This thesis concerns the development and utilisation of techniques for characterisation and transdermal delivery of various systems for pharmaceutical applications. The degree of dissociation of drug molecules and the mobilities of the different species formed are essential factors affecting the rate of drug delivery by iontophoresis. Hence, determination of drug mobility parameters and equilibrium constants are important for the development of iontophoretic systems. With capillary zone electrophoresis using a partial filling technique and methyl-β-cyclodextrin as chiral selector, the enantiomers of orciprenaline were separated. The association constants between the enantiomers of the drug and the selector were also evaluated. Precision conductometry studies were performed for the hydrochloride salts of lidocaine and 5-aminolevulinic acid in aqueous propylene glycol and water as media, respectively. Iontophoresis is a technique for drug delivery where charged molecules are transported into and through skin by application of a weak direct electrical current. The drugs 5-aminolevulinic acid and its methyl ester were used as model compounds and incorporated in two different drug delivery vehicles, a sponge phase and carbopol gel. The bicontinuous structure of the sponge phase, constituted of monoolein and a mixture of propylene glycol and water, makes it interesting for use in iontophoretic delivery, since ions can move more or less freely in the aqueous as well as in the lipid domains. Furthermore, all three components are known for their penetration enhancing abilities. Hydrogels like carbopol gels are interesting media with respect to iontophoretic studies, since devices for iontophoresis often utilize hydrogels as contact interfaces between the skin and the electrodes. The results indicate that the transport achieved iontophoretically using the gel (1 % active substance) was comparable with the passive delivery of clinically used formulations (16 % - 20 % active substance).

Different multiple-injection methodologies and the partial filling technique (PFT) have been utilized for separation of pharmaceuticals by capillary elec-trophoresis. In multiple-injection capillary zone electrophoresis (MICZE), the samples and all standards, used for construction of the calibration curve, are analyzed within a single run. Four different modes of MICZE have been described by means of equations, which were experimentally verified. The developed equations facilitate the transfer from conventional single-injection CZE to one or more of these MICZE-modes, depending on the selectivity between the analyte and the injection marker. The applicability of two of these modes was then demonstrated by quantification of buserelin and salbutamol, re-spectively in commercially available pharmaceutical products. The content of buserelin in an injection solution was determined to 0.94 mg/ml, which only deviated slightly from the declared concentration (1 mg/ml). An alter-native mode of MICZE, offering a higher number of sequential sample injec-tions, was then utilized for single-run determination of salbutamol in 15 tab-lets, with a labelled content of 8 mg. The average content of the tablets was determined to 7.8 mg, with an intra-tablet variation of 3 % or less. Moreover, UV- and mass-spectrometric detection of enantiomeric amines, resolved by non-aqueous capillary electrophoresis (NACE), was demon-strated. Separation of enantiomeric amines was achieved using the chiral selector (-)-2,3:4,6-di-O-isopropylidene-2-keto-L-gulonic acid, (-)-DIKGA. Introduction of the non-volatile (-)-DIKGA into the mass-spectrometer was avoided by using the PFT, where the capillary is only partially filled with electrolyte containing the chiral selector.

Neste trabalho são apresentados processos de microfabricação de estruturas contendo microcanais e sistemas de manipulação hidrodinâmica e eletroosmótica de fluídos. Foram desenvolvidos processos de microfabricação utilizando toner sobre poliéster, toner sobre vidro, toner como resiste, além de métodos alternativos de perfuração de lâminas e selagem de microestruturas em vidro, desenvolvimento de microestruturas para eletroforese capilar e espectrometria de massas com ionização por eletronebulização. A caracterização dos materiais e processos permitiu uma ampla visão das potencialidades e alternativas dos processos de microfabricação, tendo sido demonstrado que os dispositivos produzidos em toner-poliéster são quimicamente resistentes às substâncias tipicamente utilizadas em eletroforese capilar. Neste trabalho, um detector condutométrico sem contato foi implementado em microestruturas de toner-poliéster e a separação eletroforética de alguns metais alcalinos é demonstrada. A microestrutura foi projetada no formato padrão em cruz, tendo o canal de separação 22 mm de comprimento, 12 µm de profundidade e largura típica. A cela condutométrica foi construída sobre o canal de separação utilizando-se fita adesiva de cobre (1 mm de largura) como eletrodos. O sinal aplicado na cela foi de 530 kHz e 10 Vpp . A separação de K+, Na+ e Li+ na concentração de 100 µmol L-1 foi efetuada em torno de 0,8 min, utilizando-se 1 kV como potencial de separação. Foram desenvolvidos microchips para análise por espectrometria de massas com introdução de amostra por eletronebulização, sendo determinado cluster do íon cloreto em concentração de 1 mmol L+. Também solução com 1 mmol/L de glucosamina em água/metanol 1: 1 (v/v), sob corrente de 100 nA gerou sinal estável e livre de descarga corona. Utilizando detecção amperométrica, obteve-se eletroferogramas mostrando a separação de iodeto (10 mmol L-1) e ascorbato (40 mmol L-1) em potencial de separação de 4,0 kV (800 V cm-1 potencial de detecção de 0,9 V (vs. Ag/AgCI), injeção com 1,0 kV/1°s, tampão borato de sódio 10 mmol L+ com CTAH 0,2 mmol L-1, pH 9,2. Obteve-se eficiência de 1,6.104 pratos/m e foi possível obter limites de detecção de 500 nmol L-1 (135 amol) e 1,8 µmol L-1 (486 amol) para iodeto e ascorbato, respectivamente. O processo de fabricação utilizando toner como material estrutural para microchips em vidro foi bem estabelecido, assim como os modos de detecção fotométrico e condutométrico foram demonstrados. Foram obtidos eletroferogramas par detecção condutométrica sem contato de solução 200 µmol L-1 de K+, Na+ e U+, em tampão histidina/ácido lático 30 mmol L-1 9:1 (v/v) água:metanol, injeção eletrocinética de 2,0 kV/5,0 s, potencial de separação de 1 kV, 530 kHz de frequência e tensão de 2,0 Vpp. Também foi implementado um sistema de detecção fotométrico para microchip operando em 660 nm, tendo sido utilizado para a detecção de azul de metileno 1,0 mmol L-1 em tampão de corrida de barato de sódio 20 mmol L-1 (pH 9,2), com o detector posicionado a 40 mm do ponto de injeção e com injeção eletrocinética a 2,0 kV por 12 s com picos bem resolvidos em menos de 1 min. / Microfabrication processes and devices for hydrodynamic and electroosmotic manipulation were developed based on toner-polyester, toner-glass and toner-as-resist techniques. Additionally, techniques to perforate glass slides and sealing of glass devices were introduced. Microdevices for capillary electrophoresis and electrospray for mass spectrometry were developed using these techniques. The characterization of the materiais and the processes demonstrated that the devices obtained by the toner-polyester process are compatible with the media used for capillary electrophoresis. The detection of alkaline ions with capillary electrophoresis with contactless conductivity detection was demonstrated. The typical cross shape microstructure was designed with a 22-mm long and 12-µm deep separation channel. The conductivity cell was implemented with 1-mm wide adhesive copper stripes. The applied signal was 530kHz and 10Vpp . The separation of 100µmo1L-1 K+, Na+, and Li+ was accomplished in 0.8 min under a voltage of 1 kV. Another toner-polyester microchip was developed to demonstrate its usefulness for electrospray/mass spectrometry. Solutions of 1 mmol L-1 potassium chloride and 1 mmol L-1 glucosamine in water/methanol 1:1 (v/v) were introduced with stable current of 100 nA without corona discharge. Capillary electrophoresis with amperometric detection was also demonstrated. The separation of iodide (10 mmol L-1) and ascorbate (40 mmol L-1) was carried out at 4.0 kV (800 V cm-1) with detection potential of 0.9 V (vs. Ag/AgCl), electrokinetic injection at 1.0 kV/10 s, running buffer of sodium borate 10 mmol L-1 with CTAH 0.2 mmol L-1 , pH 9.2. The efficiency was 1.6.104 plates/m and the limits of detection were 500 nmol L-1 (135 9mol) and 1.8 µmol L-1 (486 amol) for iodide and ascorbate, respectively. The toner-glass process was proposed and conductivity and photometric detections were demonstrated for the devices generated by this new technique. The separation of 200 pmol L-1 K+, Na+, and Li+ was achieved in buffer histidine/lactic acid 30 mmol L-1 water/methanol 9: 1 (v/v), electrokinetic injection at 2.0 kV/5.0 s, separation potential of 1 kV, and contactless conductivity detection at 530 kHz and 2.0 Vpp. The photometric detection of methylene blue at 660 nm was carried out in sodium borate 20 mmol L-1 (pH 9.2).

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No. of bitstreams: 1 patriciamendoncadecastrobarra.pdf: 3313806 bytes, checksum: 1fadfbcc46e82ff9e1237f3a851d8791 (MD5) Previous issue date: 2014-01-31 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Um método alternativo para a análise simultânea dos ácidos graxos (AG) majoritários cis-trans: esteárico (C18:0), elaídico (C18:1t), oleico (C18:1c), palmítico (C16:0), linoleico (C18:2cc ) e linolênico (C18:3ccc ), foi proposto utilizando a técnica de eletroforese capilar de zona (CZE) com detecção indireta. A metodologia foi otimizada através do planejamento composto central rotacional 23 (23-CCD) e após estudos, juntamente com a Análise de Componentes Principais (PCA), chegou-se a um eletrólito com condições ótimas, composto por: 15,0 mmol L-1 de tampão NaH2PO4/Na2HPO4 com pH ≈ 6,86, 4,0 mmol L-1 de SDBS, 8,3 mmol L-1 de Brij 35, 45% v/v de acetonitrila e 2,1 % de 1-octanol. A quantificação dos AG em amostras reais foi realizada através do cálculo de fator de resposta (FR) com a posterior verificação da linearidade dos modelos matemáticos propostos. O método de CZE foi aplicado com sucesso e nenhuma diferença significativa foi encontrada dentro de intervalo de confiança de 95% quando comparado com a metodologia oficial por cromatografia à gás (GC) para as amostras alimentícias do tipo: azeite de oliva, óleo de soja, gordura vegetal hidrogenada, manteiga, margarina, requeijão e biscoito recheado. Além da utilização da metodologia para análise de amostras alimentícias, foi também aplicada na determinação AG no fígado de ratos Wistar de três grupos, que consumiram dietas, cuja fração lipídica era composta por óleo de soja utilizado por diferentes períodos de tempo em processos de fritura por imersão. Após 45 dias consumindo essas dietas, os ratos foram submetidos à eutanásia e o teor de AG no fígado foi monitorado por CZE. Os resultados obtidos foram comparados com o método oficial por cromatografia à gás e não foram observadas diferenças significativas no intervalo de confiança de 95% e/ou 99%. Os resultados dos eletroferogramas do fígado dos ratos foram avaliados por PCA, sendo possível obter um reconhecimento de padrão e discriminar o grupo final dos grupos intermediários e controle e, indicando que o tempo de exposição total de óleo submetido a processos de fritura pode ser considerado relevante para a avaliação da qualidade do óleo. Tendo observada diferença no teor de AG no fígado dos grupos de animais, buscou-se verificar possíveis biomarcadores para o metabolismo celular e, considerando que alguns elementos atuam como cofatores para a atividade enzimática, determinados minerais foram avaliados em amostras de fígado e fêmur dos ratos Wistar envolvidos no estudo supracitado, utilizando a técnica de espectroscopia de emissão atômica com plasma indutivamente acoplado (ICP OES). Sabe-se que todos os animais envolvidos neste estudo receberam a mesma quantidade de selênio e de mesma origem (Mix mineral). Entretanto, os animais pertencentes ao grupo final apresentaram uma diminuição considerável no teor de selênio quando comparados aos grupos controle e intermediário. Logo, pode-se dizer que a interação do selênio com outros componentes da dieta (AG) foi alterada, uma vez que os animais deste trabalho foram afetados distintamente pela dieta, apresentando uma grande dispersão nos valores de selênio no grupo final. A diminuição no teor de selênio do grupo final pode estar relacionada com a diminuição da função antioxidante, já que os processos de oxidação geram também compostos altamente reativos, que em excesso aumentam o estresse oxidativo de estruturas celulares. / An alternative method for the simultaneous analysis majority of cis/trans fatty acids (FA): stearic (C18:0), elaidic (C18:1 9t), oleic (C18:1 9c), palmitic (C16:0), linoleic (C18: 2 9c 12c) and linolenic (C18: 3 9c 12c 15c ) by Capillary Zone Electrophoresis (CZE) with indirect detection was proposed. The methodology was optimized through central composite rotational design 23 (CCD - 23) and after studies, along with the Principal Component Analysis (PCA) , the electrolyte with optimum conditions, consisted of 15.0 mmol L- 1 NaH2PO4/Na2HPO4 buffer at pH ≈ 6.86, 4.0 mmol L -1 SDBS , 8.3 mmol L -1 Brij 35, 45 % v/v acetonitrile and 2.1 % 1-octanol . The FA quantification in real samples was performed by calculating the response factor (Rf) with the subsequent linearity of the proposed mathematical models verification. The CZE method was successfully applied and no significant difference was found within the range of 95 % when compared with the official method by gas chromatography (GC) for food samples such as: olive oil, soybean oil, hydrogenated vegetable fat, butter, margarine, spreadable cheese and filled cookie. In addition to the use of the methodology for analysis of food samples was also applied to FA determination in the Wistar liver rat three groups and fed diets whose lipid fraction was composed of soybean oil used for different stages of time for deep frying processes. After 45 days consuming these diets, the rats were euthanized and the content of FA in liver was monitored by CZE. The results were compared with the official method by GC and no significant differences were observed in the range of 95 % and/or 99 % of confidence interval. The results of the rat liver electropherograms were evaluated by PCA, it was possible to obtain a recognition pattern and to discriminate the final group of the intermediate and control groups, indicating that total exposure time subjected to oil deep frying processes may be relevant for evaluating the quality of the oil. After detecting differences in the content of FA in the liver of animal groups, we required to verify potential biomarkers for cellular metabolism and, whereas some elements act as cofactors for enzyme activity, certain minerals were evaluated in samples of Wistar liver and femur rat involved in the study mentioned above, using the technique of Inductively Coupled Plasma Optic Emission Spectroscopy (ICP OES). It is known that all animals involved in this study received the same amount of selenium from the same origin (Mineral Mix). However, the animals belonging to the late group showed a significant decrease in selenium content in comparison with to the control and intermediate groups. Therefore, it is possible observe that the selenium interaction with other dietary components (FA) has been changed, since the animals in this study were distinctly affected by diet, with a broad dispersion in the values of selenium in the final group . The decrease in final group selenium content may be related to the decrease of antioxidant function , since oxidation processes also generate highly reactive compounds which excessive increase the oxidative stress of cell structures .

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No. of bitstreams: 1 adrianaferreirafaria.pdf: 20849316 bytes, checksum: 511836cab839ebb39105bd682c987241 (MD5) Previous issue date: 2010-08-13 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O objetivo principal da tese em questão foi o desenvolvimento, otimização e validação de metodologias analíticas para análise de formulações farmacêuticas utilizadas no tratamento da tuberculose, as quais contêm como princípio ativo etambutol (ETB), isoniazida (ISO), rifampicina (RIF) e pirazinamida (PIR), isolados ou em associação. Inicialmente, duas metodologias analíticas para análise de ETB em comprimidos foram desenvolvidas: a primeira permitiu a análise simultânea de ETB e da impureza 2-amino-1-butanol por eletroforese capilar de zona (CZE) utilizando um eletrólito de corrida contendo 60,0 mmol L-1 de tampão ácido acético/acetato de sódio e 5,0 mmol L-1 de Cu2+, polaridade normal (+25 kV) e detecção UV em 262 nm. A segunda foi realizada por espectrofotometria com diluição dos padrões e amostras em 5,0 mmol L-1 de tampão ácido acético/acetato de sódio e 0,5 mmol L-1 de Cu2+ e detecção UV em 262 nm. A formação do complexo CuETB foi necessária em ambas as metodologias desenvolvidas, uma vez que ETB apresenta baixa absortividade molar. Algumas figuras de mérito, tais como, linearidade, repetitividade, exatidão, limite de detecção e limite de quantificação foram avaliadas para as duas metodologias. Os dois métodos foram comparados e não apresentaram diferenças significativas no intervalo de 95% usando o teste t não pareado com variância agrupada. Em um segundo momento, um novo método para análise simultânea de dose fixa combinada (DFC), contendo ETB, ISO, RIF e PIR por CZE com detecção UV foi desenvolvido, otimizado e validado. A condição experimental consistiu em um eletrólito contendo 50,0 mmol L-1 de tampão ácido acético/acetato de sódio e 12,5 mmol L-1 de Cu2+ e polaridade normal (+22 kV). Essa metodologia foi aplicada com sucesso na análise de 2-DFC (ISO e PIR) em comprimidos e 4-DFC (ETB, ISO, RIF e PIR) em sache. Finalmente, uma metodologia por CZE utilizando detecção UV para a análise simultânea de ISO, suas impurezas e produtos de degradação, como o ácido isonicotínico, isonicotinato de etila, 4-cianopiridina e isonicotinamida foi otimizada. A condição experimental consistiu em um eletrólito contendo 50,0 mmol L-1 de tampão ácido acético/acetato de sódio e 12,5 mmol L-1 de Cu2+, polaridade normal (+20 kV) e detecção UV em 270 nm. É importante ressaltar que planejamentos de experimentos, tais como, planejamento fatorial completo 32 e Box-Behnken 33 foram utilizados como ferramenta auxiliar no desenvolvimento e otimização das metodologias analíticas (CZE e espectofotométrica) e na avaliação da robustez para a análise de 4-DFC. / The main focus of this thesis was the development, optimization and validation of analytical methodologies for analysis of pharmaceutical preparations used for tuberculosis treatment, which contain as active ingredients, ethambutol (ETB), isoniazid (ISO), rifampicin (RIF) and pyrazinamide (PYR) isolated or in association. Two analytical methodologies for ETB analysis in tablets were developed initially: the first one took into account the capillary zone electrophoresis (CZE) approach for simultaneous analysis of ETB and its impurity 2-amino-1-butanol using an electrolyte consisting of 60.0 mmol L-1 of acetic acid/acetate buffer, 5.0 mmol L-1 of Cu2+, under normal polarity (+25 kV) and UV detection at 262 nm. The second one was performed by spectrophotometry using 5.0 mmol L-1 of acetic acid/acetate buffer and 0.5 mmol L-1 of Cu2+ for standards and sample dilutions under UV detection at 262 nm. The CuETB complex was necessary for the two methodologies, since ETB presents low molar absorptivity. Some merit figures such as linearity, repeatability, accuracy, limit of detection and limit of quantification were evaluated for both methodologies. These methodologies were compared and they did not present significant differences at the 95% confidence level using the parametric independent sample t test. In a second phase a novel method for the simultaneous analysis of fixed dose combination (FDC) containing ETB, ISO, RIF and PYR by CZE under UV detection was developed, optimized and validated. The experimental condition optimized consisted of an electrolyte containing 50.0 mmol L-1 of acetic acid/acetate buffer and 12.5 mmol L-1 of Cu2+ under normal polarity (+22 kV). This methodology was successfully applied to the analysis of 2-FDC (ISO and PYR) in tablet samples and 4-FDC (ETB, ISO, RIF and PYR) in a sachet sample. Finally, a CZE methodology using UV detection for simultaneous analysis of ISO, its impurities and degradation products such as isonicotinic acid, ethyl isonicotinate, 4-cyanopyridine and isonicotinamide was ptimized. The optimized experimental condition consisted of an electrolyte containing 50.0 mmol L-1 of acetic acid/acetate buffer and 12.5 mmol L-1 of Cu2+ under normal polarity (+20 kV) and UV detection at 270 nm. It is important to stress that design of experiment such as 32 full factorial design and 33 Box-Behnken design were used as auxiliary tools in the development and optimization of the analytical methodologies (CZE and spectrophotemetric) and robustness evaluation for 4-FDC analysis.

Submitted by Renata Lopes (renatasil82@gmail.com) on 2016-01-15T17:27:17Z No. of bitstreams: 1 renatadejesuscoelhocastro.pdf: 4626733 bytes, checksum: 62bb62e16069cf7f8e81f59effe1bd20 (MD5) / Approved for entry into archive by Adriana Oliveira (adriana.oliveira@ufjf.edu.br) on 2016-01-25T17:42:17Z (GMT) No. of bitstreams: 1 renatadejesuscoelhocastro.pdf: 4626733 bytes, checksum: 62bb62e16069cf7f8e81f59effe1bd20 (MD5) / Made available in DSpace on 2016-01-25T17:42:17Z (GMT). No. of bitstreams: 1 renatadejesuscoelhocastro.pdf: 4626733 bytes, checksum: 62bb62e16069cf7f8e81f59effe1bd20 (MD5) Previous issue date: 2015-08-07 / CAPES - Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / A produção de leite e carne de bovinos no Brasil está baseada na utilização de pastagens, por constituírem alimento de menor custo para o produtor. No Brasil, as pastagens são a principal fonte de alimentação do rebanho, sendo responsável por quase 90% da carne bovina e pela maior parte do leite produzido no país. Sendo assim, surge a importância de um monitoramento rigoroso para se obter uma forragem de qualidade, visto que, a dieta desses ruminantes está diretamente relacionada com a qualidade do leite. Nos últimos anos, tem-se dado o interesse de conhecer substâncias químicas presentes nas forragens que possam contribuir tanto para uma boa qualidade do leite, como por exemplo: o ácido linoléico (C18:2 cis-9 cis-12) e o ácido α-linolênico (C18:3 cis-9 cis-12 cis-15), que são ácidos graxos precursores do ácido linoléico conjugado (CLA) quanto para seleção de cultivares resistentes ao estresse causado pela presença das cigarrinhas das pastagens, como por exemplo: ácidos orgânicos (ácidos cítrico, málico, aspártico e succínico). O objetivo deste trabalho foi utilizar a técnica de Eletroforese Capilar de Zona (CZE) como uma ferramenta para auxiliar na seleção de materiais com uma maior concentração dos precursores do CLA e materiais mais resistentes à presença das cigarrinhas das pastagens. Para análise dos precursores de CLA, o eletrólito utilizado constituiu-se de 17% de metanol, 33% de ACN, 12 mmol L- 1 de tetraborato de sódio, 12 mmol L-1 de Brij e pH próximo de 9,5 e as condições de análise foram: injeção hidrodinâmica (25 mbar por 5 s), voltagem de + 26 kV, temperatura no interior do capilar de 27 ºC, detecção direta no UV em 200 nm, capilar de TSU com 75 μm d.i. e 34 cm de comprimento total. O método de eletroforese capilar de zona (CZE) se mostrou eficaz para análise dos precursores de CLA, em 4 minutos. Foi realizada a comparação do método proposto com o método oficial por cromatografia a gás (CG) em seis amostras de braquiárias, sendo que, os resultados não apresentaram diferenças significativas em um intervalo de confiança a 95 %. Para análises dos ácidos orgânicos, o eletrólito utilizado constituiu-se de 20 mmolL-1 de ácido ftálico, 15 mmolL-1 de TRIS e 0,8 mmolL-1 de CTAB. As condições de análise foram: injeção hidrodinâmica (0,3 psi por 2 s), voltagem de - 15 kV, temperatura no interior do capilar de 25 ºC, detecção direta no UV em 240 nm, capilar de TSP com 75 μm d.i. e 375 μm d.e., 50,2 cm de comprimento total e 40,0 cm de comprimento efetivo. O método CZE se mostrou eficaz para análises dos ácidos orgânicos (ácido cítrico, ácido málico, ácidos aspártico e ácido succinico) relacionados aos estresses das plantas, pois foi possível a identificação em menos de 5 minutos, possibilitando assim um grande número de análises em um curto tempo. / Bovine milk and meat production in Brazil is based on the use of pastures, as they are less food costing for the producers. In Brazil, pastures are the leading power supply of the herd, which are responsable for almost 90% of bovine meat and the biggest part of milk produced in the country. By this way, rises a rigorous monitoring to get quality forage, considering that the rumminant diet is related to the milk quality. Last years, it has arised the interest of knowing chemical substances present in the feed that could contribute to a good milk quality, such as: linoleic acid (C18: 2 cis-cis-9-12) and α- linolenic acid (C18: 3 cis-cis-9-cis-12 15), which are precursors of fatty acids conjugated linoleic acid (CLA); and to selection of cultivars resistent to stress caused by the sharpshooters pastures presence, such as the organic acids (citric, malic, succinic and aspartic acid). This paper aimed use the Capillary Zone Electrophoresis (CZE) technique as a tool to assist the selection of materials with a higher concentration of CLA precursors and more resistant materials to the presence of sharpshooters pastures. For analysis of CLA precursors, the electrolyte used consisted of 17% methanol, 33% ACN, 12 mmol L-1 sodium tetraborate, 12 mmol L-1 and Brij pH of 9.5, and analysis conditions were: hydrodynamic injection (25 mbar for 5 sec) + 26 kV voltage, 27 ° C temperature capillary within, direct UV detection at 200 nm, TSU capillary with 75 μm d.i and 34 cm length. The CZE method is effective for CLA precursors analysis, in 4 minutes. A comparing of the proposed method to the official one was made, by using gas chromatography (GC) at six samples brachiaria, and the results showed no significant differences at a confidence interval of 95%. For analysis of organic acids, the electrolyte used consisted of 20 mmol L-1 phthalic acid, 15 mmol L-1 TRIS and 0.8 mmol L-1 CTAB. The analysis conditions were: hydrodynamic injection (0.3 psi/3 seconds), - 15 kV voltage, 25 ° C temperature capillary within, UV direct detection at 240 nm, TSP capillary with 75 μm d.i. and 375 μm d.e., 50,2 cm lenght and 40.0 cm effective length. The CZE method is effective for organic acids (citric acid, malic acid, aspartic acid and succinic acid) analysis related to the plants, as it was possible its identification in less than 5 minutes, allowing a large number of analyzes in a short time.

The diploma thesis is dealing with design of analytical method for determination of caseins in cow‘s milk. Capillary electrophoresis was used as separative method. The developed procedure was verified on real samples. Theoretical part gives basic information about caseins. Their chemical and physical properties, structure, way of manufacturing, usage in food industry and non-food applications are described. There are further some nutritional factors mentioned, whose application in cattle nutrition can affect milk composition, especially milk protein concentration. Literary part contains summary of capillary electrophoresis methods used for casein determination. Experimental part describes used separative system, procedure of casein isolation from milk and casein content in individual milk samples. Separation of milk caseins was performed in an untreated fused-silica capillary with dimensions of 50 um i.d. x 96 cm total length (71 cm effective length). Phosphate buffer (10 mM, pH 2,5) served as the background electrolyte. Casein from milk was obtained by isoelectric precipitation at pH 4,6 after adding 10% acetic acid. Product was washed with acetone to remove fat and dried. Total casein yield was the highest in bio-milk (27,04 g/l), the lowest in skim long-life milk (22,04 g/l). The diploma thesis has arisen on the basis of cooperation of Institute of Food Science and Biotechnology, Faculty of Chemistry, Brno University of Technology and Research Institute for Cattle Breeding, Ltd., Department of Animal Nutrition Physiology in Pohořelice.

The diploma thesis, Determination of the caseins by the capillary electrophoresis, is dealing with a creating of the analytical method for a determination of the caseins in cow‘s milk. The capillary electrophoresis was used as a separative method. The developed procedure was optimalized and verified on standards of the caseins and the real samples. The theoretical part gives an information about the compound of cow's milk, caseins, their usage in the food industry and the non-food technologies. There are mentioned some further nutritional factors, whose application in the cattle nutrition can affect the milk composition. The experimental part describes the procedure of the caseins isolation from milk, the used separative system and the determination of the caseins in the lyophilisated samples of milk. The separation was performed in two untreated fused-silica capillaries. Both of them had the same internal diameter: 50 m. The total length of the first capillary was 96 cm (71 cm of effective length) and of the second used capillary 125 cm (effective length 100 cm) The phosphate buffer (10 mM, pH 2,5) served as a basic electrolyte. The diploma thesis has arisen on the basis of the cooperation of the Institute of Food Science and Biotechnology, the Faculty of Chemistry, Brno University of Technology and the Research Institute for Cattle Breeding, Ltd., Department of Animal Nutrition Physiology in Pohořelice.

From viewpoint of environmental analysis in the whole world became popular in the latest years the question of drugs’ breakthrough to the component of environment. These contami-nants belong to the biological active compounds, with different physical-chemical and biolog-ical properties and evince great tendency to bioaccumulation. They penetrate to the environ-ment because of their increasing of usage and wrong techniques of liquidation. The most often use drugs are preparations against pain – analgetics, and from this category there are non-steroidal anti-inflammatory drugs. The frequent usage of this compounds relate to their easy accessibility. From this large group of compounds was chosen for monitoring: diclofenac, ibuprofen, ketoprofen, salicylic acid, naproxen and acetaminophen; because they include in favorite preparations. The monitoring matrix was the wastewater from two waste water treat-ment plants (WWTP). The sampling was performed in inflow and outflow because the com-paring of concentration of selected contaminants and discovering of efficiency of removing the polutants reliance on treatment technology. For extraction of selected contaminants was used solid phase extraction (SPE) and for determination was used capillary zone electrophore-sis (CZE) with diode array detection (DAD). There was identified and quantified all of se-lected analgetics in inflow and so in outflow of WWTP. That means, this polutants infuse into surface water and then into other components of environment.

High-performance electromigration separation methods, capillary zone electrophoresis (CZE) and capillary isoelectric focusing (CIEF), have been applied to physico-chemical characterization of new synthetic low-molecular mass markers of isoelectric points. Amphoteric compounds on the basis of aminomethylnitrophenols, their derivatives and other structurally related substances were analyzed by CZE in a series of background electrolytes in a wide pH range, 1.86 - 11.18. From the measured pH dependencies of effective electrophoretic mobilities of analytes (beforehand corrected to reference temperature of 25 řC), their isoelectric points (pI) were determined. In addition, using the non-linear regression analysis of the above dependencies, acid-base dissociation constants (pKa) of ionogenic groups of selected analytes were calculated. Subsequently, the analytes with sharply defined isoelectric points were analyzed by CIEF. CIEF confirmed applicability of these compounds as markers of isoelectric points for calibration of pH gradient in CIEF in the determination of pI of amphoteric compounds, especially peptides and proteins. The determined pKa values of ionogenic groups in particular compounds will be utilized in the development of new pI markers with desired pI values.