Desenvolvimento da técnica de MALDI Imaging utilizando tecidos animais

Currently the study of important molecular compounds present in low abundance in some tissues has been a challenge for proteomic analysis classic. An analysis requires more exploratory investigation of small regions of a tissue or a group of cells. MALDI Imaging Technology (MSI) is an application of mass spectrometry facing the chemical analysis of intact tissues. Thus, advances in mass spectrometry MALDI being obtained by the integration of histology, the best methods and automation are the main tools of data analysis. This tool has become essential to analyze the spatial distribution of peptides and proteins throughout the tissue sections, providing an enormous amount of data with minimum sample preparation. Thus, the aim of this study was to develop the technique of MALDI Imaging using tissue from glioblastoma multiforme (GBM), a form of most common malignant tumor in the brain. For this we used the printer chemical ChIP-1000 (Chemical Inkjet Printer, Shimadzu) and mass spectrometer type Maldi-ToF-ToF (Axima Performance, Shimadzu), a search of the identifications were performed in databases such as SwissProt. We identified more than forty proteins with diverse functions such as proteins F-actin-capping and Thymosin to the structure and organization cellular and proteins such several Tumor necrosis factor receptor development-related pathology. The development of this technique will permit to carry-out proteomic analysis directly into the tissue, enabling earlier diagnosis of diseases, as well as the identification and characterization of potential biomarkers of disease.

Analysis of fluoxetine and norfluoxetine in human plasma by HPLC-UV using a high purity C18 silica-based SPE sorbent

This paper reports on the development and validation of a simple and sensitive method that uses solid phase extraction (SPE) and liquid chromatography with ultraviolet detection to analyze fluoxetine (FLX) and norfluoxetine (NFLX) in human plasma samples. A lab-made C18 SPE phase was synthesized by using a sol–gel process employing a low-cost silica precursor. This sorbent was fully characterized by nuclear magnetic resonance (NMR), Fourier-transform infrared spectroscopy (FT-IR), and scanning electron microscopy (SEM) to check the particles' shape, size and C18 functionalization. The lab-made C18 silica was used in the sample preparation step of human plasma by the SPE-HPLC-UV method. The method was validated in the 15 to 500 ng mL 1 range for both FLX and NFLX using a matrix matched curve. Detection limits of 4.3 and 4.2 ng mL 1 were obtained for FLX and NFLX, respectively. The repeatability and intermediary precision achieved varied from 7.6 to 15.0% and the accuracy ranged from 14.9 to 9.1%. The synthesized C18 sorbent was compared to commercial C18 sorbents. The average recoveries were similar (85–105%), however the lab-made C18 silica showed fewer interfering peaks in the chromatogram. After development and validation, the method using the lab-made C18 SPE was applied to plasma samples of patients under FLX treatment (n ¼ 6). The concentrations of FLX and NFLX found in the samples varied from 46.8–215.5 and 48.0–189.9 ng mL 1 , respectively.

Peptídeos cíclicos hepatotóxicos: MALDI-TOF como uma ferramenta analítica

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

Análise química de elementos potencialmente tóxicos em baterias e pilhas por FAAS e ICP OES

Pós-graduação em Ciência dos Materiais - FEIS

Development of a new clean methodology with ultrasound-assisted extraction for analysis of sodium in pet foods

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Rapid and sensitive ultra-high-pressure liquid chromatography method for quantification of antichagasic benznidazole in plasma: application in a preclinical pharmacokinetic study

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

Direct determination of chromium in empty medicine capsules by tungsten coil atomic emission spectrometry

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

The ultra-sonication of minerals in swine feed

A sample preparation method based on ultrasound assisted-extraction (UAE) of Ca, Mg and P from swine feed has been described. The experiment was performed to cover the variables influencing the sonication process and, the method validation using standard reference material. Final solutions obtained upon sonication were analyzed by flame atomic absorption spectrometry (for Ca and Mg) and by UV-vis spectrophotometry (for P). The best conditions for metal extraction were as follows: sample mass: 100 mg in 20 mL 0.10 mol/L HCl, a particle size: <60 μm, sonication time: 5 cycles of 10 s and ultrasound power: 102 W. The UAE method was applied in digestibility assays in different piglet feeds and their results showed that it is highly comparable (P > 0.05) to the other methods used for such purposes, as block digestion, and offered a Ca, Mg and P method of quantification limit of 10.6, 12.4 and 14 mg/kg, respectively. The major advantages of the UAE method compared to other methods are the high treatment rate, low reagent usage in the extracts and, it does not generate toxic residues that might negatively affect human health and the environment, accompanied by good precision and accuracy.

Rapid determination of 12 antibiotics and caffeine in sewage and bioreactor effluent by online column-switching liquid chromatography/tandem mass spectrometry

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

Sulfamethoxazole and ciprofloxacin removal using a horizontal-flow anaerobic immobilized biomass reactor

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.

Desenvolvimento de detector eletroquímico para cromatografia líquida de alta eficiência baseado em um eletrodo modificado com nanopartículas metálicas para determinação de ácidos carboxílicos em vinhaça de cana-de-açúcar

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

Aperfeiçoamento e avaliação de um novo sistema de digestão assistida por aquecimento condutivo em frasco fechado para preparar amostras de carne “in natura” para análise elementar

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Avaliação do Clinpro XT® na redução da permeabilidade da dentina radicular e resistência a desafios ácidos, na obliteração de túbulos dentinários e rugosidade superficial da dentina

Pós-graduação em Odontologia - FOAR

Interaction of giant extracellular Glossoscolex paulistus hemoglobin (HbGp) with ionic surfactants: A MALDI-TOF-MS study

The giant extracellular hemoglobin of Glossoscolex paulistus (HbGp) is constituted by approximately 144 subunits containing heme groups with molecular masses in the range of 16-19 kDa forming a monomer (d) and a trimer (abc), and around 36 non-heme structures, named linkers (L). Matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF-MS) analysis was performed recently, to obtain directly information on the molecular masses of the different subunits from HbGp in the oxy-form. This technique demonstrated structural similarity between HbGp and the widely studied hemoglobin of Lumbricus terrestris (HbLt). Indeed, two major isoforms (d(1) and d(2)) of identical proportions with masses of 16,355+/-25 and 16,428+/-24 Da, respectively, and two minor isoforms (d(3) and d(4)) with masses around 16.6 kDa were detected for monomer d of HbGp. In the present work, the effects of anionic sodium dodecyl sulfate (SDS) and cationic cethyltrimethyl ammonium chloride (CTAC) on the oligomeric structure of HbGp have been studied by MALDI-TOF-MS in order to evaluate the interaction between ionic surfactants and HbGp. The data obtained with this technique show an effective interaction of cationic surfactant CTAC with the two isoforms of monomer d, d(1) and d(2), both in the whole protein as well as in the pure isolated monomer. The results show that up to 10 molecules of CTAC are bound to each isoform of the monomer. Differently, the mass spectra obtained for SDS-HbGp system showed that the addition of the anionic surfactant SDS does not originate any mass increment of the monomeric subunits, indicating that SDS-HbGp interaction is, probably, significantly less effective as compared to CTAC-HbGp one. The acid pI of the protein around 5.5 is, probably, responsible for this behavior. The results of this work suggest also some interaction of both surfactants with linker chains as well as with trimers, as judged from observed mass increments. Our data are consistent with a recent spectroscopic study showing a strong interaction between CTAC and HbGp at physiological pH [P.S.Santiago, et al, Biochim. Biophys. Acta. 1770 (2007) 506-517.]. (C) 2007 Elsevier B.V. All rights reserved.

Optimization of in situ derivatization SPME by experimental design for GC-MS multi-residue analysis of pharmaceutical drugs in wastewater

This paper presents the development of a procedure, which enables the analysis of nine pharmaceutical drugs in wastewater using gas chromatography-mass spectrometry (GC-MS) associated with solid-phase microextraction (SPME) for the sample preparation. Experimental design was applied to optimize the in situ derivatization and the SPME extraction conditions. Ethyl chloroformate (ECF) was employed as derivatizing agent and polydimethylsiloxane-divinylbenzene (PDMS-DVB) as the SPME fiber coating. A fractional factorial design was used to evaluate the main factors for the in situ derivatization and SPME extraction. Thereafter, a Doehlert matrix design was applied to find out the best experimental conditions. The method presented a linear range from 0.5 to 10 mu g/L, and the intraday and interday precision were lower than 16%. Applicability of the method was verified from real influent and effluent samples of a wastewater treatment plant, as well as from samples of an industry wastewater and a river.