210 resultados para HPLC-UV bioanalytical methodology


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This work presents recent results concerning a design methodology used to estimate the positioning deviation for a gantry (Cartesian) manipulator, related mainly to structural elastic deformation of components during operational conditions. The case-study manipulator is classified as gantry type and its basic dimensions are 1,53m x 0,97m x 1,38m. The dimensions used for the calculation of effective workspace due to end-effector path displacement are: 1m x 0,5m x 0,5m. The manipulator is composed by four basic modules defined as module X, module Y, module Z and terminal arm, where is connected the end-effector. Each module controlled axis performs a linear-parabolic positioning movement. The planning path algorithm has the maximum velocity and the total distance as input parameters for a given task. The acceleration and deceleration times are the same. Denavit-Hartemberg parameterization method is used in the manipulator kinematics model. The gantry manipulator can be modeled as four rigid bodies with three degrees-of-freedom in translational movements, connected as an open kinematics chain. Dynamic analysis were performed considering inertial parameters specification such as component mass, inertia and center of gravity position of each module. These parameters are essential for a correct manipulator dynamic modelling, due to multiple possibilities of motion and manipulation of objects with different masses. The dynamic analysis consists of a mathematical modelling of the static and dynamic interactions among the modules. The computation of the structural deformations uses the finite element method (FEM).

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This work presents a methodology for the development of Teleoperated Robotic Systems through the Internet. Initially, it is presented a bibliographical review of the Telerobotic systems that uses Internet as way of control. The methodology is implemented and tested through the development of two systems. The first is a manipulator with two degrees of freedom commanded remotely through the Internet denominated RobWebCam (http://www.graco.unb.br/robwebcam). The second is a system which teleoperates an ABB (Asea Brown Boveri) Industrial Robot of six degrees of freedom denominated RobWebLink (http://webrobot.graco.unb.br). RobWebCam is composed of a manipulator with two degrees of freedom, a video camera, Internet, computers and communication driver between the manipulator and the Unix system; and RobWebLink composed of the same components plus the Industrial Robot. With the use of this technology, it is possible to move far distant positioning objects minimizing transport costs, materials and people; acting in real time in the process that is wanted to be controller. This work demonstrates that the teleoperating via Internet of robotic systems and other equipments is viable, in spite of using rate transmission data with low bandwidth. Possible applications include remote surveillance, control and remote diagnosis and maintenance of machines and equipments.

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Industrial applications demand that robots operate in agreement with the position and orientation of their end effector. It is necessary to solve the kinematics inverse problem. This allows the displacement of the joints of the manipulator to be determined, to accomplish a given objective. Complete studies of dynamical control of joint robotics are also necessary. Initially, this article focuses on the implementation of numerical algorithms for the solution of the kinematics inverse problem and the modeling and simulation of dynamic systems. This is done using real time implementation. The modeling and simulation of dynamic systems are performed emphasizing off-line programming. In sequence, a complete study of the control strategies is carried out through the study of several elements of a robotic joint, such as: DC motor, inertia, and gearbox. Finally a trajectory generator, used as input for a generic group of joints, is developed and a proposal of the controller's implementation of joints, using EPLD development system, is presented.

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The objective of this work was to determine the coefficients of sorption and desorption of picloram in Ultisol (PVA) and Oxisol (LVA), displaying different physical and chemical characteristics. Samples of soil were collected at the 0 20 cm depth in degraded pasture areas in Viçosa-MG. Firstly, the equilibrium time between the herbicide in solution and the herbicide which was sorbed in the soil was determined by the Batch Equilibrium method. The time required was 24 hours. Sorption and desorption studies were carried out under controlled laboratory conditions; the sorption evaluation consisted in adding 10.0 mL of herbicide solutions at different concentrations to tubes containing 2.00 g of soil, with vertical rotary agitation being maintained during the pre-determined equilibrium time. After centrifugation, supernatant extract cleaning and filtration, herbicide concentration was determined by high performance liquid chromatography (HPLC) with UV detection at 254 nm. Desorption was evaluated using the samples in the tubes after the sorption tests. The Freundlich model was used for interpretation of the sorption process. Ultisol showed higher adsorption coefficient (Kf a) compared with Oxisol, which may be attributed to the lower pH of the soil and its higher organic matter content. Desorption process occurred in both soils; the LVA allowed greater release of the previously sorbed molecules.

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The induction of nicotinamide adenine dinucleotide phosphate-malic enzyme (NADP-ME) in etiolated maize (Zea mays) seedlings by UV-B and UV-A radiation, and different levels of photosynthetically active radiation (PAR, 400-700 nm) was investigated by measuring changes in activity, protein quantity and RNA levels as a function of intensity and duration of exposure to the different radiations. Under low levels of PAR, exposure to UV-B radiation but not UV-A radiation for 6 to 24 h caused a marked increase in the enzyme levels similar to that observed under high PAR in the absence of UV-B. UV-B treatment of green leaves following a 12-h dark period also caused an increase in NADP-ME expression. Exposure to UV-B radiation for only 5 min resulted in a rapid increase of the enzyme, followed by a more gradual rise with longer exposure up to 6 h. Low levels of red light for 5 min or 6 h were also effective in inducing NADP-ME activity equivalent to that obtained with UV-B radiation. A 5-min exposure to far-red light following UV-B or red light treatment reversed the induction of NADP-ME, and this effect could be eliminated by further treatment with UV-B or red light. These results indicate that physiological levels of UV-B radiation can have a positive effect on the induction of this photosynthetic enzyme. The reducing power and pyruvate generated by the activity of NADP-ME may be used for respiration, in cellular repair processes and as substrates for fatty acid synthesis required for membrane repair.

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R,S-sotalol, a ß-blocker drug with class III antiarrhythmic properties, is prescribed to patients with ventricular, atrial and supraventricular arrhythmias. A simple and sensitive method based on HPLC-fluorescence is described for the quantification of R,S-sotalol racemate in 500 µl of plasma. R,S-sotalol and its internal standard (atenolol) were eluted after 5.9 and 8.5 min, respectively, from a 4-micron C18 reverse-phase column using a mobile phase consisting of 80 mM KH2PO4, pH 4.6, and acetonitrile (95:5, v/v) at a flow rate of 0.5 ml/min with detection at lex = 235 nm and lem = 310 nm, respectively. This method, validated on the basis of R,S-sotalol measurements in spiked blank plasma, presented 20 ng/ml sensitivity, 20-10,000 ng/ml linearity, and 2.9 and 4.8% intra- and interassay precision, respectively. Plasma sotalol concentrations were determined by applying this method to investigate five high-risk patients with atrial fibrillation admitted to the Emergency Service of the Medical School Hospital, who received sotalol, 160 mg po, as loading dose. Blood samples were collected from a peripheral vein at zero, 0.5, 1.0, 1.5, 2.0, 3.0, 4.0, 6.0, 8.0, 12.0 and 24.0 h after drug administration. A two-compartment open model was applied. Data obtained, expressed as mean, were: CMAX = 1230 ng/ml, TMAX = 1.8 h, AUCT = 10645 ng h-1 ml-1, Kab = 1.23 h-1, a = 0.95 h-1, ß = 0.09 h-1, t(1/2)ß = 7.8 h, ClT/F = 3.94 ml min-1 kg-1, and Vd/F = 2.53 l/kg. A good systemic availability and a fast absorption were obtained. Drug distribution was reduced to the same extent in terms of total body clearance when patients and healthy volunteers were compared, and consequently elimination half-life remained unchanged. Thus, the method described in the present study is useful for therapeutic drug monitoring purposes, pharmacokinetic investigation and pharmacokinetic-pharmacodynamic sotalol studies in patients with tachyarrhythmias.

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Bioanalytical data from a bioequivalence study were used to develop limited-sampling strategy (LSS) models for estimating the area under the plasma concentration versus time curve (AUC) and the peak plasma concentration (Cmax) of 4-methylaminoantipyrine (MAA), an active metabolite of dipyrone. Twelve healthy adult male volunteers received single 600 mg oral doses of dipyrone in two formulations at a 7-day interval in a randomized, crossover protocol. Plasma concentrations of MAA (N = 336), measured by HPLC, were used to develop LSS models. Linear regression analysis and a "jack-knife" validation procedure revealed that the AUC0-¥ and the Cmax of MAA can be accurately predicted (R²>0.95, bias <1.5%, precision between 3.1 and 8.3%) by LSS models based on two sampling times. Validation tests indicate that the most informative 2-point LSS models developed for one formulation provide good estimates (R²>0.85) of the AUC0-¥ or Cmax for the other formulation. LSS models based on three sampling points (1.5, 4 and 24 h), but using different coefficients for AUC0-¥ and Cmax, predicted the individual values of both parameters for the enrolled volunteers (R²>0.88, bias = -0.65 and -0.37%, precision = 4.3 and 7.4%) as well as for plasma concentration data sets generated by simulation (R²>0.88, bias = -1.9 and 8.5%, precision = 5.2 and 8.7%). Bioequivalence assessment of the dipyrone formulations based on the 90% confidence interval of log-transformed AUC0-¥ and Cmax provided similar results when either the best-estimated or the LSS-derived metrics were used.

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We determined the antioxidant status of the aqueous humor after extracapsular lens extraction in 14 mongrel dogs weighing about 10 kg. The animals were examined by slit lamp biomicroscopy, applanation tonometry and indirect ophthalmoscopy. One eye was submitted to conventional extracapsular lens extraction and the other was used as control. Samples of aqueous humor were obtained by anterior chamber paracentesis before and at days 1, 2, 3, 7 and 15 after surgery. Total antioxidant status was determined as the capacity of aqueous humor to inhibit free radical generation by 2,2-azobis(2-amidopropane) chlorine. Ascorbic acid concentration was measured by HPLC with UV detection. Protein content was determined with the biuret reagent. Statistical analysis was performed by ANOVA followed by the Tukey-Kramer test. Protein concentration increased from 0.61 to 22 mg/ml 24 h after surgery. These levels were maintained and returned to normal at day 7. Total antioxidant capacity was reduced from 50 to about 30 min until day 3 and at day 7 it was equal to control. Ascorbic acid levels were reduced from 252 to about 110 µM and then returned to control values at day 15. Considering the importance of ascorbic acid concentration in aqueous humor for the maintenance of the antioxidant status of the anterior segment of the eye, the decrease of antioxidant defenses suggests that the surgical procedures promote an oxidative stress condition in the eye.

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In this work the separation of multicomponent mixtures in counter-current columns with supercritical carbon dioxide has been investigated using a process design methodology. First the separation task must be defined, then phase equilibria experiments are carried out, and the data obtained are correlated with thermodynamic models or empirical functions. Mutual solubilities, Ki-values, and separation factors aij are determined. Based on this data possible operating conditions for further extraction experiments can be determined. Separation analysis using graphical methods are performed to optimize the process parameters. Hydrodynamic experiments are carried out to determine the flow capacity diagram. Extraction experiments in laboratory scale are planned and carried out in order to determine HETP values, to validate the simulation results, and to provide new materials for additional phase equilibria experiments, needed to determine the dependence of separation factors on concetration. Numerical simulation of the separation process and auxiliary systems is carried out to optimize the number of stages, solvent-to-feed ratio, product purity, yield, and energy consumption. Scale-up and cost analysis close the process design. The separation of palmitic acid and (oleic+linoleic) acids from PFAD-Palm Fatty Acids Distillates was used as a case study.

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This study aims to analyze the influence of dehydration and different preparation methods during home processing related toalpha-carotene, beta-carotene and total carotenoids stability in carrots. Vitamin A values were evaluated after different treatments. Thus, carrots were submitted to steam cooking, water cooking with and without pressure, moist/dry cooking and conventional dehydration. Determination of alpha- and beta-carotenes was made by High-Performance Liquid Chromatography (HPLC) (conditions were developed by us) using spectrophotometric detection visible-UV at 470 nm; a RP-18 column and methanol: acetonitrile: ethyl acetate (80: 10: 10) as mobile phase. Total carotenoids quantification was made by 449 nm spectrophotometer. The retention of the analyzed carotenoids ranged from 60.13 to 85.64%. Water cooking without pressure promoted higher retention levels of alpha- and beta-carotene and vitamin A values, while water cooking with pressure promoted higher retention levels of total carotenoids. Dehydration promoted the highest carotenoid losses. The results showed that, among the routinely utilized methods under domestic condition, cooking without pressure, if performed under controlled time and temperature, is the best method as it reduces losses in the amount of alpha- and beta-carotene, the main carotenoids present in the carrots. Despite the significant carotenoid losses, carrots prepared through domestic methods, remain a rich source of provitamin A.

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The effects of microwave heating on the oxidative stability of refined canola, corn and soybean oils were determined by absorptivity in the UV spectrum and by chemical analysis (peroxide and acid values). Samples were heated in a microwave oven (800 W, 2,450 MHz) for 0 to 36 min. Microwave heating produced oxidative degradation in the three oils. Absorptivity at 232 and 270 nm increased gradually with an increase in microwave exposure time (0-36 min) for canola, corn and soybean oils. Values of absorptivity at 232 nm increased from 4.812, 3.568 and 4.183 to 10.579, 12.874 and 15.950 after 36 min of heating canola, corn and soybean oil, respectively. The absorptivity at 232nm, due to the formation of conjugated dienes, was a good index for measuring the degradation of microwaved samples. UV scanning (220 - 320 nm) detected alterations in the spectrum of microwaved samples. Acid value also increased within 36 min of heating for all oils. Peroxide value showed a significant difference (P<0.05) in the initial stage of heating (0-6 min) for all oils. After this period it could not be correlated with absorptivity at 232 nm, due to the instability of hydroperoxides at high temperatures.

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Milk and egg matrixes were assayed for aflatoxin M1 (AFM1) and B1 (AFB1) respectively, by AOAC official and modified methods with detection and quantification by thin layer chromatography (TLC) and high performance thin layer chromatography (HPTLC). The modified methods: Blanc followed by Romer, showed to be most appropriate for AFM1 analysis in milk. Both methods reduced emulsion formation, produced cleaner extracts, no streaking spots, precision and accuracy improved, especially when quantification was performed by HPTLC. The use of ternary mixture in the Blanc Method was advantageous as the solvent could extract AFM1 directly from the first stage (extraction), leaving other compounds in the binary mixture layer, avoiding emulsion formation, thus reducing toxin loss. The relative standard deviation (RSD%) values were low, 16 and 7% when TLC and HPTLC were used, with a mean recovery of 94 and 97%, respectively. As far as egg matrix and final extract are concerned, both methods evaluated for AFB1 need further studies. Although that matrix leads to emulsion with consequent loss of toxin, the Romer modified presented a reasonable clean extract (mean recovery of 92 and 96% for TLC and HPTLC, respectively). Most of the methods studied did not performed as expected mainly due to the matrixes high content of triglicerides (rich on saturated fatty acids), cholesterol, carotene and proteins. Although nowadays most methodology for AFM1 is based on HPLC, TLC determination (Blanc and Romer modified) for AFM1 and AFB1 is particularly recommended to those, inexperienced in food and feed mycotoxins analysis and especially who cannot afford to purchase sophisticated (HPLC,HPTLC) instrumentation.

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The 10-HDA content in Brazilian samples (São Paulo State) of royal jelly (RJ) was analyzed using an HPLC method based on the work by BLOODWORTH et al. [2]. The chromatographic conditions were: isocratic system, reversed phase C18-H column, auto sampler, diode array UV-VIS detector adjusted to 225nm, mobile phase composed by methanol/water (45:55) at pH= 2.5 adjusted with phosphoric acid; a-naphtol was used as internal standard, and the running time was 30min. By statistical analysis of the results, the 10-HDA contents of the samples analyzed seem to have two ranges: 1.8% and 3% (w/w), that would be useful to qualify the RJ. This is the first data regarding 10-HDA content of Brazilian RJ.

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The Graphite furnace atomic absorption spectrometry (GF AAS) was the technique chosen by the inorganic contamination laboratory (INCQ/ FIOCRUZ) to be validated and applied in routine analysis for arsenic detection and quantification. The selectivity, linearity, sensibility, detection, and quantification limits besides accuracy and precision parameters were studied and optimized under Stabilized Temperature Platform Furnace (STPF) conditions. The limit of detection obtained was 0.13 µg.L-1 and the limit of quantification was 1.04 µg.L-1, with an average precision, for total arsenic, less than 15% and an accuracy of 96%. To quantify the chemical species As(III) and As(V), an ion-exchange resin (Dowex 1X8, Cl- form) was used and the physical-chemical parameters were optimized resulting in a recuperation of 98% of As(III) and of 90% of As(V). The method was applied to groundwater, mineral water, and hemodialysis purified water samples. All results obtained were lower than the maximum limit values established by the legal Brazilian regulations, in effect, 50, 10, and 5 µg.L-1 para As total, As(III) e As(V), respectively. All results were statistically evaluated.

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Os corantes sintéticos são usualmente adicionados a alimentos industrializados para conferir e restaurar a cor obtendo-se a qualidade estética desejada. Em função destes aspectos e do potencial toxicológico que alguns corantes podem apresentar, o controle de qualidade destes compostos é de fundamental importância. Neste trabalho, foi estudado o potencial das metodologias matemáticas como o princípio da aditividade, espectrofotometria derivativa e técnica multivariada (Regressão por Mínimos Quadrados Parciais - PLSR) na determinação simultânea de dois corantes alimentícios: Amarelo Crepúsculo (AC) e Amarelo Tartrazina (AT), extraídos com lã natural. Estas metodologias foram avaliadas e comparadas em função das suas capacidades de previsão, sendo que o modelo PLSR otimizado (faixa espectral de 305 a 645 nm, empregando-se 1ª derivada como transformação dos dados e 2 componentes principais) apresentou o menor valor de Raiz Quadrada da Soma dos Erros de Previsão (RMSEP) (AT = 0,191 e AC = 0,102). A faixa de concentração estudada foi de 1,0 a 16,0 mg.L-1 para AC e de 2,0 a 22,0 mg.L-1 para AT. O conjunto de validação externa apresentou erros relativos médios de 1,97% para AC e 1,39% para AT. A aplicação desta metodologia em amostras reais mostrou que em todas as amostras analisadas as concentrações destes corantes estavam de acordo com os limites estabelecidos pela legislação brasileira