Plants depend on a wide variety of vectors, both biotic and abiotic, to move their seeds to locations away from parent plants. The stages between seed production and seedling establishment in the plant life cycle are under the influence of a complex set of ecological and evolutionary factors (Wang and Smith 2002). In ecological time scales, seed dispersal has been shown to influence both plant population and community dynamics (Howe and Miriti 2004, Seidler and Plotkin 2006). The evolutionary effects are seen in the way these interactions influence the fruiting season, fruit crop size, fruit size, nutritive value of fruit or seed and other strategies adopted by the plant to ensure effective dispersal of their seeds (Howe and Smallwood 1982, Schupp 1993). Most seed dispersal research has focussed on arboreal frugivores such as birds, primates and bats. In comparison, terrestrial frugivores such as ruminants, elephants and rodents are poorly researched. Large-bodied terrestrial frugivores are key to the dispersal of several large-seeded plants and are also capable of dispersing seeds to long distances (> 1 km). Given that large mammals have witnessed tremendous declines in their abundances and ranges in historical times, plants that depend upon them for long distance dispersal might have limited ability to move across fragmented landscapes under changing climates. However, very little is understood about seed dispersal by large mammals or about seed dispersal characteristics of tropical dry forests which support among the highest biomass of large terrestrial mammals. Tropical dry forests are among the most endangered tropical ecosystems (Janzen 1988). It has also been forecast that tropical dry forests in the Indian subcontinent will witness significant changes in temperature and rainfall regimes in the coming decades (Ravindranath et al. 2006). However, compared to tropical moist forests which have received most of the scientific interest, theory, methodology and base line information about seed dispersal in dry forests is poorly developed. To address some of these lacunae, I examined seed dispersal at both population and community-scales in the tropical dry forests of Mudumalai Wildlife Sanctuary from 2005 -2007. Mudumlai is located within a relatively large contiguous stretch of tropical dry forests in southern India and has an intact and abundant frugivore assemblage. In addition, the Mudumalai Forest Dynamics Plot (MFDP), a large 50 hectare plot has been monitored for recruitment, mortality and diameter growth of woody plants for the last twenty years. The plot is an invaluable resource for seed dispersal studies since it provides us an opportunity to link seed dispersal with plant population and community dynamics. In the first part of the thesis (Chapters 3 and 4), I focus on evaluating the effectiveness of ruminants as seed dispersers and then examine factors that influence frugivory by ruminants. This work was carried out at the population level, for the tree Phyllanthus emblica. The fruits of P. emblica are important non-timber forest produce from Asian dry tropics and understanding the quantity of fruit consumed by frugivores has implications for sustainable harvest practices of this resource (Shahabuddin and Prasad 2004). In the second part of the thesis (Chapters 5 and 6), I characterized seed dispersal at the community-scale for woody plants on the MFDP. Community-wide characterization of seed dispersal at sites harboring an intact faunal assemblage is critical to identify plants dependent on endangered dispersers. The second half of the thesis focuses on examining the associations between dispersal mode and fruit characteristics, and also between dispersal mode and spatial patterning of woody plants. Evaluating the effectiveness of ruminants as seed dispersers: Disperser effectiveness is defined as the contribution a disperser makes to the future reproduction of a plant. Evaluating disperser effectiveness involves addressing both quantitative aspects such as frequency of visits, quantity of fruit consumed, diversity of species dispersed, and, qualitative components such as the treatment given in mouth and gut and the suitability of seed deposition sites for germination (Schupp 1993, Dennis and Westcott 2006). Information on quantity and quality of seed dispersal services provided by ruminants is very limited. This is because techniques such as tree watches or fruit traps adapted from avian frugivore studies are ill suited to study terrestrial frugivores, and conventional camera traps provide little quantitative information on the quantity of fruit consumed by frugivores. The quantity of dispersal services provided by ruminants for P. emblica was assessed using a novel camera trap technique. Time-delay was programmed into the camera trap units (4 pictures taken 2 minutes apart) to tell apart frugivores from visitors (by comparing number of fruit remaining in the time-delay picture sequence). During a two year study using this technique, six terrestrial mammals were identified as frugivores of P. emblica. Additionally, seven mammals and one bird species visited fruiting trees but did not consume fallen fruit. Two ruminants, the Indian chevrotain Moschiola indica and chital Axis axis, were the most frequent frugivores of P. emblica and these ruminants accounted for over 95% of fruit removal, while murid rodents accounted for less than 1%. The quality of dispersal services provided by ruminants was examined using a combination of feeding trials (with captive animals) and germination experiments for large-seeded species dispersed by ruminants in tropical dry forests, including P. emblica. Ruminants regurgitated well-protected seeds larger than 5 mm, after retaining them in the rumen for 2–50 h. Though germination success was lowered after rumen retention, it was not too low (> 15%), and would still ensure dispersal if suitable micro-habitats could be found. Given the daily home-range of chital (14–20 ha in tropical dry forests (Mishra 1982)), and their seed retention times (2–50 h), movement of chital during the 1–2 d period when seeds remain in the rumen can potentially disperse seeds to distances over 1 km. Thus, our research shows that ruminants account for most of the fruit removal and are the principal primary dispersers of P. emblica. Factors influencing frugivory by ruminants: Fruit removal, the quantity of fruits consumed by frugivores, is highly variable within plant populations. Understanding factors influencing this selection operated by frugivores helps link frugivore behavior with plant demography. The outcome of such selection operated by frugivores has been examined mostly for interactions involving avian frugivores. The extension of this theory to terrestrial frugivores, which are presented with fruit on the ground, is not well understood. In addition, factors influencing fruit removal have been examined mostly with respect to changing spatial contexts such as crop size and fruiting neighborhoods. The influence of varying temporal contexts on frugivore choice, and their possible demographic and evolutionary consequences is poorly understood. We examined if temporal variation in fruit availability across a hierarchy of nested temporal levels (interannual, intraseasonal, 120 h, 24 h) altered fruit removal of P. emblica on the MFDP. The interactions between P. emblica and its primary disperser (ruminants) was mediated by another frugivore (a primate, langur), which made large quantities of fruit available on the ground to ruminants. The direction and strength of crop size and neighborhood effects on this interaction varied with changing temporal contexts. Fruit availability was higher in the first of the two study years, and at the start of the season in both years. Fruit persistence on trees, determined by primate foraging, was influenced by crop size and conspecific neighborhood densities only in the high fruit availability year. Fruit removal by ruminants was influenced by crop size in both years and neighborhood densities only in the high availability year. In both years, these effects were stronger at the start of the season. Intraseasonal reduction in fruit availability diminished inequalities in fruit removal by ruminants and the influence of crop size and fruiting neighborhoods. Temporal asymmetry in frugivore-mediated selection could reduce the potential for co-evolution between frugivores and plants by diluting selective pressures. Langurs determined the length of time fruit persisted on individual trees, and, also influenced the quantity and duration of fruit availability for the primary dispersers, ruminants. Such inter-dependencies formed between disparate animal consumers add additional levels of complexity to plant–frugivore mutualistic networks and can have potential reproductive consequences for specific individuals within populations. Seed dispersal within a tropical dry forest community: Megafauna are considered to be important seed dispersers, but their contribution has mostly been examined in isolation, with limited reference to other frugivores within communities. Community-wide characterization of seed dispersal at sites harboring an intact fauna is essential to evaluate the role of megafauna in dispersal, examine dispersal syndromes and assess the vulnerability of plant communities to disperser declines and climate change. Dispersal modes were inventoried for 92% of species and fruit traits for 84% of the woody plant community on the MFDP. Fruit–frugivore interactions were identified using a combination of camera traps, tree watches and opportunistic observations. Though 60% of the species were animal-dispersed, mechanically-dispersed species were more dominant and constituted nearly 70% of the stand. Bird-dispersed species were very rare and constituted less than 10% of the stand cumulatively. In comparison, bird-dispersed species account for ~ 50% of the stand in moist forests of tropical Asia. Three broad dispersal modes were identified – mechanical, mammal and bird-mammal, and these displayed strong associations with fruit traits. Among the 11 frequently-observed frugivore groups, the three largest frugivores, deer, bear and elephant, together dispersed 40% of the community. There was low overlap in fruit diets between these three megafaunal groups, though diets of deer and bear overlapped with different groups of small and medium-sized frugivores. Deer and bear displayed significant associations with fruit traits, while elephant did not. Among the fleshy fruits, large fruits and large-seeded fruits had fewer dispersers compared to smaller, multi-seeded fruits. Although the largest fleshy fruits and seeds were often dispersed by megafauna, few were exclusively dependent on megafauna. The three megafaunal groups were the only potential long distance dispersers for 25% of the community. For a better understanding of the impacts of megafaunal extinctions, even rare events of dispersal by megafauna should be considered since these could prove important for plants lacking other long distance dispersers. Given that these megafauna and several medium-sized frugivores are rare in degraded areas of tropical Asia (Corlett 2007, Karanth et al. 2010), one fourth of the plant community at Mudumalai could have limited ability to disperse across fragmented landscapes in response to changing climates. Seed dispersal and spatial patterns in tropical dry forests: To address the relevance of seed dispersal to plant community structure, the linkages between dispersal mode and spatial aggregation of trees were examined for the woody plant community on the MFDP. For plants, seed arrival patterns vary depending upon their dispersal mode and form the primary matrix upon which further forces shape spatial patterns. Spatial patterns were examined via the pair correlation function (statistic) and statistics derived from cluster models (Matern process). Considering only trees that had 10 individuals (48 species), it was found that tropical dry forest trees exhibited extensive spatial aggregation. The degree of spatial aggregation at Mudumalai was remarkably similar to wet forests in tropical Asia. Species with limited-dispersal (mechanically-dispersed species) were more aggregated than those with more frequent long-distance dispersal (animal-dispersed species) at smaller spatial scales (< 50 m). Animal-dispersed species had larger cluster radius, with bird-dispersed species having the largest radii. These results are in concordance with Siedler & Plotkin (2006) and Li et al. (2009) who find that mechanically-dispersed species are more aggregated than animal-dispersed species. These results demonstrate that seed dispersal has a strong influence on the spatial patterning of plant communities. Conclusion: At the population level, our results imply that P. emblica is mainly dependent on ruminants for seed dispersal. In tropical dry forest sites such as Mudumalai, up to 15% of the species are dispersed by ruminants, with langurs mediating the ruminant-plant interaction similar to P. emblica. The observed fruit removal patterns for P. emblica reiterate the fact that all fruiting individuals in a population are not equally attractive to frugivores, especially when fruits are abundant. Alterations of such complex fruit-frugivore interactions through human-mediated changes of fruit or frugivore densities (through fruit harvests or hunting of large mammalian frugivores such as ruminants), can feed back into the seed dispersal loop and affect several downstream ecological and evolutionary processes. At the community-scale, the dispersal profile of woody plants in Mudumalai’s tropical dry forests differed considerably from tropical moist sites, with a greater representation of mammal and mechanically-dispersed species. However, the linkages between seed dispersal and spatial patterns of trees at Mudumalai are similar to tropical moist forests. Extensions of the models developed from this site harbouring an intact faunal assemblage to other tropical dry forests would enable rapid characterization of dispersal profiles and identification of plant species dependent on endangered fauna for seed dispersal that should be prioritized for restoration programs.

Phyllanthus amarus, planta amplamente utilizada no tratamento de diversas doenças, em especial para hepatite B, apresenta em sua constituição lignanas de interesse farmacológico, destacando-se a filantina e hipofilantina. Com relação à atividade anticancerígina, a nirantina se destaca como potencial efeito citostático e citocida. Distintas condições ambientais podem afetar a constituição química da planta e interferir nos teores destas substâncias. O presente trabalho teve por objetivo estudar a influência do déficit hídrico sobre a produção de filantina, hipofilantina e nirantina em dois acessos de Phyllanthus amarus, cultivados em vasos em casa de vegetação, no município de Piracicaba-SP. Foram realizados 5 tratamentos: capacidade de campo, 3 dias sem irrigação, 6 dias sem irrigação, 9 dias sem irrigação e 12 dias sem irrigação, avaliando-se a altura das plantas, a degradação lipídica, o teor de umidade do solo e determinação do teor dos princípios ativos por CGMS, nos vários tratamentos. Os resultados indicaram que com o deficit hídrico houve diminuição da altura das plantas tanto para o acesso 7 quanto para o acesso 14. A diminuição na disponibilidade de água (umidade com base em peso) passou de 100% na capacidade de campo, para 2% aos 12 dias causando redução na altura de 22% para o acesso 14 e de 1,3% para o acesso 7. Para o teste de peroxidação lipídica, obteve-se respostas significativas nas plantas a partir do terceiro dia do déficit hídrico, sendo mais moderado entre 3 e 6 dias e mais severo a partir do nono dia. Observou-se para os teores das lignanas testadas sob condições de deficiência hídrica, que o acesso 14 apresentou aumento de 140% para filantina e 118% para nirantina e 40% para a hipofilantina. Já para o acesso 7, estes aumentos foram menos significativos e se pronunciaram em condições de déficit hídrico moderado. / Phyllanthus amarus, widely used plant in the treatment of diverse illnesses, in special for hepatitis B, presents in its constitution lignans of pharmacologic interest, being distinguished phyllanthin and hypophyllanthin. With relation to the anticarcinogenic activity, the niranthin if detaches as potential to stop and to decrease the effect. Distinct ambient conditions can affect the chemical constitution of the plant and intervene with texts of these substances. The present work had for objective to study the influence of the drought on the phyllanthin production, hypophyllanthin and niranthin in two accesses of Phyllanthus amarus, cultivated in pots in a greenhouse, in the city of Piracicaba-SP. Five treatments had been carried through: capacity of field, 3 days without irrigation, 6 days without irrigation, 9 days without irrigation and 12 days without irrigation, evaluating itself it height of the plants, the lipidic degradation, the text of humidity of the ground and determination of the text of the active principles for GCSM, in the some treatments. The results had indicated that with the drought it in such a way had reduction of the height of the plants for access 7 how much for access 14. The reduction in the water availability (humidity on the basis of weight) passed of 100% in the field capacity, for 2% to the 12 days causing reduction in the height of 22% for access 14 and 1,3% for access 7. For the test of lipidic peroxidation, one got significant answers in the plants from the third day of the hídrico deficit, being more moderate between 3 and 6 days and more severe from the nineth day. It was observed for texts of the lignans tested under conditions of drought that access 14 presented increase of 140% for phyllanthin and 118% for niranthin and 40% for the hypophyllanthin. Already for access 7, these increases had been less significant and if they had pronounced in conditions of moderate drought.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The vegetation to be employed in techniques proposed by the natural engineering or soil bioengineering needs to fill out some requisites related to ecological, phytossociological and of reproduction aspects. The techniques in natural engineering use the plants in different angles in relation to the land. For this reason, the present work aims to evaluate the effects of different angles of stem cuttings planting of Phyllanthus sellowianus Müll. Arg., Salix humboldtiana Willd. and Sebastiania schottiana (Müll. Arg.) Müll. Arg., in order to determine the most suitable form to the planting of these species. The experiment was conducted in the Laboratory of Silviculture, in the Federal University of Santa Maria (UFSM), Santa Maria, Rio Grande do Sul, in the period of August to November of 2007. The treatments consisted in the three species of stem cuttings planting, in three different angles of planting in relation to the land (10°, 30° and 90°). The stem cuttings with 30 cm of length, with variable diameter from 1.0 to 4.5 cm, were collected in the central region of the state of the Rio Grande do Sul, in the final of August month. Two thirds of the stem cutting s bases were inserted in inert substrate (sand). The experimental design was completely randomized, with 30 repetitions for treatment. After 30 and 60 days, the following parameters were observed: the number of sprouted stem cuttings, number and length of the sprouts, and after 90 days, besides these parameters, were determined the survival, the length, the diameter, and the dry mass of the shoots, and also the length, the diameter, the dry mass and the volume of the roots. The data were statistically analyzed through the statistical programs and Excel. Sebastiania schottiana did not present sprouting, not even rooting, which perhaps could be explained by the stem cuttings time of collection, where that should be tested in other periods of year. The planting angle showed influence on some variables for Phyllanthus sellowianus and Salix humboldtiana. Both the species presented high values of survival, with a tendency for major results for stem cuttings planted in angle of 10° and 90° for Phyllanthus sellowianus (100%) and in angle of 30° for Salix humboldtiana (87%). Significant correlations were observed between the variables of the air part and of the radicial system and the area of transverse section of the stem cuttings, for the different planting angles. Both the two species presented the majority of the roots located on the basis of the stem cuttings, except for the ones planted in angle of 30°, which had the roots distributed along the whole buried portion. The realization of new studies is suggested with other species and also with another angles of planting, as well as the use of bigger diametric breadth. / A vegetação a ser empregada em técnicas propostas pela engenharia natural ou bioengenharia de solos necessita preencher alguns requisitos relacionados a aspectos ecológicos, fitossociológicos e de reprodução. As técnicas de engenharia natural utilizam as plantas em diferentes ângulos em relação ao terreno. Por esta razão, o presente trabalho tem por objetivo avaliar os efeitos de diferentes ângulos de plantio de estacas de Phyllanthus sellowianus Müll. Arg., Salix humboldtiana Willd. e Sebastiania schottiana (Müll. Arg.) Müll. Arg., a fim de determinar a forma mais adequada para o plantio destas espécies. O experimento foi conduzido no Laboratório de Silvicultura, na Universidade Federal de Santa Maria (UFSM), Santa Maria, Rio Grande do Sul, no período de agosto a novembro de 2007. Os tratamentos consistiram no plantio de estacas das três espécies, em três ângulos diferentes de plantio em relação ao terreno (10°, 30° e 90°). As estacas com 30 cm de comprimento, com diâmetro variável de 1,0 a 4,5 cm, foram coletadas no final de agosto, na região central do estado do Rio Grande do Sul. Dois terços da base das estacas foram inseridos em substrato inerte (areia). O delineamento experimental foi o inteiramente casualizado, com 30 repetições por tratamento. Após 30 e 60 dias, foram observados os seguintes parâmetros: número de estacas brotadas, número e comprimento dos brotos, e, aos 90 dias, além destes parâmetros, foram determinados a sobrevivência, o comprimento, o diâmetro, a massa seca dos brotos e o comprimento, a massa seca e o volume das raízes. Os dados foram analisados estatisticamente através de programas estatísticos e Excel. Sebastiania schottiana não apresentou brotação, nem enraizamento, o que talvez possa ser explicado pela época de coleta das estacas, devendo ser testada em outras épocas do ano. O ângulo de plantio mostrou influência sobre algumas variáveis para Phyllanthus sellowianus e Salix humboldtiana. Ambas as espécies apresentaram altos valores de sobrevivência, com tendência de maiores resultados para estacas plantadas em ângulo de 10° e 90° para Phyllanthus sellowianus (100%) e em ângulo de 30° para Salix humboldtiana (87%). Observaram-se correlações significativas entre as variáveis da parte aérea e do sistema radicial e a área da seção transversal das estacas, para os diferentes ângulos de plantio. As duas espécies apresentaram a maioria das raízes localizadas na base das estacas, exceto para as estacas plantadas em ângulo de 30°, as quais tiveram as raízes distribuídas ao longo de toda a porção enterrada. Sugere-se a realização de novos estudos com outras espécies e também com outros ângulos de plantio, assim como a utilização de maior amplitude diamétrica.

Phyllanthus amarus, planta amplamente utilizada no tratamento de diversas doenças, em especial para hepatite B, apresenta em sua constituição lignanas de interesse farmacológico, destacando-se a filantina e hipofilantina. Com relação à atividade anticancerígina, a nirantina se destaca como potencial efeito citostático e citocida. Distintas condições ambientais podem afetar a constituição química da planta e interferir nos teores destas substâncias. O presente trabalho teve por objetivo estudar a influência do déficit hídrico sobre a produção de filantina, hipofilantina e nirantina em dois acessos de Phyllanthus amarus, cultivados em vasos em casa de vegetação, no município de Piracicaba-SP. Foram realizados 5 tratamentos: capacidade de campo, 3 dias sem irrigação, 6 dias sem irrigação, 9 dias sem irrigação e 12 dias sem irrigação, avaliando-se a altura das plantas, a degradação lipídica, o teor de umidade do solo e determinação do teor dos princípios ativos por CGMS, nos vários tratamentos. Os resultados indicaram que com o deficit hídrico houve diminuição da altura das plantas tanto para o acesso 7 quanto para o acesso 14. A diminuição na disponibilidade de água (umidade com base em peso) passou de 100% na capacidade de campo, para 2% aos 12 dias causando redução na altura de 22% para o acesso 14 e de 1,3% para o acesso 7. Para o teste de peroxidação lipídica, obteve-se respostas significativas nas plantas a partir do terceiro dia do déficit hídrico, sendo mais moderado entre 3 e 6 dias e mais severo a partir do nono dia. Observou-se para os teores das lignanas testadas sob condições de deficiência hídrica, que o acesso 14 apresentou aumento de 140% para filantina e 118% para nirantina e 40% para a hipofilantina. Já para o acesso 7, estes aumentos foram menos significativos e se pronunciaram em condições de déficit hídrico moderado. / Phyllanthus amarus, widely used plant in the treatment of diverse illnesses, in special for hepatitis B, presents in its constitution lignans of pharmacologic interest, being distinguished phyllanthin and hypophyllanthin. With relation to the anticarcinogenic activity, the niranthin if detaches as potential to stop and to decrease the effect. Distinct ambient conditions can affect the chemical constitution of the plant and intervene with texts of these substances. The present work had for objective to study the influence of the drought on the phyllanthin production, hypophyllanthin and niranthin in two accesses of Phyllanthus amarus, cultivated in pots in a greenhouse, in the city of Piracicaba-SP. Five treatments had been carried through: capacity of field, 3 days without irrigation, 6 days without irrigation, 9 days without irrigation and 12 days without irrigation, evaluating itself it height of the plants, the lipidic degradation, the text of humidity of the ground and determination of the text of the active principles for GCSM, in the some treatments. The results had indicated that with the drought it in such a way had reduction of the height of the plants for access 7 how much for access 14. The reduction in the water availability (humidity on the basis of weight) passed of 100% in the field capacity, for 2% to the 12 days causing reduction in the height of 22% for access 14 and 1,3% for access 7. For the test of lipidic peroxidation, one got significant answers in the plants from the third day of the hídrico deficit, being more moderate between 3 and 6 days and more severe from the nineth day. It was observed for texts of the lignans tested under conditions of drought that access 14 presented increase of 140% for phyllanthin and 118% for niranthin and 40% for the hypophyllanthin. Already for access 7, these increases had been less significant and if they had pronounced in conditions of moderate drought.

Constituiu objetivo deste trabalho o desenvolvimento tecnológico de formas farmacêuticas obtidas a partir de Phyllanthus niruri L. (Euphorbiaceae), incluso no Projeto de Pesquisa de Plantas Medicinais da Central de Medicamentos. Para tanto, foram desenvolvidas metodologias analíticas capazes de avaliar a qualidade da matéria-prima, das ações de transformação, dos produtos intermediários e do produto final. Estudaram-se técnicas de preparação de soluções extrativas e de produto seco por aspersão. O estudo da metodologia adaptada para quantificação de flavonóides na matéria-prima, demonstrou desvio da linearidade de resposta com o aumento da massa da amostra, limitando o seu emprego as condições fixadas neste trabalho. O teor de flavonóides totais do material vegetal foi de 0,80 g % (m/m). A solução extrativa empregada no desenvolvimento do produto seco foi selecionada através da realização do planejamento fatorial 2 3, onde foram avaliados os fatores: método de extração (decocção e infusão), relação droga solvente (2,5 e 7,5:100) e tempo de extração (5 e 15 min), utilizando como parâmetro de decisão o teor de flavonóides totais. A solução extrativa selecionada foi obtida através de decocção durante 15 minutos das partes aéreas moídas na proporção 7,5:100 (m/V). O emprego da mesma metodologia quantitativa aplicada a matéria-prima e ao produto intermediário mostrou a existência de erro sistemático proporcional com o aumento da concentração das amostras. Entretanto, alcançou-se comportamento linear, com ausência de erros sistemáticos, empregando-se amostras de menor concentração. O produto seco por aspersão foi obtido em torre de secagem, a partir da solução extrativa selecionada, adicionada de dióxido de silício coloidal a 30% do resíduo seco. Tanto a operação de secagem quanto a adição de dióxido de silício coloidal não causaram interferência nas análises qualitativas e quantitativas, obtendo-se uma taxa de recuperação, calculado como flavonóides totais, de 99%. / Phyllanthus niruri L. is a medicinal plant used in Brazil mainly against genitourinary disorders. Despite of its wide use in the folk medicine and in the pharmaceutical industry no attempts were done to provide technological knowledge in order to obtain pharmaceutical dosage forms with adequate and constant quality. The governmental program on medicinal, managed by the Brazilian Drug Agency CEME, developed pharmacological, chemical and botanical studies for this plant in a feasible dosage form, the technological development spray-dried product starting from Phyllanthus niruri was the aim of this work. For such purpose quality control and production methods for the plant raw material, aqueous extracts and spray-dried product were proposed and studied. The total flavonoid assay method for the plant raw material showed a lack of linearity at extremes high and low concentration, but, however, linear at the middle values. The extractive solution was selected through a 2 3 factorial design, considering the extraction method (decoction and infusion), plant to solvent proportion (2.5 and 7.5:100 w/v) and extraction time (5 and 15 minutes). The total flavonoids content was employed as decision parameter. The chose extractive solution was obtained by 15 min decoction of a plant to solvent ratio of 7.5 :100 (w/v). The assay method was not influenced by the amount of the plant used in the extract, but by the aliquot size. The spray dried product was prepared using colloidal silicon dioxide as drying excipient. The spray drying process and the excipient did not interfere on the qualitative and quantitative profile of the product.The total flavonoid recovery was around 99 % (w/w). The final product showed a low residual humidity content, but limited stability when exposed to high moisture atmospheres.

Constituiu objetivo deste trabalho o desenvolvimento tecnológico de formas farmacêuticas obtidas a partir de Phyllanthus niruri L. (Euphorbiaceae), incluso no Projeto de Pesquisa de Plantas Medicinais da Central de Medicamentos. Para tanto, foram desenvolvidas metodologias analíticas capazes de avaliar a qualidade da matéria-prima, das ações de transformação, dos produtos intermediários e do produto final. Estudaram-se técnicas de preparação de soluções extrativas e de produto seco por aspersão. O estudo da metodologia adaptada para quantificação de flavonóides na matéria-prima, demonstrou desvio da linearidade de resposta com o aumento da massa da amostra, limitando o seu emprego as condições fixadas neste trabalho. O teor de flavonóides totais do material vegetal foi de 0,80 g % (m/m). A solução extrativa empregada no desenvolvimento do produto seco foi selecionada através da realização do planejamento fatorial 2 3, onde foram avaliados os fatores: método de extração (decocção e infusão), relação droga solvente (2,5 e 7,5:100) e tempo de extração (5 e 15 min), utilizando como parâmetro de decisão o teor de flavonóides totais. A solução extrativa selecionada foi obtida através de decocção durante 15 minutos das partes aéreas moídas na proporção 7,5:100 (m/V). O emprego da mesma metodologia quantitativa aplicada a matéria-prima e ao produto intermediário mostrou a existência de erro sistemático proporcional com o aumento da concentração das amostras. Entretanto, alcançou-se comportamento linear, com ausência de erros sistemáticos, empregando-se amostras de menor concentração. O produto seco por aspersão foi obtido em torre de secagem, a partir da solução extrativa selecionada, adicionada de dióxido de silício coloidal a 30% do resíduo seco. Tanto a operação de secagem quanto a adição de dióxido de silício coloidal não causaram interferência nas análises qualitativas e quantitativas, obtendo-se uma taxa de recuperação, calculado como flavonóides totais, de 99%. / Phyllanthus niruri L. is a medicinal plant used in Brazil mainly against genitourinary disorders. Despite of its wide use in the folk medicine and in the pharmaceutical industry no attempts were done to provide technological knowledge in order to obtain pharmaceutical dosage forms with adequate and constant quality. The governmental program on medicinal, managed by the Brazilian Drug Agency CEME, developed pharmacological, chemical and botanical studies for this plant in a feasible dosage form, the technological development spray-dried product starting from Phyllanthus niruri was the aim of this work. For such purpose quality control and production methods for the plant raw material, aqueous extracts and spray-dried product were proposed and studied. The total flavonoid assay method for the plant raw material showed a lack of linearity at extremes high and low concentration, but, however, linear at the middle values. The extractive solution was selected through a 2 3 factorial design, considering the extraction method (decoction and infusion), plant to solvent proportion (2.5 and 7.5:100 w/v) and extraction time (5 and 15 minutes). The total flavonoids content was employed as decision parameter. The chose extractive solution was obtained by 15 min decoction of a plant to solvent ratio of 7.5 :100 (w/v). The assay method was not influenced by the amount of the plant used in the extract, but by the aliquot size. The spray dried product was prepared using colloidal silicon dioxide as drying excipient. The spray drying process and the excipient did not interfere on the qualitative and quantitative profile of the product.The total flavonoid recovery was around 99 % (w/w). The final product showed a low residual humidity content, but limited stability when exposed to high moisture atmospheres.

Constituiu objetivo deste trabalho o desenvolvimento tecnológico de formas farmacêuticas obtidas a partir de Phyllanthus niruri L. (Euphorbiaceae), incluso no Projeto de Pesquisa de Plantas Medicinais da Central de Medicamentos. Para tanto, foram desenvolvidas metodologias analíticas capazes de avaliar a qualidade da matéria-prima, das ações de transformação, dos produtos intermediários e do produto final. Estudaram-se técnicas de preparação de soluções extrativas e de produto seco por aspersão. O estudo da metodologia adaptada para quantificação de flavonóides na matéria-prima, demonstrou desvio da linearidade de resposta com o aumento da massa da amostra, limitando o seu emprego as condições fixadas neste trabalho. O teor de flavonóides totais do material vegetal foi de 0,80 g % (m/m). A solução extrativa empregada no desenvolvimento do produto seco foi selecionada através da realização do planejamento fatorial 2 3, onde foram avaliados os fatores: método de extração (decocção e infusão), relação droga solvente (2,5 e 7,5:100) e tempo de extração (5 e 15 min), utilizando como parâmetro de decisão o teor de flavonóides totais. A solução extrativa selecionada foi obtida através de decocção durante 15 minutos das partes aéreas moídas na proporção 7,5:100 (m/V). O emprego da mesma metodologia quantitativa aplicada a matéria-prima e ao produto intermediário mostrou a existência de erro sistemático proporcional com o aumento da concentração das amostras. Entretanto, alcançou-se comportamento linear, com ausência de erros sistemáticos, empregando-se amostras de menor concentração. O produto seco por aspersão foi obtido em torre de secagem, a partir da solução extrativa selecionada, adicionada de dióxido de silício coloidal a 30% do resíduo seco. Tanto a operação de secagem quanto a adição de dióxido de silício coloidal não causaram interferência nas análises qualitativas e quantitativas, obtendo-se uma taxa de recuperação, calculado como flavonóides totais, de 99%. / Phyllanthus niruri L. is a medicinal plant used in Brazil mainly against genitourinary disorders. Despite of its wide use in the folk medicine and in the pharmaceutical industry no attempts were done to provide technological knowledge in order to obtain pharmaceutical dosage forms with adequate and constant quality. The governmental program on medicinal, managed by the Brazilian Drug Agency CEME, developed pharmacological, chemical and botanical studies for this plant in a feasible dosage form, the technological development spray-dried product starting from Phyllanthus niruri was the aim of this work. For such purpose quality control and production methods for the plant raw material, aqueous extracts and spray-dried product were proposed and studied. The total flavonoid assay method for the plant raw material showed a lack of linearity at extremes high and low concentration, but, however, linear at the middle values. The extractive solution was selected through a 2 3 factorial design, considering the extraction method (decoction and infusion), plant to solvent proportion (2.5 and 7.5:100 w/v) and extraction time (5 and 15 minutes). The total flavonoids content was employed as decision parameter. The chose extractive solution was obtained by 15 min decoction of a plant to solvent ratio of 7.5 :100 (w/v). The assay method was not influenced by the amount of the plant used in the extract, but by the aliquot size. The spray dried product was prepared using colloidal silicon dioxide as drying excipient. The spray drying process and the excipient did not interfere on the qualitative and quantitative profile of the product.The total flavonoid recovery was around 99 % (w/w). The final product showed a low residual humidity content, but limited stability when exposed to high moisture atmospheres.

Submitted by Emanuel Varela Cardoso (emanuel.varela@ufcg.edu.br) on 2018-09-26T18:52:42Z No. of bitstreams: 1 ANA PAULA TRINDADE ROCHA – TESE (PPGEP) 2006.pdf: 2605471 bytes, checksum: 29c8f1076e79512504affafce03e4657 (MD5) / Made available in DSpace on 2018-09-26T18:52:42Z (GMT). No. of bitstreams: 1 ANA PAULA TRINDADE ROCHA – TESE (PPGEP) 2006.pdf: 2605471 bytes, checksum: 29c8f1076e79512504affafce03e4657 (MD5) Previous issue date: 2006-12-22 / CNPq / O presente trabalho teve como objetivo estudar o recobrimento contínuo de grânulos de quebra pedra em leito de jorro. Os grânulos foram obtidos a partir do processo de granulação por via úmida do extrato seco de quebra pedra, utilizando como ligante o PVPK30. Utilizou-se um recobridor em leito de jorro de configuração tronco-cilíndrica, construído em acrílico com base cônica de 60o, diâmetro da coluna de 15 cm, altura de 45 cm e diâmetro de entrada do ar de 3,0 cm. A suspensão polimérica a base de Eudragit foi atomizada sobre o leito de partículas, através do bico atomizador de duplo-fluido, localizado na parte superior da coluna. Trabalhou-se com uma carga de grânulos de 0,4 kg formada por partículas cuja distribuição granulométrica variou de 1,7 a 3,35 mm. Inicialmente, foi realizado o levantamento das curvas características e a partir dos resultados destas, juntamente com os resultados dos testes preliminares de recobrimento elaborou-se um planejamento experimental 23 com 6 pontos axiais e três repetições no ponto central, totalizando 17 experimentos de recobrimento. As variáveis operacionais foram: pressão de atomização, vazão de suspensão de recobrimento e temperatura do ar de secagem. Os resultados experimentais permitiram investigar as variáveis respostas: eficiência de recobrimento, crescimento relativo das partículas, índice de aglomerados e taxa de evaporação. A eficiência do processo variou de 40,78 a 93,19 %, o crescimento relativo de 7,31 a 15,86 %, o índice de aglomerado de 0 a 38,82% e a taxa de evaporação de 3,16 g/min a 10,9 g/min. O planejamento experimental forneceu modelos estatisticamente significativos para o índice de aglomerados e crescimento de partículas. Quanto à taxa de evaporação o modelo obtido além de estatisticamente significativo, foi também preditivo. Foram realizados ensaios de dissolução para avaliar o filme de recobrimento nos experimentos com maior eficiência, com menor eficiência e no ponto central. O principio ativo avaliado nesses testes foi o teor de flavonóides totais. Os resultados reforçam a importância do estudo da dissolução in vitro e que o processo de revestimento em leito de jorro é uma alternativa viável para modificação do perfil de liberação de formas farmacêuticas granulares. / The present work aimed to study the continuous coating process of stone breaker (Phylanthus niruri L.) granules in spouted bed. The granules were obtained from the granulation process, through humid via of stone breaker’s dry extract, in which the PVP-K30 was utilized as a binder. It was used a cone-cylindrical spouted bed coater, built of Plexiglass, whose dimensions were: angular base of 60o, column diameter of 5cm, column height of 45cm and inlet orifice diameter of 3.0 cm. The aqueous polymeric suspension of Eudragit was sprayed on the particles bed through the double-fluid atomizer nozzle located at the top the bed. The experiment carried out granules of 0.4kg formed by particles whose granulometric distribution varied from 1.7 to 3.35 mm. Initially, the characteristic curves had been analyzed and then, after these results were taken into consideration along with the coating preliminary tests, a 23 factorial design - with 3 central points and 6 axial points, 17-run full factorial design - was employed to evaluate the effects of the operational variables on the coating process. The independent variables studied were the air pressure of atomization, flow rate of suspension and the air inlet temperature. The responses analyzed were the process efficiency, the particles growth, the agglomeration index and the evaporation rate. The process efficiency varied from 40.78 to 93.19%, the relative growth from 7.31 to 15.86%, the agglomeration index from 0 to 38.82% and the evaporation rate from 3.16 to 10.9 g/min. The factorial design provided statistically significant models to the agglomeration index and the particles growth. As for the evaporation rate, besides being statistically significant, the model obtained was also predictive. Dissolution tests were carried out to analyze the coating film in the runs with lower and higher efficiency and with central point. The active ingredient evaluated in these tests was the total flavonoids content. The results point out the importance of the in vitro dissolution study and also that the coating process in spouted bed is a viable alternative to the profile modification in the granular pharmaceutic form’s release.

Orientador: Maria Angela de Almeida Meireles / Tese (doutorado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-10T22:47:19Z (GMT). No. of bitstreams: 1 Silva_DanielleCottadeMelloNunesda_D.pdf: 3730764 bytes, checksum: e92936b597dcfe4aa9d9a2a86ce46790 (MD5) Previous issue date: 2008 / Resumo: O presente estudo teve como objetivo avaliar o rendimento global de extração, a atividade biológica e os perfis de composição de extratos de lavanda brasileira (Aloysia gratissima), quebra pedra (Phylanthus amarus) e ginseng brasileiro (Pfaffia paniculata), obtidos por extração supercrítica (SFE), hidrodestilação e por extração com solvente a baixa pressão (LPSE) utilizando metanol, etanol e hexano. A lavanda brasileira apresentou o rendimento global de extração (X0) variando entre 2,0 ± 0,1% e 4,0 ± 0,2% (b.u), sendo que o maior rendimento de extrato foi obtido na condição de operação de 50 °C e 300 bar (4,0 ± 0,2%). O rendimento do óleo volátil de lavanda brasileira obtido por hidrodestilação foi de 1,2 ± 0,1%. Os rendimentos dos extratos obtidos por extração a baixa pressão foram: 9,4 ± 0,1% (metanol), 3,3 ± 0,2% (etanol) e 0,8 ± 0,2 %(hexano). Os compostos majoritários presentes no óleo volátil e no extrato SFE de lavanda brasileira foram pinocanfona (13,5 - 16,3%), ß-pineno (10,5 - 12%) e pinocarvil acetato (7,3 - 8,3%), guaiol (6,6 - 8,7%) e bulnesol (3,7 - 4,1%). No óleo volátil a concentração referente a 40% de redução do radical DPPH (Sc40 (óleo 1) (1,8 ± 0,1) × 103 mg/L)) e no extrato SFE a concentração referente a 50% de redução do radical DPPH (Sc50 (SFE) (1,3 ± 0,3) × 103 mg/L) de lavanda brasileira demostraram fraco potencial antioxidante pelo método DPPH (1,1-difenil-2-pirilhidrazina). Já pelo método ORAC o óleo volátil (203 ± 15 micromoles de Trolox/ góleo (µM TE/góleo)) e o extrato SFE (225±18 micromoles de Trolox/ gSFE (µM TE/gSFE)) apresentaram poder antioxidante comprável a rutina o composto de referência. Pelo método da reação acoplada do ß-caroteno/ácido linolênico, a atividade antioxidante (AA) dos extratos de lavanda brasileira variou de acordo com o solvente utilizado (metanol, etanol e hexano) e de acordo com a condição SFE empregada. A atividade antioxidante dos extratos de lavanda brasileira obtidos com solventes orgânicos variou de -5 a 62%; o extrato obtido a 50 °C e 300 bar por SFE apresentou AA, variando entre 119 e 188%; os demais extratos não apresentaram AA pelo método do ß-caroteno/ácido linolênico. A quebra pedra apresentou X0 variando de 0,80± 0,01 % a 3,60 ± 0,04% (b.u) de acordo com as condições de extração (pressão e temperatura), sendo que o maior rendimento de extração foi obtido na condição de operação de 50 °C e 250 bar (3,60 ± 0,04%). Os rendimentos dos extratos obtidos por extração a baixa pressão foram: 10,20 ± 0,04% (metanol), 3,40 ± 0,04% (etanol) e 1,90 ± 0,04% (hexano). Por meio da cromatografia em camada delgada foram confirmadas as presenças de óleo volátil, pigmentos, flavonóides e triterpenos nos extratos SFE e nos extratos LSPE de quebra pedra. Pelo método DPPH de atividade antioxidante o extrato SFE de quebra pedra apresentou fraca AA comparado à atividade dos compostos de referência (butilhidroxitolueno e rutina). De maneira geral, os extratos SFE de quebra pedra apresentaram AA superior à do antioxidante controle ß-caroteno pelo método da reação acoplada do ß-caroteno/ácido linolênico. A atividade antioxidante dos extratos de quebra pedra obtidos com solventes orgânicos variou de 17 a 217%; o extrato SFE obtido a 40 °C e 200 bar apresentou AA em torno de 100% em uma, duas e três horas de reação; os demais extratos SFE (40 °C e 250 bar e 40 °C e 300 bar) apresentaram AA variando de 21 a 66%; os extratos de quebra pedra podem ser considerados antioxidantes naturais com boa estabilidade e efeito prolongado. O ginseng brasileiro apresentou X0 variando de 0,50 ± 0,02% a 1,40 ± 0,07% (b.s) de acordo com as condições de extração (pressão e temperatura), sendo que o maior rendimento de extração foi obtido na condição de operação de 30 °C e 250 bar (1,40 ± 0,07%). Os rendimentos dos extratos obtidos por extração a baixa pressão foram: 8,2 ± 0,8% (metanol), 2,2 ± 0,7% (etanol) e 0,6 ± 0,1% (hexano). Por meio das duas técnicas analíticas utilizadas para os extratos de ginseng brasileiro pode-se verificar a presença desde compostos de baixa massa molecular até compostos com estruturas complexas de alta massa molecular (saponinas, ácidos, triterpenos, esteróis) nos extratos SFE. Os compostos majoritários identificados nos extratos SFE de ginseng brasileiro foram ácido hexadecanóico (0,5 - 27,1%), ß-sitosterol e/ou estigmasterol (1,1 - 20,1%), fitol (6,0 - 30,8%) e hexatriacontane (0,7 - 30,6%). Os compostos majoritários da fração retida na coluna de adsorção, instalada na saída da corrente de solvente foram o para-dietilbenzeno (25,7 - 59,3 %) e o meta-dietilbenzeno (3,4 - 27,1 %). Pelo método DPPH de atividade antioxidante o extrato SFE de ginseng brasileiro apresentou AA 25 vezes menos eficaz que a AA desempenhada pela rutina antioxidante natural. Pelo método da reação acoplada do ß-caroteno/ácido linolênico a AA foi observada para o extrato metanólico, variando de 94 a 117%. Os extratos SFE que apresentaram maior potencial antioxidante foram obtidos a 50 °C e 200 bar e 50 °C e 300 bar, a porcentagem de inibição da oxidação destes extratos variou entre 143 e 246% e manteve-se estável com o tempo de reação. O óleo volátil de lavanda brasileira e os extratos SFE de lavanda brasileira, quebra pedra e ginseng brasileiro, não apresentaram efeito inibitório (atividade antiinflamatória) sobre a lipoxigenase de soja nas concentrações estudadas (4-10 g/L) / Abstract: This present work has as an objective evaluate the global yield of the extraction, biological activity, and the composition profiles of the Brazilian lavender (Aloysia gratissima), quebra pedra (Phylanthus amarus) and Brazilian ginseng (Pfaffia paniculata) extracts, obtained by supercritical extraction (SFE), hydrodistillation and by low pressure solvent extraction (LPSE) using methanol, ethanol e hexane. The brazilian lavender showed a global extraction yield (X0) varying between 2,0 ± 0,1% and 4,0 ± 0,2% (b.u), and the higher extract yield was obtained on 50 °C and 300 bar (4,0 ± 0,2%) of operational conditions. The global yield of the brazilian lavender volatile oil obtained by hydrodistillation was 1,2 ± 0,1%. The global extracts yield obtained by low pressure extraction were: 9,4 ± 0,1% (methanol), 3,3 ± 0,2% (ethanol) and 0,8 ± 0,2 %(hexane). The mainly components present in the volatile oil and in the SFE brazilian lavender extract were pinocamphone (13,5 - 16,3%), ß-pinene (10,5 - 12%) and pinocarvyl acetate (7,3 - 8,3%), guaiol (6,6 - 8,7%) e bulnesol (3,7 - 4,1%). In the volatile oil the relating concentration to 40% of the radical reduction DPPH (Sc40 (oil) (1,8 ± 0,1) × 103 mg/L)) and in the SFE extract the concentration relating to 50% of the radical reduction (Sc50 (SFE) (1,3 ± 0,3) × 103 mg/L) of brazilian lavender demonstrate a weak antioxidant potential by the DPPH (1,1-diphenyl-2-picrylhydrazyl) method. Although by the ORAC method the volatile oil (203 ± 15 µM TE/ goil (µM TE/goil)) and the SFE extract (225±18 µM TE/ gSFE) showed an antioxidant power similar to the rutin the reference component. By the coupled reaction of the ß-carotene/ linolenic acid method, the antioxidant activity (AA) of the brazilian lavender extracts varied according to the utilized solvent (methanol, ethanol and hexane) and according to the SFE condition employed. The brazilian lavender extracts antioxidant activity obtained with organic solvents varied from -5 to 62%; the obtained extract on 50 °C and 300 bar by SFE presented AA, varying between 119 and 188%; the others extracts did not present an AA by the ß-carotene /linolenic acid method. The quebra pedra presented X0 varying between 0,80± 0,01 % to 3,60 ± 0,04% (b.u) according to the extraction conditions (pressure and temperature), the higher extraction yield was obtained on 50 °C and 250 bar (3,60 ± 0,04%) of operational conditions. The global extracts yield obtained by low pressure extraction were: 10, 20 ± 0,04% (methanol), 3,40 ± 0,04% (ethanol) e 1,90 ± 0,04% (hexane). By the thin layer cromatography were confirmed the presence of the volatile oils, pigments, flavonoids and triterpenes in the quebra pedra SFE and LSPE extracts. By the DPPH antioxidant activity method the quebra pedra SFE extract showed a weak AA related to the reference components (t-butyl hydroxyl toluene e rutin). Generally, the quebra perdra SFE extracts presents an AA superior to the ß-carotene control antioxidant by the ß-carotene/linolenic acid coupled reaction method. The quebra pedra extracts antioxidant activity obtained with organic solvent varied from 17 to 217%; the SFE extract obtained on 40 °C and 200 bar presented an AA around 100% in one, two and three reaction hours; the others SFE extracts (40 °C and 250 bar and 40 °C e 300 bar) presented an AA varying from 21 to 66%; the quebra pedra extracts can be considered natural antioxidants with good stability and prolonged effect. The Brazilian ginseng presented X0 varying from 0,50 ± 0,02% to 1,40 ± 0,07% (b.s) according to the extraction conditions (pressure and temperature), the higher extraction yield was obtained on the 30 °C and 250 bar (1,40 ± 0,07%) operation condition. The global extracts yield obtained by low pressure extraction were: 8,2 ± 0,8% (methanol), 2,2 ± 0,7% (ethanol) e 0,6 ± 0,1% (hexane). Trough two analytical techniques utilized for the Brazilian ginseng extracts it can verifies the presence from low molecular mass components until complex structures with high molecular mass. (saponins, acids, triterpenes, sterols) in the SFE extracts. The mainly components identified in the SFE brazilian ginseng extracts were hexadecane acid - 27,1%), ß-sitosterol e/ou stigmasterol (1,1 - 20,1%), phytol (6,0 - 30,8%) e hexatriacontane (0,7 - 30,6%). The mainly retained fraction on the adsorption column, installed on the solvent outlet were (25,7 - 59,3 %) for the p-dietilbenzene and (3,4 - 27,1 %) for the m-dietilbenzene. By the DPPH antioxidant activity method the SFE Brazilian ginseng extract presented an AA 25 times less efficient than the AA performed by the natural antioxidant rutin. By the ß-carotene/ linolenic acid coupled reaction the AA was observed for the metanolic extract, varying form 94 to 117%. The SFE extracts that presented higher antioxidant potential were obtained on 50 °C and 200 bar and 500 °C and 300 bar, the inhibition percentage of these extracts oxidation varied between 143 and 246% and keeps it stable with the reaction time. The brazilian lavender volatile oil and the brazilian lavender, quebra pedra and brazilian ginseng SFE extracts did not present an inhibited effect (anti-inflammatory activity) on the soybean lipoxigenase on the studied concentrations / Doutorado / Doutor em Engenharia de Alimentos