Fátima Bento

Simplified 2,4-dinitrophenylhydrazine spectrophotometric assay for quantification of carbonyls in oxidized proteins.

This work proposes a modification of the 2,4-dinitrophenylhydrazine (DNPH) spectrophotometric assay commonly used to evaluate the concentration of carbonyl groups in oxidized proteins. In this approach NaOH is added to the protein solution after the addition of DNPH, shifting the maximum absorbance wavelength of the derivatized protein from 370 to 450nm. This reduces the interference of DNPH and allows the direct quantification in the sample solution without the need for the precipitation, washing, and resuspension steps that are carried out in the traditional DNPH method. The two methods were compared under various conditions and are statistically equivalent.

Direct electroanalytical method for alternative assessment of global antioxidant capacity using microchannel electrodes.

A new electroanalytical method for the characterization of global antioxidant capacity is proposed based on chronoamperometric responses monitored at microchannel band electrodes. This approach does not require any titrating species, biological elements, or precalibration curves. A thin-layer regime is established at the working electrode according to the geometry of the device and hydrodynamic flow rate. Under these conditions, the currents are directly proportional to the total concentration of antioxidants and do not depend on their respective diffusion coefficients. Measurements were performed with synthetic solutions and mixtures of four antioxidants used as sample tests: trolox, ascorbic acid, gallic acid, and caffeic acid. Operating potentials were selected at the formal potentials of some reactive oxygen species to simulate their oxidative attacks. The very good agreement obtained between simulations and experimental data validated this new electroanalytical procedure. These results pave the way for the concept of innovative sensor-type microfluidic devices for alternative determination of antioxidant capacity.

Enhanced electrochemical sensing of polyphenols by an oxygen-mediated surface

We report a straightforward heat treatment in air of commercial screen-printed carbon electrodes (SPCE) at different temperatures and times (ht-SPCE) that produces considerable electrocatalytic effects. The active area and the presence of oxygen groups on the ht-SPCE surface increased upon thermal treatment, more than doubling and by 20%, respectively. The increase of oxygen-containing carbon surface groups results in strong interactions and substantial improvement in Faradaic currents, up to a 10 fold increase in the voltammetric response of relevant polyphenols (hydroquinone, catechol, pyrogallol and dopamine). Moreover, ht-SPCE displayed higher selectivity towards the oxidation of polyphenol mixtures than the untreated SPCE. Finally, good linear ranges were obtained by voltammetric determination of dopamine with detection and quantification limits 5 times lower than the values obtained for the untreated SPCE. The versatility of oxidation by air makes this activation procedure very attractive and easy to implement, which can be further used for numerous applications in the (bio)sensor field.

Radical scavenging activity of antioxidants evaluated by means of electrogenerated HO radical

A method is proposed and tested concerning the characterization of antioxidants by means of their reaction with electrogenerated HO radicals in galvanostatic assays with simultaneous O2 evolution, using a Pt anode fairly oxidized. The consumption of a set of species with antioxidant activity, ascorbic acid (AA), caffeic acid (CA), gallic acid (GA) and trolox (T), is described by a first order kinetics. The rate of the processes is limited by the kinetics of reaction with HO radicals and by the kinetics of charge transfer. Information regarding the scavenger activity of antioxidants is obtained by the relative value of the rate constant of the reaction between antioxidants and HO radicals, k(AO,HO)/k(O2). The number of HO radicals scavenged per molecule of antioxidant is also estimated and ranged from 260 (ascorbic acid) to 500 (gallic acid). The method is applied successfully in the characterization of the scavenger activity of ascorbic acid in a green-tea based beverage.

Evaluation of total polyphenol content of wines by means of voltammetric techniques: Cyclic voltammetry vs differential pulse voltammetry

Taking advantage of the low oxidation potential of polyphenolic compounds, voltammetric techniques, such as cyclic voltammetry (CV) and differential pulse voltammetry (DPV) are used rather indiscriminately. In this work, we report Total Polyphenols results (TPP) obtained by these two techniques from a set of nine samples of red and Tawny Port wine. The CV and DPV voltammograms display significant correlations with the physical-chemical parameters used to characterize red and Tawny Port wines, particularly with polyphenols. Although data obtained from CV and DPV for a single polyphenol are directly proportional, important deviations are found between voltammetric results from wines. Results from CV tend to be larger than those from DPV. This difference, that can reach 50% of the TPP value, was related to the presence of total sulphur dioxide. In view of the present study, the polyphenol quantification in wines should be performed by DPV to minimize the interference of SO2.