Made available in DSpace on 2014-06-11T19:26:41Z (GMT). No. of bitstreams: 0 Previous issue date: 2013-02-25Bitstream added on 2014-06-13T18:54:57Z : No. of bitstreams: 1 silva_pss_me_botfca.pdf: 1572244 bytes, checksum: cb40193cc7c83afdae5bc11bdedce824 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Avaliou-se a influência da sazonalidade no rendimento e composição química dos óleos essenciais de Lychnophora pinaster. O material vegetal foi coletado em Carrancas e Ingaí, Minas Gerais, no verão, outono, inverno e primavera de 2012. Foram utilizadas três populações nativas, denominadas Serra Branca, Serra do Salto (provenientes de Carrancas) e Serra da Arnica (Ingaí). Foram amostrados 25 acessos por população e os óleos essenciais das folhas extraídos por hidrodestilação e a análise da composição química conduzida em CG-EM. A sazonalidade não exerceu influência no rendimento médio de óleo essencial das populações Serra Branca e Serra do Salto e destas, Serra do Salto apresentou em todas as estações do ano maior rendimento médio (0,43% a 0,61%). Para Serra da Arnica observou-se estatisticamente influência da sazonalidade no rendimento médio de óleo essencial no inverno (0,79%). Os óleos essenciais das três populações apresentaram as mesmas substâncias, cujas majoritárias foram o trans-cinamato de metila e o trans-cariofileno. Serra Branca apresentou em todas as estações a maior proporção relativa média de trans-cariofileno (16,01% a 23,1%), alfa-humuleno (4,64% a 6,7%), óxido de cariofileno (4,79% a 5,76%) e cedr-8-(15)-en-9-alfa-ol (1,98% a 2,73%). Nesta população, os acessos que mais se correlacionaram com o acúmulo das substâncias com maior proporção relativa média, nas quatro estações foram: Pop 101, Pop 103 e Pop 125 (óxido de cariofileno), Pop 113 e Pop 123 (trans-cariofileno e alfa-humuleno) e Pop 112 e Pop 119 (trans-cinamato de metila). Serra do Salto apresentou a... / The influences of seasonality on the yield and the chemical composition of the essential oils from Lychnophora pinaster were evaluated. The plant material was collected in the regions of Carrancas and Ingaí, State of Minas Gerais, Brazil, during the whole year of 2012. Three native populations were sampled: Serra Branca, Serra do Salto (Carrancas), and Serra da Arnica (Ingaí) – a total sample of 75 plants (25 for each population). The essential oils were extracted by hydro-distillation and the analyses of chemical composition were made by GC-MS. The seasonality did not present a significant effect on the average essential oil yield of the Carrancas populations – Serra do Salto showed the highest values on essential oil production (0.43% to 0.61%). For the population of Serra da Arnica, the seasonality presented significant influence on essential oil yield and the winter sample showed the highest values in this population (0.79%). Chemically all populations presented the same composition and the main substances were trans-methyl cinnamate and trans- caryophyllene. The Serra Branca population presented the highest relative proportion of trans-caryophyllene (16.01% to 23.1%), alpha-humulene (4.64% to 6.7%), caryophyllene oxide (4.79% to 5.76%) and cedr-8-(15)-en-9-alpha-ol (1.98% to 2.73%). In this population the plants with highest correlation with the accumulation of main substances during the whole year were Pop-101, Pop-103... (Complete abstract click electronic access below)

Made available in DSpace on 2015-05-14T16:53:03Z (GMT). No. of bitstreams: 0 Previous issue date: 2014-07-08Bitstream added on 2015-05-14T16:59:30Z : No. of bitstreams: 1 000824410.pdf: 976765 bytes, checksum: 5590643000ccb76d948b31c347eb1bba (MD5) / Devido à importância do bioma cerrado, inclusive para a manutenção de outros biomas interligados, nas últimas décadas pesquisadores das mais variadas áreas vêm propondo alternativas de uso das espécies do cerrado como forma de preservar a fauna e flora local. Uma das formas para auxiliar na preservação dessas espécies esta o estudo sobre constituintes fitoquímicos e propriedades medicinais de plantas desse bioma usadas na medicina tradicional. O objetivo desse estudo é a confirmação do potencial terapêutico bem como da toxicidade dessas plantas usadas na medicina tradicional local, bem como estudar o potencial medicinal de plantas ainda não investigadas. A espécie Dipteryx alata Vogel (baru) é uma das espécies nativas do cerrado que apresenta grande potencial nutricional e farmacológico a ser investigado, pois segundo a medicina tradicional ela é utilizada no combate à bronquite, diarreia, disenteria, dor, dor de garganta, gripe, picada de cobra, tosse e como cicatrizante, entretanto, apenas a atividade contra picada de cobra foi cientificamente comprovada. Desta forma, na tentativa de aliar o valor nutricional da polpa dos frutos do baru a um possível efeito terapêutico, foram realizados estudos sobre os fitoconstituintes presentes nesta parte da planta, a fim de guiar estudos sobre as possíveis propriedades biológicas valorizando o fruto como um alimento nutracêutico, em áreas de ocorrência da planta que apresentam elevados índices de doenças negligenciadas. Os resultados dos estudos fitoquímicos mostraram a presença de alcaloides, taninos, flavonoides, glicosídeos cardiotônicos e saponinas em consideráveis concentrações e antraquinonas em baixa concentração. A presença desses fitoconstituintes forneceu suporte para a realização de ensaios para a determinação do potencial antiparasitário. A atividade antimicrobiana também foi avaliada devido ao uso medicinal da planta para infecções do trato ... / Due to the importance of the cerrado biome, including the maintenance of other interconnected biomes in recent decade’s researchers from different fields have proposed alternative use of the cerrado species as a way to preserve the local fauna and flora. One way to help the preservation of species is the study of the medicinal properties of plants of this biome used in traditional medicine. The objective of these studies is to confirm the therapeutic potential and the toxicity of these plants used in traditional local medicine as well as studying the medicinal potential of plants yet to be investigated. The specie Dipteryx alata Vogel (baru) is a native species of the cerrado with great nutritional and pharmacological potential to be investigated, because according to traditional medicine it is used to combat bronchitis, diarrhea, dysentery, pain, sore throat, flu, snakebite, cough and as cicatrizant, however, only the activity against snakebite has been scientifically proven. Therefore, trying to combine the nutritional value of baru fruits pulp to the potential therapeutic effect, studies on the determination of phytochemicals present were performed to guide studies on possible biological properties valuing the fruit as a nutraceutical food in areas of occurrence of the plant that show high negligible diseases indices. The results of the phytochemical studies showed the presence of alkaloids, tannins, flavonoids, saponins and cardiotonic glycosides in considerable amounts and anthraquinones at low concentration. The presence of these phytochemicals has given support for conducting tests to determine the antiparasitic potential. Antimicrobial activity was also evaluated due to the medicinal use of the plant for the urinary tract infections. In addition, the cytotoxicity of the pulp was also evaluated on LLC- MK 2 cells to support the safe use of the fruit. Antimicrobial assays were performed using the bacterial strains Proteus sp., E. ...

Orientador: André Gonzaga dos Santos / Coorientador: Marcos Antônio Corrêa / Banca: Dulce Helena Siqueira Silva / Banca: Iguatemy Lourenço Brunetti / Resumo: A erva-baleeira, Cordia verbenacea DC., é uma espécie de planta originária do Brasil, encontrada principalmente na Floresta Tropical Atlântica. É largamente utilizada na medicina tradicional por apresentar, principalmente, ação anti-inflamatória. Dados recentes da literatura mostraram que esta ação está relacionada à presença de compostos fenólicos, flavonoides e sesquiterpenos, além de ser constatada a atividade antioxidante em diferentes extratos de C. verbenacea. Epécies reativas de oxigênio e nitrogênio são produzidas constantemente no organismo, podendo causar danos ou benefícios aos sistemas biológicos. Estão altamente relacionadas com o processo de envelhecimento do organismo, e, consequentemente, do envelhecimento cutâneo. A busca e pesquisa de cosméticos com ativos naturais com propriedades antioxidantes tem aumentado significativamente. Este estudo teve como objetivo avaliar a ação do extrato hidroetanólico 70 % de folhas de C. verbenacea (EtOH 70 %) em diferentes espécies reativas de oxigênio de importância biológica, para uma possível aplicação na prevenção do envelhecimento cutâneo. Inicialmente, foram comparados os extratos etanólico, hidroetanólico 70 % e aquoso de folhas de C. verbenacea a fim de selecionar o extrato com maior teor de compostos fenólicos totais e flavonoides totais, além de maior atividade antioxidante. O extrato EtOH 70 % foi selecionado para avaliação da atividade antioxidante in vitro frente a diferentes espécies reativas (radicais modelo ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Erva-baleeira, Cordia verbenacea DC., is a native Brazilian plant species, mainly found at the Atlantic Forest. It is widely used in traditional medicine mostly for its anti-inflammatory properties. Literature data shows that this activity is related to the presence of phenolic compounds, flavonoids and sesquiterpenes, it was also verified the antioxidant activity for different extracts of C. verbenacea. Reactive oxygen and nitrogen species are constantly produced in the organism and might cause damage or benefits on biological systems. They are highly related to the ageing process of the body and, consequently, to skin ageing. The research for natural cosmetics drugs with antioxidant properties has had a significant increase and, therefore, the aim of this study was to evaluate the antioxidant activity of an hydroethanolic extract of C. verbenacea leaves (EtOH 70 %) on different reactive oxygen species of biological importance. Initially, the ethanolic, hydroethanolic (70 %) and aqueous extracts of C. verbenacea leaves were compared to select the extract with a higher total phenolic content and total flavonoid content, as well as higher antioxidant activity. The EtOH 70 % extract was selected for in vitro evaluation of its antioxidant activity against different reactive species (radicals model or bioprocess model involving these species), but also to evaluate its cytotoxicity, the chromatographic profiles in TLC and HPLC and to develop a phytocosmetic contaning the extract, ... (Complete abstract click electronic access below) / Mestre

Orientador: André Gonzaga dos Santos / Coorientador: Christiane Pienna Soares / Banca: Dulce Helena Siqueira Silva / Banca: Denise Crispim Tavares / Resumo: Produtos derivados de fontes vegetais tem demonstrado seu potencial no combate a diversas doenças, inclusive o câncer, à exemplo dos alcaloides derivados de Catharantus roseus (L.)G. Don, vincristina e vimblastina. A diversidade estrutural das moléculas encontradas em espécies vegetais concebem perspectivas de uso in natura bem como servem de inspiração para o desenvolvimento de outras entidades químicas que serão destinadas ao combate de doenças. O material de estudo pertence a espécie Baccharis trimera (Less.) DC, popularmente conhecida como carqueja e com amplo uso tradicional, principalmente para o alívio de afecções digestivas. O estudo teve por objetivo o fracionamento cromatográfico de derivados vegetais de partes aéreas de B. trimera e a avaliação da citotoxicidade e estudo de morte em linhagens de células humanas normal e tumorais tratadas com o OE, suas frações e componentes, além da flavona eupatorina isolada do extrato acetato de etila. O potencial citotóxico foi avaliado através do método da sulforrodamina B e os estudos de morte através de ensaios de Anexina-V, Hoechst/iodeto de propídeo e ativação de caspase-3. As linhagens celulares humanas utilizadas foram de câncer de mama (MCF-7), hepatocarcinoma (HepG2) e a linhagem de mama normal (MCF-10). As amostras avaliadas foram o óleo essencial, suas frações e componentes (α-humuleno, trans-cariofileno e óxido de cariofileno), além da eupatorina. Os constituintes majoritários do óleo essencial identificados por CG-E... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Natural products diversity is an important source to find potential treatments against various diseases, including cancer. Vincristine and vimblastine, alkaloids derived from Catharantus roseus (L.) L. Don, are some of the examples. The structural diversity of molecules from plant species allow the in natura usage and also the development of other chemical entities that will be used to diseases control. Baccharis trimera (Less.) DC., popularly known as carqueja, has an extensive traditional use, especially for digestive disorders. The aims of this study were the chromatographic fractionation of aerial parts from carqueja, in vitro evaluation of the cytotoxic potential through sulforhodamine B assay and cellular death by Annexin-V assay, Hoechst/propidium iodide method and caspase-3 activation, in normal and tumor cell lines treated with essential oil, its fractions and components, and the flavone eupatorin isolated from ethyl acetate extract. The cell lines used were human breast cancer (MCF-7), hepatocellular carcinoma (HepG2) and normal breast line (MCF-10a). The evaluated samples were: essential oil, its fractions and components (α-humulene, β- caryophyllene and caryophyllene oxide) and eupatorin. The major constituents of the essential oil identified by GC-MS were: β-caryophyllene (18.9%), bicyclogermacrene (15.6%), germacrene D (10.5%), δ-cadinene (6.6%). Other components of the essential oil, such as α-humulene (2.3%) and caryophyllene oxide (2.9%) were also identified.... (Complete abstract click electronic access below) / Mestre

Orientador: Marcia Ortiz Mayo Marques / Banca: Elizabeth Orika Ono / Banca: Paulo Roberto Hrihorowitsch Moreno / Resumo: Avaliou-se a influência da sazonalidade no rendimento e composição química dos óleos essenciais de Lychnophora pinaster. O material vegetal foi coletado em Carrancas e Ingaí, Minas Gerais, no verão, outono, inverno e primavera de 2012. Foram utilizadas três populações nativas, denominadas Serra Branca, Serra do Salto (provenientes de Carrancas) e Serra da Arnica (Ingaí). Foram amostrados 25 acessos por população e os óleos essenciais das folhas extraídos por hidrodestilação e a análise da composição química conduzida em CG-EM. A sazonalidade não exerceu influência no rendimento médio de óleo essencial das populações Serra Branca e Serra do Salto e destas, Serra do Salto apresentou em todas as estações do ano maior rendimento médio (0,43% a 0,61%). Para Serra da Arnica observou-se estatisticamente influência da sazonalidade no rendimento médio de óleo essencial no inverno (0,79%). Os óleos essenciais das três populações apresentaram as mesmas substâncias, cujas majoritárias foram o trans-cinamato de metila e o trans-cariofileno. Serra Branca apresentou em todas as estações a maior proporção relativa média de trans-cariofileno (16,01% a 23,1%), alfa-humuleno (4,64% a 6,7%), óxido de cariofileno (4,79% a 5,76%) e cedr-8-(15)-en-9-alfa-ol (1,98% a 2,73%). Nesta população, os acessos que mais se correlacionaram com o acúmulo das substâncias com maior proporção relativa média, nas quatro estações foram: Pop 101, Pop 103 e Pop 125 (óxido de cariofileno), Pop 113 e Pop 123 (trans-cariofileno e alfa-humuleno) e Pop 112 e Pop 119 (trans-cinamato de metila). Serra do Salto apresentou a... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The influences of seasonality on the yield and the chemical composition of the essential oils from Lychnophora pinaster were evaluated. The plant material was collected in the regions of Carrancas and Ingaí, State of Minas Gerais, Brazil, during the whole year of 2012. Three native populations were sampled: Serra Branca, Serra do Salto (Carrancas), and Serra da Arnica (Ingaí) - a total sample of 75 plants (25 for each population). The essential oils were extracted by hydro-distillation and the analyses of chemical composition were made by GC-MS. The seasonality did not present a significant effect on the average essential oil yield of the Carrancas populations - Serra do Salto showed the highest values on essential oil production (0.43% to 0.61%). For the population of Serra da Arnica, the seasonality presented significant influence on essential oil yield and the winter sample showed the highest values in this population (0.79%). Chemically all populations presented the same composition and the main substances were trans-methyl cinnamate and trans- caryophyllene. The Serra Branca population presented the highest relative proportion of trans-caryophyllene (16.01% to 23.1%), alpha-humulene (4.64% to 6.7%), caryophyllene oxide (4.79% to 5.76%) and cedr-8-(15)-en-9-alpha-ol (1.98% to 2.73%). In this population the plants with highest correlation with the accumulation of main substances during the whole year were Pop-101, Pop-103... (Complete abstract click electronic access below) / Mestre

CAPES / O Capim Annoni-2 (Eragrostis plana Nees), considerado uma planta daninha invasora e de difícil erradicação, tem a capacidade de dominar o meio em que se encontra, sobrepondo-se a espécies nativas e agroeconômicas. Parte desta capacidade está relacionada a efeitos alelopáticos causados por aleloquímicos liberados por esta espécie. Assim, a identificação desses aleloquímicos pode ser uma boa alternativa para produção de bio-herbicidas. Neste estudo, determinou-se o potencial alelopático de extratos brutos de éter de petróleo, acetato de etila e metanol provenientes da raíz de E. plana Nees coletadas no verão na área experimental do curso de Agronomia da UTFPR - câmpus Pato Branco, como também um estudo fitoquímico para elucidação estrutural de possíveis aleloquímicos. O potencial alelopático foi avaliado a partir de bioensaios de germinação e crescimento de radícula e hipocótilo, utilizando-se duas espécies receptoras (Ipomoea grandifolia e Euphorbia hetetophylla) e três concentrações dos extratos brutos (200, 400 e 600 ppm). Como resultados observou-se que os extratos brutos de éter de petróleo e acetato de etila atuam principalmente na cinética do processo de germinação, retardando-o, enquanto que o extrato bruto de metanol atuou, sobretudo na diminuição da germinação, inibindo 75,4% em comparação com o branco (água destilada), causando até deformidades e mortes de plântulas, estes dados para a espécie receptora E. heterophylla, a mais afetada. Quanto ao bioensaio de crescimento de hipocótilo e radícula, o extrato bruto de metanol foi o menos eficaz, destacando-se o extrato bruto de éter de petróleo a 600 ppm contra a espécie receptora I. grandifolia, causando uma diminuição no crescimento da radícula e hipocótilo de 66,3% e 65,5%, respectivamente, e o extrato bruto de acetato de etila também a 600 ppm contra a espécie receptora E. heterophylla, causou uma diminuição de 63,6% e 72% do comprimento do hipocótilo e radícula, respectivamente, ambos em comparação com seus respectivos brancos (água destilada). Através do estudo fitoquímico foi proposta a estrutura de um diterpeno, por meio do fracionamento e purificação do extrato bruto de éter de petróleo, e um composto, cuja estrutura não foi ainda completamente elucidada, este a partir do extrato bruto de metanol. Os efeitos alelopáticos apresentados pelas raízes do Capim Annoni-2, foram bastante efetivos, podendo estes serem atribuídos as substâncias fitotóxicas isoladas nesse estudo. / The Grass Annoni-2 (Eragrostis plana Nees), considered a noxious invasive and difficult to eradicate plant has the capacity to dominate the environment they are in, superseding Agrieconomic and native species. Part of this ability is related to allelopathic effects caused by allelochemicals released by this species. Thus, the identification of these allelochemicals can be a good alternative for the production of bio-herbicides. In this study, we determined the allelopathic potential of crude extracts of petroleum ether, ethyl acetate and methanol from the root of E. plana Nees collected in the summer in the experimental area of the course of Agronomy UTFPR - Pato Branco campus, as well as a phytochemical study for structural elucidation of potential allelochemicals. The allelopathic potential was assessed from bioassays of germination and growth of radicle and hypocotyl, using two receptor species (Ipomoea grandifolia and Euphorbia hetetophylla) and three concentrations of crude extracts (200, 400 and 600 ppm). As a result it was found that crude extracts of petroleum ether and ethyl acetate act mainly on the kinetics of the germination process by slowing it, while the crude methanol extract worked, especially in reducing the germination inhibiting 75.4% in comparison with blank (distilled water), causing deformities, and even death of seedlings these data to the recipient E. heterophylla, most affected. Regarding the growth bioassay hypocotyl and radicle, crude methanol extract was the least effective, highlighting the crude extract of petroleum ether at 600 ppm against receiving I. grandifolia species, causing a decrease in the growth of radicle and hypocotyl 66.3% and 65.5%, respectively, and the crude ethyl acetate extract also receiving 600 ppm against E. heterophylla caused a decrease of 63.6% and 72% of the length of the hypocotyl and radicle respectively, both compared with their white (distilled water). Through phytochemical study was proposed structure of a diterpene, through the fractionation and purification of the crude extract of petroleum ether, and a compound whose structure has not been fully elucidated, this from crude methanol extract. The allelopathic effects shown by the roots of the grass Annoni-2, were quite effective, these can be attributed to the isolated phytotoxic substances in this study.

A geografia brasileira e suas dimensões continentais tornam o país apto para cultivo e desenvolvimento de diferentes espécies vegetais, ocupando o ranking de terceiro maior produtor mundial de frutas e o segundo de soja. As tendências globais apontam o crescimento do mercado de bebidas não alcoólicas, em particular a base de frutas e extratos hidrossolúveis de soja. Diante destes fatos, este trabalho buscou a caracterização de polpas e bebidas de pitanga (Eugenia uniflora L.), amora (Morus nigra) e mirtilo (Vaccinium spp.), provenientes de plantações comerciais e do bioma Floresta com Araucária na região sudoeste do Paraná, através de análises físico-químicas, microbiológicas, reológicas, fitoquímicas, antimicrobianas e sensorial. Entre as análises físico-químicas de conteúdo mineral, proteínas, sólidos, lipídios e glicídios, o pH e a relação sólidos solúveis totais/acidez titulável total (SS/ATT) foram os descritores que melhor explicaram a diferença entre as polpas e as bebidas (Análise de Componentes Principais). A prospecção de fitoquímicos nos extratos das polpas dos frutos revelou a presença de diversos metabólitos secundários, principalmente flavonas, flavonóis, xantonas e flavanonóis. Os extratos polares apresentaram maior riqueza desses constituintes. Os extratos foram também analisados por meio de infravermelho, possibilitando a observação de que, o etanol 70% é o solvente mais indicado na pesquisa de antioxidantes, em comparação com o éter de petróleo e o éter etílico, por apresentar maior poder de extração de grupamentos polares, bem como pela presença de picos de absorção de O-H em todas as polpas. Não houve diferença estatística (p  0,05) nas capacidades antioxidantes das polpas, contudo o maior teor de fenólicos totais obtido ocorreu na polpa de mirtilo. Os testes de atividade antimicrobiana indicaram poder fungistático da polpa de mirtilo para a levedura Cândida tropicalis, com concentração inibitória mínima de 833,33 μg mL-1. As curvas de escoamento demonstraram comportamento não newtoniano com características pseudoplásticas, com excelente ajuste ao modelo de Ostwald-de-Waele. O efeito da temperatura sobre a viscosidade aparente das polpas foi verificado através de equação do tipo Arrhenius e analisado em relação à energia de ativação, cujo maior valor foi obtido para a polpa de amora. A análise sensorial das bebidas preparadas apontou para aceitação das mesmas perante os provadores, indicando potencial para a industrialização. / The Brazilian geography and its continental dimensions make the country suitable for cultivation and growth of different plant species, occupying the ranking of third largest world producer of fruits and the second of soybean. Global trends point to the growing market of non-alcoholic beverages, particularly, the ones with fruits and soybean water extracts base. Before these facts, this research sought the characterization of pulps and beverages of pitanga (Eugenia uniflora L.), mulberry (Morus nigra) and blueberry (Vaccinium spp.), from commercial plantations and the biome Araucaria Forest in the southwest of Parana, through the physico-chemical, microbiological, rheological, phytochemical, anti-microbiological and sensory analysis. Among the physico-chemical analysis of mineral content, protein, solids, lipids and carbohydrates, the pH and the relation total soluble solids/ total tritable acidity (SS/ATT) were the descriptor that best explained the difference between pulps and beverages (Principal Component Analysis). The phytochemical prospection in the extract of the fruits’ pulps revealed the presence of several secondary metabolites, mainly flavones, flavonols, xanthones and flavanonols. The polar extract presented higher richness of these constituents. The extracts were also analyzed through infrared, allowing the observation that ethanol 70% is the most suitable solvent in antioxidants search, compared with petroleum ether and ethyl ether, because of its greater potential for polar groups extraction and the presence of O-H absorption peaks in all pulps. There was not statistic differences (p  0,05) in the antioxidants capacities of the pulps, however, the highest total phenolics obtained occurred in the mulberry pulp. The antimicrobiological activity tests indicated fungus power of the blueberry pulp for the Cândida tropicalis yeast, with minimum inhibitory concentration of 833,33 μg mL-1. The flow curves demonstrated a non Newtonian behavior with pseudoplastic characteristics, with excellent adjust to the Ostwald-de-Waele model. The temperature effect on the viscosity of pulps was verified through the equation of Arrhenius sort and analyzed in relation to activation energy, which highest value was obtained for the mulberry pulp. The sensory analysis of the prepared beverages pointed to the acceptability of the tasters, indicating potential to industrialization.

CAPES / O Capim Annoni-2 (Eragrostis plana Nees), considerado uma planta daninha invasora e de difícil erradicação, tem a capacidade de dominar o meio em que se encontra, sobrepondo-se a espécies nativas e agroeconômicas. Parte desta capacidade está relacionada a efeitos alelopáticos causados por aleloquímicos liberados por esta espécie. Assim, a identificação desses aleloquímicos pode ser uma boa alternativa para produção de bio-herbicidas. Neste estudo, determinou-se o potencial alelopático de extratos brutos de éter de petróleo, acetato de etila e metanol provenientes da raíz de E. plana Nees coletadas no verão na área experimental do curso de Agronomia da UTFPR - câmpus Pato Branco, como também um estudo fitoquímico para elucidação estrutural de possíveis aleloquímicos. O potencial alelopático foi avaliado a partir de bioensaios de germinação e crescimento de radícula e hipocótilo, utilizando-se duas espécies receptoras (Ipomoea grandifolia e Euphorbia hetetophylla) e três concentrações dos extratos brutos (200, 400 e 600 ppm). Como resultados observou-se que os extratos brutos de éter de petróleo e acetato de etila atuam principalmente na cinética do processo de germinação, retardando-o, enquanto que o extrato bruto de metanol atuou, sobretudo na diminuição da germinação, inibindo 75,4% em comparação com o branco (água destilada), causando até deformidades e mortes de plântulas, estes dados para a espécie receptora E. heterophylla, a mais afetada. Quanto ao bioensaio de crescimento de hipocótilo e radícula, o extrato bruto de metanol foi o menos eficaz, destacando-se o extrato bruto de éter de petróleo a 600 ppm contra a espécie receptora I. grandifolia, causando uma diminuição no crescimento da radícula e hipocótilo de 66,3% e 65,5%, respectivamente, e o extrato bruto de acetato de etila também a 600 ppm contra a espécie receptora E. heterophylla, causou uma diminuição de 63,6% e 72% do comprimento do hipocótilo e radícula, respectivamente, ambos em comparação com seus respectivos brancos (água destilada). Através do estudo fitoquímico foi proposta a estrutura de um diterpeno, por meio do fracionamento e purificação do extrato bruto de éter de petróleo, e um composto, cuja estrutura não foi ainda completamente elucidada, este a partir do extrato bruto de metanol. Os efeitos alelopáticos apresentados pelas raízes do Capim Annoni-2, foram bastante efetivos, podendo estes serem atribuídos as substâncias fitotóxicas isoladas nesse estudo. / The Grass Annoni-2 (Eragrostis plana Nees), considered a noxious invasive and difficult to eradicate plant has the capacity to dominate the environment they are in, superseding Agrieconomic and native species. Part of this ability is related to allelopathic effects caused by allelochemicals released by this species. Thus, the identification of these allelochemicals can be a good alternative for the production of bio-herbicides. In this study, we determined the allelopathic potential of crude extracts of petroleum ether, ethyl acetate and methanol from the root of E. plana Nees collected in the summer in the experimental area of the course of Agronomy UTFPR - Pato Branco campus, as well as a phytochemical study for structural elucidation of potential allelochemicals. The allelopathic potential was assessed from bioassays of germination and growth of radicle and hypocotyl, using two receptor species (Ipomoea grandifolia and Euphorbia hetetophylla) and three concentrations of crude extracts (200, 400 and 600 ppm). As a result it was found that crude extracts of petroleum ether and ethyl acetate act mainly on the kinetics of the germination process by slowing it, while the crude methanol extract worked, especially in reducing the germination inhibiting 75.4% in comparison with blank (distilled water), causing deformities, and even death of seedlings these data to the recipient E. heterophylla, most affected. Regarding the growth bioassay hypocotyl and radicle, crude methanol extract was the least effective, highlighting the crude extract of petroleum ether at 600 ppm against receiving I. grandifolia species, causing a decrease in the growth of radicle and hypocotyl 66.3% and 65.5%, respectively, and the crude ethyl acetate extract also receiving 600 ppm against E. heterophylla caused a decrease of 63.6% and 72% of the length of the hypocotyl and radicle respectively, both compared with their white (distilled water). Through phytochemical study was proposed structure of a diterpene, through the fractionation and purification of the crude extract of petroleum ether, and a compound whose structure has not been fully elucidated, this from crude methanol extract. The allelopathic effects shown by the roots of the grass Annoni-2, were quite effective, these can be attributed to the isolated phytotoxic substances in this study.

The undeniable efficacy of medicinal plants and wide range of biological activities attributed to plant secondary metabolites are an indication that plants can serve as an excellent pool of bioactive compounds with useful therapeutic properties. The South African flora is recognised as one of the richest centres of plant diversity in the world. From this enormous biodiversity a large number of species has the potential to yield pharmacologically active compounds. C. album is an indigenous plant belonging to the Cape fynbos biome with potentially useful bioactivities. The aim of this study was to evaluate the bioactivity of C. album by screening plant extracts for antibacterial, anti-mycobacterial, antifungal, antioxidant and anti-HIV activity. For rapid and effective screening for the presence of bioactive compounds, a bioassay-guided fractionation methodology was followed. Extracts from plant material obtained from two different geographic regions, the Cape and Highveld, were prepared by liquid extraction in a ratio of 150g fresh plant material per litre solvent, either acetone or ethanol. Qualitative analysis of the crude extracts by TLC and RP-HPLC documented the multi-component plant constituents as a fingerprint, revealing a highly complex, but similar profile of extracted components in both plant groups. Preliminary identification and structural information of the bioactive components present in the active C. album extracts was obtained by a combination of preparative TLC and LC/MS. The development of resistance to all available classes of antibiotic agents, their decreased effectiveness and the re-emergence of previously uncommon infections has necessitated the search for antimicrobial substances with novel antimicrobial mechanisms. The antimicrobial activity, including the antibacterial (Gram-positive and Gram-negative), anti-mycobacterial and antifungal activity of the crude extracts were evaluated. The TLC-bioautographic method used to screen the plant extracts for antimicrobial activity, as well as the localisation of compounds with antibacterial and antifungal activity, indicated the presence of a number of inhibitory compounds with activity against all the microorganisms tested. Evaluation of the inhibitory strength of each extract by the serial microdilution assay indicated that the C. album extracts effectively inhibited all the microorganisms, with the minimum inhibitory concentrations in the low mg/ml range. The significant antimicrobial activity exhibited against all the microorganisms, especially against the Gram-negative bacteria, Mycobacterium tuberculosis and Candida albicans, could suggest the potential use of the extracts or their active constituents as therapeutic agents for the treatment of infectious diseases. The need for natural antioxidants in the health care sector and food industry, due to the role that free radicals play in the pathology of a variety of human diseases and radical-induced deterioration of food products, supported the evaluation of the free radical scavenging activity of C. album extracts against relevant free radical species. The antioxidant activity of the extracts measured using the TLC-DPPH method, revealed the presence of a number of compounds with antioxidant activity. Quantification of the radical scavenging activity by the DPPHspectrophotometric assay revealed that the acetone extracts had a higher radical scavenging activity compared to the ethanol extracts, a pattern that was also found with the fluorescencemicroplate based oxygen radical absorbance assay (ORAC), specific for peroxyl radicals. The observed antioxidant activity were correlated with the total polyphenol content of the crude extracts, determined by the Folin-Ciocalteau procedure, but not with the reducing capacity evaluated by a Fe3 + - Fe2 + reduction method. HIV/AIDS has gained significant interest due to the high mortality rate and the rapid spread of the disease. The appearance of HIV strains resistant to certain antiretroviral drugs, in addition to the high cost, severe metabolic side effects and therapeutic failure of currently available antiretroviral agents, served as motivation for evaluation of C. album for anti-HIV properties and to evaluate potential cytotoxicity of plant extracts in mammalian cell cultures. The effects of the crude extracts on the in vitro HIV-1 subtype C (the predominant HIV-1 form in South Africa) replication and cytopathic effect on CEMnkrCCR5 lymphoid cells were determined. Viability assays using tetrazolium salts and viability dyes allowed the assessment of the host cell responses in the cytotoxicity and anti-HIV screening. Assays were performed at the maximum non-toxic concentration of 50 μg/ml. Some of the plant extracts exhibited significant reduction of the virusinduced cytopathic effect and induced a significant increase in cellular viability. The effect of the extracts on HIV activity was also investigated by determining the viral p24 core protein level, an indication of the replication fitness of the virus; and a significant decrease in p24 antigen level, was found. An attempt to clarify the main active compounds and the structural elements conferring the bioactivity in the analysed systems, revealed the presence of phenolic compounds, primarily coumarins and flavonoids, which are thought to be responsible for the observed antibacterial and antioxidant activities. The results of this study indicate that C. album possess strong bioactivity that warrants further investigation. / Prof. I.A. Dubery Dr. D. Meyer

The Forensic Chemistry Laboratory of Johannesburg (FCL JHB) is tasked with the chemical analysis of a variety of samples to assist in determining the cause of death where unnatural cause is suspected. Some of the samples submitted to the laboratory have a herbal or muti connotation, but a large portion of these cases turn out to have no herbal components present as only pharmaceutical or agricultural products are detected in these samples. This study combined, for the first time, forensic investigation, chemistry and botany to create a unique platform needed for the identification of poisonous plants and their components in forensic exhibits and viscera. The research was focussed on the poisonous plants previously detected at the laboratory, as well as the requests received for the analysis of muti/toxic plant components. The selection of plants included Nicotiana glauca, Datura stramonium / Datura ferox, Callilepis laureola, Boophone disticha / Ammocharis coranica, Abrus precatorius, Ricinus communis, Nerium oleander / Thevetia peruviana and Bowiea volubilis. All these species are known to have caused fatalities, hence their choice. Nicotiana glauca has been implicated in the deaths of at least 15 people since 2001. It was previously detected by GC-MS (EI) in plant exhibits, but could not be detected in a viscera matrix. A selective extraction method for alkaloids was used to extract botanical and viscera samples. Anabasine was successfully detected on the HPLC-MS (EI) system but this detection technique was not considered sensitive enough. A very sensitive HPLC-MS method was developed on the ZMD detector by using electrospray technology. This method outperformed both electron impact detectors (GC and HPLC) and could detect 1ng/ml anabasine with relative ease in full scan mode. Datura stramonium and D. ferox have not been previously positively linked to any human poisoning or death due to exposure to botanically derived products at the FCL JHB. Atropine and scopolamine were successfully ionised in ESI positive mode and could be detected at 10 pg/ml and 100 pg/ml level respectively. The identities of the compounds were confirmed by characteristic ISCID fragmentation patterns. The developed method was successfully applied to a suspected heart attack case. The results proved conclusively that the deceased was given D. ferox seeds as part of his meal and an overdose of atropine and scopolamine contributed to his death. Callilepis laureola is reputed to be one of the more commonly used medicinal plants in South Africa, and although its use has been indicated by the specific mention of a possible nephrotoxin and/or hepatotoxin as causative agent, it has not been detected in any of the forensic chemistry laboratories in South Africa. This was mainly due to the absence of a reliable method for the analysis of the main toxic component of C. laureola, atractyloside, by mass spectrometry. A sensitive and very selective HPLC-ESI-MS method was developed that could detect atractyloside, carboxyatractyloside and their monodesulfated analogues in botanical and viscera matrices. The method was successfully applied to a variety of forensic samples and proved that C. laureola may play an important role in herbal poisonings. In a selection of suspected herbal poisonings where the cause of poisoning was unknown, 30% of the samples tested positive for the presence of atractyloside, carboxyatractyloside or their monodesulfated analogues. The bulbs of Boophone disticha are rich in isoquinoline alkaloids and some of the alkaloids were detected by GC-EI-MS and LC-EI-MS, but the detection of these alkaloids in viscera samples was not successful. A routine method used for the screening for drugs of abuse in forensic samples, were successfully used for the analysis of the bulb extracts of B. disticha and the bulb scales of A. coranica. The chromatographic profile of these two plants appeared very similar at a first glance, but a closer evaluation of the mass spectra highlighted significant differences between the two plants. Six alkaloids from B. disticha were isolated and characterised by LC-MS and NMR and these compounds were detected in suspected herbal poisoning cases. It has been shown that B. disticha is one of the commonly used plants to “clean the system” but frequently results in the death of the patient. Abrus precatorius contains one of the most toxic compounds known to mankind, namely abrin that collectively refers to a group of glycoproteins. The seeds of A. precatorius also contain two indole alkaloids, abrine and hypaphorine. The two alkaloids were fractionated and characterised by LC-MS and NMR. Due to the fact that the instrumentation of the FCL JHB is not suited to the detection of proteins, an LC-ESI-MS method was developed for the detection of the two alkaloids in plant and viscera matrix as markers for A. precatorius. The presence of these two alkaloids was indicated on the TMD system (EI spectra) in a suspected herbal poisoning case. The LC-ESI-MS method was applied to the analysis of the samples and the absence of abrine and hypaphorine were proven in the samples. Ricinus communis is similar to A. precatorius in that it also contains a group of extremely poisonous glycoproteins, collectively refered to as ricin. The analysis of R. communis seeds encountered the same problems as the analysis of A. precatorius seeds, and the analysis was again focused on the detection of the minor piperidine alkaloid ricinine. The LC-ESI-MS method developed for abrine was modified to detect ricinine and functioned well in botanical and viscera matrices. This method will enable the forensic analyst to detect ricinine in very low levels when the presence of ricinoleic acid in samples indicates the use of a R. communis-based product. Nerium oleander is a common decorative garden plant that is used medicinally. The plant is rich in cardenolides with oleandrin the main compound. A reversed-phase chromatographic method with ESI mass spectral detection was developed to separate and detect 11 cardiac glycosides. The compounds were adequately separated to allow unambiguous identification, and displayed very stable cationisation with sodium. An extraction method was developed to extract the cardiac glycosides from the leaves of N. oleander and Thevetia peruviana and was also evaluated in a viscera matrix. The extraction method functioned well and extracted a variety of compounds that produced unique chromatographic fingerprints, allowing for the easy differentiation between the two plants. The method is ideally suited for the detection of oleandrin in high concentrations (full scan mode), low concentrations (selected masses) or trace levels (SIM analysis of ion clusters). The method is able to distinguish between extracts derived from N. oleander and T. peruviana and was able to detect and confirm neriifolin, odoroside and neritaloside in N. oleander leaf extracts. Analysis of forensic case exhibits were also successfully done with this method and performed well with liquid and solid matrices. With the new method oleandrin could be detected at trace levels in viscera samples that did not produce positive results in the past. Bowiea volubilis is widely used as a medicinal plant, but is also an extremely toxic plant. It is freely available at traditional healer markets, and is one of the most highly traded plants on the Durban market. Despite the high usage of the plant, it has not been detected by any of the forensic laboratories in South Africa. Bovoside A, a bufadienolide, is reported to be the main cardiac glycoside in the bulb of B. volubilis. The cardiac glycoside method was successfully applied to the analysis of the bulb extract of B. volubilis and bovoside A was identified as the main bufadienolide present in the bulb. Bovoside A was fractionated and characterised by LC-MS. Four extracts of botanical origin could be successfully distinguished from each other by monitoring the main masses of bovoside A, oleandrin and thevetin A and thevetin B. These marker compounds were well separated from each other and made the identification of the botanical extracts quite easy, and the identity of each extract was confirmed by the mass spectrum of each peak. / Prof. F.R van Heerden