Gilberto Batista de Souza

Microwave-assisted digestion procedures for biological samples with diluted nitric acid: identification of reaction products.

Microwave-assisted sample preparation using diluted nitric acid solutions is an alternative procedure for digesting organic samples. The efficiency of this procedure depends on the chemical properties of the samples and in this work it was evaluated by the determination of crude protein amount, fat and original carbon. Soybeans grains, bovine blood, bovine muscle and bovine viscera were digested in a cavity-microwave oven using oxidant mixtures in different acid concentrations. The digestion efficiency was evaluated based on the determination of residual carbon content and element recoveries using inductively coupled plasma optical emission spectrometry (ICP OES). In order to determine the main residual organic compounds, the digests were characterized by nuclear magnetic resonance ((1)H NMR). Subsequently, studies concerning separation of nitrobenzoic acid isomers were performed by ion pair reversed phase liquid chromatography using a C18 stationary phase, water:acetonitrile:methanol (75:20:5, v/v/v)+0.05% (v/v) TFA as mobile phase and ultraviolet detection at 254 nm. Sample preparation based on diluted acids proved to be feasible and a recommendable alternative for organic sample digestion, reducing both the reagent volumes and the variability of the residues as a result of the process of decomposition. It was shown that biological matrices containing amino acids, proteins and lipids in their composition produced nitrobenzoic acid isomers and other organic compounds after cleavage of chemical bonds.

Study of the protein-bound fraction of calcium, iron, magnesium and zinc in bovine milk

Abstract Two approaches were used to study the interaction of Ca, Fe, Mg and Zn with bovine milk proteins by inductively coupled plasma optical emission spectrometry (ICPOES). Selective separations in bovine milk samples were accomplished employing an acid protein precipitation using 100 g l−1 trichloroacetic acid (TCA), and an enzymatic protein hydrolysis using 50 g l−1 pepsin (PEP) solution, respectively. The results were compared with total mineral contents determined after microwave-assisted acid digestion. The results obtained by enzymatic and acid precipitation evidenced the different interaction forms of Ca, Fe, Mg and Zn in the system formed by milk components. Iron was not solubilized by the TCA treatment, but was recovered completely after the enzymatic treatment. Quantitative recoveries of Ca, Mg and Zn were obtained using both approaches, showing that these analytes were bound to milk compounds affected by either treatment. Calcium, Mg and Zn are mainly associated with colloidal calcium phosphate and Fe is bound to the backbone of the casein polypeptide chain, cleaved by pepsin enzyme. The proposed approaches could be used to assess the complexity of these chemical interactions.

Oxygen bomb combustion of biological samples for inductively coupled plasma optical emission spectrometry

Abstract A rapid sample preparation method is proposed for decomposition of milk powder, corn bran, bovine and fish tissues, containing certified contents of the analytes. The procedure involves sample combustion in a commercial stainless steel oxygen bomb operating at 25 bar. Most of the samples were decomposed within 5 min. Diluted nitric acid or water-soluble tertiary amines 10% v/v were used as absorption solutions. Calcium, Cu, K, Mg, Na, P, S and Zn were recovered with the bomb washings and determined by inductively coupled plasma optical emission spectrometry (ICP-OES). Ethanol mixed with paraffin was used as a combustion aid to allow complete combustion. A cooling step prior releasing of the bomb valve was employed to increase the efficiency of sample combustion. Iodine was also determined in milk samples spiked with potassium iodide to evaluate the volatilization and collection of iodine in amine CFA-C medium and the feasibility of its determination by ICP-OES with axial view configuration. Most of the element recoveries in the samples were between 91 and 105% and the certified and found contents exhibited a fair agreement at a 95% confidence level.

A polyvalent flow injection system for multielemental spectrophotometric analysis of plant materials

Abstract A polyvalent flow injection system consisting of a single manifold suitable to perform different determinations after only minor adaptations was conceived. Its use is particularly attractive for laboratories processing a large number of samples including several analytes. The system was applied to spectrophotometric determination of iron, copper, manganese and zinc (micronutrients) as well as calcium, magnesium and phosphorus (macronutrients) in plants. With the proposed system, slight variations in the coefficients of the calibration equations (usually

Determination of chloride in milk using sequential injection automated conductimetry

A new conductimetric sequential injection procedure is proposed for the automatic chloride determination in milk. The preparations of milk samples are time consuming procedures. The use of a sequential injection system coupled to a dialysis camera permitted easy automation and improved process control over the parameters, giving high throughput analyses. In this work, a fast and reliable automatedsampleaddition method isdescribed. Anuntreated sample ofmilk (142ml)was injectedwithastandardreferencesolution (750ml) in a carrier stream and dialysed for the conductimetric chloride determination. Potential interferences and ideal work conditions were appraised. Data acquisition and device control were achieved by computer. Results were precise (r.s.d. <1.0%) in agreement with both the oAcial titration procedure and with the milk standard reference materials. The system was designed for conductimetric samples addition analysis, and no major problems were observed. # 1999 Elsevier Science Ltd. All rights reserved.

Potentiometric determination of urea by sequential injection using Jack bean meal crude extract as a source of urease.

A sequential injection system was proposed to accomplish the potentiometric determination of urea. This procedure used an ammonium tubular selective electrode to assess ammonium concentration produced by enzymatic hydrolysis of urea from Jack bean meal (Canavalia ensiformis DC) crude extract. A gaseous diffusion device was coupled to the flow set-up allowing on-line sampling and suitable selectivity for determinations. A detection limit of 6.0x10(-4) mol urea l(-1), a relative standard deviation of 1.9% (n=10) and a sampling rate of 20 samples h(-1) were observed when 172 Sumner units (SU) of urease and 900 mul of sample were used. Results agreeing with a comparative method were obtained by the proposed procedure and the use of the crude extract solution combined with the sequential injection approach improved the performance, producing reproducible results and low costs in comparison with procedures using commercial enzymes.

Yield, Quality Components, and Nitrogen Levels of Silage Corn Fertilized with Urea and Zeolite

The zeolite and urea mixture may be use to improve nitrogen (N)–use efficiency of silage corn. The objective of this study was to evaluate dry-matter yield and nutritional levels of N of silage corn fertilized with urea and zeolite mixture. The experimental design was a 2 × 4 × 4 factorial randomized block design with three replications. Treatments included two types of stilbite zeolite (natural and concentrated), four levels of nitrogen (0, 50, 100, and 200 kg ha−1), and four ratios of zeolite (25%, 50%, and 100% of N level). Treatments were applied 60 days after planting with the topdressing fertilization. The use of concentrated (650 g kg−1 of stilbite) or natural (470 g kg−1 of stilbite) zeolite with urea increased silage corn dry-matter production and leaf N concentrations.

Fermentation parameters, quality and losses in sugarcane silages treated with chemical additives and a bacterial inoculant

The objective of this trial was to evaluate chemical additives and a bacterial inoculant on the inhibition of alcoholic fermentation and reduction of losses in sugarcane silages. Treatments were (doses on a fresh forage basis): without additive (control); urea (10 g/kg); urea (5 g/kg) + sodium benzoate (0.5 g/kg); sodium benzoate (1 g/kg); urea + ammonium sulfate in a 9:1 relation (10 g/kg); Lactobacillus buchneri (5 × 104 cfu/g). Silages were produced in 10.16- × 30-cm PVC tubes, provided with tight lids adapted with Bunsen valves for gas losses quantification. Minisilos were opened 139 days after ensiling. Ethanol content (227 g/kg dry matter - DM) and total DM loss (30%) were high in the control silage. All additives, except benzoate, decreased ethanol concentration in silages. Inoculation with L. buchneri increased acetic acid content in the silage, resulting in a 41% reduction in ethanol content and the lowest gas loss among treatments (15.2%). There was synergistic effect between additives for the combined use of urea and benzoate. Silage treated with urea + ammonium sulfate has higher content of total digestible nutrients than the silage treated with urea exclusively.

Production, nutritional status and chemical properties of soils with addition of cattle manure, reactive natural phosphate and biotite schist in Massai cultivar

The objective of this work was to evaluate the effects of cattle manure, reactive natural phosphate and biotite schist on the soil fertility, yield and nutritional status of Megathyrsus spp. cv. Massai. The experiment was conducted under field conditions, in a dystrophic Red Yellow Latosol (Oxisol). It was used a randomized block experimental design with the following treatments: three natural reactive rock phosphate from Algeria (Djebel-Onk) doses (0, 100 and 200 kg ha-1 of P2O5), three biotite schist doses (0, 150 and 300 kg ha-1 of K2O) and three cattle manure doses (0, 20 and 40 t ha-1). The application of natural phosphate increased dry matter yield, however, application of cattle manure and biotite schist did not influence this variable. Foliar levels of nitrogen, potassium and magnesium (cattle manure), phosphorous and boron (natural rock phosphate) and potassium (biotite schist) were influenced by the applied fertilizer doses. Only the levels of phosphorous in the soil and in the plant and levels of magnesium and boron in the plant show interaction with dry matter yield of Massai cultivar.

Desenvolvimento e validação do método QuEChERS na determinação de resíduos de medicamentos veterinários em leite e carne de búfalo

DEVELOPMENT AND VALIDATION OF THE QuEChERS METHOD FOR THE DETERMINATION OF VETERINARY DRUG RESIDUES IN BUFFALO MILK AND MEAT. Analytical methods were developed and validated to determine residues of veterinary drugs in buffalo milk and meat, using the QuEChERS method and gas chromatography-mass spectrometry. Both milk and meat, at 2 g of sample, 4 mL of acetonitrile, 0.8 g of MgSO4 and 0.2 g of NaCl, were used in the liquid-liquid partition, whereas 50 mg of C18, 50 mg of PSA and 150 mg of MgSO4 were employed in the dispersive solid-phase extraction (d-SPE). The methods showed sensitivity, precision and accuracy. The quantitation limits were in agreement with the maximum residue limit established by the Codex Alimentarius, FAO and WHO.