Os compostos fenólicos presentes em uvas e vinhos apresentam atividades antioxidantes benéficas à saúde humana, além de indicarem a qualidade desses produtos. Assim, esta tese teve como objetivo desenvolver, validar e aplicar métodos para a análise dos compostos fenólicos e avaliar a atividade antioxidante de uvas e vinhos tintos finos provenientes de uma nova região vitivinícola brasileira (Campanha Gaúcha - RS, Brasil). Ao total, foram utilizadas 103 amostras de vinhos finos tintos obtidos a partir de diferentes processos de vinificação (experimentais e comerciais), variedades (10), safras (2004 a 2015) e regiões (97 das microrregiões Ocidental, Central e Meridional da Campanha Gaúcha e seis de outras regiões do RS). Além disso, foram utilizadas oito amostras de uvas finas tintas da Campanha Gaúcha, das safras 2014 e 2015. O primeiro método, por cromatografia líquida de alta eficiência com detector de arranjo de diodos (HPLC-DAD), com propósito de dosar dois marcadores de compostos fenólicos, trans-resveratrol e quercetina, mostrou-se adequado para diferenciar 12 vinhos tintos do RS. O segundo método, por HPLC-DAD e espectrometria de massas de alta resolução, possibilitou o doseamento de seis fenóis bioativos (trans-resveratrol, quercetina, miricetina, quempferol, trans-Ɛ-viniferina e ácido trans-cinâmico) em 87 amostras da Campanha Gaúcha. Os teores mais elevados de compostos fenólicos foram encontrados em: variedades Arinarnoa, Marselan, Rebo, Syrah, Tempranillo e Teroldego; microrregião da Campanha Central; safra de 2013. Os vinhos foram classificados mediante análises de agrupamento hierárquico (HCA) e de componentes principais (PCA), conforme seus respectivos processos de vinificação, variedades e microrregiões. O terceiro e último método abordado foi com o intuito de analisar um maior número de compostos fenólicos, o que foi possível através da técnica de extração em fase sólida por troca catiônica forte. Esse método foi utilizado para análises de uvas e seus vinhos experimentais correspondentes da Campanha Gaúcha, por HPLC-DAD e por cromatografia líquida de ultra eficiência acoplado a espectrômetro de massas (UPLC-MS). Uvas com alto teor de galato de (-)-epicatequina originaram vinhos com elevada concentração de fenóis biativos. Tais amostras também foram analisadas quanto ao índice de fenóis totais e quanto à atividade antioxidante in vitro, demonstrando uma correlação positiva entre uvas e vinhos das mesmas variedades. Nesse mesmo estudo, foi possível atribuir atividades antioxidantes in vivo (modelo Caenorhabditis elegans) superiores nas amostras com teores mais elevados de astilbina e flavanóis, como a (+)-catequina, a (-)-epicatequina e a (-)-epigalocatequina. Assim, esses compostos, principalmente a astilbina, poderão ser utilizados como modelos para modificações moleculares na área de química medicinal. Além disso, o estudo dos compostos fenólicos pode contribuir para a melhoria da qualidade de vinhos da Campanha Gaúcha, trazendo benefícios diretos à vitivinicultura nacional e indiretos a toda a sociedade. / Phenolic compounds found in grapes and wines show antioxidant activities with health benefits for humans, and also indicate the quality of these products. Therefore, this thesis aimed to develop, validate and investigate methods for phenolic compound analysis, and evaluate the antioxidant activity of grapes and wines from a new Brazilian viticulture region (Campanha Gaúcha, Southern Brazil). In total, 103 red wine samples were used. These samples were obtained from different winemaking processes (experimental and commercial), varieties (10), vintages (2004 to 2015), and regions (97 from occidental, central e meridional from Campanha Gaúcha and six from other Southern Brazilian regions). Eight grape samples, 2014 and 2015 vintages, from Campanha Gaúcha were also used. The first method, using high-performance liquid chromatography with a diode array detector (HPLC-DAD) was used to quantify two phenolic compound markers, trans-resveratrol, and quercetin, and was adequate for differentiating 12 red wines from Southern Brazil. The second method, using HPLC-DAD and high-resolution mass spectrometry, enabled the quantification of six bioactive phenols (trans-resveratrol, quercetin, myricetin, kaempferol, trans-Ɛ-viniferin, and trans-cinnamic acid) in 87 samples from Campanha Gaúcha. The highest levels of phenolic compounds were found in the Arinarnoa, Marselan, Rebo, Syrah, Tempranillo, and Teroldego varieties; central Campanha Gaúcha micro-region varieties; and the 2013 vintage. The wines were classified according to their winemaking processes, varieties, and micro-regions using hierarchical cluster and principal component analyses. The third and final method aimed to analyze a large number of phenolic compounds using a solid phase extraction technique employing strong cation exchange. This method was used to analyze grapes and their corresponding wines from Campanha Gaúcha using HPLC-DAD and ultra-performance liquid chromatography coupled with mass spectrometry. Grapes with high levels of (-)-epicatechin galate corresponded to wines more concentrated in bioactive phenols. The samples were also analyzed using the total phenol content index and in vitro antioxidant activity, showing a positive correlation between grapes and wines of the same varieties. In the same study, it was possible to attribute the in vivo antioxidant activities (Caenorhabditis elegans model) to samples with higher levels of astilbin and flavanols such as (+)-catechin, (-)-epicatechin, and (-)-epigalocatechin. Therefore, such compounds, mainly astilbin, may be used for a molecular modification model in medicinal chemistry. Moreover, the study of phenolic compounds may contribute to the improvement of the quality of wines from Campanha Gaúcha, and can bring several direct benefits for national viticulture and indirect benefits to all society.

Os compostos fenólicos presentes em uvas e vinhos apresentam atividades antioxidantes benéficas à saúde humana, além de indicarem a qualidade desses produtos. Assim, esta tese teve como objetivo desenvolver, validar e aplicar métodos para a análise dos compostos fenólicos e avaliar a atividade antioxidante de uvas e vinhos tintos finos provenientes de uma nova região vitivinícola brasileira (Campanha Gaúcha - RS, Brasil). Ao total, foram utilizadas 103 amostras de vinhos finos tintos obtidos a partir de diferentes processos de vinificação (experimentais e comerciais), variedades (10), safras (2004 a 2015) e regiões (97 das microrregiões Ocidental, Central e Meridional da Campanha Gaúcha e seis de outras regiões do RS). Além disso, foram utilizadas oito amostras de uvas finas tintas da Campanha Gaúcha, das safras 2014 e 2015. O primeiro método, por cromatografia líquida de alta eficiência com detector de arranjo de diodos (HPLC-DAD), com propósito de dosar dois marcadores de compostos fenólicos, trans-resveratrol e quercetina, mostrou-se adequado para diferenciar 12 vinhos tintos do RS. O segundo método, por HPLC-DAD e espectrometria de massas de alta resolução, possibilitou o doseamento de seis fenóis bioativos (trans-resveratrol, quercetina, miricetina, quempferol, trans-Ɛ-viniferina e ácido trans-cinâmico) em 87 amostras da Campanha Gaúcha. Os teores mais elevados de compostos fenólicos foram encontrados em: variedades Arinarnoa, Marselan, Rebo, Syrah, Tempranillo e Teroldego; microrregião da Campanha Central; safra de 2013. Os vinhos foram classificados mediante análises de agrupamento hierárquico (HCA) e de componentes principais (PCA), conforme seus respectivos processos de vinificação, variedades e microrregiões. O terceiro e último método abordado foi com o intuito de analisar um maior número de compostos fenólicos, o que foi possível através da técnica de extração em fase sólida por troca catiônica forte. Esse método foi utilizado para análises de uvas e seus vinhos experimentais correspondentes da Campanha Gaúcha, por HPLC-DAD e por cromatografia líquida de ultra eficiência acoplado a espectrômetro de massas (UPLC-MS). Uvas com alto teor de galato de (-)-epicatequina originaram vinhos com elevada concentração de fenóis biativos. Tais amostras também foram analisadas quanto ao índice de fenóis totais e quanto à atividade antioxidante in vitro, demonstrando uma correlação positiva entre uvas e vinhos das mesmas variedades. Nesse mesmo estudo, foi possível atribuir atividades antioxidantes in vivo (modelo Caenorhabditis elegans) superiores nas amostras com teores mais elevados de astilbina e flavanóis, como a (+)-catequina, a (-)-epicatequina e a (-)-epigalocatequina. Assim, esses compostos, principalmente a astilbina, poderão ser utilizados como modelos para modificações moleculares na área de química medicinal. Além disso, o estudo dos compostos fenólicos pode contribuir para a melhoria da qualidade de vinhos da Campanha Gaúcha, trazendo benefícios diretos à vitivinicultura nacional e indiretos a toda a sociedade. / Phenolic compounds found in grapes and wines show antioxidant activities with health benefits for humans, and also indicate the quality of these products. Therefore, this thesis aimed to develop, validate and investigate methods for phenolic compound analysis, and evaluate the antioxidant activity of grapes and wines from a new Brazilian viticulture region (Campanha Gaúcha, Southern Brazil). In total, 103 red wine samples were used. These samples were obtained from different winemaking processes (experimental and commercial), varieties (10), vintages (2004 to 2015), and regions (97 from occidental, central e meridional from Campanha Gaúcha and six from other Southern Brazilian regions). Eight grape samples, 2014 and 2015 vintages, from Campanha Gaúcha were also used. The first method, using high-performance liquid chromatography with a diode array detector (HPLC-DAD) was used to quantify two phenolic compound markers, trans-resveratrol, and quercetin, and was adequate for differentiating 12 red wines from Southern Brazil. The second method, using HPLC-DAD and high-resolution mass spectrometry, enabled the quantification of six bioactive phenols (trans-resveratrol, quercetin, myricetin, kaempferol, trans-Ɛ-viniferin, and trans-cinnamic acid) in 87 samples from Campanha Gaúcha. The highest levels of phenolic compounds were found in the Arinarnoa, Marselan, Rebo, Syrah, Tempranillo, and Teroldego varieties; central Campanha Gaúcha micro-region varieties; and the 2013 vintage. The wines were classified according to their winemaking processes, varieties, and micro-regions using hierarchical cluster and principal component analyses. The third and final method aimed to analyze a large number of phenolic compounds using a solid phase extraction technique employing strong cation exchange. This method was used to analyze grapes and their corresponding wines from Campanha Gaúcha using HPLC-DAD and ultra-performance liquid chromatography coupled with mass spectrometry. Grapes with high levels of (-)-epicatechin galate corresponded to wines more concentrated in bioactive phenols. The samples were also analyzed using the total phenol content index and in vitro antioxidant activity, showing a positive correlation between grapes and wines of the same varieties. In the same study, it was possible to attribute the in vivo antioxidant activities (Caenorhabditis elegans model) to samples with higher levels of astilbin and flavanols such as (+)-catechin, (-)-epicatechin, and (-)-epigalocatechin. Therefore, such compounds, mainly astilbin, may be used for a molecular modification model in medicinal chemistry. Moreover, the study of phenolic compounds may contribute to the improvement of the quality of wines from Campanha Gaúcha, and can bring several direct benefits for national viticulture and indirect benefits to all society.

Orientadores: Maria Aparecida Azevedo Pereira da Silva, Flavia Maria Netto / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-10T07:33:51Z (GMT). No. of bitstreams: 1 Biasoto_AlineCamaraoTelles_M.pdf: 1388304 bytes, checksum: 23ab6b8b00ef4435f7099f4b17af8def (MD5) Previous issue date: 2008 / Resumo: No Estado de São Paulo, a produção de vinhos de mesa de uvas americanas e/ou híbridas, supera grandemente a de uva Vitis vinífera. No entanto, pouco conhecimento se tem sobre a tecnologia de fabricação utilizada para a elaboração desses vinhos no Estado de São Paulo, seu nível de segurança, perfil sensorial e aceitabilidade entre os consumidores paulistas. Assim, o objetivo da presente pesquisa foi caracterizar o processo de fabricação das vinícolas paulistas, avaliar o nível de implantação de sistemas básicos da qualidade e as condições higiênico-sanitárias destas vinícolas, caracterizar o perfil sensorial, a aceitação e parâmetros físico-químicos dos vinhos tintos por elas produzidos. Para a realização desse trabalho, após convite, foram selecionadas para participar da pesquisa 10 vinícolas artesanais e industriais do Estado de São Paulo. Todas encaminharam um de seus vinhos tintos secos para análises físico-químicas e sensoriais, os quais, em sua maioria, eram elaborados a partir de uvas americanas e/ou híbridas, e pertenciam à safra 2006. Sete dessas dez vinícolas foram selecionadas para participar da pesquisa de campo, por meio da qual, caracterizou-se as etapas de elaboração dos vinhos, desde o cultivo das uvas até o engarrafamento da bebida. Durante a visita, foi também preenchida a lista de verificação (check list) da legislação vigente brasileira, de forma a avaliar o cumprimento das Boas Práticas de Fabricação (BPF) e dos Procedimentos Operacionais Padronizados (POP¿s), bem como das condições higiênico-sanitárias dessas vinícolas. Através de Análise Descritiva Quantitativa (ADQ®), uma equipe de 12 julgadores selecionados e treinados desenvolveu a terminologia sensorial descritiva e o perfil sensorial dos vinhos. A intensidade de cada descritor foi avaliada em seis repetições para cada amostra, através de uma escala não estruturada de nove centímetros, ancorada em seus extremos com os termos de intensidade "fraco" e "forte". Os dados descritivos foram analisados por Análise de Variância (ANOVA), teste de médias Tukey (p= 5%) e Análise de Componentes Principais (ACP). Na sequência, 120 consumidores de vinhos tintos secos avaliaram através de escala hedônica híbrida, cada uma das dez amostras com relação a aparência, impressão global, aroma e sabor. Os dados afetivos foram analisados através de Mapa de Preferência Interno (MDPREF), ANOVA e teste de Tukey (p= 5%) para comparação das médias. Os vinhos foram também caracterizados quanto ao: teor de açúcares redutores, sólidos solúveis (ºBrix), compostos fenólicos totais, acidez total, acidez volátil, dióxido de enxofre livre, pH, graduação alcoólica, extrato seco e extrato seco reduzido, relação álcool/ extrato seco reduzido (p/p) e metanol. Os resultados físico-químicos foram também analisados por ANOVA e teste de médias de Tukey (p = 5%). Os parâmetros químicos dos vinhos avaliados, em geral, enquadraram-se nos Padrões de Identidade e Qualidade (PIQ¿s) estabelecidos pela legislação brasileira. Entretanto, de um modo geral, as vinícolas visitadas demonstraram estar em condições higiênico-sanitárias insatisfatórias; apenas um estabelecimento foi classificado no Grupo 1, atendendo a mais de 76% dos itens da lista de verificação. As amostras apresentaram perfis sensoriais bastante distintos entre si, diferindo significativamente (p = 0,05) com relação à maioria dos 33 descritores desenvolvidos através da análise descritiva, quais sejam: cor roxa, cor vermelha/rubi, viscosidade aparente e translucidez; notas aromáticas de uva, suco de uva, doce, amadeirado, alcoólico, frutas secas, frutas vermelhas, ácido, fermento, cogumelo, enxofre, chá, floral cítrico, floral não cítrico e vegetativo; notas de sabores descritas como uva, suco de uva, gosto doce, amadeirado, alcoólico, frutas secas, frutas vermelhas, ácido acético, gosto azedo, gosto amargo, semente e fermentado; além de sensações bucais de corpo e adstringência. Com relação aos seus perfis sensoriais, os vinhos segmentaram-se principalmente em função do varietal das uvas utilizadas na elaboração dos mesmos, a despeito das bebidas terem sido elaboradas por diferentes vinícolas, processos, etc. Vinhos elaborados a partir de uvas Bordô apresentaram notas mais intensas de aromas e sabores frutados (uva, suco de uva e frutas vermelhas) aroma e gosto doce, e aroma floral não cítrico (notadamente aroma de rosas); notas sensoriais realmente esperadas em vinhos de mesa de uvas americanas e/ou híbridas. Destacaram-se na preferência dos consumidores um vinho elaborado com o varietal Seibel 2 (¿Corbina¿), e vários contendo uvas Bordô / Abstract: In the State of Sao Paulo, Brazil, the production of table wine from American and/or hybrid grapes strongly surpasses the production from Vitis Vinifera grape varieties. However, in this State, there is little knowledge regarding the technology used to manufacture these wines, their level of food safety, sensory profiles and acceptability among the Brazilian consumers. Thus, the objectives of the present research were to characterize the manufacturing processes used by Sao Paulo¿s wineries, analyzing their quality control systems, sanitary conditions and their wines sensory profile, acceptance and chemical parameters. Thus, ten wineries joined the research, sending for analysis, a sample of their dry red table wine, from the 2006's crop. The majority of the wines were produced from hybrid or American grapes. Seven of the ten wineries were visited and all stages of their production plant was inspected, from the grapes plantation to the wine bottling. A checklist was filled out in order to verify the wineries performance regarding procedures of ¿Good Manufacturing Practices¿ (GMP) and Standard Operational Procedures (POP¿s), as well as sanitary conditions. Using Quantitative Descriptive Analysis, 12 trained panelists developed the wines lexicon and sensory profiles. Panelists rated the intensity of each attribute, using a 9 cm non-structure scale, anchored in its left and right extremes with terms ¿weak¿ and ¿strong¿ respectively. Each wine was evaluated in six replications by each panelist. The resulting data was evaluated using analysis of variance (ANOVA), Tukey test (p = 5%), and the Principal Component Analysis (PCA). Sequentially, 120 consumers evaluated the overall liking of all ten wines, using a hybrid hedonic scale. Additionally, the consumers evaluated how much they like/disliked the sample¿s appearance, aroma and flavor. The affective data was analyzed through the Internal Preference Map (MDPREF), ANOVA and Tukey test (p= 5%). Finally, the wines were analyzed with respect to reducing sugars, pH, total acidity, volatile acidity, total phenols, free sulphur dioxide, pH, methanol, soluble solids (oBrix), total solids, alcohol content and methanol content. The physical-chemical results were analyzed by ANOVA and Tukey test (p = 5%). Overall, the wines chemical parameters complied with the Brazilian legislation for red table wine. Nevertheless, several wineries showed substandard sanitary conditions. The wines possessed very distinct sensory profiles, differing significantly among them (p = 5%) in most of the 33 descriptors, such as: purple color, red/ruby color, visual viscosity; aroma and/or flavor notes of grape, sweet, woody, alcohol, dried fruit, berry, sour, yeasty, mushroom, sulfur, tea, floral citrus, rose and vegetative among others. Despite the fact that production had taken place at different wineries using different processes, wines produced from the same varietals showed similar sensory profiles, The wines produced with Ives (Bordô) grapes showed highest intensities of fruity aroma and flavor (grape, juice grape and berry fruit), sweet aroma and taste and rose aroma; all sensory attributes expected in wines produced from American and/or hybrid grapes. Among the ten wines evaluated, the consumers preferred one sample produced from Seibel 2 variety (¿Corbina¿) and the wines made from Ives (Bordô) grapes / Mestrado / Consumo e Qualidade de Alimentos / Mestre em Alimentos e Nutrição

Il est généralement admis que la qualité des vins et des eaux de vie dépend de leur vieillissement au contact du bois de chêne. La sélection du bois de chêne et sa maturation constituent des étapes essentielles dans la fabrication de barriques. Compte tenu des nombreux facteurs impliqués dans le choix de la matière première et dans la phase de séchage naturel du bois de chêne, il est très complexe de déterminer l’impact de la durée de séchage et l’incidence des facteurs de sélection combinés. Nous avons pu mettre en œuvre un plan d’essai qui nous a permis d’étudier l’évolution de la composition chimique du bois de chêne pour quatre stades de séchage : bois frais, 12 mois, 18 mois et 24 mois. Pour ce plan d’essai, l’échantillonnage a été réalisé en prenant en compte trois facteurs de sélection d’intérêt en tonnellerie : l’âge du bois, le type de grain et la classe d’Indice Polyphénolique mesurée par Oakscan®. En plus de la partie extractible (~10%), trois polymères constituent la majeure partie (~90%) du bois de chêne : la cellulose, les hémicelluloses et les lignines. Ces composés sont susceptibles de subir des hydrolyses ou des réactions chimiques au cours des différents processus de tonnellerie et notamment lors de la chauffe, libérant ainsi des composés aromatiques ou certains précurseurs aromatiques ayant un véritable intérêt sensoriel pour le vin vieilli en fût ou en contact avec du bois de chêne. Aucune étude n’a, à ce jour, montré un lien entre les proportions de ces composés dans le bois de chêne et l’impact au niveau chimique et organoleptique dans des vins élevés au contact de bois de chêne. Nous avons pu montrer que leurs proportions évoluent de façon significative au cours du séchage du bois de chêne et nos résultats soulignent l’incidence des différents facteurs de sélection. Respectivement, les proportions en composés extractibles, lignines, hémicelluloses et cellulose apparaissent majoritaires aux stades bois frais, 12 mois, 18 mois et 24 mois de séchage. La mise en place d’un plan d’essai sur un vin de Merlot à partir d’un deuxième échantillonnage de bois de chêne, selon les mêmes modalités que le plan d’essai sur bois, nous a permis d’évaluer l’impact du temps de séchage du bois sur la composition chimique d’un vin élevé au contact de morceaux de bois de chêne. Les résultats ont montré entre autres une baisse de près de 8% de teneur en ellagitanins moléculaires du vin entre les modalités 12 et 24 mois de séchage mais aussi des impacts sur les concentrations en composés volatiles du vin : les aldéhydes furaniques sont corrélés positivement à des temps de séchage longs (18 et 24 mois), tandis que les aldéhydes phénoliques le sont avec un temps de maturation plus court (12 mois). Nos résultats soulignent aussi l’influence des facteurs de sélection sur la composition chimique du vin. Sur le même vin de Merlot, la réalisation d’analyses sensorielles a mis en évidence des différences significatives perçues par notre panel entre les modalités de séchage 12 et 24 mois, tandis que les modalités 18 et 24 mois n’ont pas été distinguées lors de tests triangulaires. Des profils sensoriels ont également été établis pour tenter d’associer les différences perçues à un ou plusieurs descripteurs. Pour cette épreuve, les résultats soulignent la difficulté pour notre panel de dégustateurs de s’accorder sur l’existence d’une différence globale et significative entre les modalités 12 et 24 mois. Cependant, ils contribuent à expliquer les différences mises en évidence lors des tests triangulaires pour certaines modalités de séchage pour les descripteurs grillé/fumé, amertume, astringence, rondeur en bouche et perception sucrée. / It is widely accepted that alcoholic beverages quality depends on their ageing in premium quality oak wood. Oak wood selection and maturation are essential steps in the course of barrel fabrication. Given the existence of many factors involved in the choice of raw material and in natural seasoning of oak wood, it is very difficult to determine the real impact of seasoning and selection factors on oak wood composition. A sampling was done to study the evolution of oak wood chemical composition during four seasoning steps: non matured, 12 months, 18 months and 24 months. For this sampling, three selection factors were taken into account: age, grain type and the Polyphenolic Index measured by Oakscan®. Besides extractables (~10%), three polymers constitute the main part of oak wood: cellulose, hemicelluloses and lignins. These compounds may undergo hydrolysis or chemical reactions during cooperage processes, especially during heat treatment, which release some aromatic compounds or aromatic precursors having a genuine sensorial interest on wine aged in barrel or in contact with oak products. To date, no studies revealed a link between the proportions of these compounds in oak wood and the chemical and sensorial impact in wines ageing with oak wood. Our study showed that the proportions of these compounds evolved significantly during oak wood seasoning and the results highlighted the impact of selection factors. Respectively, extractables, lignins, hemicelluloses and cellulose proportions were mostly for non-matured, 12 months, 18 months and 24 seasoning months. The development of a test plan with a Merlot wine from a second oak wood sampling, using similar modalities as the previous test plan allowed the evaluation of oak wood seasoning impact on the chemical composition of a wine ageing with oak wood pieces. Results showed a lowering of 8% in ellagitannins content of wine between 12 and 24 months modalities. An impact on volatile composition in wine has also been established: furanic aldehydes were positively correlated with a long maturation time (18 and 24 months), whereas phenolic aldehydes were positively correlated with a shorter seasoning time (12 months). Our results highlighted also the impact of selection factors on wine chemical composition. Sensorial analysis on this Merlot wine led to significant differences detected by the panel between 12 and 24 seasoning modalities, but not between 18 and 24 months modalities with triangular tests. Sensorial profiles were also established to attempt to associate the differences detected to one or more descriptors. For this test, results highlighted the difficulty for the panel to make a significant difference between 12 and 24 months modalities. However they contribute to explain the differences demonstrated with triangular tests for some seasoning modalities: toasted, bitterness, astringency, roundness and sweet perception.

Flavonoids are plant secondary metabolites, which belong to several groups varying in their chemical structure. Anthocyanins and tannins are important flavonoid components of wine that are responsible for its color, taste and other sensory properties. The concentration of anthocyanins in wine is affected by grape variety, processing technology, and climatic conditions. In this Thesis, we studied the changes in color and in related chemical composition, using three non-commercial samples of red wine: Svatovavřinecké (year 2010 and 2012), and home-made wine (prepared without addition of SO2). These changes in color were determined using standard colorimetric method (CIELab) and also a simplified two-parametric spectrophotometric method (tint/color density). The content of anthocyanins was followed using analytical RP-HPLC method. In paralel, simplified oenologic methods for estimation of phenolic compounds were used. Generally the wine samples changed color to darker tint. Chemically, this was caused by polymerisation reactions between anthocyanins and phenolic compounds. This led to the formation of stable pigments characterised by a higher absorption maximum at longetr wavelength, hence a darker tint. Key words: anthocyanins, color, red wine, phenolic compounds, malvidin-3-glucosid, polymeric reactions,...

Submitted by Maurício Bonatto Machado de Castilhos (mbonattosp@yahoo.com.br) on 2016-03-21T14:52:28Z No. of bitstreams: 1 Tese Bonatto_corrigida_final.pdf: 3969473 bytes, checksum: b52bf638d9c0de021358f0dd1af6792a (MD5) / Approved for entry into archive by Sandra Manzano de Almeida (smanzano@marilia.unesp.br) on 2016-03-22T16:52:25Z (GMT) No. of bitstreams: 1 castilhos_mbm_dr_sjrp.pdf: 3969473 bytes, checksum: b52bf638d9c0de021358f0dd1af6792a (MD5) / Made available in DSpace on 2016-03-22T16:52:25Z (GMT). No. of bitstreams: 1 castilhos_mbm_dr_sjrp.pdf: 3969473 bytes, checksum: b52bf638d9c0de021358f0dd1af6792a (MD5) Previous issue date: 2016-03-15 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / A produção brasileira de vinhos encontra-se em constante especialização e tem como objetivo incrementar a qualidade dessas bebidas a fim de atender e superar as expectativas dos consumidores. Diante desse contexto, algumas entidades governamentais pesquisam o melhoramento genético de uvas almejando produzir matérias-primas com características singulares a fim de elaborar vinhos com qualidade ímpar e de caráter regional. Adicionalmente, cientistas da área pesquisam variações no processo de vinificação e, através dessas técnicas, promovem variações no perfil químico dos vinhos, respondendo positivamente no perfil sensorial descritivo e de aceitação. Esse estudo pesquisou os efeitos da pré-secagem das uvas e o emprego do chapéu submerso e explorou o comportamento químico e sensorial de vinhos elaborados com uvas americanas e híbridas. Os resultados mostraram que os vinhos elaborados pelo processo de pré-secagem das uvas apresentaram forte degradação de compostos fenólicos como antocianinas e flavonóis, sendo os flavan-3-óis (galoilados ou não) e proantocianidinas os que apresentaram maior resistência ao calor produzido pela secagem. De uma forma geral, tais vinhos foram descritos como amargos, adstringentes, encorpados, pungentes e persistentes por apresentarem elevada concentração desses compostos fenólicos. Os vinhos elaborados pelo processo de chapéu submerso não apresentaram diferenças significativas nos conteúdos de antocianinas, piranoantocianinas e flavonóis quando comparados aos vinhos elaborados pelo processo tradicional de vinificação, sendo descritos com elevada intensidade de cor e matiz violeta, além de apresentarem, em alguns casos, aroma foxado e frutado evidente. Apresentaram elevada aceitação em alguns casos, sendo considerados como alternativa ao tratamento tradicional. / Brazilian wine production is in constant specialization and aims to enhance the quality of red wines in order to meet and exceed consumer expectations. Thus, some governmental entities develop genetic improvement programs aiming at producing grapes with singular features that will be employed in the production of regional wines, i.e., wines whose features indicate their origin of production. Moreover, scientists from enological area study the variations in winemaking aiming at promoting such improvement on the chemical properties of wines, enhancing their acceptance sensory profile. Therefore, this study aimed at analyzing the effects of grape pre-drying and submerged cap in order to explore the chemical and sensory behavior of red wines produced from Vitis labrusca grapes and their hybrids. The results showed that the wines produced from pre-drying process presented relevant degradation of the phenolic compounds, mainly anthocyanins and flavonols; and flavan-3-ols (galloylated or not) and proanthocyanidins were the compounds that seemed to be less affected by the heat produced by the pre-drying procedure. In general, these wines were described as bitter, astringent, full-bodied, pungent and persistent to palate due to their high concentration of these aforementioned compounds. Wines produced form submerged cap winemaking presented no significant differences from the wines elaborated by traditional process concerning anthocyanin, pyranoanthocyanin and flavonol contents, and were described as colorful with intense violet hue and strong foxy and fruity aroma. These wines presented higher acceptance in some cases and were considered as an alternative to the traditional winemaking treatment.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Diabetes mellitus is a major public health problem throughout the world. The chronic hyperglycemia present in the diabetic state is associated with increase of oxidative stress, which has been appointed to play a central role in the development and progression of diabetic complications. Changes in the platelet morphology and function observed in diabetes have been considered as important aggravating to the pathogenesis of vascular complications in this endocrinopathia. The extracellular nucleotides ATP, ADP and nucleoside adenosine are known to regulate various physiological processes in the vascular system, including platelet aggregation, vascular tone and cardiac functions. The control of extracellular levels of these molecules and consequent purinergic signaling induced by them is controlled by a variety of enzymes such as NTPDases (Nucleotide Triphosphates Diphosphohydrolase), E-NPPs (Ecto-Nucleotide Pyrophosphatase Phosphodiesterases), 5'-nucleotidase and adenosine deaminase (ADA). On the other hand, studies have revealed a number of beneficial actions of red wine and grape juice, which can be attributed to high concentrations of polyphenols presents in these beverages, mainly of resveratrol, which presented antioxidant and antiplatelet properties. In this context, the objective of this study was to investigate the effect of resveratrol, of red wine and of grape juice in the biomarkers of oxidative stress in liver and kidney, in the ectoenzimas activities in platelets and platelet aggregation of streptozotocin-induced diabetic rats. In addition, tests were performed in vitro with the polyphenols resveratrol, quercetin, rutin, caffeic acid and gallic acid on the activity of ectonucleotidases, ADA and platelet aggregation in diabetic and control rats. The results showed an increase in the levels of lipid peroxidation in liver and kidney of diabetic rats and treatment with resveratrol (10 and 20 mg/kg) prevented this increase. The activities of catalase (CAT), superoxide dismutase (SOD) and aminolevulinic acid dehydratase (-ALA-D) and the levels of non protein thiols (NPSH) and vitamin C presented a significant decrease in diabetic rats. However, the treatment with resveratrol (10 and 20 mg/kg) was able to prevent the decrease in the antioxidant defenses in diabetic rats. In relation to NTPDase, E-NPP, 5'-nucleotidase and ADA, there was an increase in activity of these ectoenzimas in platelets of diabetic rats accompanied by an increase in platelet aggregation in these animals. Treatment with resveratrol (10 and 20 mg/kg), grape juice and red wine increased the activity of NTPDase, E-NPP and 5'-nucleotidase and prevented the increase in ADA activity in diabetic rats. Platelet aggregation significantly decreased in diabetic rats treated with grape juice and red wine. In the in vitro tests, resveratrol, caffeic acid, and gallic acid increased ATP, ADP and AMP hydrolysis, while quercetin and rutin decreased the hydrolysis of these nucleotides in platelets of diabetic rats. The ADA activity and platelet aggregation were reduced in platelets of diabetic rats in the presence of all polyphenols tested in vitro. The results described here suggest that resveratrol could have a protector effect against hepatic and renal damage induced by oxidative stress in the diabetic state, which was evidenced by the capacity of this polyphenol to modulate the antioxidant defense and to decrease the lipid peroxidation in these tissues. In addition, resveratrol, red wine and grape juice as well as caffeic acid, gallic acid, quercetin and rutin were able to modulate the ectoenzimas activities and decrease platelet aggregation could contribute to the prevention and reduction of platelet abnormality and consequently vascular complications in diabetic state. / O diabetes mellitus (DM) é um dos maiores problemas de saúde pública no mundo. A hiperglicemia crônica presente no estado diabético está associada com um aumento do estresse oxidativo, o qual tem sido apontado por ter um papel central no desenvolvimento e na progressão de complicações diabéticas. Alterações na morfologia e função das plaquetas observadas no diabetes têm sido consideradas agravantes importantes para a patogênese das complicações vasculares desta endocrinopatia. Os nucleotídeos extracelulares ATP, ADP e o nucleosídeo adenosina regulam diversos processos fisiológicos no sistema vascular, incluindo a agregação plaquetária, o tônus vascular e as funções cardíacas. O controle dos níveis extracelulares destas moléculas e a conseqüente sinalização purinérgica por elas induzida é realizada por uma variedade de enzimas como as NTPDases (Nucleotídeo Trifosfato Difosfoidrolase), E-NPPs (Ecto-Nucleotídeo Pirofosfatase/Fosfodiesterases), 5 -nucleotidase e a adenosina deaminase (ADA). Estudos têm revelado uma série de ações benéficas do vinho tinto e do suco de uva, que podem ser atribuídas às altas concentrações de polifenóis que estes possuem, principalmente de resveratrol o qual apresenta propriedades antioxidantes e antiplaquetárias. Neste contexto, o objetivo do presente estudo foi verificar o efeito do resveratrol, do vinho tinto e do suco de uva no perfil oxidativo em fígado e rim, na atividade de ectoenzimas em plaquetas e na agregação plaquetária de ratos diabéticos induzidos com estreptozotocina. Além disso, foram realizados testes in vitro com os polifenóis resveratrol, quercetina, rutina, ácido caféico e ácido gálico na atividade das ectonucleotidases, ADA e agregação plaquetária de ratos controles e diabéticos. Os resultados obtidos demonstraram um aumento nos níveis de peroxidação lipídica em fígado e rim de ratos diabéticos e o tratamento com resveratrol (10 e 20 mg/kg) preveniu este aumento. A atividade da superóxido dismutase (SOD), catalase (CAT) e -aminolevulinato ácido desidratase (-ALA-D) e os níveis de tióis não protéicos (NPSH) e vitamina C foram significativamente decrescidos no fígado e rim de ratos diabéticos. Contudo o tratamento com resveratrol (10 e 20 mg/kg) preveniu o decréscimo nas defesas antioxidantes de ratos diabéticos. Em relação à NTPDase, E-NPP ,5 -Nucleotidase e ADA, observou-se um aumento na atividade destas ectoenzimas em plaquetas de ratos diabéticos acompanhado de um aumento na agregação plaquetária neste animais. O tratamento com resveratrol (10 e 20 mg/kg), suco de uva e vinho tinto aumentou a atividade da NTPDase, E-NPP and 5′-nucleotidase e preveniu o aumento na atividade da ADA em ratos diabéticos. A agregação plaquetária decresceu significativamente em ratos diabéticos tratados com suco de uva e vinho tinto. Nos testes in vitro, resveratrol, ácido caféico e ácido gálico aumentaram a hidrólise do ATP, ADP e AMP, enquanto a rutina e a quercetina decresceram a hidrólise destes nucleotídeos em plaquetas de ratos diabéticos. A atividade da ADA e agregação plaquetária foram reduzidas na presença de todos os polifenóis testados in vitro. Assim, os resultados descritos aqui sugerem que o resveratrol tem efeito protetor contra danos hepáticos e renais induzidos pelo estresse oxidativo no estado diabético, o qual foi evidenciado pela capacidade deste polifenol de modular as defesas antioxidantes e decrescer a peroxidação lipídica nestes tecidos. Além disso, o resveratrol, o vinho tinto e o suco de uva, bem como o ácido cafeíco, o ácido gálico, a quercetina e a rutina são capazes de modular a atividade das ectoenzimas e decrescer a agregação plaquetária, podendo prevenir e reduzir anormalidades plaquetárias e consequentemente complicações vasculares no estado diabético.

This Ph.D. thesis deals with the deep analysis of polyphenols effects toward vascular disorders. This work provides a number of experimental results of studying both the effects of natural bioactive compounds in red wines and potatoes, and their application to the experiments which includes experimental animal models with spontaneous hypertension. Methods and equipment used in experimental studies allowed us to make several new statements regarding the universal nature of the relationships between the antioxidant capacity and the polyphenolic content in examined foodstuffs. Besides this it is also recognized that food is a source of minerals which also contribute to the total antioxidant capacity and therefore may have influence the endogenous antioxidant enzyme system by providing the essential cofactors. Experimentally we have found that the concentrations of magnesium known for its therapeutic action in wine and red wine extract are comparable to the mineral waters recommended for the treatment and prevention of cardiovascular and metabolic diseases. Moreover, synergistic interactions between selected minerals have been found. The key issue of bioavailability of polyphenols for supporting the idea of the beneficial effects of diet rich in fruits and vegetables toward vascular disease prevention...

Les tanins condensés et les pigments polymérisés sont deux grandes familles de macromolécules qui jouent des rôles importants sur la qualité organoleptique du vin rouge. Leurs structures oligo-polymériques ainsi que leurs évolutions durant le vieillissement du vin rouge sont mal connues. L’objectif de cette étude est d’approfondir notre connaissance sur leurs structures et leurs évolutions durant le vieillissement. Durant ce travail, nous avons caractérisé, pour la première fois dans le règne végétal, une nouvelle sous-famille de tanins condensés nommés : les tanins condensés couronnes, qui ont des propriétés spécifiques. Ils sont plus polaires que des tanins condensés de type B. Durant le vieillissement, les concentrations des tanins de type B diminuent alors que celles des tanins couronnes restent plutôt stables. Leurs concentrations sont plus élevées dans les vins issus de cépage Syrah que dans les vins issus de cépage Cabernet Sauvignon et Merlot. Au niveau sensoriel, une forte corrélation entre la concentration des tanins couronnes et l’intensité d’astringence ressentie par les dégustateurs a été montrée. Un premier test d’activité biologique du tétramère couronne montre une activité inhibitrice intéressante de l’agrégation du peptide β-amyloïde impliquée dans la maladie d’Alzheimer. En même temps, une nouvelle méthode de quantification des tanins condensés liés par pont éthylidène via un marqueur spécifique en utilisant une détection par fluorescence a été développée. Concernant les pigments polymérisés avec différents types de liaisons inter-flavonoïdes, leurs mécanismes de dépolymérisation chimique en milieu acide ont été clarifiés et une méthode de quantification sur un système UPLC-Q-TOF a été développée. Les pigments oligo-/polymériques contribuent plus à la couleur du vin vieux que les pigments mono-/dimériques. La concentration des structures oligo-polymériques ayant des liaisons de type A est plus stable que celle des pigments polymérisés par des liaisons de type B ou par le pont éthylidène durant le vieillissement du vin. De plus, durant nos investigations sur les pigments polymérisés, les structures de type A-F(type A)-F(n) et de type F(n)-A-F(type A) ainsi que les formes acétylées d’anthocyane trimère et les formes acétylées des anthocyanes dimères liées par pont éthylidène ont été montrées pour la première fois. / Condensed tannins and polymeric pigments are two families of macromolecules which play important roles on the organoleptic quality of red wine. Their oligo-polymeric structures and their evolutions during red wine aging are poorly known. The objective of this study is to deepen our knowledge of their structures and their evolutions during red wine aging. During our work, we characterized, for the first time in plant kingdom, a new family of condensed tannins named : crown condensed tannins, which have specific properties. They are more polar than B-type tannins. During red wine aging, the concentration of B-type tannins decreased while the crown tannins remained stable. Their concentrations are higher in Syrah wines than in wines made from Cabernet Sauvignon and Merlot grapes. From a sensory point of view, a strong correlation between the concentration of crown tannins and the intensity of astringency rated by the tasters was obtained. The first biologic activity assay of the crown tetramer showed an interesting inhibitory activity on aggregation of β-amyloid peptide involved in Alzheimer's disease. At the same time, a new quantification method of ethylidene bridge linked condensed tannins via a specific marker using fluorescence detection has been developed. Concerning polymeric pigments with different types of inter-flavonoids linkages, their acidic depolymerization mechanisms have been clarified for the first time and a quantification method using a UPLC-Q-TOF system has been developed. Oligo-/polymeric pigments contributed more to the color of old wine than mono-/dimeric pigments. The concentration of the pigmented oligo-polymers with A type linkages is more stable than that of polymeric pigments by B-type linkages or by ethylidene bridge during red wine aging. In addition, during our study of polymeric pigments, A-F(A type)-F(n) and F(n)-A-F(A type) structures as well as the acetylated form of trimeric anthocyanin and acetylated forms of dimeric anthocyanin linked by ethylidene bridge have been shown for the first time.

<p>Fenolna jedinjenja su zaslužna za<br />osnovna senzorna svojstva srvenih vina,<br />pre svega boju i trpkost. Superoksid (O2^&bull;-) i hidroksil radikali (^&bull;OH) izazivaju<br />ozbiljna o&scaron;tećenja tkiva degradaciju<br />proteina, nerastvornih lipida, ugljenih<br />hidrata i nukleinskih kiselina. U ovom<br />radu ispitan je uticaj tehničkotehnolo&scaron;kih<br />uslova proizvodnje i odnosa<br />čvrste i tečne faze u kljuku, na boju vina,<br />sadržaj fenolnih jedinjenja i<br />antioksidativni potencijal. Izmena odnosa<br />čvrste i tečne faze u kljuku vr&scaron;ena je<br />vraćanjem jednog dela ili celokupne<br />količine &scaron;epurine i povećavanjem<br />sadržaja semenki dodavanjem 100, 200 i<br />300 % semenki u odnosu na prirodni<br />sadržaj. Stabilnost boje, sadržaja fenolnih<br />jedinjenja i antioksidativnog potencijala<br />ispitivana je pre i nakon tretmana vina sa<br />dva organska i dva neorganska sredstva<br />za bistrenje i stabilizaciju. Utvrđeno je da<br />povećanje sadržaja semenki u kljuku<br />pojačava antioksidatovno delovanje vina.<br />U vinu Cabernet sauvignon utvrđena je<br />vi&scaron;a vrednost antiradikalske aktivnosti<br />prema DPPH radikalima (AA_(DPPH&bull;)),<br />koja se nije promenila pod delovanjem<br />sredstava za bistrenje i stabilizaciju vina.<br />Vino Merlot ispoljilo je veću<br />antiradikalsku aktivnost na hidroksil<br />radikale (AA_(&bull;OH)).</p> / <p>Phenolic compounds are responsible for<br />basic sensory properties of red wines,<br />primarily color and astrigency.<br />Superoxide (O2^&bull;-) and hydroxyl radicals<br />(^&bull;OH) causes serious damage to the tissue<br />degradation of proteins, ansolubility<br />lipids, carbohydrates and nucleic acids.<br />The paper examined the impact of<br />technical and technological conditions of<br />production and relations of liquid and<br />solid phases in pomase on the color of<br />wine, phenolic compounds content and<br />antioxidant potential. Changing relations<br />between solid and liquid phases in<br />pomase carried out the return of a part, or<br />the whole amount of stem and increasing<br />seeds content adding +100, +200 and<br />+300 % of seeds. Color stability,<br />phenolic compounds content and<br />antioxidative potential was examined<br />before and after treatment of wine with<br />two organic and two inorganic agens for<br />fining and stabilization. It was found that<br />increasing the seeds content in pomase<br />increases antioxidative action of wines.<br />In the wine Cabernet Sauvignon was<br />found higher value antiradical activity to<br />DPPH radicals (AA_(DPPH&bull;)), which are not<br />changed under the fining action and<br />stabilization of wine. Merlot wine<br />exhibited a higher antioxidant potential<br />of the hydroxyl radicals (AA_(&bull;OH)).</p>