I present a simulation model that should be a useful tool for risk assessment of the impact of insecticide inhibitors of cholinesterase (ChE) applied in irrigated agricultural fields on non-target wildlife. I developed the model as a compartment model based on difference equations (Δt = 1 hour) and programmed with Stella® VII software. Conceptually the model is compartmentalized into six submodels describing the dynamics of (1) insecticide application, (2) insecticide movement into floodable soil, (3) irrigation and rain, (4) insecticide dissolution in water, (5) foraging and insecticide intake from water, and (6) ChE inhibition and recovery. To demonstrate application of the model, I simulate historical, current, and “worst-case” scenarios, that examined the impact of ChE-inhibiting insecticides on white-winged doves (WWDO - Zenaida asiatica) in the Lower Rio Grande Valley of Texas (LRGV), USA. To my knowledge, there are no field data verifying that the cause of ChE deprivation in WWDO is due to the ingestion of ChE-inhibiting insecticide residues dissolved in drinking water. I parameterized the model to represent a system composed of fields of cotton, sorghum, corn, citrus, and brushland that encompasses the activity range of a WWDO in the LRGV. I simulated situations representing the typical scenario of WWDO using irrigated crop fields in the absence and in the presence of rain. I also simulated “worst case” scenarios in which methyl parathion was applied at high rates and high frequency. Based on results of the simulations, I conclude that it is unlikely that WWDO are seriously exposed to ChE-inhibiting insecticides by drinking contaminated water. Only in rare cases, for example, when a rain event occurs just after the application of insecticides, are levels of ChE inhibition likely to approach diagnostic levels (20 %). The present simulation model should be a useful tool to predict the effect of ChE-inhibiting insecticides on the ChE activity of different species that drink contaminated water from irrigated agricultural fields. It should be particularly useful in identifying specific situations in which the juxtaposition of environmental conditions and management schemes could result in a high risk to non-target wildlife.

I present a simulation model that should be a useful tool for risk assessment of the impact of insecticide inhibitors of cholinesterase (ChE) applied in irrigated agricultural fields on non-target wildlife. I developed the model as a compartment model based on difference equations (Δt = 1 hour) and programmed with Stella® VII software. Conceptually the model is compartmentalized into six submodels describing the dynamics of (1) insecticide application, (2) insecticide movement into floodable soil, (3) irrigation and rain, (4) insecticide dissolution in water, (5) foraging and insecticide intake from water, and (6) ChE inhibition and recovery. To demonstrate application of the model, I simulate historical, current, and “worst-case” scenarios, that examined the impact of ChE-inhibiting insecticides on white-winged doves (WWDO - Zenaida asiatica) in the Lower Rio Grande Valley of Texas (LRGV), USA. To my knowledge, there are no field data verifying that the cause of ChE deprivation in WWDO is due to the ingestion of ChE-inhibiting insecticide residues dissolved in drinking water. I parameterized the model to represent a system composed of fields of cotton, sorghum, corn, citrus, and brushland that encompasses the activity range of a WWDO in the LRGV. I simulated situations representing the typical scenario of WWDO using irrigated crop fields in the absence and in the presence of rain. I also simulated “worst case” scenarios in which methyl parathion was applied at high rates and high frequency. Based on results of the simulations, I conclude that it is unlikely that WWDO are seriously exposed to ChE-inhibiting insecticides by drinking contaminated water. Only in rare cases, for example, when a rain event occurs just after the application of insecticides, are levels of ChE inhibition likely to approach diagnostic levels (20 %). The present simulation model should be a useful tool to predict the effect of ChE-inhibiting insecticides on the ChE activity of different species that drink contaminated water from irrigated agricultural fields. It should be particularly useful in identifying specific situations in which the juxtaposition of environmental conditions and management schemes could result in a high risk to non-target wildlife.

No disponible a texto completo / Tesis para optar al grado de Magíster en Finanzas / Los ADRs permitieron en la pasado ampliar la opciones de inversión, al poder invertir en paises extranjeros donde existian barreras de entrada. Usando datos de ADRs, investigamos en diferentes paises que ocurria con el precio de estos activos cuando ocurría un shock financiero, como lo es una devaluación o la devaluación en un pais vecino con estrechos vinculos economicos. Tomando datos de Argentina, Brasil, México, Rusia y algunos paises de la crisis asiática, usamos el metodo de los eventos para determinar si existian retornos anormales positivos o negativos para una cartera de ADRs de cada pais. Los resultados fueron mixtos, porque en paises de economías solidas y desarrolladas, hubo mayormente retornos positivos, mientras que en economías emergentes y con mayor riesgo pais, hubo por lo general retornos negativos. Tambien se encontró un efecto contagio en la crisis asiatica.

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No. of bitstreams: 2 Dissertação - Priscila Bianca Rodrigues da Rocha - 2016.pdf: 3342071 bytes, checksum: 4810677b5ff246d6d0dd384d773187fd (MD5) license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Previous issue date: 2015-03-31 / Fundação de Amparo à Pesquisa do Estado de Goiás - FAPEG / The extract of Centella asiatica L. Urban (CA) plant has been used both for cosmetic purposes and for the treatment of cutaneous diseases, due to its cicatrizing and antiproliferative activities. The main active component of this plant, asiaticoside (AS), has an important effect on scleroderma (ES), a disease that affects the skin causing its thickening and hardening. These characteristics make difficult the localized topical treatment of ES. Thus, nanostructured lipid carriers (NLC) containing the CA extract were developed and characterized for the topical application of AS. The NLC were produced with a 1: 3 mixture of solid and liquid lipids (stearic acid and oleic acid, respectively), 0.25% sodium taurodeoxycholate and 1% soy lecithin by the microemulsion technique. For formulations with higher amounts of lipid, concentrations of 2 to 4% of the lipid mixture were evaluated. After the development of high lipid particles, different concentrations of CA dry extract (0.926, 1.274 and 1.853%) and glycolic extract (1.282, 2.418 and 3.336%) were added to the formulations. All NLC were characterized for mean diameter, polydispersity index (PdI), zeta potential, encapsulation efficiency (EE%) and drug loading (DL%). In vitro release and permeation studies of the unencapsulated extract (aqueous dispersions and commercial cream) and encapsulated were evaluated using Franz cells. The NLC obtained had a mean size of 130 nm and a uniform size distribution, with a PdI of 0.2, and an encapsulation efficiency of more than 90% for NLC containing dry extract and 40% for NLC developed with glycolic extract. The AS release profile from the developed formulations showed a controlled release, no more than 45% in 48 hours of study. Encapsulation of the dry extract in NLC resulted in an increase in AS retention in the stratum corneum 2-fold compared to the unencapsulated extract in aqueous dispersion, but was similar to the retained amount of AS when the commercial formulation was applied topically. NLC developed with glycolic extract were able to increase the permeation of AS to the deeper layers of the skin, when compared to the carriers containing the CA dry extract and the commercial cream. The results suggest that these systems significantly increase the permeation of AS and, therefore, may present potential benefits for the treatment of cutaneous diseases such as ES. / O extrato da planta Centella asiatica L. Urban (CA) tem sido utilizado tanto para fins cosméticos quanto para tratamento de doenças cutâneas, devido as suas atividades cicatrizantes e antiproliferativas. O principal componente ativo desta planta, o asiaticosídeo (AS), tem um importante efeito na esclerodermia (ES), doença que acomete a pele causando o seu espessamento e endurecimento. Estas manifestações dificultam o tratamento tópico localizado da ES. Assim, carreadores lipídicos nanoestruturados (CLN) contendo o extrato da planta CA foram desenvolvidos e caracterizados visando a aplicação tópica do AS. Os CLN foram produzidos com uma mistura de 1:3 de lipídeos sólido e líquido (ácido esteárico e ácido oleico, respectivamente), 0,25% de taurodeoxicolato de sódio e 1% de lecitina de soja pela técnica da microemulsão. Para produção de formulações com maior quantidade de lipídeo, concentrações de 2 a 4% da mistura de lipídeos foram avaliadas. Após o desenvolvimento das partículas com alto teor lipídico, diferentes concentrações do extrato seco de CA (0,926, 1,274 e 1,853%) e do extrato glicólico (1,282, 2,418 e 3,336%) foram adicionadas às formulações. Todos os CLN foram caracterizados quanto ao diâmetro médio, índice de polidispersão (PdI), potencial zeta, eficiência de encapsulação (EE%) e carga de fármaco (CF%). Estudos de liberação e permeação in vitro do extrato não encapsulado (dispersões aquosas e creme comercial) e encapsulado foram avaliados utilizando células de “Franz”. Os CLN obtidos apresentaram tamanho médio de 130 nm e distribuição de tamanho uniforme, com PdI de 0,2, além valores de eficiência de encapsulação superiores a 90% para os CLN contendo extrato seco e 40% para os CLN desenvolvidos com extrato glicólico. O perfil de liberação do AS a partir das formulações desenvolvidas mostrou uma liberação controlada, não mais que 45% em 48 horas de estudo. A encapsulação do extrato seco nos CLN resultou em aumento na retenção de AS no estrato córneo de 2 vezes, comparado ao extrato não encapsulado em dispersão aquosa, mas foi semelhante a quantidade retida de AS quando a formulação comercial foi aplicada topicamente. Os CLN desenvolvidos com extrato glicólico foram capazes de aumentar a permeação do AS para as camadas mais profundas da pele, em relação aos carreadores contendo o extrato seco da CA e o creme comercial. Os resultados sugerem que estes sistemas aumentam significativamente a permeação do AS e, por isso, podem apresentar potenciais benefícios para o tratamento de patologias cutâneas como a ES.

Ce mémoire de thèse confidentiel porte sur la valorisation phytochimique et biologique de plantes tropicales. Il s’inscrit dans le cadre d’une procédure de validation des acquis de l’expérience et retrace 14 ans de travaux de recherche appliquée pour la mise sur le marché de nouveaux actifs végétaux destinés à l’industrie dermo cosmétique ainsi que des recherches fondamentales. Ce travail a permis le développement de nouveaux actifs, à forte valeur ajoutée, issus de plantes de Madagascar par la valorisation de nombreux métabolites secondaires tels que les polyphénols (flavonoïdes, tannins, dérivés d’acide chlorogénique) et les triterpènes pentacycliques. Cette activité centrale de recherche s’accompagne d’activités essentielles liées à la maitrise de la ressource végétale en lien avec les actions de développement durable, de la recherche phytochimique, du développement de procédé et du positionnement biologique. Il est illustré entre autre le développement d’extraits issus de Buddleja axillaris, Vernonia appendiculata et les derniers travaux sur la plante Centella asiatica. / This confidential memory thesis focuses on phytochemical and biological enhancement of tropical plants. It is part of a process of validation of experience and retraces 14 years of applied research for the development of new plant extract for cosmetics industry and elemental research.This work enabled the development of new actives, with high value, from Madagascar plants by enhancement of secondary metabolites such as polyphenols (flavonoids, tannins, chlorogenic acid derivatives) and pentacyclic triterpenes. This important activity goes along with essential activities related to the mastery of plant resources in line with the sustainable development actions, phytochemical research, process development and biological positioning. It is illustrated, among other developing, extracts from Buddleja axillaris, Vernonia appendiculata and the latest work on the plant Centella asiatica.

Saponinas triterpênicas incluem uma ampla variedade de moléculas com diversas aplicações farmacológicas. As saponinas das folhas de Quillaja brasiliensis, espécie nativa do Sul do Brasil, são estrutural e funcionalmente semelhantes às encontradas nas cascas da espécie chilena Quillaja saponaria, as quais são utilizadas atualmente como adjuvantes em formulações comerciais de vacinas. O perfil de acúmulo da fração imunoadjuvante de saponinas triterpênicas de Q. brasiliensis (QB-90) foi avaliado em resposta a diferentes fatores de estresse. O conteúdo de QB-90 em discos foliares aumentou significativamente com a aplicação de diferentes agentes de estresse osmótico. Também pôde ser observado aumento nos teores de saponinas pela exposição de discos foliares a ácido salicílico, ácido jasmônico, ultrassom e luz ultravioleta C. Experimentos com plântulas indicaram um aumento significativo nos teores de QB-90 com o aumento moderado de irradiância luminosa e pela aplicação de dano mecânico nas folhas. Os resultados obtidos apoiam o papel de defesa induzida que estes metabólitos apresentam in planta. Com o objetivo de avançar os estudos de atividade adjuvante de Q. brasiliensis e ampliar a extensão de antígenos testados, uma vacina contra poliovírus foi avaliada em camundongos. Resultados demonstraram que as formulações de vacinas contendo saponinas da espécie brasileira aumentaram níveis de anticorpos específicos IgG, IgG1 e IgG2a, sugerindo um estímulo de ambos tipos de resposta imune (Th1 e Th2). A resposta celular foi confirmada por ensaios de DTH e pela avaliação dos níveis de mRNA das citocinas IL-2 e IFN- em esplenócitos murinos. A resposta imune de mucosas foi demonstrada pelos aumentos nos níveis de IgA em bile, fezes e lavados vaginais. O segundo objeto de estudo desta Tese é a espécie Centella asiatica (L.) Urban, que também apresenta saponinas triterpênicas bioativas em suas partes aéreas. Uma enzima responsável pela glicosilação de ácido asiático e ácido madecássico foi parcialmente purificada. Usando técnicas de proteômica e dados de sequências de cDNA, a sequência completa do clone que codifica uma glicosiltransferase foi obtida. O produto gênico recombinante, UGT73AD1, foi funcionalmente expresso em Escherichia coli, purificado por cromatografia de afinidade por metal imobilizado e funcionalmente caracterizado. UGT73AD1 foi identificada como sendo uma glicosiltransferase triterpênica de ácidos carboxílicos, provavelmente envolvida na biossíntese de monodesmosídeos da espécie. / Triterpene saponins include a large variety of molecules that find several applications in pharmacology. The saponins from leaves of Quillaja brasiliensis, a native species from Southern Brazil, show structural and functional similarities to those of Quillaja saponaria barks, which are currently used as adjuvants in vaccine formulations. The accumulation patterns of an immunoadjuvant fraction of leaf triterpene saponins (QB-90) in response to stress factors were examined. The content of QB-90 in leaf disks was significantly increased by application of different osmotic stress agents. Higher yields of bioactive saponins were also observed upon exposure to salicylic acid, jasmonic acid, ultrasound and UV-C light. Experiments with shoots indicated a significant increase in QB-90 yields with moderate increases in white light irradiance and by mechanical damage applied to leaves. These results support a general induced defense role for these metabolites in planta. Aiming at advancing the studies of Q. brasiliensis adjuvant activity and amplifying the range of antigens tested, an inactivated virus-based vaccine against poliovirus was evaluated in mice. Results demonstrated that saponin formulations were able to enhance poliovirus- specific total IgG, IgG1 and IgG2a, suggesting stimulation of both Th1 and Th2 immune responses. Cellular response stimulation was confirmed by DTH assays and by evaluating the enhancement of IL-2 and IFN-γ cytokine mRNA expression in mice splenocytes. Mucosal immune responses demonstrated that the QB-90-adjuvanted vaccine enhanced IgA titers in bile, feces and vaginal washings. Centella asiatica (L.) Urban, a species accumulating triterpene bioactive saponins in its shoots, was also subject of study in this Thesis. An enzyme capable of glucosylating asiatic and madecassic acids was partially purified. Using proteomic methods and cDNA sequence data, a full-length cDNA clone encoding the glucosyltransferase was obtained. The recombinant gene product, UGT73AD1, was functionally expressed in Escherichia coli, purified by immobilized metal-affinity chromatography and functionally characterized. UGT73AD1 was identified as a triterpene carboxylic acid glucosyltransferase, likely involved in monodesmoside biosynthesis in C. asiatica.

Saponinas triterpênicas incluem uma ampla variedade de moléculas com diversas aplicações farmacológicas. As saponinas das folhas de Quillaja brasiliensis, espécie nativa do Sul do Brasil, são estrutural e funcionalmente semelhantes às encontradas nas cascas da espécie chilena Quillaja saponaria, as quais são utilizadas atualmente como adjuvantes em formulações comerciais de vacinas. O perfil de acúmulo da fração imunoadjuvante de saponinas triterpênicas de Q. brasiliensis (QB-90) foi avaliado em resposta a diferentes fatores de estresse. O conteúdo de QB-90 em discos foliares aumentou significativamente com a aplicação de diferentes agentes de estresse osmótico. Também pôde ser observado aumento nos teores de saponinas pela exposição de discos foliares a ácido salicílico, ácido jasmônico, ultrassom e luz ultravioleta C. Experimentos com plântulas indicaram um aumento significativo nos teores de QB-90 com o aumento moderado de irradiância luminosa e pela aplicação de dano mecânico nas folhas. Os resultados obtidos apoiam o papel de defesa induzida que estes metabólitos apresentam in planta. Com o objetivo de avançar os estudos de atividade adjuvante de Q. brasiliensis e ampliar a extensão de antígenos testados, uma vacina contra poliovírus foi avaliada em camundongos. Resultados demonstraram que as formulações de vacinas contendo saponinas da espécie brasileira aumentaram níveis de anticorpos específicos IgG, IgG1 e IgG2a, sugerindo um estímulo de ambos tipos de resposta imune (Th1 e Th2). A resposta celular foi confirmada por ensaios de DTH e pela avaliação dos níveis de mRNA das citocinas IL-2 e IFN- em esplenócitos murinos. A resposta imune de mucosas foi demonstrada pelos aumentos nos níveis de IgA em bile, fezes e lavados vaginais. O segundo objeto de estudo desta Tese é a espécie Centella asiatica (L.) Urban, que também apresenta saponinas triterpênicas bioativas em suas partes aéreas. Uma enzima responsável pela glicosilação de ácido asiático e ácido madecássico foi parcialmente purificada. Usando técnicas de proteômica e dados de sequências de cDNA, a sequência completa do clone que codifica uma glicosiltransferase foi obtida. O produto gênico recombinante, UGT73AD1, foi funcionalmente expresso em Escherichia coli, purificado por cromatografia de afinidade por metal imobilizado e funcionalmente caracterizado. UGT73AD1 foi identificada como sendo uma glicosiltransferase triterpênica de ácidos carboxílicos, provavelmente envolvida na biossíntese de monodesmosídeos da espécie. / Triterpene saponins include a large variety of molecules that find several applications in pharmacology. The saponins from leaves of Quillaja brasiliensis, a native species from Southern Brazil, show structural and functional similarities to those of Quillaja saponaria barks, which are currently used as adjuvants in vaccine formulations. The accumulation patterns of an immunoadjuvant fraction of leaf triterpene saponins (QB-90) in response to stress factors were examined. The content of QB-90 in leaf disks was significantly increased by application of different osmotic stress agents. Higher yields of bioactive saponins were also observed upon exposure to salicylic acid, jasmonic acid, ultrasound and UV-C light. Experiments with shoots indicated a significant increase in QB-90 yields with moderate increases in white light irradiance and by mechanical damage applied to leaves. These results support a general induced defense role for these metabolites in planta. Aiming at advancing the studies of Q. brasiliensis adjuvant activity and amplifying the range of antigens tested, an inactivated virus-based vaccine against poliovirus was evaluated in mice. Results demonstrated that saponin formulations were able to enhance poliovirus- specific total IgG, IgG1 and IgG2a, suggesting stimulation of both Th1 and Th2 immune responses. Cellular response stimulation was confirmed by DTH assays and by evaluating the enhancement of IL-2 and IFN-γ cytokine mRNA expression in mice splenocytes. Mucosal immune responses demonstrated that the QB-90-adjuvanted vaccine enhanced IgA titers in bile, feces and vaginal washings. Centella asiatica (L.) Urban, a species accumulating triterpene bioactive saponins in its shoots, was also subject of study in this Thesis. An enzyme capable of glucosylating asiatic and madecassic acids was partially purified. Using proteomic methods and cDNA sequence data, a full-length cDNA clone encoding the glucosyltransferase was obtained. The recombinant gene product, UGT73AD1, was functionally expressed in Escherichia coli, purified by immobilized metal-affinity chromatography and functionally characterized. UGT73AD1 was identified as a triterpene carboxylic acid glucosyltransferase, likely involved in monodesmoside biosynthesis in C. asiatica.

Saponinas triterpênicas incluem uma ampla variedade de moléculas com diversas aplicações farmacológicas. As saponinas das folhas de Quillaja brasiliensis, espécie nativa do Sul do Brasil, são estrutural e funcionalmente semelhantes às encontradas nas cascas da espécie chilena Quillaja saponaria, as quais são utilizadas atualmente como adjuvantes em formulações comerciais de vacinas. O perfil de acúmulo da fração imunoadjuvante de saponinas triterpênicas de Q. brasiliensis (QB-90) foi avaliado em resposta a diferentes fatores de estresse. O conteúdo de QB-90 em discos foliares aumentou significativamente com a aplicação de diferentes agentes de estresse osmótico. Também pôde ser observado aumento nos teores de saponinas pela exposição de discos foliares a ácido salicílico, ácido jasmônico, ultrassom e luz ultravioleta C. Experimentos com plântulas indicaram um aumento significativo nos teores de QB-90 com o aumento moderado de irradiância luminosa e pela aplicação de dano mecânico nas folhas. Os resultados obtidos apoiam o papel de defesa induzida que estes metabólitos apresentam in planta. Com o objetivo de avançar os estudos de atividade adjuvante de Q. brasiliensis e ampliar a extensão de antígenos testados, uma vacina contra poliovírus foi avaliada em camundongos. Resultados demonstraram que as formulações de vacinas contendo saponinas da espécie brasileira aumentaram níveis de anticorpos específicos IgG, IgG1 e IgG2a, sugerindo um estímulo de ambos tipos de resposta imune (Th1 e Th2). A resposta celular foi confirmada por ensaios de DTH e pela avaliação dos níveis de mRNA das citocinas IL-2 e IFN- em esplenócitos murinos. A resposta imune de mucosas foi demonstrada pelos aumentos nos níveis de IgA em bile, fezes e lavados vaginais. O segundo objeto de estudo desta Tese é a espécie Centella asiatica (L.) Urban, que também apresenta saponinas triterpênicas bioativas em suas partes aéreas. Uma enzima responsável pela glicosilação de ácido asiático e ácido madecássico foi parcialmente purificada. Usando técnicas de proteômica e dados de sequências de cDNA, a sequência completa do clone que codifica uma glicosiltransferase foi obtida. O produto gênico recombinante, UGT73AD1, foi funcionalmente expresso em Escherichia coli, purificado por cromatografia de afinidade por metal imobilizado e funcionalmente caracterizado. UGT73AD1 foi identificada como sendo uma glicosiltransferase triterpênica de ácidos carboxílicos, provavelmente envolvida na biossíntese de monodesmosídeos da espécie. / Triterpene saponins include a large variety of molecules that find several applications in pharmacology. The saponins from leaves of Quillaja brasiliensis, a native species from Southern Brazil, show structural and functional similarities to those of Quillaja saponaria barks, which are currently used as adjuvants in vaccine formulations. The accumulation patterns of an immunoadjuvant fraction of leaf triterpene saponins (QB-90) in response to stress factors were examined. The content of QB-90 in leaf disks was significantly increased by application of different osmotic stress agents. Higher yields of bioactive saponins were also observed upon exposure to salicylic acid, jasmonic acid, ultrasound and UV-C light. Experiments with shoots indicated a significant increase in QB-90 yields with moderate increases in white light irradiance and by mechanical damage applied to leaves. These results support a general induced defense role for these metabolites in planta. Aiming at advancing the studies of Q. brasiliensis adjuvant activity and amplifying the range of antigens tested, an inactivated virus-based vaccine against poliovirus was evaluated in mice. Results demonstrated that saponin formulations were able to enhance poliovirus- specific total IgG, IgG1 and IgG2a, suggesting stimulation of both Th1 and Th2 immune responses. Cellular response stimulation was confirmed by DTH assays and by evaluating the enhancement of IL-2 and IFN-γ cytokine mRNA expression in mice splenocytes. Mucosal immune responses demonstrated that the QB-90-adjuvanted vaccine enhanced IgA titers in bile, feces and vaginal washings. Centella asiatica (L.) Urban, a species accumulating triterpene bioactive saponins in its shoots, was also subject of study in this Thesis. An enzyme capable of glucosylating asiatic and madecassic acids was partially purified. Using proteomic methods and cDNA sequence data, a full-length cDNA clone encoding the glucosyltransferase was obtained. The recombinant gene product, UGT73AD1, was functionally expressed in Escherichia coli, purified by immobilized metal-affinity chromatography and functionally characterized. UGT73AD1 was identified as a triterpene carboxylic acid glucosyltransferase, likely involved in monodesmoside biosynthesis in C. asiatica.

In recent years, the use of natural medicinal properties of plants has again come to the forefront of interest of the public. Because of their therapeutic properties, these plants are commonly used in alternative treatments for minor ailments. This thesis deals with the determination of polyphenolic acids in extracts from aronia, echinacea, tetradenia and centella. One of the aims of the thesis was to compare the amount of polyphenolic acids in individual plants with their antioxidant properties. The theoretical part includes a general introduction to flavonoids, introduction to polyphenolic acids, introduction to the principles of capillary zone electrophoresis and a summary of selected foreign studies for the determination of flavonoids. An important chapter of the theoretical part is a description of individual plants with an explanation of their possible therapeutic use. The experimental part is devoted to the process of optimization of electrophoretic determination of selected polyphenolic acids and evaluation of their content. It was found that in the extracts six representatives of polyphenolic acids occur in minimal amounts, often below the limit of quantification or detection of the method.

No / To analyze the fatty acid contents of Buddleja asiatica Lour, both the non-volatile oil and fat obtained from the n-hexane soluble sub-fraction were subjected to GC/MS using BSTFA (N,O-bis(trimethylsilyl) trifloroacetamide) derivatization. The oil showed the presence of six fatty acids including palmitic acid (46.75 %), linoleic acid (37.80 %), stearic acid (10.98 %), arachidic acid, margaric acid and lignoceric acid (< 3 %). Analysis of the fat revealed nine fatty acids including lignoceric acid (43.12 %), behenic acid (26.39 %), arachidic acid (9.29 %) and stearic acid (5.3 %). Cerotic acid, montanic acid, melissic acid and palmitic acid were found in low amounts (< 5 %) while trycosylic acid (4.83 %) was the only fatty acid with odd number of carbon atoms. The oil showed a low thermal stability. / Higher Education Commission Pakistan