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The nutrition and feeding
Argon was used as inert gas at a constant flow of 1 L min-1 throughout the heating program, except during the atomization step, when the gas flow was interrupted. The absorbance signals were measured in peak area. The samples were cryogenically ground in a spex - freezer model Mill 6750 cryogenic mill. The slurries of fish feces and feeds were sonicated in a unique ultrasonic cell disruptor. Preparation of the graphite tube coated internally with tungsten. The inner walls of the pyrolytic graphite tubes with integrated platform used for determining manganese were coated with tungsten. This was made by injecting al" of 25 µL of a solution containing 1000 mg L-1 of the sodium tungstate modifier into the atomizer, which was then submitted to the stages of the heating program described.
Biological material for preparing the voor standard slurries. A feed devoid of some metal nutrients was prepared (in the case, devoid of calcium, iron, cobalt, copper, manganese, selenium and zinc). This feed was formulated with dehydrated starch, albumin and premix potassium and magnesium oxides. A lot of Nile tilapia, oreochromis niloticus juveniles, was fed with this feed in separate aquarium. The collected excrements were similar to the diets and contained all the metal nutrients. After the collection, the feces were treated as described in the previous item, however, the cryogenic milling was performed after washing exhaustingly with pure nitric acid.10 mol L-1, ultrapure water and drying was applied as already mentioned. Slurry sample preparation, after cryogenic grinding, 5 mg of the biological material samples (fish feed or feces) were transferred directly to the containers of the spectrometer autosampler, to which 5 mL of pure nitric acid 14 kompas mol L-1, 50 mL of Triton X-100.
The biological material slurry samples were then sonicated for 40 s directly in the autosampler containers. Apparatus, a provecto Analítica model dgt 100 plus microwave oven (Campinas, Brazil) was used to mineralize the samples. For the manganese determinations, a shimadzu model aa-6800 atomic absorption spectrometer was used, equipped with a background absorption corrector by self-reversal method (SR) system, and a pyrolytic graphite tube with integrated platform and automatic asc-6100 sampler. A shimadzu hollow cathode manganese lamp operated with a 10 mA current was also used. The wavelength applied was 279.5 nm and the spectral resolution was.5.
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Stock solutions of the analytes were prepared from reagents of spectroscopic purity (Johnson matthey, royston, hertfordshire, uk). The remaining solutions utilized, including the concentrated acid solutions used for mineralizing the samples, were analytical grade. All bottles for storing samples and standard solutions, glassware and containers of the atomic absorption spectrometer's autosampler were immersed in 10 v/v nitric acid for 24 h, rinsed with ultrapure water and dried before being used. The fish feces and feed samples were dried at 50 c in an oven with forced air circulation for 48 h and then cryogenically ground. A mass of approximately.0 g of the sample, together with a magnetic bar, were put into a polycarbonate flask, which was then closed and immersed in liquid nitrogen.
The impact between the sample and the magnetic bar, subjected to an oscillating magnetic field (20 impacts s-1 pulverized the sample. The sample grinding program consisted of an initial stage of 2 min of prefreezing, 1 min of pulverization, and again 1 min of freezing, followed by a second stage comprising two cycles of two pulverization and freezing stages, making a total of 8 min. This procedure yielded particles with a granulometry of less than.15,16. A portion of the samples was also mineralized in a microwave oven, as follows. Portions of 50 mg of cryogenically ground samples were transferred directly to the teflon flasks of the microwave oven, and.5 mL of pure nitric acid 14 mol L-1 plus.50 mL of hydrogen peroxide 30 m/m were added. Thereafter, the following power/timer program was run: step 1, 300W/3 min; step 2, 0 W/2 min; step 3, 450 W/5 min, step 4, 550 W/5 min; step 5, 650 W/5 min; step 6 (ventilation 0 W/5 min.16 After cooling, the resulting acid digestates were diluted.
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Considering also that the atomizer can act as a chemical reactor, the possibility of making solid sampling presents some advantages over the conventional digestion procedures. Besides eliminating the stage of total previous decomposition of sample, it diminishes the sample preparation time, decreases the analyte losses for excessive manipulation or retention on insoluble products, reduces the possibility of sample contamination, and mainly minimizes the action of dangerous acids on the analyst. Taking into account theses acpects, this work describes the development of a method to determine manganese in slurries of fish feed and feces samples by gfaas that eliminates the sample's mineralization step and allows for an estimate of its bioavailability. Experimental, reagents, standard solutions and samples, high purity deionized water (18.2 mw cm-1) obtained with an Elga ionic system (purelab option, usa pure nitric acid (Merck. hydrogen peroxide (Merck) and Triton X-100 products (Merck) were used throughout this work. The solution containing tungsten, which was employed to coat the inside of the graphite tube and used as a permanent modifier, was prepared by diluting a stock solution containing 1000 mg L-1 of sodium tungstate (Merck) with ultrapure water. The Pd(II) solution, also employed as a chemical modifier, was made in the same way, utilizing palladium nitrate (Merck) instead.
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An inadequate supply of supplements manganese usually results in retardation of growth. The importance of manganese has been recognized in broodstock nutrition. The manganese content of the diet influences its level and that of other trace elements in gonads.1-5 The absence of manganese in a fish meal diet significantly influenced the mineral composition of common carp gonads. The eggs produced by broodstock of brook trout and rainbow trout fed fish meal diets lacking manganese contained only low levels of this trace metal and subsequently hatchability was poor.6-9. So, the development of new methodologies that allow the reliable quantification of the metallic nutrients such as manganese, present at low concentrations in feeds, becomes fundamental in fish nutrition studies. In this context, the determination of metallic analytes in slurries by graphite furnace atomic absorption spectrometry shows that it is a robust technique. It provides several advantages, such as, high sensitivity, detection limits extremely low, the use of small sample volumes, determination of a wide variety of trace elements, etc.
Keywords: slurries samples, gfaas, chemical modifier, manganese determination. Resumo, neste trabalho, um método simples, rápido e sensível, é proposto para determinação de manganês em amostras de fezes e rações de peixes por espectrometria de absorção atômica em forno de grafite (gfaas) utilizando-se a introdução direta de suspensões das amostras no tubo de grafite. Os limites de detecção (LOD) e de quantificação (LOQ) calculados em relação a 20 leituras do branco das suspensões (0,50 m/v de fezes ou ração isentas de manganês) foram de 28 e 92 µg kg1 para as suspensões padrão de fezes e de. O método proposto foi aplicado em estudos de biodisponibilidade de manganês em diferentes amostras de rações de peixes e os resultados mostraram-se de acordo com os resultados obtidos utilizando-se amostras previamente mineralizadas por digestões ácidas em forno de microondas. Introduction, manganese is important for fish and hair is widely distributed in fish and animal tissue. The mitochondria have a greater concentration of manganese than cytoplasm or other cell organelles. Manganese is necessary for the normal functioning of brain and for proper lipid and carbohydrate metabolism. Manganese activates specific enzymes such as glycosyltransferase and non-specific enzymes such as kinases, transferases, hydrolases and decarboxylases. The activation of leucine aminopeptidase by manganese has been demonstrated in sole.
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Article, manganese determination by gfaas in feces and fish feed slurries. Vanessa rosa loureiroI; mayra. Padilhai, iinstituto de biociências, departamento de química e bioquímica, universidade Estadual paulista, cp botucatu-sp, brazil. Iifaculdade mannen de medicina veterinária e zootecnia, departamento de melhoramento e nutrição animal, Universidade Estadual paulista, cp botucatu-sp, brazil. Iiiinstituto de biociências, departamento de física e biofísica, universidade Estadual paulista, cp botucatu-sp, brazil. Abstract, this paper presents a simple, fast and sensitive method to determine manganese in samples of feces and fish feed by graphite furnace atomic absorption spectrometry (gfaas) by the direct introduction of slurries into the graphite tube. The limits of detection (LOD) and quantification (LOQ) calculated for 20 readings of the blank of the standard slurries (0.50 m/v of feces or feed devoid of manganese) were 28 and 92 µg kg-1 for the standard feces slurries and 34 and 110 µg kg-1. The proposed method was applied in bioavailability studies of manganese in different fish feeds and their results proved compatible with those obtained for samples mineralized by acid digestion using microwave oven.