection of antibodies against infectious bronchitis virus

A Saccharomyces cerevisiae-expressed nucleocapsid (N) polypeptide of the M41 strain of infectious bronchitis virus (IBV) was used as antigen in a recombinant yeast-expressed N protein-based enzyme-linked immunosorbent assay (Y-N-ELISA). The Y-N-ELISA was rapid, sensitive, and specific for detecting chicken serum antibodies to IBV, and it compared favorably with a commercial ELISA.

Development of an electrochemical sensor for potassium ions based on KSr(2)Nb(5)O(15) modified electrode

In the work described by this paper, we studied the development of a selective potassium ion sensor constituted of a carbon paste electrode modified (CPEM) with a novel KSr(2)Nb(2)O(15). The material KSr(2)Nb(2)O(15) is an oxide with the tetragonal tungsten bronze structure (TTB) type are in forefront both in the area of research as well as in industrial applications. The sensor response to potassium ions was linear in the concentration range 1.26 x 10(-5) at 1.62 x 10(-3) mol L(-1) (E (mV) = 32.7 + 51.1 log [K(+)]). The sensor based KSr(2)Nb(2)O(15), of the TTB-type presented very good potentiometric response, with a slope of 51.1 mV/dec (at 25 degrees C) and detection limit for the potassium ions of 7.27 x 10(-5) mol.L(-1)

Development of Nanostructured Electrochemical Sensor Based on Polymer Film Nickel-Salen for Determination of Dissolved Oxygen

An amperometric oxygen sensor based on a polymeric nickel-salen (salen = N,N '-ethylenebis(salicydeneiminato)) film coated platinum electrode was developed. The sensor was constructed by electropolymerization of nickel-salen complex at a platinum electrode in acetonitrile/tetrabuthylamonium perchlorate by cyclic voltammetry. The voltammetric behavior of the modified electrode was investigated in 0.5 mol L-1 KCl solution in the absence and presende of molecular oxygen. A significant increased of cathodic peak current (at -0.20 vs. SCE) of the modified electrode with addition of oxygen to the solution was observed. This result shows that the nickel-salen film on the surface of the electrode promotes the reduction of oxygen. The reaction can be brought about electrochemically where in the nickel(II) complex is first reduced to a nickel(I) complex at the electrode surface. The nickel(I) complex then undergoes a catalytic oxidation by the oxygen molecular in solution back to the nickel(II) complex, which can then be electrochemically re-reduced to produce an enhancement of the cathodic current. The plot of the cathodic current versus the dissolved oxygen concentration for chronoamperometry (potential fixed = -0.20 V) at the sensor was linear in the concentration range of 3.95 to 9.20 mg L-1 with concentration limit of 0.17 mg L-1 O-2. The modified electrode proposed is useful for the quality control and routine analysis of dissolved oxygen in commercial water and environmental water samples. The results obtained for the levels of dissolved oxygen are in agreement with the results obtained with an O-2 commercial sensor. (C) 2011 Published by Elsevier Ltd.

A soy-based product fermented by Enterococcus faecium and Lactobacillus helveticus inhibits the development of murine breast adenocarcinoma

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Fatigue and Fluoride Corrosion on Streptococcus mutans Adherence to Titanium-Based Implant/Component Surfaces

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

DEVELOPMENT OF A CHEMICALLY MODIFIED ELECTRODE BASED ON CARBON PASTE AND FUNCTIONALIZED SILICA-GEL FOR PRECONCENTRATION AND VOLTAMMETRIC DETERMINATION OF MERCURY(II)

A mercury-sensitive chemically modified electrode (CME) based on modified silica gel-containing carbon paste was developed. The functional group attached to the silica gel surface was 3-(2-thiobenzimidazolyl)propyl, which is able to complex mercury ions. This electrode was applied to the determination of mercury(II) ions in aqueous solution. The mercury was chemically preconcentrated on the CME prior to voltammetric determination by anodic stripping in the differential-pulse mode. A calibration graph covering the concentration range from 0.08 to 2 mg l-1 was constructed. The precision for six determinations of 0.122 and 0.312 mg l-1 Hg(II) was 3.2 and 2.9% (relative standard deviation), respectively. The detection limit for a 5-min preconcentration period was 0.013 mg l-1. A study for foreign ions was also made.

Development of biosensors based on hexacyanoferrates

Ferric and copper hexacyanoferrates (PB and CuHCF, respectively) were electrodeposited on glassy carbon electrodes providing a suitable catalytic surface for the amperometric detection of hydrogen peroxide. Additionally glucose oxidase was immobilized on top of these electrodes to form glucose biosensors. The biosensors were made by casting glucose oxidase-Nafion layers onto the surface of the modified electrodes. The operational stability of the films and the biosensors were evaluated by injecting a standard solution (5 mu M H2O2 for PB, 5 mM H2O2 for CuHCF and 1.5 mM glucose for both) over 5-10 h in a now-injection system with the electrodes polarized at - 50 (PB) and -200 mV (CuHCF) versus Ag/AgCl, respectively. The glucose biosensors demonstrated suitability for glucose determination: 0.0-2.5 mM (R-2 = 0.9977) for PB and 0.0-10 mM (R-2 = 0.9927) for CuHCF, respectively. The visualization of the redox catalyst modifiers (PB and CuHCF films) was presented by scanning electron micrographs. (C) 2000 Elsevier B.V. B.V. All rights reserved.

The role of oxygen vacancies on the microstructure development and on the electrical properties of SnO2-based varistors

Variations on the microstructure development and on the electrical properties of SnO2-based varistors are discussed on the basis of the oxygen vacancies created or annihilated by the presence of different additives. Electron paramagnetic resonance (EPR) analysis of sintered samples evidenced a substantial increase in the paramagnetic oxygen vacancies concentration when Nb2O5 is added to the SnO2 center dot Co3O4 system. on the other hand, the observed diminution in the concentration of such species after the addition of Fe2O3 indicates solid solution formation. The quantification of paramagnetic oxygen vacancies allowed to confirm the proposed substitutions taking place in the lattice during sintering. These findings are supported by scanning electron microscopy, by density measurements and by current density versus electric field curves. The characterization of secondary phases through EDS assisted SEM and TEM is also reported in this work.

Development of an open case-based decision-support system for diagnosis in oral pathology

Making diagnoses in oral pathology are often difficult and confusing in dental practice, especially for the lessexperienced dental student. One of the most promising areas in bioinformatics is computer-aided diagnosis, where a computer system is capable of imitating human reasoning ability and provides diagnoses with an accuracy approaching that of expert professionals. This type of system could be an alternative tool for assisting dental students to overcome the difficulties of the oral pathology learning process. This could allow students to define variables and information, important to improving the decision-making performance. However, no current open data management system has been integrated with an artificial intelligence system in a user-friendly environment. Such a system could also be used as an education tool to help students perform diagnoses. The aim of the present study was to develop and test an open case-based decisionsupport system.Methods: An open decision-support system based on Bayes' theorem connected to a relational database was developed using the C++ programming language. The software was tested in the computerisation of a surgical pathology service and in simulating the diagnosis of 43 known cases of oral bone disease. The simulation was performed after the system was initially filled with data from 401 cases of oral bone disease.Results: the system allowed the authors to construct and to manage a pathology database, and to simulate diagnoses using the variables from the database.Conclusion: Combining a relational database and an open decision-support system in the same user-friendly environment proved effective in simulating diagnoses based on information from an updated database.

Broiler breeder trace mineral nutrition and feeding practices on embryo progeny development

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

The contribution of biofuels to the sustainability of development in Latin America and the Caribbean

Includes bibliography