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Sambucus Nigra L UNIVERSIDADE FEDERAL DO RIO GRANDE DO SUL FACULDADE DE FARMÁCIA PROGRAMA DE PÓS-GRADUAÇÃO EM CIÊNCIAS FARMACÊUTICAS Análise Botânica, Química e Biológica Comparativa entre Flores das Espécies Sambucus nigra L. e Sambucus australis Cham. & Schltdl. e Avaliação Preliminar da Estabilidade MARINA SCOPEL PORTO ALEGRE, 2005 2 UNIVERSIDADE FEDERAL DO RIO GRANDE DO SUL FACULDADE DE FARMÁCIA PROGRAMA DE PÓS-GRADUAÇÃO EM CIÊNCIAS FARMACÊUTICAS Análise Botânica, Química e Biológica Comparativa entre Flores das Espécies Sambucus nigra L. e Sambucus australis Cham. & Schltdl. e Avaliação Preliminar da Estabilidade Dissertação apresentada por Marina Scopel para obtenção do GRAU DE MESTRE em Ciências Farmacêuticas. Orientador: Profa. Dra. Amélia T. Henriques Co-orientador: Profa. Dra. Lílian A. Mentz 3 Dissertação apresentada ao Programa de Pós-Graduação em Ciências Farmacêuticas, em nível de Mestrado da Faculdade de Farmácia da Universidade Federal do Rio Grande do Sul e aprovada em 29 de agosto de 2005, pela Banca Examinadora constituída por: Profa. Dra. Edna Sayuri Suyenaga Universidade Luterana do Brasil Prof. Dr. Leandro Machado Rocha Universidade Federal Fluminense Profa. Dra. Mara Ritter Universidade Federal do Rio Grande do Sul Prof. Dr. Martin Steppe Universidade Federal do Rio Grande do Sul S422a Scopel, Marina Análise Botânica, Química e Biológica Comparativa entre Flores das Espécies Sambucus nigra L. e Sambucus australis Cham. & Schltdl. e Avaliação Preliminar de sua Estabilidade / Marina Scopel – Porto Alegre: UFRGS, 2005. – 263p.: il., gráf., tab. Dissertação (mestrado). UFRGS. Faculdade de Farmácia. Programa de Pós-Graduação em Ciências Farmacêuticas. 1. Sambucus nigra. 2. Sambucus australis. 3. Cromatografia líquida de alta eficiência. 4. Sabugueiro. 5. Caprifoliaceae. I. Henriques, Amélia T. II. Mentz, Lílian A. III. Título. CDU: 547.9:582.973 Bibliotecárias responsáveis: Heloisa do Canto Canabarro, CRB 10/1036 Cláudia da Silva Gonçalves, CRB 10/1012 4 Este trabalho foi subvencionado pela CAPES e desenvolvido nos Laboratórios de Farmacognosia e Fitoquímica, de Controle de Qualidade e de Botânica da Faculdade de Farmácia desta Faculdade, bem como no Departamento de Produtos Naturais e Alimentos da Faculdade de Farmácia da Universidade Federal do Rio de Janeiro. 5 6 “O maior prazer que a vida proporciona é a superação das dificuldades, passar de uma etapa do sucesso para a outra, criar novos desejos e satisfazê-los”. SAMUEL JOHNSON 7 8 Aos alicerces Assis e Beatriz, a irmã Cláudia, a tia Cuca e especialmente à nossa luz, meu afilhado, João Victor Dedico. 9 10 AGRADECIMENTOS A Deus por olhar por mim. À professora Dra. Amélia T. Henriques por ter acreditado no meu potencial e por sua orientação. À professora Dra. Lílian A. Mentz pela co-orientação e a professora Dra. Eliane Nunes, Msc. Márcia Vignoli, Msc. Giovana Vendruscolo pelo exaustivo e paciente trabalho botânico realizado. Ao professor Dr. André A. Souto pelos ensinamentos na iniciação científica (PUCRS) e às professoras Dras. Clarice M. Azevedo e Renata P. Limberger pelo incentivo ao ingresso no programa de pós-graduação desta universidade. Às companheiras de luta Ana Lúcia Arigony e Camila Sebben, aos amigos do laboratório Carolina Passos, Eduardo Konrath, Fabiane Farias, Jean Andrade, Rafaela Marin, Raquel Giordani, Roger Dresh, Rogério Petersen, Valquiria Reis, aos novos integrantes Dimas Jr., Graziele Ramos, Letícia Pagliosa, Tiago Souza e Maria T. Kreinecker e especialmente às pacientes Ana Aboy, Melissa Schwanz e Miriam Apel. Além das amigas e vizinhas Msc. Ana Cristina Stein, Ana Paula Bernardi, Carolina Nör, Daniela Albring, Daniela Fritz, Juliana Haas, Kenia Correa, Simone Cargnin e aos Profs. Elfrides Schapoval, Gilsane von Poser e José Ângelo Zuanazzi. Aos mestres Cláudia A. S. Pires, Nael Hamid, Paulo Artur Coelho, Tatiana Castilhos, pelo alegre acolhimento e momentos felizes vividos no laboratório. As amigas, parceiras em momentos de descontração, ouvidoras e apoiadoras constantes Cassiana Viau, Clarissa Xavier, Fernanda Ponticelli, Kátia Verenzuck, Lisiane Fanton e ao amigo Paulo Mendes. E àquelas pessoas que, de alguma forma, contribuíram para a realização deste trabalho ou estiveram presentes ao meu lado nesta jornada. 11 12 SUMÁRIO LISTA DE FIGURAS ........................................................................................ xvii LISTA DE TABELAS ....................................................................................... xxi LISTA DE EQUAÇÕES ................................................................................... xxiii LISTA DE ABREVIATURAS ............................................................................ xxv RESUMO ......................................................................................................... xxvii ABSTRACT ..................................................................................................... xxix INTRODUÇÃO GERAL ................................................................................... 1 OBJETIVOS GERAIS ...................................................................................... 5 CAPÍTULO 1 DESCRIÇÃO E ANALISE BOTÂNICA ...................................... 9 1.1 Introdução .................................................................................................. 11 1.2 Objetivos .................................................................................................... 13 1.3 Revisão da Literatura ................................................................................ 14 1.3.1 Sambucus nigra L. x Sambucus australis Cham. & Schltdl. ................... 14 1.4 Materiais e Métodos .................................................................................. 17 1.4.1 Material vegetal ...................................................................................... 17 1.4.2 Análise macroscópica ............................................................................. 19 1.4.3 Análise microscópica .............................................................................. 19 1.4.4 Testes histoquímicos .............................................................................. 20 1.5 Resultados ................................................................................................. 22 1.5.1 Análise macroscópica ............................................................................. 22 1.5.2 Análise microscópica .............................................................................. 31 CAPÍTULO 2 AVALIAÇÃO QUÍMICA QUALI E QUANTITATIVA ................... 69 2.1 Introdução .................................................................................................. 71 2.2 Objetivos .................................................................................................... 78 2.3 Revisão da Literatura ................................................................................ 79 2.3.1 Estudos químicos em Sambucus nigra L. e Sambucus australis Cham & Schltdl........................................................................................................... 79 2.3.2 Análises cromatográficas e espectrofotométricas................................... 82 2.3.3 Estudo de estabilidade............................................................................ 83 2.4 Materiais e Métodos................................................................................... 85 13 2.4.1 Material vegetal....................................................................................... 85 2.4.2 Determinação de perda por dessecação................................................. 85 2.4.3 Determinação de materiais estranhos (contaminantes).......................... 86 2.4.4 Determinação de cinzas totais................................................................ 86 2.4.5 Análise qualitativa.................................................................................... 86 2.4.5.1 Avaliação do perfil cromatográfico utilizando cromatografia em camada delgada............................................................................................... 86 2.4.5.2 Avaliação do perfil cromatográfico utilizando cromatografia líquida de alta eficiência.................................................................................................... 87 2.4.6 Análise quantitativa................................................................................. 90 2.4.6.1 Validação do método por cromatografia líquida de alta eficiência....... 91 2.4.7 Determinação do teor de flavonóides totais............................................ 95 2.4.8 Estudo preliminar da estabilidade acelerada........................................... 97 2.4.9 Análises das amostras coletadas, adquiridas no comércio ou fornecidas pelo Laboratório de Farmacognosia (UFRGS)............................... 98 2.5 Resultados e Discussão............................................................................. 100 2.5.1 Determinação da perda por dessecação................................................. 100 2.5.2 Determinação de materiais estranhos (contaminantes).......................... 101 2.5.3 Determinação de cinzas totais................................................................ 102 2.5.4 Análise qualitativa.................................................................................... 103 2.5.5 Análise quantitativa................................................................................. 112 2.5.5.1 Validação do método por cromatografia líquida de alta eficiência......
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