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Fernanda Anselmo Moreira Janayne Gagliano Análise do potencial farmacológico de Merostachys pluriflora Munro ex. E. G. Camus., uma espécie de bambu nativo da Mata Atlântica. Analysis of the pharmacological potential of Merostachys pluriflora Munro ex. E. G. Camus., a specie of native bamboo from Atlantic Forest. VERSÃO CORRIGIDA São Paulo 2016 Janayne Gagliano Análise do potencial farmacológico de Merostachys pluriflora Munro ex. E. G. Camus., uma espécie de bambu nativo da mata atlântica. Analysis of the pharmacological potential of Merostachys pluriflora Munro ex. E. G. Camus., a specie of native bamboo from Atlantic Forest. Dissertação apresentada ao Instituto de Biociências da Universidade de São Paulo, para a obtenção de Título de Mestre em Botânica, na Área de Recursos Econômicos Vegetais. Orientadora: Profª Dr.ª Cláudia Maria Furlan SÃO PAULO 2016 Gagliano, Janayne Análise do potencial farmacológico de Merostachys pluriflora Munro ex. E. G. Camus., uma espécie de bambu nativo da Mata Atlântica. 102 páginas. Dissertação (Mestrado) – Instituto de Biociências da Universidade de São Paulo. Departamento de Botânica. Palavra-Chave: bambus, Mata Atlântica, substâncias fenólicas, antioxidantes, antimicrobiano, anti-HIV-1 Universidade de São Paulo. Instituto de Biociências. Departamento de Botânica. Comissão Julgadora: ______________________ _____________________ Prof(a). Dr(a). Prof(a). Dr(a). _________________________ Profa. Dra. Cláudia Maria Furlan Orientadora A aquele que foi meu melhor amigo, meu pai. Agradecimentos Foram tantas pessoas queridas envolvidas durante esse período, que espero não esquecer ninguém em meus agradecimentos! À minha querida família, em especial meus pais Eduardo e Neide, pelo apoio de uma vida inteira, por sempre acreditarem em mim e me incentivarem incessantemente a seguir os meus sonhos, pois sem eles eu não teria chego até aqui. Aos meus irmãos, Aline e Renan, por todo apoio, conselhos, amizade, muitíssimo obrigada meus amores! Ao meu noivo Rogério, por todo seu companheirismo, paciência, apoio, e por acreditar no meu potencial, obrigada amor! À minha mãe de coração, Maria, por ter me conduzido ao caminho das plantas, por todo o seu amor e conhecimento sobre elas, obrigada minha querida! À minha querida orientadora Cláudia, por todo aprendizado, por sua dedicação, apoio, paciência; e principalmente por ter me aceito como sua aluna, foi um privilégio ter trabalhado com você durante esses dois anos, muito obrigada Clau! Aos professores da fitoquímica, que direta ou indiretamente, contribuíram para a realização deste trabalho. Ao professor Marcelo por toda ajuda com as análises de RMN, a professora Deborah por todas as valiosas contribuições em minha banca de qualificação, aos professores, Maria Luiza e Antonio Salatino, por suas aulas incrivelmente inspiradoras, muito obrigada a todos! A todos os amigos que fiz no laboratório de fitoquímica, em especial, Cris, Cinthia, Fê Anselmo, Fê Rezende, Wilton, Priscila, Alice, Kátia, Dalila, muito obrigado por todos os almoços, cafés, conversas, risadas, ajudas na bancada, valeu turma!!! Aos funcionários do laboratório de fitoquímica, Mourisa e Aline, por toda ajuda com as análises das amostras, pelas dicas, paciência; ao Leandro, por todo o suporte na organização dos laboratórios, muito obrigada a todos! Aos meus queridos amigos da graduação, Ani e Leo, por toda amizade durante esses anos, pelo companheirismo, por todos os momentos que estivemos juntos, obrigada amigos! À minha querida amiga Aline Lopes, por ter me apresentado ao “mundo dos bambus”, muito obrigada! Às pesquisadoras do IBt, Maria Tereza Guaratini, por toda ajuda na escolha e coleta da espécie de estudo; à Regina Shirasuna, pela identificação da mesma, muito obrigada! Ao Instituto de Biociências da Universidade de São Paulo por conceder toda a infraestrutura para a realização deste projeto; e a CAPES pela viabilização financeira, através da bolsa concedida. Índice Índice Capítulo 1 - Introdução Geral ............................................................................................................................ 3 1. Introdução .................................................................................................................................................... 3 1.1. Bambus: filogenia, distribuição geográfica e ecologia ............................................................................ 3 1.2. Bambus: uso na medicina tradicional ..................................................................................................... 6 1.3. Espécie vegetal de estudo ....................................................................................................................... 8 1.4. Área de coleta ......................................................................................................................................... 9 2. Justificativa e Hipóteses ............................................................................................................................. 10 3. Objetivos .................................................................................................................................................... 11 4. Referências Bibliográficas........................................................................................................................... 12 Capítulo 2 - Caracterização química de Merostachys pluriflora ..................................................................... 15 1. Introdução .................................................................................................................................................. 17 1.1. Os metabólitos vegetais ........................................................................................................................ 17 2. Materiais e Métodos .................................................................................................................................. 22 2.1. Preparo das amostras............................................................................................................................ 22 2.1.1. Extração e partição .................................................................................................................... 22 2.2.Quantificações de metabólitos primários ................................................................................................. 23 2.2.1. Amido ......................................................................................................................................... 23 2.2.2. Carboidratos solúveis ................................................................................................................. 24 2.2.3. Lipídeos ...................................................................................................................................... 25 2.3. Quantificações de substâncias fenólicas ............................................................................................... 26 2.3.1. Doseamento de fenóis totais ..................................................................................................... 26 2.3.2. Doseamento de flavonas/flavonóis ........................................................................................... 26 2.3.3. Doseamento de proantocianidinas (Taninos condensados) ...................................................... 26 2.3.4. Doseamento de taninos totais ................................................................................................... 27 2.3.5. Caracterização e quantificação das substâncias fenólicas via CLAE-DAD .................................. 27 2.4. Fracionamento via CLAE-DAD semi-preparativo ................................................................................... 28 2.5. Identificação via Ressonância Magnética Nuclear (RMN) dos constituintes fenólicos isolados .......... 28 3. Resultados e Discussão............................................................................................................................... 29 3.1. Rendimentos dos extratos e fases ........................................................................................................ 29 3.2. Quantificações de metabólitos primários ............................................................................................. 30 3.2.1. Amido e carboidratos solúveis ................................................................................................... 30 3.2.2. Lipídeos ....................................................................................................................................... 31 3.3. Quantificação de substâncias fenólicas .................................................................................................... 34 i Índice 3.3.1. Fenóis totais e flavonoides .......................................................................................................... 34 3.3.2. Taninos totais e proantocianidinas ............................................................................................. 36 3.3.3. Caracterização e quantificação de substâncias fenólicas via CLAE-DAD .................................... 37 3.4. Fracionamento e identificação dos constituintes fenólicos via RMN ...................................................... 46 4. Conclusões ................................................................................................................................................... 51 5. Referências bibliográficas ...........................................................................................................................
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