Les lipopolysaccharides (LPS) sont les constituants majeurs de la membrane externe des bactéries à Gram négatif. Ils sont composés d’un polysaccharide (PS) relié à une région lipidique : le lipide A. Ces structures sont très variables d’une bactérie à l’autre. Elles sont aussi susceptibles de subir des modifications post-traductionnelles par des enzymes en réponse à des stimuli extérieurs. Chaque bactérie possède sa propre signature représentée par la structure unique de ses LPS. Les LPS sont reconnus par le récepteur MD-2 :TLR4. Ce complexe discrimine la structure des lipides A et déclenche de façon structure-dépendante ses voies de signalisation. Les LPS sont très étudiés pour leurs activités néfastes, notamment dans le déclenchement de la cascade inflammatoire qui mène au choc septique. Cependant ils présentent aussi de multiples activités bénéfiques. Dans le cadre de cette thèse, nous avons exploré deux types d’activités bénéfiques des LPS chez l’homme.L’analyse structurale des LPS par spectrométrie de masse est une étape indispensable pour comprendre leurs activités biologiques. Nous proposons deux nouvelles micro-méthodes d’analyse appliquées directement sur les membranes bactériennes. Ceci nous permet d’identifier plus rapidement les molécules de LPS au plus proche de leurs configurations natives, en évitant les artefacts dus à l’extraction. Cette méthode puissante pourra également être développée comme outil de détection rapide de pathogènes.Ensuite, nous avons étudié la relation structure-activité de LPS naturellement peu toxiques mais actifs au niveau cutané. Ce sont des composés du lysat bactérien de Vitreoscilla filiformis contenus dans des crèmes dermatologiques et cosmétiques visant à traiter la dermatite atopique. Les lipides A possèdent des chaînes courtes d’acides gras et les groupes phosphates peuvent être substitués par des groupements phosphoéthanolamines. Les activités biologiques induites par ces LPS prennent place comme des mécanismes de défense pour améliorer la réponse immune et lutter contre les pathogènes.En outre, le pouvoir adjuvant des LPS est l’une de leurs activités bénéfiques les plus étudiées. Les LPS sont naturellement trop toxiques pour être utilisés tels quels. Nous avons, caractérisé des LPS détoxifiés par modifications chimiques, établi et effectué différents tests de « screening » permettant d’évaluer leur adjuvanticité et leur non-pyrogénicité.Enfin, pour des applications thérapeutiques chez l’homme, les LPS doivent être produits en grandes quantités. Nous exposons la compatibilité du procédé de production de la société LPS-BioSciences à l’échelle industrielle et la réglementation des médicaments. Une amélioration de la méthode d’extraction est proposée pour extraire des quantités de l’ordre du gramme. De plus, le procédé de production est réalisable en respectant les bonnes pratiques de fabrication des médicaments. / Lipopolysaccharides (LPS) are the main components of the outer membrane of Gram negative bacteria. They are composed of a polysaccharide moiety linked to a lipid one, the lipid A. These structures are different from one bacterium to the other. They are also able to be modified by enzymatic post-translational modifications in response to external stimuli. Each bacterium possesses its own footprint shown by the unique structure of its LPS. LPS are recognized by the MD-2:TLR4 receptor. This complex distinguishes the lipid A structures and it actives some signalling pathways with a structure dependant manner. LPS are studied for their harmful effects, especially for their implication in the inflammatory cascade leading to septic shock. However, LPS keep multiple beneficial activities. In the context of this thesis, we have explored two kinds of beneficial activities for human.Structural analysis of LPS by mass spectrometry is an indispensable step to understand their biological activities. We propose two new micromethods of analysis directly applied on bacterial membranes. We can identify quickly the LPS molecules as close as possible to their native configurations. Thus, we can check the LPS structures before their extraction. This powerful method could be developed as a rapid tool for pathogens detection.Then, we studied the structure-activity relationships of naturally low-toxic, but active LPS at the skin level. They are compounds of the Vitreoscilla filiformis bacterial lysate. This lysate is used in dermatological and cosmetic creams to treat atopic dermatitis. Lipids A are composed of short fatty-acid chains (10 and 12 carbons), and the phosphate groups can be substituted by ethanolamine-phosphate. The biological activities induced by these LPS take place as defense mechanisms to improve the immune response against pathogens.Moreover, the adjuvant capability of LPS is another well studied beneficial activity. LPS are naturally too toxic to be used as vaccine adjuvant. We characterized chemically detoxified LPS and we established and realized different « screening » tests to evaluate their adjuvanticity and non-pyrogenicity.Finally, for human therapeutic applications, LPS must be produced on a large scale. We exposed the compatibility between the LPS-BioSciences process of production at the industrial scale and the medical regulation. One improvement of the extraction method is proposed in order to extract about one gram of LPS. The process is compatible with the good manufacturing practices.

<p>The medication errors associated with intravenous (IV) administration may cause severe patient harm. To address this issue, smart infusion pumps now include a built-in dose error reduction system (DERS) to help ensure the safety of IV administration in clinical settings. However, a drug limit alert triggered by DERS may be overridden by the practitioners which can potentially cause patient harm, especially for high-risk medications. Most analytical measures used to estimate the associated risk of harm are frequency-based and only consider the overall drug performance rather than the severity impact from individual alerts. Unlike these other measures, the IV medication harm index attempts to quantify risk of harm for individual alerts. However, it is not known how well these measures describe the risk associated with alert-overridden scenarios. The goal of this research was (1) to quantitatively measure the risk for simulated individual alert-overridden infusions, (2) to compare these assessments against the risk scores obtained among four different analytical methods, and (3) to propose better risk quantification methods with a higher correlation to risk benchmarks than traditional measures, such as the IV Harm index. </p> <p>In this study, 25 domain experts (20 pharmacists and 5 nurses) were recruited to assess the risk (adjusted for risk benchmarks) for representative scenarios created based on hospital alert data. Four analytical methods were applied to quantify risk for the scenarios: the linear mixed models (Method A), the IV harm index (Method B), Huang and Moh’s matrix-based ranking method matrix-based method (Method C), and the analytical hierarchy process method, adjusted by linear mixed models (Method D). Method A used seven alert factors (identified as key risk factors) to build models for risk prediction, and Methods B and C used two out of seven factors to obtain risk scores. Method D used pairwise comparison surveys to calculate the risk priorities. The quantified scores from the four methods were evaluated in comparison to the risk benchmarks.</p> <p>Risk assessment results from the domain experts indicated that overdosing scenarios with continuous and bolus dose field limit types had significantly higher risks than those of bolus dose rate type. About the soft limit type, the expected risk in the group with a large soft maximum limit was significantly higher than the group with a small soft maximum limit. This significant difference could be found in the adult intensive care unit (AICU), but not in adult medical/surgical care unit (AMSU). The comparisons between four analytical methods and risk benchmarks showed that the risk scores from Method A (<i>ρ</i> = 0.94) and Method D (<i>ρ </i>= 0.87) were highly correlated to the risk benchmarks. The risk scores derived from Method B and Method C did not have a positive correlation with the benchmarks.</p> <p>This study demonstrated that the traditional IV harm index should include more risk factors, along with their interaction effects, for increased correlation with risk benchmarks. Furthermore, the linear mixed models and the adjusted AHP method allow for better risk quantification methods where the quantified scores most correlated with the benchmarks. These methods can provide risk-based analytical support to evaluate alert overrides of four high-risk medications, propofol, morphine, insulin, and heparin in the settings of adult intensive care unit (AICU) and adult medical/surgical care unit (AMSU). We believe that healthcare systems can use these analytical methods to efficiently identify the riskiest medication-care unit combinations (e.g. propofol in AICU), and reduce medication error/harm associated with infusions to enhance patient safety.</p> <p> </p>

Cette thèse a pour objectif le développement d’une méthode de pré-dimensionnement des systèmes mécatroniques prenant en compte l’aspect vibratoire sans passer par des techniques de conception coûteuses en temps de calculs et de mise en œuvre, telles que la CAO 3D et la méthode des éléments finis.Dans la phase amont de choix d’architecture du processus de conception des systèmes mécatroniques, des modèles analytiques simples sont nécessaires à l’architecte du système mécatronique afin de lui permettre de faire des choix d‘architecture prenant en compte les contraintes multi-physiques, notamment les vibrations. Dans ce but, une bibliothèque de modèles analytiques d’éléments mécaniques flexibles simples a été développée dans cette thèse en utilisant le langage de modélisation Modelica.Pour démontrer les possibilités de cette approche, une étude des réponses vibratoires de certains systèmes mécatroniques a été réalisée. Cette approche de pré-dimensionnement a ainsi été appliquée dans un premier temps à un système mécatronique simple formé d’une plaque rectangulaire supportant des composants tels que des moteurs et des cartes électroniques, puis dans un second temps à une éolienne représentant un système mécatronique complet.Les résultats obtenus lors des simulations ont été comparés avec ceux obtenus par la méthode des éléments finis ainsi qu’avec les résultats d’études présentes dans la littérature scientifique. Ces simulations nous ont permis de prouver que les modèles compacts développés fournissent à l’architecte du système mécatronique des résultats très précis avec un besoin en ressources informatiques faibles. / This thesis focuses on the development of a method for the preliminary design of mechatronic systems, taking into account the vibratory aspect, without going through costly design techniques, such as 3D CAD and finite element method.In an early stage of the design process of mechatronic systems, simple analytical models are necessary to the architect engineer in Mechatronics, for important conceptual decisions related to multi-physics coupling and vibration. For this purpose, a library of flexible elements, based on analytical models, was developed in this thesis, using the Modelica modeling language.To demonstrate the possibilities of this approach, we conducted a study of the vibration response of some mechatronic systems. Therefore, the pre-sizing approach was applied in a first phase to a simple mechatronic system, formed with a rectangular plate supporting electrical components such as electric motors and electronic cards, and in a second phase the approach was applied to a wind turbine, considered as a complete mechatronic system. Simulation results were compared with the finite elements method and other studies found in the scientific literature. Simulation results have enabled us to prove that the developed compact models assist the mechatronic architect to find results of simulation with an important accuracy and a low computational cost.

No description available.

No description available.

The role of biological markers (biomarkers) in public health, pediatric medicine and clinical toxicology cannot be understated. Clinically validated biomarkers used in newborn screening (NBS) serve to detect individuals at risk for a disease in the population, pre-symptomatically diagnose affected neonates early in life and/or accurately predict disease progression and treatment responses to therapy. However, there is urgent need for the discovery of more specific biomarkers that can improve screening accuracy in a high throughput, cost-effective yet ethical manner. The major objectives of this thesis were to develop innovative nontargeted metabolite profiling methodologies based on multisegment injection-capillary electrophoresis-mass spectrometry (MSI-CE-MS) for early detection of treatable genetic diseases, as well as comprehensive surveillance of drugs of abuse (DoA) in high risk subjects. Chapter II introduces a multiplexed MSI-CE-MS strategy for confirmatory testing of known biomarkers for various inborn errors of metabolism from a dried blood spot (DBS) that was rigorously validated using proficiency test specimens from Centres for Disease Control and Prevention (CDC) and authentic neonatal samples from Newborn Screening Ontario (NSO) with quality assurance. Additionally, MSI-CE-MS together with temporal signal pattern recognition revealed for the first time a novel class of pathognomonic marker elevated in galactosemia, namely N-galactated amino acids. Chapter III describes an untargeted metabolomic study to discover biomarkers of cystic fibrosis (CF) to reduce the high false positive rate and widespread carrier identification by current two-tiered NBS algorithms that rely on genetic testing. A panel of metabolites from retrospective DBS specimens, including several amino acids, ophthalmic acid and an unknown peptide, allowed for differentiation of asymptomatic CF neonates from screen-positive yet unaffected carriers and transient hypertrypsinogenemic cases. Chapter IV develops and validates a high throughput MSI-CE-MS assay for rapid screening for DoA and their metabolites in human urine with improved specificity and broad spectrum coverage as compared to classic targeted immunoassays. This method can also applied to ensure compliance, drug efficacy and patient safety while detecting for potential substitution or adulteration when using high resolution MS/MS. In summary, this thesis contributes an innovative methodology and data workflow for biomarker discovery for improved neonatal screening of rare genetic diseases in the population, which was also applied for more effective drug surveillance strategies in public health given the alarming worldwide opioid crisis. / Thesis / Doctor of Philosophy (PhD)

Hydroxylated polybrominated diphenyl ethers (OH-PBDEs) are naturally produced in aquatic ecosystems e.g. by algae. Many OH-PBDEs have been observed to be highly bioactive and to cause adverse effects through several pathways, e.g. via disrupting oxidative phosphorylation (OXPHOS). The levels of some OH-PBDEs have increased in Baltic biota over the past decades. This may be associated with the nutrient enrichment of the Baltic Sea, which has favored growth of some of the OH-PBDE producers. Ceramium tenuicorne has been suggested to be a producer of OH-PBDEs in the Baltic Sea, which is supported by the results presented in this thesis. The levels of OH-PBDEs were observed to fluctuate greatly in C. tenuicorne over the summer season, and to correlate with the levels of pigments in the algae. However, the observed congener pattern in C. tenuicorne questioned theories regarding the mechanism of their biosynthesis. The results indicate a much more selective pathway for biosynthesis than previously suggested for the production of OH-PBDEs. One of the most abundant OH-PBDEs in C. tenuicorne, 6-OH-BDE137, has previously been observed to be toxic to bacteria, fungi, and crustaceans. Furthermore, Baltic gammarids seemed to change their feeding preferences towards less grazing on C. tenuicorne during the production peek of OH-PBDEs in the alga. This suggests that OH-PBDEs may serve as allelochemical defense agents for C. tenuicorne. The transport and fate of OH-PBDEs through a Baltic food chain was also studied, including C. tenuicorne, Gammarus spp., three-spined stickleback (Gasterosteus aculeatus), and perch (Perca fluviatilis). A small portion of the OH-PBDEs were observed to be methylated in the alga, or by associated bacteria. The methylated OH-PBDEs biomagnified in the food chain up to perch, in which they were converted back to the OH-PBDEs via demethylation. The OH-PBDEs and their methylated counterparts were also partially debrominated in the food chain, which resulted in high concentration of 6-OH-BDE47 in the perch. This congener is the most toxic OH-PBDE with regards to OXPHOS disruption. Another biotransformation of OH-PBDEs was identified in Baltic Sea blue mussels (Mytilus edulis). High concentrations of OH-PBDEs were conjugated with lipophilic moieties, e.g. fatty acids. This increases the residence time of the OH-PBDEs in the mussels. Mussels have been suggested to conjugate steroids with fatty acids as a means to regulate hormone levels. The conjugation of OH-PBDEs to fatty acids may occur due to intrusion into this pathway. Methods were developed to include quantification of conjugated OH-PBDEs in the analysis of mussels. OH-PBDEs were also quantified in blood from Baltic Sea grey seals (Halichoerus grypus). Seals originating from the Baltic proper were observed to be more highly exposed to 6-OH-BDE47 than seals from the Gulf of Bothnia. However, the levels of OH-PBDEs were generally low. A major effort was invested into securing these results, including development of a new analytical method. Blood obtained from dead seals is a difficult matrix for quantification of OH-PBDEs, and previous attempts using an established method yielded unsatisfactory results. / <p>At the time of the doctoral defense, the following paper was unpublished and had a status as follows: Paper 4: Manuscript.</p>

[pt] O desenvolvimento e validação de um método analítico para determinação de vitamina K é de fundamental importância considerando a crescente preocupação mundial com questões referentes à qualidade de vida, nutrição, saúde e meio ambiente, além da importância deste micronutriente na saúde. Desenvolveu-se um método baseado na cromatografia eletrocinética capilar micelar (MEKC) para a determinação das vitaminas K1 e K3. As condições ótimas de análise por MEKC foram obtidas a partir de estudo univariado de parâmetros experimentais e instrumentais (tampão borato 3,1 g.L(-1), pH igual a 8,5; CTAB 18,2 g.L(-1); acetonitrila 20 por cento v/v, 25 graus Celsius, 30 kV, Tigual a 15 s). Realizou-se um estudo com uma cela de caminho óptico alongado de modo a melhorar o limite de detecção (LOD) e o limite de quantificação (LOQ) do método. As curvas analíticas de ambas as vitaminas apresentaram comportamento linear e homoscedástico com valores de r próximos a 0,99. Os valores de LOD e de LOQ para a vitamina K1 e para a vitamina K3 ficaram na ordem de 10(-5) g.L(-1) e 10(-4) g.L(-1), respectivamente. A precisão do método apresentou valores entre 2 e 7 por cento para a área dos picos. As recuperações obtidas para a vitamina K1 foram de 101 mais ou menos 2 por cento e 103 mais ou menos 2 por cento para as amostras de suplemento vitamínico e chá verde, respectivamente. Para a vitamina K3, a recuperação obtida foi de 99 mais ou menos 3 por cento para a amostra de chá. Testes comparativos realizados entre o método proposto por MEKC e o método de referência baseado em HPLC demonstraram uma boa exatidão (tcalculado menor que ttabelado). A incerteza associada ao método desenvolvido apresentou valores entre 2 e 30 por cento, sendo as fontes mais relevantes aquelas associadas à construção da curva analítica e ao preparo das soluções. O método analítico desenvolvido pode contribuir para a realização de medições confiáveis e sensíveis de diferentes formas de vitamina K. / [en] The development and validation of an analytical method for determination of vitamin K is of paramount importance considering the growing worldwide concern with questions regarding the quality of life, nutrition, health and environment and the importance of this micronutrient in health. It is developed a method based on capillary micellar electrokinetic chromatography (MEKC) for the determination of vitamin K1 and K3. The optimal condition for analysis by MEKC were obtained by means of experimental and instrumental univariate study (borate buffer 3.1 gL(-1), pH equal 8.5; CTAB 18.2 gL(-1), 20 per cent acetonitrile v/v 25 celsius degrees, 30 kV, T equal 15 s). A study with a longer optical path-length capillary cell was performed to improve the detection limit (LOD) and the limit of quantification (LOQ) of the method. Analytical curves of both vitamins presented linear and homoscedastic behavior, with r values near 0.99. The values of LOD and LOQ for vitamin K1 and vitamin K3 were on the order of 10(-5) g.L(-1) and 10(-4) g.L(-1), respectively. The precision of the method was between 2 and 7 per cent for the peak area. Recoveries for vitamin K1 were 101 more or less 2 per cent and 103 more or less 2 per cent for samples of vitamin supplement and green tea, respectively. For vitamin K3, recovery obtained was 99 more or less 3 per cent for the sample tea. Comparative tests performed among the proposed method by MEKC and the reference method based on HPLC showed good accuracy (tcalculated less than ttabulated). The measurement uncertainty associated with the method developed showed values between 2 and 30 per cent, and the relevant sources were those associated with the analytical curve and the preparation of solutions. The developed analytical method can contribute to achieve sensitive and reliable measurements of different forms of vitamin K.

[pt] Um procedimento analítico é considerado apropriado para uma aplicação específica quando é capaz de gerar resultados confiáveis, que possibilitem a tomada de decisão com grau de confiança adequado, sendo sua adequação consolidada mediante a realização de ensaios de validação. Este trabalho propõe a utilização de um procedimento analítico alternativo, mais simples e econômico, baseado na técnica espectrofotométrica de absorção molecular, para quantificação de imidacloprid, um inseticida sistêmico, em amostras de solução aquosa. Foi demonstrado que o procedimento proposto é adequado ao uso pretendido descrevendo-se, detalhadamente, cada etapa da validação, considerando-se os limites estabelecidos para cada parâmetro de validação e aplicando-se técnicas estatísticas apropriadas na avaliação dos mesmos: análise de variância, análise de regressão, testes de significância, gráficos de controle e a estimativa da incerteza de medição. As incertezas de medição dos procedimentos de rotina e alternativo foram estimadas e comparadas às tolerâncias estipuladas, estabelecendo o procedimento alternativo como adequado. Uma comparação experimental deste procedimento com o de rotina (baseado em HPLC) foi realizada como parte do protocolo de validação. Além da avaliação do procedimento para quantificação de imidacloprid em nível de traços, foi investigada também a possibilidade de sua utilização, para a quantificação do mesmo ingrediente ativo em produtos formulados. Neste caso, foi demonstrado, através da comparação das incertezas estimadas às tolerâncias estabelecidas, que o procedimento alternativo não é adequado, por apresentar incertezas na ordem de aproximadamente 50% do valor destas tolerâncias, não possuindo assim o rigor metrológico requerido para esta aplicação. / [en] Fit for purpose analytical procedures must be sufficiently reliable to support any decision taken based on the generated results. In order to achieve that, consolidated adequacy evaluation of the analytical procedure must be obtained by performing validation experiments. In this work, an alternative and simpler spectrophotometric method for the quantification of imidacloprid in aqueous samples was compared to the HPLC-UV based reference method used in routine. The overall validation procedure started with a detailed description of each validation stage, followed by the settling of the limits for each of the validation parameters and then, applying the following statistical techniques to evaluate each of the parameters: ANOVA, regression analysis, significance tests, control charts and uncertainty estimation. The measurement of uncertainties estimation, based on ISO-GUM recommendations, was done for both analytical procedures (the alternative one and the reference one). After comparing these uncertainties with the tolerance values, the adequacy of the alternative proposed procedure was confirmed. Finally, by consolidating the validation, the experimental comparison of quantification methods was conducted. Besides evaluating the analytical procedure for trace-level imidacloprid quantification in water samples, the proposed method was also evaluated as the analytical procedure for imidacloprid based formulated products. In this case, it was demonstrated that the spectrophotometric method did not present the requested metrological requirements for such application, since the estimated uncertainties of the alternative procedure were about 50 % of the tolerance values.

A hipertensão arterial é um fator de risco de alta prevalência para as doenças cardiovasculares, principalmente no mundo industrializado, aumentando o problema de saúde em virtude do aumento da longevidade e da prevalência de fatores contribuintes como obesidade, sedentarismo e dietas inadequadas. Estima-se que 10% das 55 milhões de mortes que acontecem a cada ano, são consequências da hipertensão arterial. Dois terços desses eventos ocorrem nos países em desenvolvimento, incluindo o Brasil, afetando principalmente a população de menor nível socioeconômico. Diuréticos como hidroclorotiazida associados a betabloqueadores como o metoprolol são exemplos de fármacos utilizados no controle da hipertensão. O presente trabalho teve como objetivo desenvolver, validar e comparar métodos analíticos para a identificação e quantificação simultânea do tartarato de metoprolol e da hidroclorotiazida por eletroforese capilar (CE) e por cromatografia líquida de ultra eficiência (UHPLC). Para a definição das melhores condições de análise e determinação simultânea dos analitos em estudo, e otimização do método, utilizou-se metodologia de superfície de resposta (ou Response Surface Methodology - RSM). O método por CE utilizou um capilar de sílica fundida com comprimento total de 30,2 cm x 50 &#181;m d.i. O tampão utilizado foi tetraborato de sódio 25,0 mmol L-1, injeção hidrodinâmica 0,5 psi/6s, tensão aplicada +15 kV, detecção em 225 nm temperatura a 25ºC. O tartarato de metoprolol e hidroclorotiazida foram separados em 1,2 e 2,1 min, respectivamente .O método por UHPLC foi realizado empregando uma coluna ZORBAX® SB C18 (50 mm x 2,1 mm x 1,8 &#181;m), fase móvel constituída por água:acetornitrila:trietilamina (83:17:0,2 v/v/v), pH 3 ajustado com ácido fosfórico, utilizou vazão de 0,9 mL min-1. A hidroclorotiazida e o tartarato de metoprolol foram separados em 0,7 e 1,0 min, respectivamente. Os métodos analíticos foram validados de acordo com os requerimentos da ANVISA, ICH e Farmacopéia Americana. Os métodos apresentaram boa linearidade com coeficiente de correlação maiores que 0.99, a precisão intra- e inter-day para os tempos de migração foi apropriada (DPR < 2%), a exatidão do método foi comprovada mediante teste de recuperação, obtendo-se valores de 100±2. Portanto, os métodos propostos demonstraram ser lineares, precisos, exatos e rápidos para quantificação simultânea da hidroclorotiazida e do tartarato de metoprolol. E podem ser considerados confiáveis para serem empregados em análise de rotina para controle de qualidade destes produtos farmacêuticos. / Hypertension is a risk factor of high prevalence for cardiovascular diseases, especially in the industrialized world, increasing health problems as a result of increased longevity and the prevalence of contributing factors such as obesity, physical inactivity and inadequate diets. It is estimated that 10% of 55 million deaths that occur each year, are consequences of hypertension. Two-thirds of those events occur in developing countries, including Brazil, affecting mainly the population from lower socioeconomic backgrounds. Diuretics like hydrochlorothiazide associated with beta-blockers such as metoprolol tartrate are examples of drugs used in the control of hypertension. The present work had as objective to develop, validate and compare analytical methods for identification and simultaneous quantification of metoprolol tartrate and hydrochlorothiazide by capillary electrophoresis (CE) and ultra performance liquid chromatography (UHPLC). For the definition of the best conditions of analysis and simultaneous determination of drugs in study, and optimization of the method, it was used response surface methodology (RSM or \"Response Surface Methodology). The CE method used a fused silica capillary with total length of 30.2 cm x 50 &#181;m d.i. The electrolyte used was sodium tetraborate 25,0 mmol L-1, hydrodynamic injection 0.5 psi/6s, applied voltage +15 kV, detection in 225 nm temperature at 25ºC. The metoprolol tartrate and hydrochlorothiazide were separated into 1.2 and 2.1 min, respectively. The UHPLC method was carried out employing a ZORBAX SB® C18 (50 mm x 2.1 mm x 1.8 &#181;m) column, mobile phase consisting of water: acetornitrila: triethylamine (83: 17: 0.2 v/v/v), 3 pH adjusted with phosphoric acid, the flow was 0.9 mL min-1. Hydrochlorothiazide and metoprolol tartrate were separated in 0.7 and 1.0 min, respectively. The analytical methods have been validated in accordance with the requirements of ANVISA, ICH and American Pharmacopoeia. The methods showed good linearity with correlation coefficient greater than 0.99, precision inter and intra day for times of migration was appropriate (DPR < 2%), the accuracy of the method has been proven by recovery test, obtaining values of 100 ± 2. Therefore, the proposed methods have shown to be linear, precise, accurate and fast for simultaneous quantification of hydrochlorothiazide and metoprolol tartrate; and can be considered reliable to be used in routine analysis for quality control of pharmaceutical products.