Caracterização Nutricional E Bioatividade De Cogumelos Silvestres: Lactarius Citriolens, Lactarius Turpis, Leucopaxillus Candidus E Morchella Conica

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Caracterização Nutricional E Bioatividade De Cogumelos Silvestres: Lactarius Citriolens, Lactarius Turpis, Leucopaxillus Candidus E Morchella Conica Caracterização nutricional e bioatividade de cogumelos silvestres: Lactarius citriolens, Lactarius turpis, Leucopaxillus candidus e Morchella conica. Vanessa Ariana Azevedo Vieira Dissertação apresentada ao Instituto Politécnico de Bragança e à Universidade de Salamanca para obtenção do Grau de Mestre em Farmácia e Química de Produtos Naturais Orientado por Isabel Cristina Fernandes Rodrigues Ferreira Anabela Lourenço Rodrigues Martins Bragança 2014 Lactarius citriolens, Lactarius turpis, Leucopaxillus candidus e Morchella conica 2 Vanessa Vieira Caracterização nutricional e bioatividade de cogumelos silvestres Índice Índice de Figuras .............................................................................................................. 5 Índice de Tabelas .............................................................................................................. 5 Agradecimentos ................................................................................................................ 7 Abreviaturas...................................................................................................................... 9 Resumo ........................................................................................................................... 13 Abstract ........................................................................................................................... 14 Enquadramento do estudo .............................................................................................. 15 1. Introdução ............................................................................................................... 19 1.1. Caracterização das amostras ............................................................................ 19 1.2. Valor nutricional de cogumelos silvestres ....................................................... 24 1.3. Moléculas bioativas em cogumelos silvestres ................................................. 31 1.4. Bioatividade de cogumelos silvestres .............................................................. 35 1.5. Objetivos .......................................................................................................... 40 2. Material e Métodos ................................................................................................. 43 2.1. Amostragem das espécies de cogumelos ......................................................... 43 2.2. Padrões e Reagentes ......................................................................................... 43 2.3. Composição química e nutricional .................................................................. 44 2.3.1. Determinação de macronutrientes ............................................................ 44 2.3.2. Determinação de açúcares livres .............................................................. 44 2.3.3. Determinação de ácidos gordos ................................................................ 45 2.3.4. Determinação de tocoferóis ...................................................................... 46 2.3.5. Determinação de ácidos orgânicos ........................................................... 46 2.3.6. Determinação de ácidos fenólicos e compostos relacionados .................. 47 2.4. Bioatividade dos extratos metanólicos das amostras ....................................... 48 3 Vanessa Vieira Lactarius citriolens, Lactarius turpis, Leucopaxillus candidus e Morchella conica 2.4.1. Preparação dos extratos ............................................................................ 48 2.4.2. Ensaios de atividade antioxidante............................................................. 48 2.4.3. Ensaio de Folin-Ciocalteu ........................................................................ 48 2.4.4. Ensaio de Ferricianeto/Azul da Prússia .................................................... 49 2.4.5. Ensaio da atividade captadora de radicais DPPH ..................................... 49 2.4.6. Ensaio de inibição de descoloração de β-caroteno/linoleato .................... 49 2.4.7. Ensaio de substâncias reativas do ácido tiobarbitúrico (TBARS) ............ 50 2.4.8. Ensaios de atividade antibacteriana .......................................................... 50 2.4.9. Ensaios de atividade antifúngica .............................................................. 51 2.4.10. Ensaios de citotoxicidade em células de fígado .................................... 52 2.5. Análises estatísticas ......................................................................................... 53 3. Resultados e Discussão ........................................................................................... 55 3.1. Nutrientes, não nutrientes e bioactividade de Morchella conica silvestre de diferentes origens ........................................................................................................ 55 3.2. Expansão do conhecimento do género Lactarius relativamente à composição química e atividade antioxidante ................................................................................ 64 3.3. O que ainda não foi descrito sobre o cogumelo silvestre Leucopaxillus candidus (Bres.) Singer .............................................................................................. 73 4. Considerações Finais .............................................................................................. 79 5. Bibliografia ............................................................................................................. 81 4 Vanessa Vieira Caracterização nutricional e bioatividade de cogumelos silvestres Índice de Figuras Figura 1. Estrutura mais comum de um corpo de frutificação ....................................... 21 Figura 2. Lactarius turpis Fr., Lactarius citriolens Pouzar, Leucopaxillus candidus (Bres.) Singer e Morchella conica Pers. ......................................................................... 21 Figura 3. Estrutura química dos aminoácidos mais abundantes em cogumelos silvestres ........................................................................................................................................ 26 Figura 4. Estrutura química do ácido palmítico, ácido oleico e ácido linoleico............. 28 Figura 5. Estrutura química da glucose, manitol e trealose ............................................ 30 Figura 6. Estrutura química dos tocoferóis e tocotrienóis .............................................. 32 Figura 7. Estrutura química do ácido hidroxibenzóico e hidroxicinâmico..................... 33 Figura 8. Estrutura química do ácido cítrico os ácidos oxálico, quínico, málico, cítrico e fumárico. ......................................................................................................................... 35 Figura 9. Perfil individual de açúcares de M. conica de Portugal e da Sérvia. ............. 58 Figura 10. Perfil individual de açúcares de L. turpis e L. citriolens .............................. 71 Figura 11. Perfil individual de ácidos orgânicos de L. citriolens e L. turpis .................. 72 Índice de Tabelas Tabela 1. Macronutrientes, açúcares livres, ácidos gordos e tocoferóis dos corpos de frutificação de M. conica expressos em massa seca (média ± DP).. .............................. 56 Tabela 2. Ácidos orgânicos e compostos fenólicos dos corpos de frutificação de M. conica expressos em massa seca (média ± DP). ............................................................. 59 Tabela 3. Atividade antioxidante dos extratos metanólicos de M. conica (média DP)... ........................................................................................................................................ 61 5 Vanessa Vieira Lactarius citriolens, Lactarius turpis, Leucopaxillus candidus e Morchella conica Tabela 4. Atividade antibacteriana e antifúngica dos extratos metanólicos de M. conica e drogas de referência (mg/mL) (média DP). .............................................................. 63 Tabela 5. Macronutrientes, açúcares livres, ácidos gordos e tocoferóis dos corpos de frutificação de L. citriolens e L. turpis expressos em massa seca (média ± DP). .......... 65 Tabela 6. Ácidos orgânicos e compostos fenólicos dos corpos de frutificação de L. citriolens e L. turpis expressos em massa seca (média ± DP). ....................................... 69 Tabela 7. Atividade antioxidante dos extratos metanólicos de L. citriolens e L. turpis (média DP). ................................................................................................................. 70 Tabela 8. Macronutrientes, açúcares livres, ácidos gordos e tocoferóis dos corpos de frutificação de Leucopaxillus candidus expressos em massa seca (média ± DP)……...74 Tabela 9. Ácidos orgânicos e compostos fenólicos dos corpos de frutificação de Leucopaxillus candidus expressos em massa seca (média ± DP). ................................. 76 Tabela 10. Atividade antioxidante dos extratos metanólicos de Leucopaxillus candidus (média DP). ................................................................................................................. 77 6 Vanessa Vieira Caracterização nutricional e bioatividade de cogumelos silvestres Agradecimentos Para a realização desta dissertação foram vários os intervenientes que colaboraram direta e indiretamente, os quais merecem o meu reconhecimento e gratidão. Às minhas orientadoras, a Doutora Isabel Ferreira e a Doutora Anabela Martins, pelo apoio, orientação e dedicação que sempre demonstraram. Agradeço
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