976 resultados para solid phase extraction


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

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

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The antibiotics sulfamethoxazole (SMTX) and ciprofloxacin (CIP) are commonly used in human and veterinary medicine, which explains their occurrence in wastewater. Anaerobic reactors are low-cost, simple and suitable technology to wastewater treatment, but there is a lack of studies related to the removal efficiency of antibiotics. To overcome this knowledge gap, the objective of this study was to evaluate the removal kinetics of SMTX and CIP using a horizontal-flow anaerobic immobilized biomass reactor. Two different concentrations were evaluated, for SMTX 20 and 40 μg L(-1); for CIP 2.0 and 5.0 μg L(-1). The affluent and effluent analysis was carried out in liquid chromatography/tandem mass spectrometry (LC-MS/MS) with the sample preparation procedure using an off-line solid-phase extraction. This method was developed, validated and successfully applied for monitoring the affluent and effluent samples. The removal efficiency found for both antibiotics at the two concentrations studied was 97%. Chemical oxygen demand (COD) exhibited kinetic constants that were different from that observed for the antibiotics, indicating the absence of co-metabolism. Also, though the antibiotic concentration was increased, there was no inhibitory effect in the removal of COD and antibiotics.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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In 2001, it was estimated that pesticide used worldwide exceeded 2.27 billion kilograms, over 35%, of which, were herbicides. Brazil is considered one of the leaders in the production of sugarcane and mainly ethanol as fuel. The monoculture of sugarcane requires the usage of a range of pesticides, among these, the herbicides diuron and tebuthiuron. The degradation products most studied (DCA and DCPU) are diuron's, especially for toxicological characteristics of this herbicide that is identified as carcinogen and suspected to be endocrine disruptor in mammals. After optimization of the chromatographic separation using HPLC-UV, the analytical curve was constructed in solvent and subsequently in the matrix (surface water). The extraction method contains the usage of SPE (solid phase extraction) (Strata-X, 200 mg/6 mL), applicating 1L of sample and elution with 5 mL of acetonitrile / methanol (50:50, v/v). Analysis by HPLC/UV was performed in gradient mode, acetonitrile/water (70/30-74/26 by 1 min, 74/26 - 78/22 till 3.2 min, returning to initial conditions and remaining this way until 10 min), 018 column (Phenomenex, 4.6 mm diameter, 250 mm long and 5pm particle size) and detection at 254 nm. Tests F and t were performed to verify the presence of the matrix effect. There was matrix effect to all analytes, ranging from -33% (DCA) and 38% (tebuthiuron). Thereby the method was optimized and validated for analysis of diuron, tebuthiuron, and DCPU DCA in surface water using HPLC/UV. The data obtained show that in order to assure the analytical reliability desired the use of the analytical curves in the matrix for the quantification of these analytes in water is required.

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Orange juice is a rich source of flavonoids, mainly the flavanones hesperidin and narirutin, associated with health benefits in humans. The objective of this study was to analyze the uptake of flavonoids in humans after the consumption of two types of orange juice, fresh squeezed (fresh juice, FJ) and commercially extracted and pasteurized (processed juice, PJ). Preliminary measurements showed that the main flavanones in PJ were approximately three-fold higher than in FJ. This study involved healthy volunteers including 12 men and 12 women, aged 27 ± 6, with a BMI of 24 ± 3 kg/m2. Volunteers drank 11.5 mL/kg body weight of fresh orange juice, and after an interval of 30 days they drank the same quantity of pasteurized orange juice. Urine was collected from each volunteer during 24 hours following juice consumption. Urine metabolites were recovered by solid phase extraction, and measured by HPLC–ESI–MS. Analyses of the urine samples showed high concentrations of glucuronic acid and sulfate conjugates of hesperetin and naringenin. The data indicate that the concentrations of the flavanone metabolites following consumption of PJ were approximately three times higher than for FJ, thus matching the relative doses of these compounds in the juices provided to the volunteers.

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

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Tetradifon, a potentially carcinogenic and mutagenic pesticide, can contribute to environmental and human contamination when applied to green bell pepper crops. In this context, in this work, a reliable and sensitive method for determination of tetradifon in Brazilian green bell pepper samples involving a differential pulse voltammetry (DPV) technique on a glassy carbon electrode is proposed. The electrochemical behavior of tetradifon as followed by cyclic voltammetry (CV) suggests that its reduction occurs via an irreversible five-electron transfer vs. Ag vertical bar AgCl, KCl 3 M reference electrode. Very well-resolved diffusion controlled voltammetric peaks have been obtained in a supporting electrolyte solution composed of a mixture of 40% dimethylformamide (DMF), 30% methanol, and 30% NaOH 0.3 mol L-1 at -1.43, -1.57, -1.73, -1.88, and -2.05 V. The proposed DPV method has a good linear response in the 3.00 - 10.0 mu mol L-1 range, with a limit of detection (L.O.D) of 0.756 mu mol L-1 and 0.831 mu mol L-1 in the absence and in the presence of the matrix, respectively. Moreover, improved L.O.D results (0.607 mu mol L-1) have been achieved in the absence of DMF from the supporting electrolyte solution. Recovery has been evaluated in five commercial green bell pepper samples, and recovery percentages ranging from 91.0 to 109 have been obtained for tetradifon determinations. The proposed voltammetric method has also been tested for reproducibility, repeatability, and potential interferents, and the results obtained for these three analytical parameters are satisfactory for electroanalytical purposes. (C) 2012 The Electrochemical Society. [DOI: 10.1149/2.024207jes] All rights reserved.

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Snake venom proteomes/peptidomes are highly complex and maintenance of their integrity within the gland lumen is crucial for the expression of toxin activities. There has been considerable progress in the field of venom proteomics, however, peptidomics does not progress as fast, because of the lack of comprehensive venom sequence databases for analysis of MS data. Therefore, in many cases venom peptides have to be sequenced manually by MS/MS analysis or Edman degradation. This is critical for rare snake species, as is the case of Bothrops cotiara (BC) and B. fonsecai (BF), which are regarded as near threatened with extinction. In this study we conducted a comprehensive analysis of the venom peptidomes of BC, BF, and B. jararaca (BJ) using a combination of solid-phase extraction and reversed-phase HPLC to fractionate the peptides, followed by nano-liquid chromatography-tandem MS (LC-MS/MS) or direct infusion electrospray ionization-(ESI)-MS/MS or MALDI-MS/MS analyses. We detected marked differences in the venom peptidomes and identified peptides ranging from 7 to 39 residues in length by de novo sequencing. Forty-four unique sequences were manually identified, out of which 30 are new peptides, including 17 bradykinin-potentiating peptides, three poly-histidine-poly-glycine peptides and interestingly, 10 L-amino acid oxidase fragments. Some of the new bradykinin-potentiating peptides display significant bradykinin potentiating activity. Automated database search revealed fragments from several toxins in the peptidomes, mainly from L-amino acid oxidase, and allowed the determination of the peptide bond specificity of proteinases and amino acid occurrences for the P4-P4' sites. We also demonstrate that the venom lyophilization/resolubilization process greatly increases the complexity of the peptidome because of the imbalance caused to the venom proteome and the consequent activity of proteinases on venom components. The use of proteinase inhibitors clearly showed different outcomes in the peptidome characterization and suggested that degradomic-peptidomic analysis of snake venoms is highly sensitive to the conditions of sampling procedures. Molecular & Cellular Proteomics 11: 10.1074/mcp.M112.019331, 1245-1262, 2012.