Made available in DSpace on 2014-12-17T15:42:30Z (GMT). No. of bitstreams: 1 OldemarRC_TESE.pdf: 3699257 bytes, checksum: 0779dc655c0cadfa72a92f99bc053514 (MD5) Previous issue date: 2014-02-21 / Conselho Nacional de Desenvolvimento Cient?fico e Tecnol?gico / The development of products whose purpose is to promote blockages in high permeability zones as well as to control the hydrate or scale formation also needs some tests in porous media before using the product in the field, where attempts and unavoidable operational errors costs would able to derail any projects. The aim of this study was to analyze and compare the Botucatu and Berea sandstones properties, involving problems related to loss permeability. It was observed that even cores of Berea, without expansible clays in their composition had their permeability reduced, as soon as the salinity of brine reached a lower limit. As expected, the same happened with the Botucatu sandstone samples, however, in this case, the sensitivity to low salinity was more pronounced. In a second phase, the research was focused on the Botucatu Sandstone behavior front of dilute polymer solutions injection, checking the main relationships between the Rock / Fluid interactions, considering the Mobility Reduction, Resistance and Residual Resistance Factors, as well as adsorption/desorption processes of these polymers, and the polymer molecules average size and porous sandstone average size ratio. The results for both phases showed a real feasibility of using the Botucatu sandstone in laboratory tests whose objective is the displacement of fluids through porous media / O desenvolvimento de produtos cuja finalidade ? a de promover bloqueios em ?reas de alta permeabilidade, assim como de controladores ou inibidores de corros?o e incrusta??o e de hidratos, tamb?m necessita de estudos no meio poroso, antes de serem empregados em opera??es de campo. Considerando esses aspectos, o presente trabalho teve como principal objetivo compreender melhor a natureza do arenito Botucatu, buscando, na medida do poss?vel: (a) tra?ar compara??es entre o seu comportamento e o comportamento do arenito Berea, o mais popular entre os pesquisadores e (b) considerando sua abund?ncia no Brasil e facilidade de explora??o, refor?ar a possibilidade de seu uso seguro como meio poroso virgem em ensaios de laborat?rio, visando an?lises e qualifica??o de materiais pass?veis de serem usados na ind?stria do petr?leo. Em plugs desse arenito, foram injetadas solu??es aquosas de v?rias salinidades, tendo como ponto de partida a ?gua do mar sint?tica, com o objetivo de se detectar poss?veis altera??es na permeabilidade do arenito devido ? expans?o de argilas ou ? libera??o de part?culas capazes de tamponar definitiva ou temporariamente as gargantas dos poros da rocha, bem como comparar seu comportamento com outros arenitos normalmente utilizados para estudos envolvendo meio poroso. Para tanto, em uma segunda fase, foram utilizados pol?meros regularmente empregados na ind?stria do petr?leo (as poliacrilamidas), onde se procurou verificar se o comportamento desses materiais em solu??o, e n?o mais somente a ?gua, quando empregados no meio poroso representado pelo arenito Botucatu, retratava com fidelidade e seguran?a os resultados reportados na literatura especializada para os meios porosos tradicionais. Os resultados para ambas as fases mostraram uma real viabilidade do emprego do arenito Botucatu em an?lises de laborat?rio cujo objetivo seja o deslocamento em meio poroso

Polímeros e copolímeros hidrossolúveis a base de acrilamida e seus derivados são sintetizados e aplicados principalmente como floculantes, auxiliando na remoção de sólidos finos presentes em águas residuais e industriais. As poliacrilamidas, como são assim chamadas, podem se apresentar em sua forma neutra ou contendo diferentes cargas iônicas (positivas ou negativas) em sua composição em função do tipo de aplicação específica. As poliacrilamidas são sintetizadas, em sua grande maioria, através de emulsões inversas via polimerização radicalar, visando à elevada massa molar do polímero formado. A formulação da poliacrilamida catiônica com fração molar de 0,6 do comonômero iônico foi otimizada através de planejamento fatorial experimental, variando-se os principais fatores de processo: concentração do pacote de surfactante para formação da emulsão inversa, tempo de micronização, concentração de VISCOPLEX® e concentração de ácido graxo. Com base nos resultados obtidos a partir do primeiro planejamento, foram estudadas as seguintes variáveis: HLB do sistema de surfactantes e a troca dos surfactantes monoméricos por surfactantes poliméricos. A otimização da formulação levou em conta principalmente o custo-benefício para produção e comercialização em escala industrial. O fator micronização foi elevado em seu máximo e os fatores VISCOPLEX® e ácido graxo foram excluídos da formulação, após conclusão dos estudos. A troca dos surfactantes monoméricos por poliméricos foi vantajosa devido ao ganho de qualidade do produto final, mesmo com maior custo de implementação. Para minimizar os custos de formulação o fator HLB foi utilizado em seu máximo, onde houve incremento do ALKONAT® L 50 para formação da emulsão inversa. / Acrylamide based hydrosoluble polymers and copolymers and their derivatives are synthesized and mainly applied as flocculants, assisting removal of fine solids present in wastewater and industrial wastewater. Polyacrylamides may be present in their neutral form or with different ionic charges (positive or negative) in their composition depending on the type of specific application. Polyacrylamides are synthesized, mostly by inverse emulsions using radical polymerization, aiming polymers with high molecular weight. Cationic polyacrylamide formulation with 0.6 mole fraction of the ionic comonomer was optimized through design of experiments (DOE), varying the major process factors: surfactant package concentration to form inverse emulsion, micronization time, VISCOPLEX® concentration and fatty acid concentration. Based on the responses from the first DOE, a second DOE was performed, where the variables surfactant system HLB and the exchange of monomeric surfactant by polymeric surfactants were investigated. The formulation optimization considered mainly cost-effective production and industrial scale. The micronization factor was set at its maximum and VISCOPLEX® and fatty acid factors were excluded from the formulation, after conclusion of studies. Exchanging monomeric surfactants by polymeric surfactants led to a final product with higher quality, even considering a higher implementation cost. In order to minimize formulation costs HLB factor was used at its maximum, increasing the content of ALKONAT® L50 to form the inverse emulsion.

Hidrogéis são membranas formadas pela reticulação de cadeias poliméricas, empregados na área farmacêutica como produtos biomédicos. Dentre os principais polímeros selecionados para a síntese de hidrogéis, destaca-se a poliacrilamida (PAAM) devido às suas propriedades como hidrofilicidade e alto grau de intumescimento. Proteínas terapêuticas e enzimas são veiculadas em hidrogéis como carreadores de fármaco ou como dispositivos para tratamento de feridas e escaras na pele. Este trabalho teve como objetivo a síntese de uma membrana à base de PAAM favorável para veiculação de proteínas. As proteínas empregadas foram papaína e albumina de soro bovino (BSA) e as etapas do processo englobaram síntese da membrana, adição das proteínas no sistema, irradiação em condições específicas e caracterização da membrana. Ao utilizar temperaturas criogênicas na síntese e na irradiação das amostras, houve predomínio de reticulação da cadeia polimérica, fato que não ocorria em temperatura ambiente. As membranas foram obtidas com incorporação dos ativos na concentração de 0,2 a 1% (p/p), obtendo-se concentração de PAAM entre 4% a 10% (p/p), as quais receberam irradiação com raios gama provenientes de uma fonte 60Co, na dose de 25 kGy. Nas condições realizadas, as membranas não apresentaram citotoxicidade nem adesão celular, o perfil de liberação das proteínas foi adequado, a papaína manteve sua bioatividade preservada apesar do decaimento biológico e, segundo estudos de carga das moléculas, a membrana possui maior afinidade com a papaína, liberando-a mais lentamente. Desta forma, o método proposto e as membranas obtidas foram apropriados para a obtenção de um biomaterial. / The use of hydrogels for biomedical purposes has been extensively investigated. Polyacrylamide (PAAM) is widely used due to properties such as hydrophilicity and swelling degree. Pharmaceutical proteins correspond to highly active substances which may be applied for distinct purposes. This work concerns the development of radio-synthesized hydrogel for protein release using papain and bovine serum albumin (BSA) as model proteins. The polymer was solubilized (1% w/v) in water and lyophilized. The proteins were incorporated into the lyophilized polymer and the hydrogels were produced by simultaneous crosslinking and sterilization using gamma radiation at 25 kGy under frozen conditions. The produced systems were characterized in terms of swelling degree, gel fraction, crosslinking density, fluid handling capacity, determination pH at point of polymer zero charge and evaluated according to protein release, bioactivity, cytotoxicity and cell adhesion. The hydrogels developed presented different properties as a function of polymer concentration and the optimized results were found for the samples containing 4-10% polyacrylamide. Protein release was controlled by the electrostatic affinity of acrylic moieties of polymer and proteins. This selection was based on the release of the proteins during the experiment period (up to 50 hours), maintenance of enzyme activity and the nanostructure developed. The system was suitable for protein loading and release and according to the cytotoxic assay and cell adhesion it was also adequate for biomedical purposes and this method was able to generate a matrix to protein release.

<p>The thesis focuses on the development of new homogeneous gels for the separation of drug enantiomers, peptides, DNA and virus by electrophoresis and electrochromatography in capillaries and microchips. This type of separation media offers high resolution and small zone broadening. Compared to particulate beds the resolution in this type of separation media is high because the eddy diffusion is zero and the resistance to mass transfer is small, since the diffusional distance between two polymer chains in the gel is considerably shorter than that between two beads in a packed bed.</p><p>The gels have been characterized in terms of plate heights, plate numbers, resolution, etc. Gels of agarose, polyvinyl alcohol, albumin and polyacrylamide have been employed for electrochromatography or electrophoresis. <i>N,N’</i>-methylene-bisacrylamide, the most widely used crosslinker in polyacrylamide gels, was exchanged for allyl-β-cyclodextrin to get a multi-purpose gel, <i>i.e.,</i> a separation medium the separation properties of which is determined not only by the polyacrylamide chains, but also by β-cyclodextrin with its complexation power.</p><p>A cost-effective, hybrid microdevice has been designed for fast electrophoretic and electrochromatographic analyses as well as for microchromatography. It consists of a fused silica capillary mounted on a supporting plate which integrates most of the compartments necessary for automation and sensitive detection at short UV wavelengths.</p>

The thesis focuses on the development of new homogeneous gels for the separation of drug enantiomers, peptides, DNA and virus by electrophoresis and electrochromatography in capillaries and microchips. This type of separation media offers high resolution and small zone broadening. Compared to particulate beds the resolution in this type of separation media is high because the eddy diffusion is zero and the resistance to mass transfer is small, since the diffusional distance between two polymer chains in the gel is considerably shorter than that between two beads in a packed bed. The gels have been characterized in terms of plate heights, plate numbers, resolution, etc. Gels of agarose, polyvinyl alcohol, albumin and polyacrylamide have been employed for electrochromatography or electrophoresis. N,N’-methylene-bisacrylamide, the most widely used crosslinker in polyacrylamide gels, was exchanged for allyl-β-cyclodextrin to get a multi-purpose gel, i.e., a separation medium the separation properties of which is determined not only by the polyacrylamide chains, but also by β-cyclodextrin with its complexation power. A cost-effective, hybrid microdevice has been designed for fast electrophoretic and electrochromatographic analyses as well as for microchromatography. It consists of a fused silica capillary mounted on a supporting plate which integrates most of the compartments necessary for automation and sensitive detection at short UV wavelengths.

The mechanism through which a charged polymer cationic polyacrylamide (C-PAM) operates to increase the rate of cornstarch hydrolysis was investigated. The main objective was to determine the major factors that affect the mechanism so that these parameters may be adjusted to achieve optimal hydrolysis rates. A combination of analytical methods including dynamic light scattering, optical imaging, and uv-vis spectroscopy were used to study polymer, starch, and enzyme interactions as a function of process conditions. It was found that C-PAM binds strongly to starch granules, increasing solubilization and decreasing onset gelatinization temperature. Granule swelling was unaffected by C-PAM. Both binding of enzyme to cornstarch, and rate of cornstarch hydrolysis were found to increase in the presence of C-PAM. By analogy to previous work on cationic polymer promoted hydrolysis of cellulose, it was proposed that the polymer reduces the charge on the starch surface through a "charge-patch" mechanism. Because both enzyme and substrate are negatively charged, the positively charged polymer reduces the charge repulsion experienced by the approaching enzyme. This leads to stronger enzyme-substrate binding, and faster hydrolysis. There is a mirror image relationship between viscosity of the medium and hydrolysis rate, which allows optimization of these parameters with enzyme and C-PAM dosage. Overall, the polymer addition reduced enzyme dose by 62% depending on the conditions used, so this method could have significant economic impact on the industrial conversion of starch to ethanol.

It is necessary to understand the erodibility and hydrological response of mine-site slope forming materials (SFMs), because of increasing awareness of the environmental impacts of mining. Steep engineered slopes in high intensity rainfall environments present a serious erosion risk. Temporary surface stabilisers, such as polyacrylamides (PAMs) and polyvinylacrylic latex (PVALs) are potentially cost effective erosion control solutions. In this study PAM and PVAL efficacy to reduce runoff, leachate and erosion was assessed at two application rates, with and without gypsum on SFMs from an iron ore mine in Guinea (West Africa). NSPASS (near-surface photogrammetry assessment of slope forming materials’ surface roughness) is a novel method that integrates digital image capture and GIS. It is shown to detect and quantify surface micro-relief changes of 2-3 mm, not visible to the naked eye. As expected, soil and non-soil SFMs were significantly different in terms of their physical and chemical properties. Phase I of the study investigated the erodibility of ten SFMs, including soil, ore and waste-rock. The results indicate that the hydrological response to rainfall of most SFMs is to generate leachate. Weathered phyllite (PHY-WEA) is the most erodible SFM by both runoff and leachate. Multiple regression analysis demonstrated that magnetic susceptibility, mineralogy and dry aggregate distribution; parameters not commonly assessed in erosion studies, are important in explaining SFM erodibility and hydrological response. Phase II evaluated critically the effectiveness of three commercially available polymer solutions (two PAMs and one PVAL) at reducing runoff, leachate and erosion from four of the most erodible SFMs identified in Phase I. The results indicate that some PAM and PVAL treatments significantly reduce runoff, leachate and erosion. Polymer efficacy is highly dependent on the physical and chemical properties of the SFM, as well as the mechanism of polymer to SFM adsorption. Increasing the application rate of select treatments lowered leachate volumes, runoff and leachate total sediment loads. Contrary to previous studies, gypsum amendments did not significantly improve polymer efficiency. This research has added to our understanding of the erodibility and hydrological response of soil and non-soil SFMs. This is the first study to evaluate critically the efficacy of PVALs in controlling erosion from mine-site SFMs. Future studies should continue to optimise NSPASS performance in monitoring changes in surface micro-relief.

Este trabalho visa estudar a viabilidade do emprego de processos oxidativos avancados na degradacao de solucoes aquosas de polimeros que sao comumente utilizados em aplicacoes industriais e estao presentes em seus efluentes. Para maior entendimento das reacoes envolvidas no processo oxidativo utilizou-se modelagem molecular nos calculos de constantes cineticas. Na parte experimental estudou-se a degradacao de polietilenoglicol (PEG), polivinilpirrolidona (PVP) e poliacrilamida (PAM) pelos processos foto-Fenton e UV/H2O2. Realizaram-se analises de carbono organico dissolvido (COD), cromatografia liquida de alto desempenho (HPLC) e cromatografia de permeacao em gel (GPC). Observou-se separacao de fases para PVP e PAM, o que contribuiu para uma remocao final de COD de ate 85%. Nas analises de HPLC foram detectados como intermediarios da reacao os acidos: oxalico, formico, malonico e acetico. Os resultados de GPC indicaram comportamentos distintos das distribuicoes de pesos moleculares em funcao do tempo de reacao para os processos UV/H2O2 e foto- Fenton. No conjunto, os resultados comprovam a viabilidade dos processos estudados na degradacao destes polimeros, embora a analise de intermediarios para PAM nao tenha sido conclusiva quanto a formacao de acrilamida. A modelagem molecular confirma que a reacao de abstracao de hidrogenio de uma cadeia polimerica pelos radicais alquilperoxila e de fato a etapa mais lenta do processo oxidativo para PEG e PAM. Para PAM os resultados da modelagem mostram que o ataque dos radicais hidroxila acontece preferencialmente no hidrogenio ligado ao carbono terciario e que a abstracao de hidrogenio do grupo lateral amida parece pouco viavel, ou acontece por um mecanismo diferente do modelado. A reacao de formacao da acrilamida de acordo com o mecanismo proposto tambem apresentou uma constante cinetica muito baixa, o que esta de acordo com os resultados experimentais, ja que nao foi detectada acrilamida como produto da degradacao de poliacrilamida pelos processos foto-Fenton e UV/H2O2, com limite de deteccao de 50 ppb. / The aim of this work is to apply advanced oxidative processes to study the degradation of industrial wastewater containing water-soluble polymers. Kinetic constants of reactions involved in the oxidation processes were calculated by molecular modeling. In the experimental part the degradation of polyvinylpirrolidone (PVP), polyacrylamide (PAM) and polyethyleneglycol (PEG) was studied by photo-Fenton and UV/H2O2 processes. Intermediates of the degradation processes were identified by HPLC and the molecular weight distribution was monitored by GPC. In the experiments with PAM and PVP phase separation was observed, and more than 85% of the DOC was removed. The following intermediate acids were identified: acetic, malonic, formic and oxalic. The molecular weight distribution curves measured by GPC indicate that distinct reaction paths exist for each process studied, although the analysis of intermediates formed during the degradation of PAM was not conclusive with respect to acrylamide formation. The kinetic constants for the reactions involving hydrogen abstraction from PEG chain by hydroxyl and alkylperoxy radicals were calculated. The molecular modeling results confirm that hydrogen abstraction from polymeric chain by alkylperoxy radicals is the determinant step in the oxidation process for PEG and PAM. Polyacrylamide possesses three sites for attack by hydroxyl radicals and the favored path is the abstraction of tertiary hydrogen atom, followed by secondary hydrogen. The abstraction of the amide hydrogen was more difficult. The mechanism proposed for acrylamide formation was not the preferential reaction path. This was expected since acrylamide was not detected in the experiments with a limit detection of 50 ppb.

Submitted by (lucia.rodrigues@ufrpe.br) on 2016-10-19T14:44:51Z No. of bitstreams: 1 Valerio Ferreira Marcelo.pdf: 1719967 bytes, checksum: 92c5571022fd389d06cae5f46fa03f1a (MD5) / Made available in DSpace on 2016-10-19T14:44:51Z (GMT). No. of bitstreams: 1 Valerio Ferreira Marcelo.pdf: 1719967 bytes, checksum: 92c5571022fd389d06cae5f46fa03f1a (MD5) Previous issue date: 2015-02-20 / The Coastal Plains cover of Rio de Janeiro to Amapá, this landscape unit has cohesive layers of impediment located near the soil surface that promote changes in the state of water in the soil, which reduces the availability of water to plants and interfere with the development of its root system. The cohesive character is the term applied to describe subsurface horizons that have a significant increase in the cohesion of its particles. Soils with the cohesive character have serious physical limitations, especially by the presence of horizons that have, when dry, a strong cohesion, restricting plant growth, but when wet, its structure becomes very unstable. The objective of this study was to evaluate the use of chemical conditioners in improving the physical quality of cohesive soils in the environment of the coastal Pernambuco, from physical and physical-hydric attributes trials. Opened a profile on acreage of cane sugar on the North Coast Plains Forest Zone and collected samples structured and unstructured the cohesive horizon. The samples were divided into three treatments T0 (control), T1 (polyacrylamide - PAM / 100 mg kg-1) and T2 (PAM + Gypsum-CaSO42H2O / 1,523 g L-1). The evaluated attributes were: Total porosity (macro, meso, micro and criptoporosidade); bulk density (Ds); water retention curve characteristic of the soil (CCRAS); soil resistance to root penetration (RP), limiting water range (LLWR). The mean physical parameters analyzed were subjected to analysis of variance (ANOVA) and the parameters that differ significantly was the Tukey test, using the SAS application. The embodiment of the toilet in soil contained in the soil core before or after application of conditioners do not affect the results of the bulk density (BD); The form of distribution of pores, showed changes in porosity in the soil under the action of conditioners; The penetration resistance was efficient attribute to evaluate the reduction of cohesion; The structural stability index was not conclusive for choosing the best treatment; The values of the IHO demonstrated the efficacy of this attribute, regarding the availability of water for the plants; Treatment with WFP increased the availability of water to plants. / Os Tabuleiros Costeiros abrangem do Rio de Janeiro ao Amapá, esta unidade de paisagem possui camadas coesas de impedimento localizadas próximas à superfície do solo que promovem alterações no estado da água no solo, que reduz a disponibilidade de água às plantas, além de interferir no desenvolvimento de seu sistema radicular. O caráter coeso é o termo aplicado para descrever horizontes subsuperficiais que apresentam aumento significativo na coesão das suas partículas. Solos que apresentam o caráter coeso têm sérias limitações físicas, sobretudo pela presença de horizontes que apresentam, quando secos, uma forte coesão, restringindo o crescimento das plantas, mas, quando úmidos, a sua estrutura torna-se muito instável. O objetivo deste estudo foi avaliar o uso de condicionadores químicos na melhoria da qualidade física de solos coesos no ambiente dos Tabuleiros Costeiros de Pernambuco, a partir de ensaios de atributos físicos e físico-hídricos. Foi aberto um perfil em área de plantio de cana-de-açúcar sobre Tabuleiro Costeiro na Zona da Mata Norte e coletado amostras estruturadas e não estruturadas do horizonte coeso. As amostras foram subdivididas em três tratamentos T0 (Controle), T1 (Poliacrilamida - PAM /100 mg kg-1) e T2 (PAM + Gesso-CaSO42H2O/1,523 g L-1). Os atributos físicos avaliados foram: Porosidade Total (macro, meso, micro e criptoporosidade); densidade do solo (Ds); curva característica de retenção de água no solo (CCRAS); resistência do solo a penetração de raízes (RP), intervalo hídrico ótimo (IHO). As médias dos parâmetros físicos analisados foram submetidas à análise de variância (ANOVA) e para os parâmetros que apresentarem diferenças significativas foi aplicado o teste de Tukey, por meio do aplicativo SAS. A realização da toalete no solo contido nos anéis volumétricos, antes ou após a aplicação dos condicionadores, não interfere nos resultados da densidade (Ds); A forma de distribuição de poros, evidenciou as modificações na porosidade no solo sob ação dos condicionadores; A resistência à penetração foi um atributo eficiente para avaliar a redução da coesão; O índice de estabilidade estrutural não foi conclusivo para a escolha do melhor tratamento; Os valores do IHO comprovam a eficiência deste atributo, com relação à disponibilidade de água para às plantas; O tratamento com PAM aumentou a disponibilidade de água às plantas.

Submitted by Franciele Moreira (francielemoreyra@gmail.com) on 2017-09-21T20:25:51Z No. of bitstreams: 2 Dissertação - Cálita Pollyanna Marques - 2017.pdf: 1468493 bytes, checksum: 90300902d8a48fc5600761dfccf90324 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2017-09-22T11:43:21Z (GMT) No. of bitstreams: 2 Dissertação - Cálita Pollyanna Marques - 2017.pdf: 1468493 bytes, checksum: 90300902d8a48fc5600761dfccf90324 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) / Made available in DSpace on 2017-09-22T11:43:21Z (GMT). No. of bitstreams: 2 Dissertação - Cálita Pollyanna Marques - 2017.pdf: 1468493 bytes, checksum: 90300902d8a48fc5600761dfccf90324 (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2017-08-30 / Fundação de Amparo à Pesquisa do Estado de Goiás - FAPEG / American Cutaneous Leishmaniasis (LTA) is caused by protozoa of the genus Leishmania and it attacks the skin and/or mucous membranes. It is an endemic zoonosis in Brazil being notified in all the Regions. There is a relation between the species of parasite and the clinical manifestations, contributing to the diagnostic complexity, which up to the present, did not exist species-specific diagnosis. In the present work, the proposal was to standardize the analysis of the PCR/RFLP products to the ITS and G6PD genes in polyacrylamide gel. In order to standardize PCR/RFLP analysis on the ITS and G6PD genes, DNA from the reference Leishmania spp. Sample was used and 5%, 6%, 7% 8% and 10% polyacrylamide were tested. The protocol design considered the size of the amplicons generated by the PCR / RFLP, correlating the concentration of the polyacrylamide. To standardize the electrophoresis time the migration of the dyes used in the DNA sample buffer was monitored. To validate the technique, we selected 521 samples from Tocantins patients fixed on slides and 43 samples of parasite isolates stored in Leishbank. The samples from the patients from Tocantins were analyzed by PCR/RFLP to the kDNA and were selected to those positive for the validation. The concentration of the matrix for analysis of the PCR and RFLP products to the ITS gene was standardized in 6% and 8%, respectively, and distinguished the species L. (L.) amazonensis, but the species L. (V.) braziliensis, L. (V.) guyanensis and L. (V.) lainsoni were not distinguished among themselves, occurring the same between L. (V.) naiffi and L. (V.) shawi. The concentration of the matrix for analysis of the PCR products to the G6PD gene was standardized in 6% and distinguished L. (V.) braziliensis from the others. The validation of the polyacrylamide gel analysis of the PCR products to the ITS target from samples of patients fixed in slides was not performed, since of the 256 samples analyzed only 4 amplified. The validation of the analysis of the products of the PCR to the G6PD target in polyacrylamide gel from 48 samples fixed in slides characterized 81% as L. (V.) braziliensis and, characterized 63% of the isolates of parasites as L. (V.) braziliensis. The PCR/RFLP analyzes performed to discriminate Leishmania spp. are complementary and thus contribute to the species-specific diagnosis of LTA. / A Leishmaniose Tegumentar Americana (LTA) é causada por protozoários do gênero Leishmania e acomete pele e/ou mucosas. É uma zoonose endêmica no Brasil sendo notificada em todas as Regiões. Há uma relação entre a espécie de parasito e as manifestações clínicas, contribuindo à complexidade diagnóstica, que até o presente, não existi o diagnóstico espécie-específico. No presente trabalho, a proposta foi padronizar a análise dos produtos da PCR/RFLP aos genes ITS e G6PD em gel de poliacrilamida. Para padronizar as análises dos produtos da PCR/RFLP aos genes ITS e G6PD, foram utilizadas DNA da amostra de Leishmania spp., de referência, sendo testada a poliacrilamida na concentração de 5%, 6%, 7% 8% e 10%. O delineamento do protocolo considerou o tamanho dos amplicons gerados pela PCR/RFLP, correlacionando-o a concentração da poliacrilamida. Para padronizar o tempo da eletroforese foi monitorada a migração dos corantes usados no tampão da amostra de DNA. Para validar a técnica, foram selecionadas 521 amostras de pacientes tocantinenses fixadas em lâminas e 43 amostras de isolados de parasitos armazenados no Leishbank. As amostras dos pacientes tocantinenses foram analisadas pela PCR/RFLP ao kDNA e foram selecionadas àquelas positivas para a validação. A concentração da matriz para analise dos produtos da PCR e RFLP ao gene ITS ficou padronizada respectivamente em 6% e 8% e distinguiu a espécie L.(L.) amazonensis, porém as espécies L. (V.) braziliensis, L. (V.) guyanensis e L. (V.) lainsoni não foram distinguidas entre si, ocorrendo o mesmo entre L. (V.) naiffi e L. (V.) shawi. A concentração da matriz para analise dos produtos da PCR ao gene G6PD ficou padronizada em 6% e distinguiu L. (V.) braziliensis das demais. A validação da análise em gel de poliacrilamida dos produtos da PCR ao alvo ITS a partir de amostras de pacientes fixadas em lâminas não ser realizada, pois das 256 amostras analisadas somente 4 amplificaram. A validação da análise dos produtos da PCR ao alvo G6PD em gel de poliacrilamida a partir de 48 amostras fixadas em lâminas caracterizou 81% como L. (V.) braziliensis e, caracterizou 63% dos isolados de parasitos como L. (V.) braziliensis. As análises pela PCR/RFLP realizadas para discriminar Leishmania spp. se complementam e, assim, contribuem para o diagnóstico espécie-específico da LTA.