Electroactive LbL films of metallic phthalocyanines and poly(<Emphasis Type="Italic">0</Emphasis>-methoxyaniline) for sensing

Multilayered nanostructured films have been widely investigated for electrochemical applications as modified electrodes, including the layer-by-layer (LbL) films where properties such as thickness and film architecture can be controlled at the molecular level. In this study, we investigate the electrochemical features of LbL films of poly(o-methoxyaniline; POMA) and tetrasulfonated phthalocyanines containing nickel (NiTsPc) or copper (CuTsPc). The films displayed well-defined electroactivity, with redox pairs at 156 and 347 mV vs SCE, characteristic of POMA, which allowed their use as modified electrodes for detecting dopamine and ascorbic acid at concentrations as low as 10⁻⁵ M. This paper is dedicated to Prof. Francisco Nart, in memoriam.     

Characterization of must and wine of six varieties of grapes by direct infusion electrospray ionization mass spectrometry

Samples of must derived from six different varieties of grapes taken during the fermentation process, as well as the respective wine samples directly after the end of the malolactic fermentation, were analyzed by direct infusion negative ion mode electrospray ionization mass spectrometry (ESI-MS). Diagnostic ions for must were different from those of wine samples, although small variations for each of the grape varieties were also detected. The addition of unfermented must or sugar to wine could also be clearly detected. The spectra were acquired in a few minutes per sample, indicating that ESI-MS can be used for high-throughput analysis of samples and should prove useful for quality control during and after the fermentation process.     

New designs for inhibitors of the NF-κB: DNA binding

We present a series of new inhibitors of the association between nuclear factor kappa B (NF-B) and the corresponding B site in DNA. They were designed using the lead compound 15-deoxy-^12,14 -prostaglandin J₂ (PGJ2), which is a natural product with demonstrated inhibitory efficiency for this system. First, the binding mode of PGJ2 to NF-B was unraveled by GOLD docking calculation. Subsequently, substitutions were made to PGJ2 to optimize its association with NF-B. Care was taken not to strongly increase the reactivity of the new compounds, and to keep the overall shape, size and hydrophilicity of the lead compound, which should render them a similar bioavailability. Molecular mechanics calculations were performed to decide on the suitability of the substitutions, and to evaluate the energies of association with NF-B. Density functional theory calculations were performed also to study the overall reactivity of the substituted drugs towards NF-B. Important general conclusions were obtained, concerning the improvement of these natural inhibitors; namely, a set of rational methodologies were deduced to improve the association between the PGJ2 derivatives and NF-B, and their efficiency demonstrated by generating a set of substituted complexes, some of them with a very much increased affinity for NF-B, opening new doors to enlarge the therapeutic capabilities of this class of drugs.     

Hydrolysis of new phthalimide-derived esters catalyzed by immobilized lipase

The last step of the production of four phthalimide-derived acids, designed to act as antiasthma drugs, was performed by enzymatic hydrolysis of the respective methyl or ethyl esters. The esters 4-ethyl-[2-(1,3-dioxo-1,3-dihydro-2-isoindoylyl)]-phenoxyacetic methyl ester (PHT-MET), 4-ethyl-[2-(1,3-dioxo-1,3-dihydro-2-isoindoylyl)]-phenoxyacetic ethyl ester, 4-(1,3-dioxo-1,3-dihydro-2-isoindoylyl)-phenoxyacetic ethyl ester, and 2-(1,3-dioxo-1, 3-dihydro-2-isoindoylyl)-phenoxyacetic ethyl ester were hydrolyzed by immobilized lipase. The enzymatic reaction could be used only to produce the desired 4-substituted compounds. The best result that was found to hydrolysis of PHT-MET, and, therefore, that ester was selected for optimization experiments in a three-phase system. Reactions were performed with solid biocatalyst (Lipozyme® RM IM), organic solvent phase (ethyl acetate), and aqueous phase (saturated Na₂CO₃ solution). To optimize the reaction conditions, an experimental design optimization procedure was used. The variables studied were the amount of enzyme, the temperature, and the volume of the aqueous solution. Time course experiments were then performed for different initial enzyme concentrations (0.5, 0.9, and 1.4 U_H/mL of solvent). The optimized reaction conditions found were 20 mg of Lipozyme (0.9 U_H/mL_solvent) and 5.0 mL of Na₂CO_3(sat) at 40°C for 6 h.     

Optimization of matrix solid-phase dispersion extraction method for the analysis of isoflavones in Trifolium pratense

A method based on matrix solid-phase dispersion (MSPD) has been developed for the determination of 12 isoflavones in Trifolium pratense L. Dried leaf samples were blended with C₁₈, placed in small columns and isoflavones extracted with dichloromethane–methanol. Analyses were performed by high performance liquid chromatography with diode array detection (HPLC-DAD) with 2-methoxyflavone as internal standard. Several dispersants, eluents and clean-up steps were tested during the optimization of the process in order to obtain the best selectivity and yields. Mean recoveries ranged from 70% to 119%, with relative standard deviations <18%. The limits of detection were between 0.006 mg/l for biochanin A and 0.108 mg/l for daidzin. The performance of the optimized method in real samples was compared with a conventional method based in solid–liquid extraction (SLE).     

Determination of Picoxystrobin and Pyraclostrobin by MEKC with On-Line Analyte Concentration

Micellar electrokinetic capillary chromatography was used for the determination of picoxystrobin and pyraclostrobin. The background electrolyte consisted of borate buffer (40 mmol L⁻¹ pH 8.5), SDS (30 mmol L⁻¹) and acetonitrile (15% in volume). Runs were made at 25 °C with 25 kV applied potential. The developed method was applied to analyte fortified urine samples. On-line analyte concentration, combined with a capillary of a longer optical path length, allowed limits of quantification of 8.6 × 10⁻⁸ mol L⁻¹ for picoxystrobin and 1.8 × 10⁻⁷ mol L⁻¹ for pyraclostrobin.

     

Qualitative properties and stabilizability of a model for blood thrombin formation

We perform a qualitative analysis of a differential equation that was originally introduced by Stortelder, Hemker and Hemker to model the formation of thrombin, and discuss issues of controllability and stabilizability. Results include a general proof of convergence to equilibrium, and of local exponential stabilizability.