O nebivolol é um fármaco anti-hipertensivo, comercializado na forma de uma mistura equimolar dos enantiômeros RSSS e SRRR-nebivolol. Esses dois enantiômeros possuem propriedades farmacológicas distintas, o que sugere uma farmacocinética diferente entre ambos. Sendo assim, foram desenvolvidos dois métodos para a análise do nebivolol por eletroforese capilar (CE), um aquiral e outro quiral. O primeiro consistiu em um método para análise de forma farmacêutica comercial de comprimidos de nebivolol. Foram definidas as seguintes condições analíticas: capilar de sílica fundida 50 ?m de diâmetro interno (d.i) e 38 cm de comprimento efetivo (c.ef), eletrólito de corrida composto por tampão acetato de sódio 50 mM, pH 4,0, temperatura de 30 °C, tensão de 30 kV e detecção a 200 nm. O método desenvolvido permitiu a análise rápida e eficiente de comprimidos comerciais de nebivolol, sendo simples, seletivo, preciso e exato, está em conformidade com o guia de validação de métodos analíticos da ANVISA (2003), e foi aplicado para quantificação de comprimidos comerciais de nebivolol. O segundo método teve como objetivo realizar a análise enantiosseletiva da ligação do nebivolol à albumina humana do soro (HSA). Após a avaliação de diversos parâmetros, foram estabelecidas as seguintes condições analíticas: capilar de sílica fundida 50 ?m d.i e 38 cm c.ef, eletrólito de corrida composto por tampão acetato de sódio 50 mM, carboxi-metil-?-ciclodextrina 12,5 mM, 1% acetonitrila, pH 4,0, temperatura de 25 oC, tensão de 25 kV e detecção a 200 nm; e como técnica de stacking foi utilizada field amplified sample injection com plug de água (5 psi, 5 s) e injeção eletrocinética 5 kV por 30 s. Nesta condição foi obtida uma resolução de 1,58 e tempo de análise inferior a 25 minutos. No estudo de ligação à HSA foi observado que há enantiosseletividade na ligação, porém, este estudo ainda precisa ser melhor delineado em relação às concentrações de proteína e analito, bem como tempo de incubação e procedimento de filtração para separação das frações livre e ligada / Nebivolol is an anti-hypertensive drug, commercialized as a racemic mixture of RSSS and SRRR-nebivolol. Both enantiomers have distintict pharmacological properties, what suggests a different pharmacokinectis between them. Therefore, it was developed two methods for the nebivolol analysis by capillary electropforesis (CE), one of them is achiral and the other is chiral. The first method aimed the analysis of tablets. The analytical conditions were determined: silica fused capillary 50 ?m internal diameter (i.d.) and 38 cm effective length (ef. l.), running electrolyte composed by 50 mM sodium acetate buffer, pH 4.0, 30 °C temperature, 0 kV applied voltage and 200 nm UV detection. The developed method allowed a quickly and efficient tablets analysis, being simple, selective, accurate and precise, and it is also in accordance with ANVISA (2003) analytical methods validation guide, and it was applied to the quantification of nebivolol in tablets. The second method aimmed to analyze the enantiosselective nebivolol binding to HSA. After the evaluation of many parameters, it was stabilished the following analytical conditions: 50 ?m i.d. and 38 cm ef.l. fused silica capillary, running electrolyte composed by 50 mM sodium acetate buffer, 12.5 mM carboxymethyl- ?-cyclodextrin, acetonytrile 1%, pH 4.0, 25 oC, 25 kV applied voltage and 200 nm UV detection; and as stacking technique it was applied field amplified sample injection with water plug(5 psi, 5 s) and 5 kV por 30 s eletrokinect injection. At this condition, it was possible to achive 1.58 resolution and less than 25 minutes of analysis. At the HSA binding study, it was observed an enantiosselectivity on the binding; however this study still needs better desing in conserne to analyte and protein concentration, as well as, incubation time and filtration proceadure to the separion of binded and free fractions.

A microextração líquido-líquido dispersiva (DLLME) é uma técnica de preparo de amostra baseada no equilíbrio de distribuição do analito entre a fase doadora (amostra) e a fase aceptora (solvente orgânico) em um sistema ternário de solventes. Foi desenvolvida em 2006 por Rezaee e colaboradores para a determinação de hidrocarbonetos policíclicos aromáticos em amostras de água e por ser uma técnica muito recente,ainda é pouca explorada para o preparo de amostras biológicas. Portanto, o objetivo deste trabalho foi avaliar a DLLME como técnica para a extração da oxibutinina (OXY), fármaco usado para o tratamento da incontinência urinária, e da N-desetiloxibutinina (DEO), seu principal metabólito ativo, em amostras de urina. A análise da OXY e DEO foi realizada por eletroforese capilar (CE), utilizando um capilar de sílica fundida de 50 ?m de diâmetro interno, com comprimento efetivo de 36,5 cm, trietilamina 50 mmol/L, pH 3 como solução de eletrólito, tensão de30 kV, temperatura de 30°C e detecção em 204 nm. Nestas condições o tempo de migração da DEO foi de 7,12 min e da OXY foi de 7,42 min, com resolução de 3,1. O procedimento utilizado para preparo da amostra foi baseado na DLLME, utilizando 5 mL de urina,na qual foi adicionado 2,5% de NaCle cujo pH foi ajustado para 11. O solvente para a extração consistiu de uma mistura de 140 ?L de tetracloreto de carbono (solvente extrator) e 260 ?L de acetonitrila (solvente dispersor), que permaneceram em contato com a amostra pordois minutos. A avaliação das características de desempenho analítico apresentou faixa linear de 90-300 ng/mL para a OXY e 187,5-750ng/mL para a DEO. A recuperação absoluta foi de 71,4 e 60,9% e o limite de quantificação foi de 90 e 187,5 ng/mL para a OXY e DEO, respectivamente. Os estudos de precisão e exatidão apresentaram coeficientes de variação e erros relativos inferiores a 15% e as amostras foram estáveis nos estudos de estabilidade. Portanto, foi possível desenvolver um método rápido, fácil e confiável para analisar e quantificar a OXY e a DEO em amostras de urina por CE, usando a DLLME como técnica de preparo de amostra. / The dispersive liquid-liquid microextraction (DLLME) is a sample preparation technique based on the equilibrium distribution between an extraction solvent and a sample solution in a ternary solvent system. It was developed by Rezaee and co-workers in 2006 for the determination of polycyclic aromatic hydrocarbons in water samples. Until now, for being a very recent technique it was little explored for the analysis of drugs in biological fluids. Therefore, the aim of this work was to evaluate the DLLME as an extraction technique for oxybutynin (OXY), a drug used to treat urinary incontinence, and N-desethyloxybutynin (DEO), its main active metabolite in urine samples. The OXY and DEO\'s analysis was performed by capillary electrophoresis (CE) using a 50 ?m ID fused-silica capillary with an effective length of 36.5 cm with a photodiode array detector set at 204 nm. It made use of triethylamine 50 mmol/L pH 3 as background electrolyte, voltage of +30 kV and temperature of 30°C. Under these conditions the migration time were 7.12 minutes for DEO and 7.42 minutes for OXY, with a resolution of 3.1. The sample preparation procedure was based on DLLME and used 5 mL of urine samples whichionic strength was increased by the addition of 2.5% NaCland pH were adjusted to 11. The extraction mixture consisted of 140 ?L of carbon tetrachloride (extraction solvent) and 260 ?L of acetonitrile (disperser solvent), which remained in contact with the sample for two minutes. The analytical performance\'s evaluation presented linear range of 90-300 ng/mL for OXY and 187.5-750ng/mL for DEO. The absolute recovery were 71.4 and 60.9% and the limit of quantification were 90.0 and 187.5 ng/mL for OXY and DEO, respectively. The accuracy and precision studies showed coefficients of variation and errors below 15% and the samples were stable at stability studies. Therefore, it was possible to develop a fast, reliable and easy method to analyze and quantify OXY and DEO in urine samples by CE, using DLLME as a sample preparation technique.

A lercanidipina (LER) é uma dihidropiridina bloqueadora de canais de cálcio disponível comercialmente como uma mistura racêmica, ou seja, uma mistura equimolar dos enantiômeros (R)-LER e (S)-LER. Os efeitos farmacológicos da LER residem essencialmente no enantiômero (S)-LER, e estudos in vitro mostram que este enantiômero apresenta cerca de 100-200 vezes maior afinidade pelos canais de cálcio que o enantiômero (R)-LER. Consequentemente, os efeitos farmacodinâmicos da LER são devidos, principalmente, ao enantiômero (S)-LER. Neste sentido, fármacos racêmicos são exaustivamente estudados, para avaliar suas propriedades farmacocinéticas e farmacodinâmicas estereosseletivas e verificar se existem vantagens na produção do enantiômero puro. Além disso, devido às diferenças na atividade dos enantiômeros, é imprescindível realizar o controle de qualidade enantiosseletivo destas formulações. Associado a isto, as moléculas pertencentes a esta classe de fármacos são fotossensíveis. Assim, o objetivo deste trabalho foi desenvolver e validar um método enantiosseletivo para análise da LER e aplicá-lo no controle de qualidade de formas farmacêuticas e também avaliar sua fotodegradação. A análise enantiosseletiva da LER e aplicação no controle de qualidade de formas sólidas foi realizado por eletroforese capilar (CE) utilizando tampão acetato de sódio 200 mmol L-1, pH 4,0 como eletrólito de corrida, adicionado de TM-?-CD (2, 3, 6-O-metil-?-ciclodextrina) 10 mmol L-1 como seletor quiral, na temperatura de 15 °C e 25 kV de tensão. Os parâmetros de desempenho analítico, linearidade, precisão, exatidão, limite de quantificação e detecção, seletividade e robustez foram avaliados e estão de acordo com os requisitos preconizados pelos guias oficiais. O método foi aplicado para análise de comprimidos comerciais contendo LER, e mostrou ser adequado para quantificação das amostras, apresentando valores de coeficiente de variação (CV, %) e erro relativo (E, %) inferiores a 5 %. Além disso, foi avaliada a fotoestabilidade dos enantiômeros da LER na mistura racêmica e também dos enantiômeros separados, quando expostos à luz UVC (254 nm) e visível, por HPLC. A separação dos enantiômeros da LER foi alcançada empregando a coluna quiral Chiralpak® AD 250 × 4,6 mm, partículas de 10 ?m e fase móvel composta por hexano-etanol-dietilamina (97:3:0,3, v/v/v) e vazão de 1,0 mL min-1. O método foi validado e aplicado nos estudos de fotodegradação em soluçãos-padrão de LER. Em relação à exposição na luz UVC (254 nm) e visível foi observada degradação significativa dos enantiômeros da LER após 0,5 h de exposição. Além disso, os produtos de degradação foram caracterizados por LC/MS/MS e foi observada que a principal fragmentação ocorre no anel dihidropiridínico, assim como para outros fármacos desta classe, formando piridinas sem efeitos farmacológicos. / Lercanidipine (LER) is a dihydropyridine calcium channel blocker commercially available as a racemic mixture, or an equimolar mixture of enantiomers (R) and LER (S)-LER. The pharmacological effects of LER essentially reside in the enantiomer (S)-LER, and in vitro studies show that this enantiomer has about 100-200 times greater affinity for calcium channels that the enantiomer (R)-LER. Consequently, the pharmacodynamic effects of LER are due mainly to the enantiomer (S)-LER. Thus, racemic drugs are extensively studied to evaluate their pharmacokinetic and pharmacodynamic properties stereoselective and check if there are advantages in the production of pure enantiomer. Moreover, due to differences in the activity of the enantiomers is essential to perform quality control enantioselective these formulations. Additonally, the molecules of this class of drugs are photosensitive. The objective of this study was to develop and validate a method for enantioselective analysis of LER and apply it in the quality control of pharmaceutical forms and also evaluate their photodegradation. The enantioselective analysis of LER and application in quality control of solid forms was performed by capillary electrophoresis (CE) using sodium acetate buffer 200 mmol L-1, pH 4.0 as background electrolyte, 10 mmol L-1TM-?-CD (2, 3, 6-O-methyl-?-cyclodextrin) as chiral selector, at temperature of 15 °C and 25 kV voltage. The analytical parameters, linearity, precision, accuracy, limit of quantification and detection, selectivity and robustness were evaluated and accordance with the requirements recommended by official guideline. The method was applied for the analysis of commercial tablets containing LER, and proved to be suitable for quantification of the samples, with of CV (%) and E (%) less than 5%. In addition, the photostability was evaluated on the LER enantiomers and racemic mixture of the enantiomers also separated when exposed to UVC (254 nm) and visible light by HPLC. Separation of enantiomers of LER was achieved using the chiral column Chiralpak ® AD 250 × 4.6 mm, particles of 10 micrometres and a mobile phase composed of hexane-ethanol-diethylamine (97:3:0.3 v/v/v ) and flow rate of 1.0 mL min-1. The method was validated and applied in studies of photodegradation of standard solutions of LER. The method was validated and applied in studies of photodegradation in soluçãos standard of LERR With relation to UVC exposure (254 nm) and visible light, significant degradation was observed after 0.5 hour exposure of the solution containing the racemic mixture and after 1 h of exposure the solution containing the isolated enantiomers of LER. Also, the degradation products were characterized by LC/MS/MS and it was observed that the major fragmentation occurs in dihydropyridine ring, as well as other drugs of this class, forming pyridines no pharmacological effects.

Um sistema completo foi desenvolvido para o novo acoplamento de uma célula eletroquímica em fluxo com um eletroforese capilar equipado com detector condutométrico sem contato (EC-CE-C4D). Este acoplamento compreendeu o design da interface EC-CE, um potenciostato protegido contra fuga de alta tensão, subsistema de controle de fluxo por bombas solenóide e um equipamento CE-C4D adaptado para a aplicação. Um software dedicado para aquisição de dados, controle e operação do sistema foi desenvolvido na ferramenta gráfica de programação LabView. O sistema foi inicialmente aplicado na eletrodeposição de traços dos metais Cd2+, Pb2+, Cu2+, Zn2+ e Tl+, seguida pela redissolução e separação destes por CE, o que resultou em fator de pré-concentração de até 55 vezes para todos os metais, exceto Zn2+. Os metais acumulados foram redissolvidos na matriz original da amostra para permitir a separação e determinação conjunta com cátions majoritários (p. ex. alcalinos e alcalinos terrosos e amônio, se acima do LD). Para amostras cuja matriz possa causar interferência, pouco antes da injeção no CE, foi realizado processo de cleanup pela redissolução dos metais no eletrólito de separação. Experimentos pra separação bidimensional com o potencial de deposição ou redissolução voltamétrica como primeira dimensão e o tempo de eletromigração como segunda dimensão foram propostos e demonstrados. A oxidação eletroquímica de espécies orgânicas na interface EC-CE surgiu como um segundo campo de aplicação para: a) derivatização de analitos neutros tornando-os carregados ou ionizaveis, p. ex., conversão de álcoois alifáticos de cadeia curta em carboxilatos que possam ser separados e quantificados por CE-C4D; b) investigação/monitoramento de processos de eletrodo pela detecção de produtos carregados ou ionizáveis de reação de eletrodo, p. ex., investigação de condições experimentais para eletrooxidação de glicerol objetivando-se a síntese de produtos com maior valor comercial. / A complete instrumental system was developed for the new coupling of an electrochemical flow cell to capillary electrophoresis with contactless conductivity detection (EC-CE-C4D) and comprising an innovative design of the EC-CE interface, a potentiostat circuit protected from high voltage leakages, a solenoid pump based flow management subsystem, and a CE-C4D equipment tailored for this application. Dedicated software for data acquisition, devices control and operation automation was developed in LabVIEW graphical programming tool. The system was first applied to the electrodeposition of traces of Cd2+, Pb2+, Cu2+, Zn2+ and Tl+ , followed by stripping and separation by CE, allowing a 55-fold preconcentration factor in 6 min for all metals but Zn2+. The accumulated metals were stripped into the original sample matrix to allow their separation together with the sample\'s major cations (e.g. alkaline and alkaline earth metals and ammonium, if above LOQ). For samples with a matrix that originates interferences, cleanup was achieved by stripping the metals into background electrolyte shortly before injection in the capillary. Two-dimensional separations with voltammetric deposition or stripping potential as one dimension and electromigration time as the second one were proposed and demonstrated. Electrochemical oxidation of organic species in the EC-CE interface emerged as a second field of application for: a) derivatization of neutral analytes into charged or ionizable species, e.g., conversion of short chain aliphatic alcohols into carboxylates that can been determined by CE-C4D; b) investigation/monitoring of electrode processes by detection of charged or ionizable reaction products of electrode reactions, e.g., investigation of experimental conditions for glycerol electro-oxidation aiming the synthesis of commercially valuable products.

In the first part of this work, magnetized nanoporous silica based microparticles (MMPs) are used for horseradish peroxidase (HRP) immobilization and applied in amperometric peroxidase-based biosensors. A homemade magnetized carbon paste electrode permits the MMPs attraction close to the electrode surface. The resulting original biosensor is applied to the investigation of enzymatic oxidation of model drug compounds namely, clozapine (CLZ) and acetaminophen (APAP) by HRP in the presence of hydrogen peroxide. The biosensor operates at a low applied potential and the signal corresponds to the electro-reduction of electroactive species enzymatically generated. The biosensor allows performing the quantitation of the two drug compounds in the micromolar concentration range. It allows also the study of thiol compounds based on the inhibition of the biosensor response. Interestingly, distinct inhibition results are observed for HRP entrapped in the silica microparticles compared to the soluble HRP.<p>We expect that this type of biosensors holds high promise in quantitative analysis and in biotransformation studies of drug compounds.<p><p>In the second part of this thesis work, HRP immobilized magnetic nanoparticles are injected on-line and magnetically retained, as a microreactor, in the capillary of a CE setup. The purpose of such a configuration is to develop an analytical tool for studying “in vitro” drug biotransformation. The advantages expected are (i) minimum sample (drug compound) and biocomponent (enzyme) consumption, (ii) high analysis throughput, (iii) selectivity and sensitivity. In order to illustrate the potential of such an instrumental configuration, it has been applied to study acetaminophen as model drug compound. The mechanistic information obtained by the HRP/H2O2 system is in agreement with literature data on acetaminophen metabolization. Horseradish peroxidase kinetic studies are realized by this setup and the apparent Michaelis constant is determined. Capillary electrophoresis permitted the identification of APAP off-line biotransformed products such as N-acetyl-p-benzoquinone imine (NAPQI), the APAP dimer and APAP polymers as inferred from literature data. The formation of the APAP dimer was further confirmed by electrospray ionization mass spectrometry.<p> / Doctorat en Sciences biomédicales et pharmaceutiques / info:eu-repo/semantics/nonPublished

The fexofenadina is an antihistamine non sedative that acts as antagonistic selective in the outlying receivers H1 of the histamine, being used in the symptomatic treatment of allergic manifestations as medication of first choice. The project in subject has the objective of to determine and to validate methods physical-chemistries, for the evaluation of fexofenadine hydrochloride in covered tablets, for liquid chromatographic of high efficiency and zone capillary electrophoresis, both with detection for diodo array detector, for ends of to accomplish dissolution studies and to compare the quantitative methods. The two quantitative methods presented linearity, accurate, precision, specificity and robust.Dissolution of tablets of 120 and 180 mg was realize utilizing distillate water, HCl 0.1 M, buffer phosphate pH 4.5 and buffer phosphate pH 6,8 with dissolution medium, using apparatus II. The dissolution profiles obtained of each medium were compared through the comparison factors: f1, f2 and ED. Different resultes were obted of the likeness the formulation.The method based on the application of ANOVA in the use of the dissolution efficiency (ED) it is more discriminative than the f-factors. If we used the factor f2 as comparison parameter, all the means don't present difference in the formulations, but only in the medium of HCl 0,1 M, the formulations showed similarity for the parameters f1, f2 and ED, allowing to affirm that the two formulations are similar and with same performance in alive. The described methods, HPLC and CEZ, availability for variance analysis, presented equivalence for quantification of fexofenadine hydrochloride. / A fexofenadina é um anti-histamínico não sedativo que age como antagonista seletivo nos receptores periféricos H1 da histamina, sendo usada no tratamento sintomático de manifestações alérgicas como medicamento de primeira escolha. O trabalho em questão tem o objetivo de determinar e validar metodologias físico-químicas, para a avaliação de cloridrato de fexofenadina em comprimidos revestidos por cromatografia líquida de alta eficiência e eletroforese capilar de zona, ambas com detecção por DAD, para fins de realizar estudos de dissolução e comparar as metodologias quantitativas. Os dois métodos quantitativos apresentaram-se lineares, específicos, exatos, precisos e robustos. A dissolução dos comprimidos de 120 e 180 mg foi realizada nos meios de água destilada, HCL 0,1 M, tampões fosfato pH 4,5 e pH 6,8, utilizando o métodos das pás. Os perfis de dissolução obtido para as formulações em cada meio foram comparados pelos fatores de comparação: f1, f2 e ED, apresentando resultados diferentes quanto à semelhança das formulações. O método baseado na aplicação de ANOVA na utilização da eficiência de dissolução (ED) é mais discriminativo que os f-fatores. Quando usamos o fator f2 como parâmetro de comparação, os meios não apresentaram diferença nas formulações, mas somente no meio de HCl 0,1 M, as formulações mostraram similaridade para os parâmetros f1, f2 e ED, podendo apresentar o mesmo desempenho in vivo. Possibilitando a realizado de estudo de biodisponibilidade apenas com a formulação de maior dosagem.Os métodos, CLAE eECZ, avaliados por análise de variância, apresentaram-se equivalentes para quantificação de cloridrato de fexofenadina.

Natural products are complex mixtures of compounds with therapeutic effects which are often reported to be due to the synergistic action of multiple and sometimes unknown components. Consequently, standardization of these products is complex and a lack of effective quality control (QC) criteria in most countries has led to marketing of commercial products with questionable quality, safety and efficacy (QSE). The aim of this study was therefore to develop qualitative and quantitative analytical methods for use in the QC of Ginkgo biloba solid oral dosage forms. Initially, a micellar electrokinetic chromatography (MEKC) method was developed for the identification of the flavonol glycosides, rutin and quercitrin as well as 3 flavonol aglycones, quercetin, kaempferol and isorhamnetin in crude extracts of 4 Ginkgo biloba solid oral dosage forms using ultraviolet (UV) detection. A reversed-flow cyclodextrin-modified MEKC method was subsequently developed for the simultaneous determination of the aforementioned flavonols as well as ginkgolide A, B, C, J and bilobalide (all positive markers) in Ginkgo commercial products. A non-aqueous capillary electrophoresis (CE) method was also developed for fingerprinting the presence of ginkgolic acids (negative markers) in Ginkgo biloba leaf extracts, which are purported to be associated with toxic properties. This method was also applied to 2 Ginkgo biloba commercial products. Since the flavonols have strong UV absorbing chromophores, a reversed phase high-performance liquid chromatographic (RP-HPLC) method was developed and validated using photo-diode-array (PDA) detection which was then successfully applied to fingerprint commercially available Ginkgo biloba solid oral dosage forms as well as quantify the relevant flavonol markers present in these extracts. Sample preparation was simple, rapid and cost efficient with minimal clean-up and the employment of a minibore column which requires low mobile phase flow rates contributed to the economy of the method. Unlike the conventional QC approach, samples were not hydrolyzed and direct determination of 2 intact flavonol glycosides, together with the usual aglycone markers was facilitated which provided maximal content information for fingerprint comparisons. On the other hand, terpene trilactones possess poor chromophores and an alternative detection method to UV was required in order to obtain suitable sensitivity. RP-HPLC with evaporative light scattering detection (ELSD) was selected for quantification of these non-volatile constituents in Ginkgo dosage forms and this method was deemed suitable for the routine QC analysis of these positive markers in commercial products. Since approximately 33 flavonoids have been identified in Ginkgo biloba leaf extracts, baseline separation using UV/PDA detection normally requires complex gradient programs and long analysis times. In addition, unequivocal identification of the flavonoids with similar UV spectra and elution times cannot be guaranteed. A liquid chromatographic tandem mass spectrometric (LC-MS-MS) method was therefore developed and validated in order to ensure accurate quantification of the selected flavonol marker compounds in Ginkgo commercial products. LC-MS-MS analysis of Ginkgo extracts revealed, in addition to rutin, the possible presence of other quercetin analogues, quercetin-3-Orhamnoside-7-O-glucoside or quercetin-3-O-glucoside-7-O-rhamnoside, previously unreported in Ginkgo biloba leaf extracts or dosage forms. In terms of evaluating the most suitable analytical method for QC, CE shows exceptional potential in the future analysis of Ginkgo biloba dosage forms while HPLC-PDA and HPLC-ELSD are currently the most affordable and practical instruments for the routine analysis of the flavonols and terpenoids, respectively. LC-MS-MS proved to be pivotal for the accurate identification and quantification of the flavonols due to interference by other flavonoid compounds with similar retention times and UV spectra to the peaks of interest. All quantitative and qualitative results revealed large discrepancies in the marker content between the products regardless of which batch was analysed and product labels disclosed little relevant information. Although currently not required by most regulatory agencies, some of the usual quality criteria applied to orthodox medicines was evaluated. In particular, dissolution analysis, disintegration, tablet hardness and weight uniformity were assessed and revealed similar inconsistencies. This thesis emphasises that implementation of effective QC criteria is long overdue and is essential to ensure consistent product QSE of commercially available Ginkgo biloba solid oral dosage forms.

Orientador: Helena Teixeira Godoy / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-03T14:50:05Z (GMT). No. of bitstreams: 1 Prado_MarceloAlexandre_D.pdf: 10679979 bytes, checksum: 271e49a76278f93376a9b985c59b1111 (MD5) Previous issue date: 2003 / Resumo: O uso de corantes artificiais pelas indústrias de alimentos em todo o mundo é bastante difundido, isto porque os corantes permitem suplementar ou repor a coloração perdida durante o processamento e ou estocagem, e assim garantir a aceitabilidade do produto frente ao consumidor, sendo utilizados ainda como um importante instrumento para garantir a uniformidade dos produtos em linhas de produção de larga escala. Do ponto de vista da saúde pública, existem diferentes opiniões quanto à inocuidade dos diversos corantes artificiais utilizados em alimentos. Muitos estudos mostram que esses aditivos podem causar uma série de males à saúde da população quando consumidos de forma incorreta, seja por abusos da indústria ou exagero no consumo. O fato é que, técnicas analíticas para a determinação desses corantes devem ser desenvolvidas, e principalmente validadas, para garantir a segurança alimentar dos produtos que ingerimos. No presente trabalho foram desenvolvidos e validados dois métodos para a determinação de corantes artificiais em bebidas alcoólicas, utilizando duas diferentes técnicas, a cromatografia a líquido de alta eficiência (CLAE) e a eletroforese capilar (EC). Os métodos foram desenvolvidos para a separação simultânea dos onze corantes artificiais permitidos para uso em alimentos no Brasil. No método por CLAE, para a separação dos corantes, utilizou-se coluna de fase reversa e eluição por gradiente, com fase móvel composta por água/metanol. Na EC a separação ocorreu utilizando um capilar de sílica, com 73 cm de comprimento efetivo, preenchido com uma solução tampão composta por fosfato (10mmol/L) e dodecil sulfato de sódio (l0mmol/L), a pH 11, com aplicação de voltagem de 25 kV. Nos dois métodos, a detecção dos corantes foi feita na região do visível e a quantificação através de curvas de calibração externa. Os limites de detecção obtidos ficaram na faixa de 0,1 a 0,4 µ tg/mL e 0,4 a 2,5 µ tg/mL, enquanto os limites de quantificação foram de 0,2 a 1,3 µ tg/mL e 1,3 a 7,1 µ tg/mL para a CLAE e EC, respectivamente. As taxas de recuperação, em dois níveis de concentração, para todos os corantes foram de 95,2 a 103,2% para a CLAE, e de 92,6 a 104,0% para a EC. Os valores de repetibilidade calculado para padrões e amostras demonstraram a boa precisão para os dois métodos desenvolvidos. As metodologias propostas e validadas foram aplicadas em 45 amostras de bebidas alcoólicas de diferentes fabricantes brasileiros, sendo: 6 aguardentes aromatizadas, 9 coolers, 7 aperitivos, 3 coquetéis, 8 licores e 12 vinhos tinto. Não houve diferença significativa entre os dados obtidos pelos dois métodos. Em todas as amostras analisadas, os teores de corantes artificiais encontrados estavam em conformidade com a legislação brasileira / Abstract: The use of synthetic dyes for the food industries in the whole world is sufficiently spread out, it is because the colors allow to supplemental or to replace the lost coloration during the processing and or storage, and thus to guarantee the acceptability of the product front to the consumer, being used still as an important instrument to guarantee the uniformity of the products in the production. Of the point of view of the public health, there are different opinions about the safety of the different synthetic dyes used in foods. Much of the studies show that these additives can be dangerous for the health of the population when consumed inadequately, either for abuses of the industry or exaggerate in the consumption. The fact is that, analytical methods for the determination of these colors must be developed, and mainly validated, to guarantee the alimentary security of the products that we ingest. In the present work they had been developed and validated two methods for synthetic dyes determination in alcoholic beverages, using two different techniques, the high performance liquid chromatography (HPLC) and capillary eletrophoresis (CE). The methods had been developed for the simultaneous separation of the eleven synthetic dyes allowed for use in foods in Brazil. In the method for HPLC, for the separation of the synthetic dyes, a reverse phase column was used with gradient elution system composed by water/methanol. In the EC method the separation occurred using a silica capillary with 73 cm of effective length, filled with buffer phosphate solution (l0mmol/L) with sulpfate dodecyl sodium (SDS) (10mmol/L), at pH 11, with application 25kV of voltage. The detection and quantification were done made in same manner for the two methods, using absorption in the visible region and external standardization, respectively. The detection limits were 0.1 to 0.4 µ g/rnL and 0.4 to 2.5 µ g/rnL, while quantification limits were 0.2 to 1.3 µ g/rnL and 1.3 to 7.1 µ g/rnL for HPLC and CE, respectively. Recovery percentage at two levels of concentration for all the synthetic dyes were of the order of 95.2 to 103.2% for HPLC, and of92.6 to 104.0% for CE. The values of repeatability calculated for standards and samples demonstrated the precision of the two methods. The proposed and validated methods were used to analyses 45 alcoholic beverage samples of different Brazilian manufacturers, being: 6 perfumed spirits, 9 coolers, 7 bitters, 3 cocktails, 8 liquors and 12 red wines. The data obtained were the two methods did not present significant difference. It was observed that the limits permitted by Brazilian Legislation for the use of these synthetic dyes were respected in the analyzed samples / Doutorado / Mestre em Ciência de Alimentos

Orientadores: Cesar Costapinto Santana, Eliana A. de Rezende Duek / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia Quimica / Made available in DSpace on 2018-08-03T15:25:35Z (GMT). No. of bitstreams: 1 Vanin_Mirela_D.pdf: 8682882 bytes, checksum: 13666989fdfc12242eada33959fa5c98 (MD5) Previous issue date: 2003 / Resumo: A utilização de materiais poliméricos biodegradáveis na área médica foi um importante avanço biotecnológico, esses materiais podem ser utilizados como implantes temporários substituindo uma função particular do organismo por um período prédeterminado e degradar "in vivo" evitando cirurgia para a retirada do implante. O material a ser escolhido depende das necessidades da aplicação e da biocompatibilidade do material com o organismo vivo. O presente trabalho teve por objetivo estudar blendas de poli(Lácido láctico) (PLLA) / polihidroxibutirato (PHB) para aplicação na área médica. As blendas foram preparadas por dois métodos: evaporação do solvente e por fusão, obtendose amostras nas formas de filmes e pinos, respectivamente. Os filmes foram caracterizados por: análise termogravimétrica (TGA), calorimetria diferencial de varredura modulada (l / Abstract: The use of polymeric biodegradable materials in medicine is an important progress in biotechnology. These materials can be used like temporary implants as substitute a specific function of the organism for a pre-determined period, and its degradation "in vivo", so that there is no need for subsequent surgery to remove the implant The choice of the material will depend on the needs demanded for a certain application and the biocompatibility material/organism The aim of this work was to study poly(ß-hydroxybuyrate) (PHB) I poly(L-lactic acid) (PLLA) blends for medical area application. The blends were prepared from two different methods: casting and melting, obtaining samples in the forms of films and pins, respectively. The films were characterized by thermogravimetric analysis (TGA), modulated dynamical scanning calorimetry (MDSC), dynamical mechanical analysis (DMA), scanning electron microscopy (SEM), wide-angle X-ray scattering (W AXS), small-angle X-ray scattering (SAXS), and the pins, besides the techniques mentioned above, were characterized by flexural mechanical test. The "in vitro" study of the PHBIPLLA blends was accomplished and their hydrolytic degradation was evaluated. Experiments of human protein adsorption (albumin - HSA, imunoglobulin G HIgG, fibrinogen - HFg) onto biomaterials surfaces (PHB, PLLA PHBIPLLA) were made. These experiments used the Fourier transform intrared spectroscopy (FT-IR) coupled with attenuated total reflectance (ATR) in a continuous flow system, and simulated the physiological conditions in real time to study the biomaterials biocompatibility. The biocompatibility study was complemented using capillary electrophoresis analysis. The results showed that the PHB/PLLA are immiscible, with phase separation; however, with some interaction among the phases. Pins showed dense fracture surfaces, like films of pure PHB and PLLA, while the blends of films showed porous fracture surface. The PHBIPLLA blends revealed better thermal and mechanical properties than pure PHB. It is suggested that the PLLA presence promoted these results. In the degradation study we could observe that PLLA is more susceptible to biological environrnent than PHB and blends, because it presents a faster degradation in a comparison for films and pins. The results also showed that the PHB got to maintain its mechanical properties for more time than PLLA, despite of PHB has mechanical properties inferior than PLLA. During the degradation the samples disclosed tendency to crystallize, probably increasing the crystal lamellar thickness. The biocompatibility study showed density adsorption protein values, HSA and HIgG, onto biomaterials surfaces very similar to those reported in the literature. lt is suggested that the PHBIPLLA blends have potentiality to be used as bioreabsorbables polymers in the medical area / Doutorado / Desenvolvimento de Processos Biotecnologicos / Doutor em Engenharia Química

The diploma thesis is focused on the development of in-line micro-electromembrane extraction (EME) coupled to capillary electrophoresis (CE) for the analysis of selected drugs in body fluids. Up to now, direct coupling of EME to CE has been demonstrated on diluted river samples only [1]. Although the published set-up has been implemented within a commercial CE it suffers from several drawbacks that can have a negative impact on the analysis of samples with higher complexity. The instrumental arrangement presented in this thesis eliminates these deficiencies. The experimental part is based on the optimization of fundamental extraction and separation conditions for the analysis of model basic drugs (nortriptyline, haloperidol, loperamide) with the use of transient isotachophoresis (tITP) principle. The extraction conditions were optimized for electro-driven transport of basic analytes from complex matrices (urine) through free liquid membrane followed by injection step utilized by electrokinetic supercharging (EKS), which focused target analytes into the CE capillary. Optimized conditions have been applied on blood in the form of dry blood spots, which are highly attractive samples in the current clinical analysis. The repeatability of the measurements was