Maria do Carmo Hespanhol da Silva

A green and sensitive method to determine phenols in water and wastewater samples using an aqueous two-phase system

A greener and more sensitive spectrophotometric procedure has been developed for the determination of phenol and o-cresol that exploits an aqueous two-phase system (ATPS) using a liquid-liquid extraction technique. An ATPS is formed mostly by water and does not require organic solvent. Other ATPS components used in this study were the polymer, polyethylene oxide, and some salts (i.e., Li(2)SO(4), Na(2)SO(4) or K(2)HPO(4)+KOH). The method is based on the reaction between phenol, sodium nitroprusside (NPS) and hydroxylamine hydrochloride (HL) in an alkaline medium (pH 12.0), producing the complex anion [Fe(2)(CN)(10)](10-) that spontaneously concentrates in the top phase of the system. The linear range was 1.50-500microgkg(-1) (R>or=0.9997; n=8) with coefficients of variation equal to 0.38% for phenol and 0.30% for o-cresol (n=5). The method yielded limits of detection (LODs) of 1.27 and 1.88microgkg(-1) and limits of quantification (LOQs) of 4.22 and 6.28microgkg(-1) for phenol and o-cresol, respectively. Recoveries between 95.7% and 107% were obtained for the determination of phenol in natural water and wastewater samples. In addition, excellent agreement was observed between this new ATPS method and the standard 4-aminoantipyrine (4-AAP) method.

Application of the response surface methodology for optimization of whey protein partitioning in PEG/phosphate aqueous two-phase system.

In order to develop a new strategy for β-lactoglobulin (β-lg) removal from whey protein, partitioning of α-lactalbumin (α-la), β-lg and glycomacropeptide (Gmp) was studied using aqueous two phase systems (ATPS). A system composed of 13% (w/w) polyethylene glycol (PEG, average molar mass 2000 g/mol) and 13% (w/w) potassium phosphate was used at 25°C. A central composite rotatable design (CCRD) associated to the response surface methodology (RSM) was applied to investigate the effects of NaCl concentration and pH on the partition of these proteins. It was found that α-la and Gmp partitioned to the top phase rich in PEG, whereas β-lg partitioned to the bottom phase rich in salt. According to the RSM, optimal conditions for β-lg removal where found where pH was equal to 6.7 and salt concentration was 0.35 mol/L. Under these conditions, the partition coefficient K(α) was 0.48 and K(Gmp) was 0.92. On the other hand, the partition coefficient K(β) was only 0.01. In such conditions β-lg preferentially concentrates in the bottom phase, while the top phase exclusively contains the proteins α-la and Gmp. Fractionation of the proteins from fresh whey was performed in a three stage cross-flow extraction system. The extraction yield for β-lg in the bottom phase was 97.3%, while the yields for α-la and Gmp in the top phase were 81.1% and 97.8%, respectively.

Thermodynamic studies of partitioning behavior of lysozyme and conalbumin in aqueous two-phase systems

The objective of this study was to determine the thermodynamic parameters (Delta(tr)G, Delta(tr)H and Delta(tr)S) associated with lysozyme and conalbumin partitioning in aqueous two-phases systems (ATPS). Influence of salt type and polyethylene glycol (PEG) concentrations on the partition coefficient of lysozyme and conalbumin from egg white was studied. The evaluated ATPS were composed of PEG 1500 and inorganic salts (sodium citrate and sodium sulfate) at a temperature of 25 degrees C and pH 7.0, with PEG 1500 g mol(-1) concentrations of 14%, 16% and 18% (mass basis). Partitioning of lysozyme in PEG-citrate ATPS was enthalpically driven, however the PEG-sulfate ATPS was entropically driven. The tested systems can be employed for the separation of these two proteins in egg white, due to the fact that lysozyme migrates toward the polymeric phase and conalbumin to the saline phase in both ATPS. A high recovery of conalbumin in the saline phase of the PEG-sulfate ATPS was determined to be enthalpically driven.

Aqueous two-phase systems: A new approach for the determination of p-aminophenol

A new method has been developed for the spectrophotometric determination of p-aminophenol (PAP) in water, paracetamol formulations and human urine samples with a recovery rate between 94.9 and 101%. This method exploits an aqueous two-phase system (ATPS) liquid-liquid extraction technique with the reaction of PAP, sodium nitroprusside and hydroxylamine hydrochloride in pH 12.0, which produces the [Fe(2)(CN)(10)](10-) anion complex that spontaneously concentrates in the top phase of the ATPS ([Formula in text]). The ATPS does not require an organic solvent, which is a safer and cleaner liquid-liquid extraction technique for the determination of PAP. The linear range of detection was from 5.00 to 500 μg kg(-1) (R ≥ 0.9990; n=8) with a coefficient of variation of 2.11% (n=5). The method exhibited a detection limit of 2.40 μg kg(-1) and a quantification limit of 8.00 μg kg(-1). The ATPS method showed a recovery that ranged between 96.4 and 103% for the determination of PAP in natural water and wastewater samples, which was in excellent agreement with the results of the standard 4-aminoantipyrine method that was performed on the same samples.

Microcalorimetric and SAXS Determination of PEO―SDS Interactions: The Effect of Cosolutes Formed by Ions

The effect of different ionic cosolutes (NaCl, Na(2)SO(4), Li(2)SO(4), NaSCN, Na(2)[Fe(CN)(5)NO], and Na(3)[Co(NO)(6)]) on the interaction between sodium dodecyl sulfate (SDS) and poly(ethylene oxide) (PEO) was examined by small-angle X-ray scattering (SAXS) and isothermal titration calorimetric techniques. The critical aggregation concentration values (cac), the saturation concentration (C(2)), the integral enthalpy change for aggregate formation (ΔH(agg)(int)) and the standard free energy change of micelle adsorption on the macromolecule chain (ΔΔG(agg)) were derived from the calorimetric titration curves. In the presence of 1.00 mmol L(-1) cosolute, no changes in the parameters were observed when compared with those obtained for SDS-PEO interactions in pure water. For NaCl, Na(2)SO(4), Li(2)SO(4), and NaSCN at 10.0 and 100 mmol L(-1), the cosolute presence lowered cac, increased C(2), and the PEO-SDS aggregate became more stable. In the presence of Na(2)[Fe(CN)(5)NO], the calorimetric titration curves changed drastically, showing a possible reduction in the PEO-SDS degree of interaction, possibility disrupting the formed nanostructure; however, the SAXS data confirmed, independent of the small energy observed, the presence of aggregates adsorbed on the polymer chain.

Sistema aquoso bifásico: uma alternativa eficiente para extração de íons

Solvent extraction has been successfully applied to metal ion preconcentration and often meant the use of toxic organic diluents. However, regulatory pressure is increasingly focusing on the use and disposal of organic solvents, and thus the development of nonhazardous alternatives is important. In this review, we examine the application of aqueous biphasic systems (ABS) to extraction of ions, analyzing their potential and limitations and suggest that ABSs could be an efficient substitute for oil/water biphasic systems. ABSs are formed by mixing certain inorganic salts and water-soluble polymers, or by mixing two water-soluble polymers.

Binding thermodynamics of synthetic dye Allura Red with bovine serum albumin.

The interaction between Allura Red and bovine serum albumin (BSA) was studied in vitro at pH 7.4. The fluorescence quenching was classified as static quenching due to the formation of AR-BSA complex, with binding constant (K) ranging from 3.26±0.09 to 8.08±0.0610(4)L.mol(-1), at the warfarin binding site of BSA. This complex formation was driven by increasing entropy. Isothermal titration calorimetric measurements also showed an enthalpic contribution. The Allura Red diffusion coefficient determined by the Taylor-Aris technique corroborated these results because it reduced with increasing BSA concentration. Interfacial tension measurements showed that the AR-BSA complex presented surface activity, since interfacial tension of the water-air interface decreased as the colorant concentration increased. This technique also provided a complexation stoichiometry similar to those obtained by fluorimetric experiments. This work contributes to the knowledge of interactions between BSA and azo colorants under physiological conditions.

Etiologia do oídio do eucalipto

Isolates of Oidium spp. from Eucalyptus pellita were compared to Erysiphe cichoracearum, obtained from Dahlia sp. and Sphaerotheca pannosa, from Rosa sp. Based on the morphological characteristics of the anamorph, it was concluded that the Oidium isolate from eucalypt was similar to the isolate from rose identified as S. pannosa. Inoculations of Oidium isolates from all studied host species showed that S. pannosa obtained from eucalypt and rose, and E. cichoracearum from Dahlia sp. were pathogenic to seedlings of Eucalyptus pellita, one of the most susceptible species to the pathogen, under greenhouse conditions.Compararam-se isolados de Oidium sp. obtidos de mudas de eucalipto (Eucalyptus spp.) com isolados de Erysiphe cichoracearum, originario de dalia, e Sphaerotheca pannosa de roseira (Rosa sp.), por meio de caracteristicas morfologicas. Com base nas caracteristicas morfologicas (fase anamorfica) do tubo germinativo, do micelio, do apressorio, de conidioforos e conidios e na presenca de corpos de fibrosina, concluiu-se que o isolado de Oidium, obtido de eucalipto e similar ao de roseira, descrito como S. pannosa. Inoculacoes dos isolados do fungo originarios de todos os hospedeiros estudados, indicaram que S. pannosa, obtido de roseira, e E. cichoracearum, de dalia (Dahlia sp.), foram patogenicos a mudas de Eucalyptus pellita, uma das especies mais suscetiveis a esta doenca, em condicoes de casa de vegetacao.

Polydiacetylene/triblock copolymer nanoblend applied as a sensor for micellar casein: A thermodynamic approach

Polydiacetylene (PDA) and triblock copolymer nanoblends were synthesized to detect micellar casein (MC), the main milk protein and an indicator of milk quality. UV-Vis spectrum showed that MC induced blue-to-red transition in nanoblends. When nanoblends and MC were separated by dialysis membrane colorimetric response (CR) was similar, whereas a remarkable CR reduction was noticed after addition of dialyzed-MC, suggesting that small molecules present in MC (salts) caused PDA color change. Interaction enthalpy variation between nanoblends and MC showed an abrupt increase that coincided with MC concentration when colorimetric transition occurred. Copolymer hydrophobic/hydrophilic balance and presence of other molecules in the system affected nanoblends CR. MC salts were found to interact with nanoblends leading to color changes. MC concentration, MC salt release, copolymer hydrophobic/hydrophilic balance, and presence of other molecules in the system affected responses of the sensors. These results contribute to future applications of PDA/copolymer nanosensors to dairy models.

Optimization of flavor ester synthesis catalysed by Aspergillus niger lipase

The performance of Aspergillus niger lipase in catalyzing of butyl butyrate synthesis was studied as a function of reaction parameters temperature (°C), substrate molar ratio and added water (% (v/v)). A face-centered design (FCD) and response surface methodology (RSM) were applied in order to optimize the esterification yield (Y). The optimal conditions for butyl butyrate synthesis were found at 40°C, substrate molar ratio of 1:2.41 butyric acid:n-butanol and added water content of 1.05% (v/v). Under these conditions, over 94.5% of esterification was obtained in 20 h. It was verified a favorable effect of alcohol excess in relation to acid. High temperatures (above 40°C) were detrimental to the synthesis of butyl butyrate. Addition of water also provided a strong increase in esterification yield (%).This high esterification yield represents an improvement to previously reported results and confirms the potential application of Aspergillus niger lipases (ANL) in processes for producing flavor ester. Key words: Lipase, Aspergillus niger, esterification, butyl butyrate, flavor ester, optimization.