Submitted by Marlene Santos (marlene.bc.ufg@gmail.com) on 2014-12-22T20:30:07Z No. of bitstreams: 2 Dissertação - Jordana Nunes de Sousa - 2014.pdf: 2730306 bytes, checksum: 3529cda6d9291d1b7887c15186b6b530 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Approved for entry into archive by Luciana Ferreira (lucgeral@gmail.com) on 2014-12-23T10:06:05Z (GMT) No. of bitstreams: 2 Dissertação - Jordana Nunes de Sousa - 2014.pdf: 2730306 bytes, checksum: 3529cda6d9291d1b7887c15186b6b530 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) / Made available in DSpace on 2014-12-23T10:06:07Z (GMT). No. of bitstreams: 2 Dissertação - Jordana Nunes de Sousa - 2014.pdf: 2730306 bytes, checksum: 3529cda6d9291d1b7887c15186b6b530 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2014-03-28 / The present work aimed to obtain extracts from the barks of Stryphnodendron adstringens (Mart) Coville (Fabaceae) and evaluating of biological activities in vitro, from them. Chapter 1 using pharmacopoeial and not pharmacopoeial methods; barks, powder and liquid extract of S. adstringens were characterized and then a HPLC method was developed and validated to idenfication of epigallocatechin gallate (EGCG) at extracts. In Chapter 2, described the cytotoxic activity of the dry extract of S. adstringens, obtained by atomization, and the EGCG standard against L929 fibroblast cells by the method of neutral red. EGCG at 200 µg/mL reduced, p ≤ 0.05, the cell viability of L929 fibroblasts to 47.70%, such as the dried extract of S. adstringens at 225 mg/mL. Chapter 3 presented the obtainment of dry extracts of S. adstringens by atomization. The influence of the proportion of adjuvant drying, the drying air temperature and flow rate of the drying air on total phenols, EGCG, tannins, total antioxidant activity and efficiency of the drying process were evaluated, the significant results (p≤0.05) were obtained by ANOVA and represented by response surface methodology. The increase in the adjuvant drying was not significantly favorable to efficiency of the drying process. Increasing the temperature of the drying air had a quadratic behavior negative to total phenols and positive to total tannins. The increasing of flow air drying showed linear and quadratic behavior. negative to total antioxidant activity. The efficiency of the drying process was not influenced by any variable. The dried extracts were analyzed for their content of volatile, water activity and the morphologies of the particles were observed by scanning electron microscopy. In Chapter 4 we describe the optimization of extraction of total phenols, tannins and epigallocatechin gallate powders of barks of S. adstringens ultrasonic using Box- planning Benhken 33. The use and validation of the desirability global statistical showed that the proportion of ratio drug solvent 4mg/mL, time extraction 30 min and the 65 % (v/v) hydroalcoholic solution are ideal to simultaneous extraction of these metabolites. The results indicate that the extracts were obtained and standardized quality, and have potential to be investigated their biological and/or insertion activities in herbal formulations / O presente trabalho teve como objetivos obter extratos a partir das cascas de Stryphnodendron adstringens (Mart) Coville (Fabaceae) e a avaliar as atividades biológicas, in vitro dos mesmos. No capítulo 1, por meio de metodologias farmacopéicas e não farmacopéicas, as cascas, o pó e o extrato líquido de S. adstringens foram caracterizados, em seguida, um método foi desenvolvido e validado por CLAE, para idenficar o galato de epigalocatequina (GEGC) nos extratos. No capítulo 2 está descrito a atividade citotóxica do extrato seco de S. adstringens, obtido por spray dryer, e do padrão de GEGC ante células de fibroblasto L929, pelo método de vermelho neutro. Na concentração de 200 μg/mL o GEGC reduziu significativamente, p≤0,05, a viabilidade celular dos fibroblastos L929 à 47,70%, assim como o extrato seco de S. adstringens nas concentrações de 225 μg/mL. No capítulo 3 está apresentada a obtenção de extratos secos de S. adstringens por atomização. Foram avaliadas a influência da proporção do adjuvante de secagem, da temperatura do ar de secagem e da vazão do ar de secagem sobre os fenóis totais, GEGC, taninos totais, atividade antioxidante total e rendimento do processo de secagem sobre os extratos secos obtidos, os resultados significativos (p≤0,05) foram obtidos por ANOVA e representados pela metodologia de superfície de resposta. O aumento do adjuvante de secagem não foi significativamente favorável apenas para o rendimento do processo de secagem. O aumento da temperatura do ar de secagem apresentou um comportamento quadrático negativo para fenóis e positivo para os taninos totais. O aumento da vazão do ar de secagem apresentou comportamento linear e quadrático negativo para a atividade antioxidante total e o rendimento do processo de secagem não sofreu influência de nenhuma variável. Os extratos secos foram caracterizados quanto aos seus teores de voláteis, atividade de água e as morfologias das partículas foram observadas por microscopia eletrônica de varredura. No capítulo 4, está descrita a otimização de extração de fenóis totais, taninos totais e galato de epigalocatequina dos pós das cascas de S. adstringens em banho ultrassônico, utilizando um planejamento BoxBenhken 33. A aplicação e validação da função estatística desejabilidade global demonstrou que a proporção de droga/solvente de 4mg/mL, o tempo de 30 min e a solução hidroalcoólica de 65% (v/v) são ideais para a extração simultânea destes metabólitos. Os resultados permitem concluir que os extratos foram obtidos e padronizados com qualidade, e que os mesmo apresentam potencial para investigação de suas atividades biológicas e/ou posterior inserção em formulações fitoterápicas.

L’objectif de la thèse est une mise au point des espèces de Stephania répertoriées au Cambodge par l’étude systématique, phytochimique et pharmacologique. Du point de vue systématique, les travaux ont permis de répertorier sept espèces et deux variétés de Stephania au Cambodge : S. cambodica, S. japonica var. discolor et var. timoriensis, S. oblata, S. pierrei, S. rotunda (espèce-type), S. suberosa, et S. venosa. La clef de détermination établie lors de ce travail de thèse permet d’identifier ces espèces. D’après les résultats phylogénétiques, seul S. pierrei est monophylétique, ce qui représente moins de 10% des espèces de Stephania. L’étude chimique de cinq espèces a été réalisée par des méthodes chromatographiques (CLHP, UCLHP) et des méthodes spectroscopiques. Différents alcaloïdes ont été proposés comme marqueurs : la palmatine, la roémérine et la tétrahydropalmatine pour S. cambodica et S. rotunda, la cépharanthine, la tétrahydropalmatine et la xylopinine pour S. suberosa, la cépharanthine pour S. pierrei, la crébanine, et la tétrahydropalmatine pour S. venosa. Neuf alcaloïdes ont été isolés à partir du tubercule de S. cambodica, dont un nouveau composé dénommé «angkorwatine ». Une méthode d’éco-extraction des alcaloïdes assistée par ultrasons a été mise au point et optimisée par un plan d’expérience pour cette espèce. Une méthode analytique par UCLHP a été développée et validée pour le contrôle qualité de S. cambodica. Les résultats montrent que les extraits hydroéthanoliques obtenus par macération à partir du tubercule de S. suberosa, S. venosa et S. rotunda présentent une activité anti-inflammatoire importante (in vitro) avec un ratio anti-inflammatoire élevé. / The thesis aims to provide update on species of the genus Stephania recorded in Cambodia by systematic, phytochemical and pharmacological studies.The systematic study allowed to list seven species and two varieties of Stephania in Cambodia: S. cambodica, S. japonica var. discolor and var. timoriensis, S. oblata, S. pierrei, S. rotunda (type species), S. suberosa and S. venosa. The presence of S. japonica var. timoriensis and S. oblata is reported for the first time in Cambodia. According to the phylogenetic results, only S. pierrei is monophyletic, which represents less than 10% of species in the genus Stephania. Chemical studies on the tubers of five species were carried out using chromatographic methods (HPLC, UHPLC) and spectroscopic methods. Different isolated alkaloids have been proposed as markers: palmatine, roemerine and tetrahydropalmatine for S. cambodica and S. rotunda, cepharanthine, tetrahydropalmatine, and xylopinine for S. suberosa, cepharanthine for S. pierrei, crebanine, and tetrahydropalmatine for S. venosa. Nine alkaloids belonging to five classes were isolated from the tuber of S. cambodica and a new compound namely "angkorwatine" was described. Ultrasound assisted extraction of alkaloids was developed and optimized by Design of Experiment for this species. This innovative extraction is a good alternative to conventional methods. An analytical UHPLC method was developed and validated for the quality control of S. cambodica. Results show that hydroethanolic extracts obtained by maceration of S. suberosa, S. rotunda and S. venosa exhibit significant anti-inflammatory activity with high anti-inflammatory ratio (selectivity index).