Recuperação e purificação de quitosanases usando adsorção em leito expandido com streamline DEAE com modelagem e simulação usando redes neurais

Expanded Bed Adsorption (EBA) is an integrative process that combines concepts of chromatography and fluidization of solids. The many parameters involved and their synergistic effects complicate the optimization of the process. Fortunately, some mathematical tools have been developed in order to guide the investigation of the EBA system. In this work the application of experimental design, phenomenological modeling and artificial neural networks (ANN) in understanding chitosanases adsorption on ion exchange resin Streamline® DEAE have been investigated. The strain Paenibacillus ehimensis NRRL B-23118 was used for chitosanase production. EBA experiments were carried out using a column of 2.6 cm inner diameter with 30.0 cm in height that was coupled to a peristaltic pump. At the bottom of the column there was a distributor of glass beads having a height of 3.0 cm. Assays for residence time distribution (RTD) revelead a high degree of mixing, however, the Richardson-Zaki coefficients showed that the column was on the threshold of stability. Isotherm models fitted the adsorption equilibrium data in the presence of lyotropic salts. The results of experiment design indicated that the ionic strength and superficial velocity are important to the recovery and purity of chitosanases. The molecular mass of the two chitosanases were approximately 23 kDa and 52 kDa as estimated by SDS-PAGE. The phenomenological modeling was aimed to describe the operations in batch and column chromatography. The simulations were performed in Microsoft Visual Studio. The kinetic rate constant model set to kinetic curves efficiently under conditions of initial enzyme activity 0.232, 0.142 e 0.079 UA/mL. The simulated breakthrough curves showed some differences with experimental data, especially regarding the slope. Sensitivity tests of the model on the surface velocity, axial dispersion and initial concentration showed agreement with the literature. The neural network was constructed in MATLAB and Neural Network Toolbox. The cross-validation was used to improve the ability of generalization. The parameters of ANN were improved to obtain the settings 6-6 (enzyme activity) and 9-6 (total protein), as well as tansig transfer function and Levenberg-Marquardt training algorithm. The neural Carlos Eduardo de Araújo Padilha dezembro/2013 9 networks simulations, including all the steps of cycle, showed good agreement with experimental data, with a correlation coefficient of approximately 0.974. The effects of input variables on profiles of the stages of loading, washing and elution were consistent with the literature

Recuperação e purificação de proteínas do soro de queijo tipo coalho usando cromatografia de troca iônica e interação hidrofóbica em leito na forma expandida

Expanded Bed Adsorption plays an important role in the downstream processing mainly for reducing costs as well as steps besides could handling cells homogenates or fermentation broth. In this work Expanded Bed Adsorption was used to recover and purify whey proteins from coalho cheese manufacture using Streamline DEAE and Streamline SP both ionic resins as well as a hydrophobic resin Streamline Phenyl. A column of 2.6 cm inner diameter with 30 cm in height was coupled to a peristaltic pump. Hydrodynamics study was carried out with the three resins using Tris-HCl buffer in concentration of 30, 50 and 70 mM, with pH ranging from 7.0 to 8.0. In this case, assays of the expansion degree as well as Residence Time Distribution (RTD) were carried out. For the recovery and purification steps, a whey sample of 200 mL, was submitted to a column with 25mL of resin previously equilibrated with Tris/HCl (50 mM, pH 7.0) using a expanded bed. After washing, elution was carried out according the technique used. For ionic adsorption elution was carried out using 100 mL of Tris/HCl (50 mM, pH 7.0 in 1M NaCl). For Hydrophobyc interaction elution was carried out using Tris/HCl (50 mM, pH 7.0). Adsorption runs were carried out using the three resins as well as theirs combination. Results showed that for hydrodynamics studies a linear fit was observed for the three resins with a correlation coefficient (R2) about 0.9. In this case, Streamline Phenyl showed highest expansion degree reaching an expansion degree (H0/H) of 2.2. Bed porosity was of 0.7 when both resins Streamline DEAE and Streamline SP were used with StremLine Phenyl showing the highest bed porosity about 0.75. The number of theorical plates were 109, 41.5 and 17.8 and the axial dipersion coefficient (Daxial) were 0.5, 1.4 and 3.7 x 10-6 m2/s, for Streamline DEAE, Streamline SP and Streamline Phenyl, respectively. Whey proteins were adsorved fastly for the three resins with equilibrium reached in 10 minutes. Breakthrough curves showed that most of proteins stays in flowthrough as well as washing steps with 84, 77 and 96%, for Streamline DEAE, Streamline SP and Streamline Phenyl, respectively. It was observed protein peaks during elution for the three resins used. According to these peaks were identified 6 protein bands that could probably be albumin (69 KDa), lactoferrin (76 KDa), lactoperoxidase (89 KDa), β-lactoglobulin (18,3 KDa) e α-lactoalbumin (14 KDa), as well as the dimer of beta-lactoglobulin. The combined system compound for the elution of Streamline DEAE applied to the Streamline SP showed the best purification of whey proteins, mainly of the α-lactoalbumina

Desenvolvimento de protótipo de coletor automático destinado à coleta de amostras de água em corpos aquáticos para análise em laboratório

The aim of this approach is to describe the design and construction of a low-cost automated water sampler prototype. In recent years, there is an increasing need on the use of automated equipments for hydro climatic variables to be use in urban and rural environments. Such devices are always used to provide measured information which is of crucial importance on the development of water resources strategies at watershed scale. Actually, many research and water public institutions have been using these kinds of equipments. In most of the cases, automated equipments are expensive and need to be imported, generating a situation of technologic dependency. The prototype is based on an electronic system which controls a peristaltic pump functioning, five solenoid valves and an ultrasonic sensor connected to a datalloger. An interface with the user allows communication with a PC, when the equipment functioning parameters can be provided. The equipment has a hydraulic module composed by a 12V peristaltic pump connected to a distribution circuit composed by five solenoid valves, one of them being used to clean the circuit before each sampling procedure. Samples are collected by four 1.95 polyethylene bottles. The sampler body was made of acrylic material, with a cylindrical shape, and dimensions 0.72 m and 0.38 m height and diameter, respectively. The weight of the equipment without samples is approximately 15 kg, which infers to its portability. The prototype development total cost budget was approximately US$ 1,560.00. Laboratory tests aimed to evaluate the equipment performance and functioning demonstrated satisfactory results

Recuperação e purificação do antígeno 503 de Leishmania i. chagasi expresso em E. coli e remoção de endotoxina utilizando adsorção em leito expandido

The growing interest and applications of biotechnology products have increased the development of new processes for recovery and purification of proteins. The expanded bed adsorption (EBA) has emerged as a promising technique for this purpose. It combines into one operation the steps of clarification, concentration and purification of the target molecule. Hence, the method reduces the time and the cost of operation. In this context, this thesis aim was to evaluate the recovery and purification of 503 antigen of Leishmania i. chagasi expressed in E. coli M15 and endotoxin removal by EBA. In the first step of this study, batch experiments were carried out using two experimental designs to define the optimal adsorption and elution conditions of 503 antigen onto Streamline chelating resin. For adsorption assays, using expanded bed, it was used a column of 2.6 cm in diameter by 30.0 cm in height coupled to a peristaltic pump. In the second step of study, the removal of endotoxin during antigen recovery process was evaluated employing the non-ionic surfactant Triton X-114 in the washing step ALE. In the third step, we sought developing a mathematical model able to predict the 503 antigen breakthrough curves in expanded mode. The experimental design results to adsorption showed the pH 8.0 and the NaCl concentration of 2.4 M as the optimum adsorption condition. In the second design, the only significant factor for elution was the concentration of imidazole, which was taken at 600 mM. The adsorption isotherm of the 503 antigen showed a good fit to the Langmuir model (R = 0.98) and values for qmax (maximum adsorption capacity) and Kd (equilibrium constant) estimated were 1.95 mg/g and 0.34 mg/mL, respectively. Purification tests directly from unclarified feedstock showed a recovery of 59.2% of the target protein and a purification factor of 6.0. The addition of the non-ionic surfactant Triton X-114 to the washing step of EBA led to high levels (> 99%) of LPS removal initially present in the samples for all conditions tested. The mathematical model obtained to describe the 503 antigen breakthrough curves in Streamline Chelanting resin in expanded mode showed a good fit for both parameter estimation and validation steps. The validated model was used to optimize the efficiencies, achieving maximum values of the process and of the column efficiencies of 89.2% and 75.9%, respectively. Therefore, EBA is an efficient alternative for the recovery of the target protein and removal of endotoxin from an E. coli unclarified feedstock in just one step.

Simulação do escoamento monofásico em um estágio de uma bomba centrífuga utilizando técnicas de fluidodinâmica computacional

Oil production and exploration techniques have evolved in the last decades in order to increase fluid flows and optimize how the required equipment are used. The base functioning of Electric Submersible Pumping (ESP) lift method is the use of an electric downhole motor to move a centrifugal pump and transport the fluids to the surface. The Electric Submersible Pumping is an option that has been gaining ground among the methods of Artificial Lift due to the ability to handle a large flow of liquid in onshore and offshore environments. The performance of a well equipped with ESP systems is intrinsically related to the centrifugal pump operation. It is the pump that has the function to turn the motor power into Head. In this present work, a computer model to analyze the three-dimensional flow in a centrifugal pump used in Electric Submersible Pumping has been developed. Through the commercial program, ANSYS® CFX®, initially using water as fluid flow, the geometry and simulation parameters have been defined in order to obtain an approximation of what occurs inside the channels of the impeller and diffuser pump in terms of flow. Three different geometry conditions were initially tested to determine which is most suitable to solving the problem. After choosing the most appropriate geometry, three mesh conditions were analyzed and the obtained values were compared to the experimental characteristic curve of Head provided by the manufacturer. The results have approached the experimental curve, the simulation time and the model convergence were satisfactory if it is considered that the studied problem involves numerical analysis. After the tests with water, oil was used in the simulations. The results were compared to a methodology used in the petroleum industry to correct viscosity. In general, for models with water and oil, the results with single-phase fluids were coherent with the experimental curves and, through three-dimensional computer models, they are a preliminary evaluation for the analysis of the two-phase flow inside the channels of centrifugal pump used in ESP systems

Análise de confiabilidade de uma bomba centrífuga: aplicação na injeção de água para recuperação de petróleo

The oil industry`s need to produce with maximum efficiency, not to mention the safety and the environment aspects, encourages the optimization of processes. It makes them look for a level of excellence in acquisition of equipment, ensuring the quality without prejudice security of facilities and peoples. Knowing the reliability of equipment and that this stands for a system is fundamental to the production strategy to seeks the maximum return on investment. The reliability analysis techniques have been increasingly applied in the industry as strategy for predicting failures likelihood ensuring the integrity of processes. Some reliability theories underlie the decisions to use stochastic calculations to estimate equipment failure. This dissertation proposes two techniques associating qualitative (through expertise opinion) and quantitative data (European North Sea oil companies fault database, Ored) applied on centrifugal pump to water injection system for secondary oil recovery on two scenarios. The data were processed in reliability commercial software. As a result of hybridization, it was possible to determine the pump life cycle and what impact on production if it fails. The technique guides the best maintenance policy - important tool for strategic decisions on asset management.

Otimização dos parâmetros de um modelo de bomba em sistemas de bombeio centrífugo submerso

One of several techniques applied to production processes oil is the artificial lift, using equipment in order to reduce the bottom hole pressure, providing a pressure differential, resulting in a flow increase. The choice of the artificial lift method depends on a detailed analysis of the some factors, such as initial costs of installation, maintenance, and the existing conditions in the producing field. The Electrical Submersible Pumping method (ESP) appears to be quite efficient when the objective is to produce high liquid flow rates in both onshore and offshore environments, in adverse conditions of temperature and in the presence of viscous fluids. By definition, ESP is a method of artificial lift in which a subsurface electric motor transforms electrical into mechanical energy to trigger a centrifugal pump of multiple stages, composed of a rotating impeller (rotor) and a stationary diffuser (stator). The pump converts the mechanical energy of the engine into kinetic energy in the form of velocity, which pushes the fluid to the surface. The objective of this work is to implement the optimization method of the flexible polyhedron, known as Modified Simplex Method (MSM) applied to the study of the influence of the modification of the input and output parameters of the centrifugal pump impeller in the channel of a system ESP. In the use of the optimization method by changing the angular parameters of the pump, the resultant data applied to the simulations allowed to obtain optimized values of the Head (lift height), lossless efficiency and the power with differentiated results.

Simulação do escoamento monofásico em um estágio de uma bomba centrífuga utilizando técnicas de fluidodinâmica computacional

Oil production and exploration techniques have evolved in the last decades in order to increase fluid flows and optimize how the required equipment are used. The base functioning of Electric Submersible Pumping (ESP) lift method is the use of an electric downhole motor to move a centrifugal pump and transport the fluids to the surface. The Electric Submersible Pumping is an option that has been gaining ground among the methods of Artificial Lift due to the ability to handle a large flow of liquid in onshore and offshore environments. The performance of a well equipped with ESP systems is intrinsically related to the centrifugal pump operation. It is the pump that has the function to turn the motor power into Head. In this present work, a computer model to analyze the three-dimensional flow in a centrifugal pump used in Electric Submersible Pumping has been developed. Through the commercial program, ANSYS® CFX®, initially using water as fluid flow, the geometry and simulation parameters have been defined in order to obtain an approximation of what occurs inside the channels of the impeller and diffuser pump in terms of flow. Three different geometry conditions were initially tested to determine which is most suitable to solving the problem. After choosing the most appropriate geometry, three mesh conditions were analyzed and the obtained values were compared to the experimental characteristic curve of Head provided by the manufacturer. The results have approached the experimental curve, the simulation time and the model convergence were satisfactory if it is considered that the studied problem involves numerical analysis. After the tests with water, oil was used in the simulations. The results were compared to a methodology used in the petroleum industry to correct viscosity. In general, for models with water and oil, the results with single-phase fluids were coherent with the experimental curves and, through three-dimensional computer models, they are a preliminary evaluation for the analysis of the two-phase flow inside the channels of centrifugal pump used in ESP systems

Análise da estrutura e transporte do zooplâncton no estuário do Rio Caravelas (Bahia-Brasil) através do uso do zooscan

Spatial-temporal dynamics of zooplankton in the Caravelas river estuary (Bahia, Brazil). The survey was conducted in order to describe the zooplankton community of the estuary Caravelas (Bahia, Brazil), to quantify and relate the patterns of horizontal and vertical transport with the type of tide (neap and spring) and tidal phase (flood and ebb). Zooplankton samples were collected with the aid of a suction pump (300L), filtered in plankton nets (300μm) and fixed in saline formalin 4%. Samples were collected at a fixed point (A1), near the mouth of the estuary, with samples taken at neap tides and spring tides during the dry and rainy seasons. Samples were collected for 13 hours, at intervals of 1 hour in 3 depths: surface, middle and bottom. Simultaneous collection of biological, we measured the current velocity, temperature and salinity of the water through CTD. In the laboratory, samples were selected for analysis in estereomicroscope, with 25 groups identified, with Copepoda getting the highest number of species. The 168 samples obtained from temporal samples were subsampled and processed on equipment ZooScan, with the aid of software ZooProcess at the end were generated 458.997 vingnettes. 8 taxa were identified automatically, with 16 classified as a semi-automatic. The group Copepoda, despite the limited taxonomic refinement ZooScan, obtained 2 genera and 1 species identified automatically. Among the seasons dry and wet groups Brachyura (zoea), Chaetognatha, and the Calanoid copepods (others), Temora spp., Oithona spp. and Euterpina acutifrons were those who had higher frequency of occurrence, appearing in more than 70% of the samples. Copepoda group showed the largest percentage of relative abundance in both seasons. There was no seasonal variation of total zooplankton, with an average density of 7826±4219 org.m-3 in the dry season, and 7959±3675 org.m-3 in the rainy season, neither between the types and phases of the tides, but seasonal differences were significant recorded for the main zooplankton groups. Vertical stratification was seen for the major zooplankton groups (Brachyura, Chaetognatha, Calanoida (other), Oithona spp, Temora spp. e Euterpina acutifrons). The scale of this stratification varied with the type (square or tide) and tidal phase (flood or ebb). The instantaneous transport was more influenced by current velocity, with higher values observed in spring tides to the total zooplankton, however, there was a variation of this pattern depending on the zooplankton group. According to the data import and export of total zooplankton, the outflow of organisms of the estuary was higher than the input. The results suggest that the estuary of Caravelas may influence the dynamics of organic matter to the adjacent coast, with possible consequences in National Marine Park of Abrolhos

Sistema de sensoriamento eletromagnético utilizado para detecção da contaminação do óleo isolante do motor no método de elevação artificial do tipo bombeio centrífugo submerso

In the artificial lift method by Electrical Submersible Pump (ESP), the energy is transmitted for the well´s deep through a flat electric handle, where it is converted into mechanical energy through an engine of sub-surface, which is connected to a centrifugal pump. This transmits energy to the fluid under the pressure form, bringing it to the surface In this method the subsurface equipment is basically divided into: pump, seal and motor. The main function of the seal is the protect the motor, avoiding the motor´s oil be contaminated by oil production and the consequent burning of it. Over time, the seal will be wearing and initiates a contamination of motor oil, causing it to lose its insulating characteristics. This work presents a design of a magnetic sensor capable of detecting contamination of insulating oil used in the artificial lift method of oil-type Electrical Submersible Pump (ESP). The objective of this sensor is to generate alarm signal just the moment when the contamination in the isolated oil is present, enabling the implementation of a predictive maintenance. The prototype was designed to work in harsh conditions to reach a depth of 2000m and temperatures up to 150°C. It was used a simulator software to defined the mechanical and electromagnetic variables. Results of field experiments were performed to validate the prototype. The final results performed in an ESP system with a 62HP motor showed a good reliability and fast response of the prototype.

Controle aplicado a poços com método de elevação e bombeio centrífugo submerso

The Electrical Submersible Pumping is an artificial lift method for oil wells employed in onshore and offshore areas. The economic revenue of the petroleum production in a well depends on the oil flow and the availability of lifting equipment. The fewer the failures, the lower the revenue shortfall and costs to repair it. The frequency with which failures occur depends on the operating conditions to which the pumps are submitted. In high-productivity offshore wells monitoring is done by operators with engineering support 24h/day, which is not economically viable for the land areas. In this context, the automation of onshore wells has clear economic advantages. This work proposes a system capable of automatically control the operation of electrical submersible pumps, installed in oil wells, by an adjustment at the electric motor rotation based on signals provided by sensors installed on the surface and subsurface, keeping the pump operating within the recommended range, closest to the well s potential. Techniques are developed to estimate unmeasured variables, enabling the automation of wells that do not have all the required sensors. The automatic adjustment, according to an algorithm that runs on a programmable logic controller maintains the flow and submergence within acceptable parameters avoiding undesirable operating conditions, as the gas interference and high engine temperature, without need to resort to stopping the engine, which would reduce the its useful life. The control strategy described, based on modeling of physical phenomena and operational experience reported in literature, is materialized in terms of a fuzzy controller based on rules, and all generated information can be accompanied by a supervisory system

Desenvolvimento de sistemas magnéticos com potencialidades terapêuticas para vetorização de fármacos

Magnetic targeting is being investigated as a means of local delivery of drugs, combining precision, minimal surgical intervention, and satisfactory concentration of the drug in the target region. In view of these advantages, it is a promising strategy for improving the pharmacological response. Magnetic particles are attracted by a magnetic field gradient, and drugs bound to them can be driven to their site of action by means of the selective application of magnetic field on the desired area. Helicobacter pylori is the commonest chronic bacterial infection. The treatment of choice has commonly been based upon a triple therapy combining two antibiotics and an anti-secretory agent. Furthermore, an extended-release profile is of utmost importance for these formulations. The aim of this work was to develop a magnetic system containing the antibiotic amoxicillin for oral magnetic drug targeting. First, magnetic particles were produced by coprecipitation of iron salts in alkaline medium. The second step was coating the particles and amoxicillin with Eudragit® S-100 by spray-drying technique. The system obtained demonstrated through the characterization studies carried out a possible oral drug delivery system, consisting in magnetite microparticles and amoxicillin, coated with a polymer acid resistant. This system can be used to deliver drugs to the stomach for treatment of infections in this organ. Another important finding in this work is that it opens new prospects to coat magnetic microparticles by the technique of spray-drying.

Modelagem de dados de falhas de equipamentos de sub-superfície em poços de petróleo da Bacia Potiguar

This master´s thesis presents a reliability study conducted among onshore oil fields in the Potiguar Basin (RN/CE) of Petrobras company, Brazil. The main study objective was to build a regression model to predict the risk of failures that impede production wells to function properly using the information of explanatory variables related to wells such as the elevation method, the amount of water produced in the well (BSW), the ratio gas-oil (RGO), the depth of the production bomb, the operational unit of the oil field, among others. The study was based on a retrospective sample of 603 oil columns from all that were functioning between 2000 and 2006. Statistical hypothesis tests under a Weibull regression model fitted to the failure data allowed the selection of some significant predictors in the set considered to explain the first failure time in the wells

Estudo de estratégias de otimização para poços de petróleo com elevação por bombeio de cavidades progressivas

The progressing cavity pump artificial lift system, PCP, is a main lift system used in oil production industry. As this artificial lift application grows the knowledge of it s dynamics behavior, the application of automatic control and the developing of equipment selection design specialist systems are more useful. This work presents tools for dynamic analysis, control technics and a specialist system for selecting lift equipments for this artificial lift technology. The PCP artificial lift system consists of a progressing cavity pump installed downhole in the production tubing edge. The pump consists of two parts, a stator and a rotor, and is set in motion by the rotation of the rotor transmitted through a rod string installed in the tubing. The surface equipment generates and transmits the rotation to the rod string. First, is presented the developing of a complete mathematical dynamic model of PCP system. This model is simplified for use in several conditions, including steady state for sizing PCP equipments, like pump, rod string and drive head. This model is used to implement a computer simulator able to help in system analysis and to operates as a well with a controller and allows testing and developing of control algorithms. The next developing applies control technics to PCP system to optimize pumping velocity to achieve productivity and durability of downhole components. The mathematical model is linearized to apply conventional control technics including observability and controllability of the system and develop design rules for PI controller. Stability conditions are stated for operation point of the system. A fuzzy rule-based control system are developed from a PI controller using a inference machine based on Mandami operators. The fuzzy logic is applied to develop a specialist system that selects PCP equipments too. The developed technics to simulate and the linearized model was used in an actual well where a control system is installed. This control system consists of a pump intake pressure sensor, an industrial controller and a variable speed drive. The PI control was applied and fuzzy controller was applied to optimize simulated and actual well operation and the results was compared. The simulated and actual open loop response was compared to validate simulation. A case study was accomplished to validate equipment selection specialist system

Desenvolvimento de uma ferramenta computacional para aplicação em métodos de elevação por bombeio centrífugo submerso

The Electrical Submersible Pump (ESP) has been one of the most appropriate solutions for lifting method in onshore and offshore applications. The typical features for this application are adverse temperature, viscosity fluids and gas environments. The difficulties in equipments maintenance and setup contributing to increasing costs of oil production in deep water, therefore, the optimization through automation can be a excellent approach for decrease costs and failures in subsurface equipment. This work describe a computer simulation related with the artificial lifting method ESP. This tool support the dynamic behavior of ESP approach, considering the source and electric energy transmission model for the motor, the electric motor model (including the thermal calculation), flow tubbing simulation, centrifugal pump behavior simulation with liquid nature effects and reservoir requirements. In addition, there are tri-dimensional animation for each ESP subsytem (transformer, motor, pump, seal, gas separator, command unit). This computer simulation propose a improvement for monitoring oil wells for maximization of well production. Currenty, the proprietaries simulators are based on specific equipments manufactures. Therefore, it is not possible simulation equipments of another manufactures. In the propose approach there are support for diverse kinds of manufactures equipments