A avaliação da atividade antioxidante de compostos fenólicos em vegetais tem sido estudada como alternativa ao uso de antioxidantes sintéticos como butil hidroxi tolueno (BHT). No presente trabalho foi avaliada a capacidade antioxidante in vitro dos extratos (etéreo, etanólico e aquoso) e frações de ácidos fenólicos (livres, ésteres de ácidos fenólicos solúveis e ácidos fenólicos insolúveis ligados) do café in natura e torrado em diferentes condições de tempo (10 e 20 min) e temperatura (140ºC, 160ºC e 180ºC). As análises in vitro foram realizadas através do sistema &#946;-caroteno e ácido linoléico (que indica a inibição da oxidação em meio emulsionado) e pelo método do Rancimat (que indica a inibição da oxidação em meio lipídico). Em ambos os métodos alguns extratos e todas as frações de ácidos fenólicos apresentaram atividade antioxidante igual ou superior ao BHT nas mesmas concentrações. Independente das condições de torrefação aplicadas, o extrato aquoso apresentou a maior atividade antioxidante em função do maior conteúdo de fenólicos. Os fatores cinéticos obtidos demonstraram também resultados maiores que o BHT. O perfil de ácidos fenólicos das frações foi caracterizado por cromatografia gasosa sendo identificados os seguintes ácidos fenólicos: salicílico, ferúlico, caféico, sinápico, clorogênico, quínico, p-cumárico, gentísico e protocatequínico. Foi realizada a avaliação sensorial da infusão de café não havendo diferença estatística entre as amostras mais torradas cujas torras estão próximas das amostras comerciais (p< 0,05). Com base nesses dados a amostra torrada a 180ºC/10 min foi utilizada na avaliação do potencial antioxidante in vivo. Para tanto, ratos Wistar foram suplementados por 30 dias com diferentes doses do extrato aquoso de café torrado (180ºC/10 min). As doses ministradas foram eficientes para evitar a oxidação dos tecidos plasmático, hepático, cerebral e cardíaco quando avaliados pelo método de TBARS (substâncias reativas ao ácido tiobarbitúrico) sendo observado efeito dose resposta em todos os tecidos exceto no cardíaco (p< 0,05). O perfil lipídico do plasma dos animais suplementados em relação ao grupo controle indicou também que o extrato aquoso de café foi eficiente para inibir a oxidação dos ácidos graxos poliinsaturados (p< 0,05). Os dados aqui apresentados sugerem o uso potencial do extrato aquoso de café como antioxidante tanto como aditivo quanto o seu consumo como bebida desde que o extrato aquoso apresentou a maior atividade antioxidante. / Evaluation of antioxidant activity of plant phenolic compounds has been studied as an alternative to the use of synthetic antioxidants, such as butil-hydroxy -toluene (BHT). In this paper we evaluated the in vitro antioxidant capacity of coffee extracts (ether, ethanol and aqueous) and fractions ( free, bound and unbound) roasted for different periods of time ( 0 and 20 min) and at varying temperatures (140&#186;C, 160&#186;C and 180&#186;C). In vitro analyses were made both by the &#946;-carotene and linoleic acid (that indicates oxidation inhibition in an emulsion) as well as by the Rancimat method (that indicates oxidation inhibition in lipid medium). All extracts and fractions presented antioxidant activity equal to or higher than BHT at the same concentrations. Aqueous extract had a higher antioxidant activity due to its higher phenolic content. Kinetic factors obtained were also higher than those of BHT. The fractions\' phenolic acid profile was characterized by gaseous chromatography, identifying the following phenolic acids ( eu acrescentaria: with antioxidant activity, ou antioxidant potential]: salicylic, ferulic coffeic, synaptic, chlorogenic quinic, p-cusmaric ,gentisic and protocatechinic . No statistic differences were obtained in sensorial evaluation of coffee infusion between more intensely roasted samples, which are close to those of commercial samples (p> 0.05). Based on these results the 180&#186;C/10 min sample was used in evalution of antioxidative potential in vivo . For this purpose Wistar rats received a daily dietary complement of different doses of roasted (180&#186;C/10 min) coffee aqueous extract. Administered doses were efficient in avoiding oxidation of plasmatic, hepatic, cerebral and cardiac tissues as evaluated by the TBARS method (substances reactive to thio-barbituric acid) a dosis-response effect being observed for all but the cardiac tissue (p> 0.05). The plasmatic lipidic profile of supplemented animals also indicated efficiency in oxidation inhibition of poli-unsaturated fatty-acids as compared to control group results (p> 0.05). The data presented suggest the potential usefulness of aqueous coffee extract as an antioxidant, when administered as an additive as well as consumed as a beverage, since the aqueous extract had the highest antioxidative activity.

Orientador: Helena Teixeira Godoy / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-19T16:32:45Z (GMT). No. of bitstreams: 1 Sousa_JessicaMariaSilva_M.pdf: 1636413 bytes, checksum: eef8c829695e9cc3aff1047919b3eaed (MD5) Previous issue date: 2012 / Resumo: O aprofundamento do estudo relativo aos compostos naturais com propriedades antioxidantes e antimicrobianas em cogumelos torna-se extremamente relevante tendo em vista a possibilidade de esses compostos serem empregados nas indústrias de alimentos, farmacêutica e de cosméticos. No entanto, não foram encontrados relatos sobre o perfil fenólico de cogumelos comestíveis produzidos no Brasil assim como também não foram encontrados trabalhos que utilizassem quatro ensaios diferentes para avaliar o potencial antioxidante dos cogumelos. Portanto, o objetivo desse trabalho foi avaliar o teor de compostos fenólicos e a capacidade antioxidante e antimicrobiana de 4 espécies de cogumelos comestíveis (A. bisporus, P. ostreatus, P. ostreatoroseus e L. edodes) produzidas no Brasil. Os compostos fenólicos totais (CFT) foram determinados pelo método de Folin-Cioacalteu e a identificação e quantificação individual dos fenólicos foi realizada por cromatografia líquida de alta eficiência (CLAE). A atividade antioxidante foi avaliada por quatro métodos: DPPH (2,2-difenil-1-picrilhidrazila), ABTS+¿ [2,2¿-azino-bis (3-etilbenzo-tiazolina-6-sulfonato)], FRAP (Ferric Reducing Antioxidant Power) e ORAC (Oxygen Radical Absorbance Capacity). O potencial antimicrobiano dos extratos foi avaliado através da concentração mínima inibitória (CIM). Todos os resultados obtidos para as diferentes análises foram analisados levando em consideração o período de colheita, os diferentes processamentos, graus de maturação e substratos de cultivo. No ensaio com o radical DPPH¿ o cogumelo que apresentou maior atividade antioxidante foi o Champignon de Paris (fresco). Já no ensaio com o radical ABTS+¿ e no ORAC os cogumelos que apresentaram maior ação antioxidante foram o Hiratake, o Shimeji (fresco), o Salmon e o Portobello. No ensaio FRAP o Portobello mostrou a maior atividade antioxidante. Os valores médios de CFT variaram entre 0,78 e 10,85 mg EAG/g de amostra liofilizada. Os cogumelos que apresentaram os maiores teores de CFT para o cultivo de verão e de inverno, respectivamente, foram o Hiratake com 10,09 mg EAG/g e 8,45 mg EAG/g, Salmon com 9,61 mg EAG/g e 9,45 mg EAG e Portobello com 9,07 mg EAG/g e 9,60 mg EAG. O processamento empregado ao cogumelo Champignon de Paris diminuiu consideravelmente o teor de CFT (de 7,04 mg EAG/g para 0,90 mg EAG/g - cultivo de verão) enquanto que no processo de secagem empregado ao cogumelo Shimeji não houve diminuição drástica no teor de CFT (de 8,83 mg EAG/g para 7,62 mg EAG/g - cultivo de verão). O grau de maturação influenciou no teor final de CFT mostrando que o Shimeji, colhido em estágio imaturo, possui menor teor de CFT (8,83 mg EAG/g ¿ cultivo de verão e 7,24 mg EAG/g ¿ cultivo de inverno) em comparação ao Hiratake, colhido em estágio mais maturo. Correlações positivas entre os resultados de CFT e os métodos de atividade antioxidante foram encontradas, e essa correlação foi maior para o ensaio com o radical livre ABTS¿+ e o ORAC (r = 0,92 e r = 0,89, respectivamente) do que para o ensaio FRAP (r = 0,45). Já entre o DPPH e os resultados de CFT foi encontrada baixa correlação negativa (r = -0,14). Dentre os compostos investigados por CLAE foram identificados e quantificados apenas os ácidos siríngico, p-cumárico e o trans-cinâmico, com predominância deste último. No ensaio antimicrobiano realizado verificou-se que o crescimento das leveduras C. albicans, C. rugosa, C. tropicalis, C. glabrata e C. utilis foi fortemente inibido pelos extratos aquosos dos cogumelos Shiitake (tora e composto), com CIMs variando entre 0,08 e 0,31 mg/mL. Estes resultados demonstram que os cogumelos comestíveis possuem grande potencial de utilização pela indústria farmacêutica e podem contribuir significativamente para uma boa condição de saúde devido à presença de antioxidantes / Abstract: The study of natural compounds with antioxidant and antimicrobial properties of mushrooms is extremely relevant to the use in the food, pharmaceutical and cosmetic industries. However, no reports were found about the phenolic profile of edible mushrooms produced in Brazil as well as were not found papers describing the use four different tests to evaluate the antioxidant potential of mushrooms. Therefore, the aim of this study is evaluate the phenolic content, antioxidant and antimicrobial activities of four species of edible mushrooms (A. bisporus, P. ostreatus, L. edodes and P. ostreatoroseus) produced in Brazil. The total phenolic compounds (TPC) were determined by Folin-Cioacalteu and the individual identification and quantification of phenolic compounds was performed by high performance liquid chromatography (HPLC). The antioxidant activity was evaluated by four methods: DPPH¿(2,2-diphenyl-1-picrylhydrazyl), ABTS+¿ [2,2 '-azino-bis (3-etilbenzo-thiazoline-6-sulfonate)], FRAP (Ferric Reducing antioxidant Power) and ORAC (Oxygen Radical Absorbance Capacity). The antimicrobial activity of extracts was evaluated by minimum inhibitory concentration (MIC). All the results obtained for the different analyzes were analyzed taking into account the period of harvesting, different processing, maturation degrees and cultivation substrates. In the assay with DPPH, the mushroom with the highest antioxidant activity was A. bisporus white (fresh). In the assays with the radical ABTS+¿ and ORAC, the mushrooms that showed the highest antioxidant activity were P. ostreatus (mature and fresh immature), P. ostreatoroseus and A. bisporus brown. In the FRAP assay the A. bisporus brown. showed the highest antioxidant activity. The average values of TPC ranged varied from 0.78 and 10.85 mg GAE/g lyophilized sample. The mushrooms with highest levels of TPC for the summer and winter cultivation, respectively, were P. ostreatus (mature) with 10.09 mg GAE/g and 8.45 mg GAE/g, P. ostreatoroseus with 9.61 mg GAE/g and 9, 45 mg GAE and A. bisporus brown with 9.07 mg GAE / g and 9.60 mg GAE. The processing used in the A. bisporus white has considerably reduced the content of TFC (from 7.04 mg GAE/g to 0.90 mg GAE/g - summer cultivation) while in the process of drying the mushroom used in the P. ostreatus (immature), no drastic decrease content of the TPC (from 8.83 mg of GAE/g to 7.62 mg GAE/g - summer cultivation). The maturation degree influenced the final content of the TPC showing P. ostreatus, harvested in an immature state, has a lower content of TPC (8.83 mg GAE/g - summer cultivation and 7.24 mg GAE/g - winter cultivation) compared to P. ostreatus, collected in more mature stage. Positive correlations between the results of TPC and methods of antioxidant activity were founded, and this correlation was higher for the test with free radical ABTS+¿ and ORAC (r = 0.92 and r = 0.89, respectively) than for the FRAP test (r = 0.45). Among the DPPH and the results of TFC was found low negative correlation (r = -0.14). Among the compounds investigated by HPLC were identified and quantified only the syringic, p-coumaric and trans-cinnamic acids, with predominance of the latter. In the assay antimicrobial it was found that the growth of yeasts C. albicans, C. rugosa, C. tropicalis, C. glabrata and C. utilis was strongly inhibited by aqueous extracts from the mushrooms Lentinula edodes (logs and compound), with MICs ranged varied from 0.08 to 0.31 mg / mL. These results demonstrate that edible mushrooms have great potential use by the pharmaceutical industry and can contribute significantly for a good health condition due to the presence of antioxidants / Mestrado / Ciência de Alimentos / Mestre em Ciência de Alimentos

Submitted by Aline Batista (alinehb.ufpel@gmail.com) on 2018-05-16T14:04:54Z No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_Cristina_Jansen.pdf: 1461357 bytes, checksum: 1eabd3351ea2e61e231bd4b28f9e3f34 (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2018-05-16T20:22:16Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_Cristina_Jansen.pdf: 1461357 bytes, checksum: 1eabd3351ea2e61e231bd4b28f9e3f34 (MD5) / Approved for entry into archive by Aline Batista (alinehb.ufpel@gmail.com) on 2018-05-16T20:22:31Z (GMT) No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_Cristina_Jansen.pdf: 1461357 bytes, checksum: 1eabd3351ea2e61e231bd4b28f9e3f34 (MD5) / Made available in DSpace on 2018-05-16T20:22:31Z (GMT). No. of bitstreams: 2 license_rdf: 0 bytes, checksum: d41d8cd98f00b204e9800998ecf8427e (MD5) Dissertacao_Cristina_Jansen.pdf: 1461357 bytes, checksum: 1eabd3351ea2e61e231bd4b28f9e3f34 (MD5) Previous issue date: 2015-03-23 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / A colmeia constitui-se em uma fonte de geração de produtos de alto valor nutritivo e com grandes potencialidades farmacológicas. A própolis é um dos produtos obtidos da colmeia, que é produzida pelas abelhas através da coleta de resinas e gomas de árvores, de plantas e de flores, materiais que são parcialmente digeridos e acrescentados de cera e pólen. As propriedades biológicas da própolis são atribuídas principalmente ao conteúdo de fitoquímicos. Dentre estes compostos, os que estão presentes em maior proporção são os compostos fenólicos, majoritariamente os da classe dos flavonoides. Porém, vários são os fatores que interferem na composição química da própolis, o que acaba refletindo no conteúdo e proporção dos diferentes fitoquímicos, e, portanto, em suas propriedades. O Brasil, sendo um país com grande diversidade de vegetação, favorece ampla variação na composição da própolis, contribuindo para dificultar a padronização deste produto no país. Assim, o objetivo deste estudo foi avaliar a composição de fitoquímicos em amostras de própolis oriundas do Sul do Rio Grande do Sul, afim de testar suas propriedades biológicas. Parte das amostras de própolis foram analisadas quanto aos principais parâmetros físico-químicos, e de outra parte foram preparados extratos hidroalcoólicos concentrados. Os extratos foram submetidos à determinação química de compostos fenólicos, flavonoides e carotenoides, e da análise de espectroscopia Ultravioleta-Visível. Também foram avaliados quanto à capacidade antioxidante, através dos métodos de DPPH, ABTS e FRAP; quanto à atividade antibacteriana frente a cepas de bactérias patogênicas (Staphylococcus aureus, Escherichia coli, Listeria monocytogenes); e quanto ao efeito citotóxico, o qual foi realizado através do método de redução do MTT em células da linhagem melanoma B16F10. Dentre os parâmetros físico-químicos, apenas uma amostra apresentou conteúdo de umidade acima do permitido pela legislação brasileira. A cor foi definida como amarela para as amostras oriundas de Rio Grande e de Canguçu, e castanho claro para as amostras oriundas de Pelotas e de Pedro Osório. Todas as amostras apresentaram alto teor dos fitoquímicos analisados (compostos fenólicos, flavonoides e carotenoides); porém, houve variação significativa no conteúdo destes compostos entre as diferentes amostras. Pelo espectro UV observou-se absorção máxima entre 290 e 400 nm, região característica da absorção dos flavonoides. As amostras de própolis de Pelotas e de Rio Grande apresentaram alta capacidade antioxidante determinada pelos métodos com o radical DPPH e FRAP, sendo comparáveis aos obtidos com o padrão quercetina. Constatou-se uma relação positiva entre as amostras com maior teor de fitoquímicos e a elevada ação antioxidante. Todas as amostras apresentaram atividade antibacteriana frente às cepas testadas; porém, os resultados foram superiores na inibição das bactérias Gram-positivas (S. aureus e L. monocytogenes) do que para a bactéria Gram-negativa (E. coli O157:H7), sendo que as amostras de Pelotas e Rio Grande tiveram alta ação bactericida. Todas as amostras de própolis analisadas demonstraram ação citotóxica em 48h no combate às células de câncer de melanoma de camundongo, nas concentrações de 250 e 500 μg.mL-1. Após 72h a própolis causou citotoxicidade na concentração de 25 μg.mL-1. Amostras oriundas de Pelotas e de Rio Grande apresentaram resultados promissores no combate à linhagem testada. / The beehive is a source of nutritious high-value products and with great pharmacological potential. Propolis is one of the products obtained from the beehive, produced by honey bees by collecting resin and gum trees, plants and flowers, which are partially digested and added of wax and pollen. Its biological properties are mainly attributed to phytochemicals. Among these, the highest contend are phenolic compounds, mainly the class of flavonoids. However, there are several factors that influence the chemical composition of propolis, which ends up reflecting on their properties. The Brazil is a country with great diversity of vegetation that enhances the variations in the composition of propolis, contributing to hinder its standardization in the country. Therefore, the aim of this study was to evaluate the composition of phytochemicals of propolis samples of South of Rio Grande do Sul, and test their biological properties. Part of the propolis samples were analyzed for major physical and chemical parameters and another part was used to obtain the etanolic extracts. The extracts were subjected to determination of phenolic compounds, flavonoids and carotenoids, and analysis of Ultraviolet-visible spectroscopy. Also it was determined the antioxidant capacity that was measured by the methods DPPH, ABTS and FRAP. The antibacterial activity was evaluated against strains of pathogenic bacteria (Staphylococcus aureus, Escherichia coli, Listeria monocytogenes). The cytotoxic effect was performed using the MTT reduction method melanoma cell line B16F10. For the physico-chemical parameters, only one sample showed humidity values above those permitted by brasilian legislation. The color was defined as yellow for the Rio Grande and Canguçu samples, and light brown to Pelotas and Pedro Osorio samples. The samples showed high concentration of the analyzed phytochemicals (phenolics, flavonoids and carotenoids), but there was significant variation between samples. The UV spectrum showed absorption maxima between 290 and 400 nm, characteristic of the region flavonoids. Propolis from Pelotas and Rio Grande showed the highest antioxidant capacity by method with DPPH radical and FRAP, comparable to those obtained with the standard quercetin. It was found a positive relationship between the samples with higher levels of phytochemicals and antioxidant effects. All samples showed antibacterial activity against all tested strains, but the results were better in the inhibition of Gram-positive bacteria (S. aureus and L. monocytogenes) than for Gram-negative (E. coli O157: H7), and samples of Pelotas and Rio Grande had high bactericidal action. The propolis samples analyzed showed cytotoxic action in 48 hours in fighting cells of mouse melanoma cancer at concentrations of 250 and 500 μg.mL-1. After 72h propolis caused decrease in carcinogenic lineage in concentrations 25μg.mL-1. Samples of Pelotas and Rio Grande showed promising results in combating tested lineage.

Made available in DSpace on 2014-08-20T13:59:15Z (GMT). No. of bitstreams: 1 dissertacao_isabel_brandao.pdf: 668952 bytes, checksum: bc23ce5c20fc53077a23108874ce5ea7 (MD5) Previous issue date: 2012-02-15 / Alternanthera tenella Colla (Amaranthaceae), popularly known as "apaga-fogo", presents bioactive compounds as betacyanin and phenolics compounds being utilized in folk medicine as diuretic, anti-inflammatory and antipyretic. The use of elicitors allies with the tissue culture is an alternative to promote changes in secondary metabolism of plants, because it can stimulate the production of biologically active compounds, but may become harmful to plant growth. This present work aimed to evaluate the influence of salicylic acid on the growth of plants of A. tenella cultivated in vitro and verify their potential to optimize the production of phenolic compounds, flavonoids and betacyanin, and to verify the antioxidant capacity of the plant extracts. For this, in a first experiment, nodal segments were inoculated in MS medium with different concentrations of salicylic acid (0, 100, 200, 300 and 400 μM) where they remained for 35 days. The plants were evaluated for growth and betacyanin concentrations and of total phenolic compounds when exposed to different concentrations of elicitors. Separately, in a second experiment, the plants grown up on the vermiculite substrat plus of MS liquid medium and after 35 days were exposed to concentration 400 μM of salicylic acid for 0, 12, 36 and 48-h. After this period were quantified the bioactive compounds (betacyanin, total phenols and total flavonoids) and verified the antioxidant capacity in a non-enzyme from plants subjected to treatments. In relation to growth variables the elicitors was shown to be visibly harmful, occurring decreased of mean already on the lowest concentration used (100 μM). The concentration of 300 μM salicylic acid promoted an increase in the quantity of betacyanin, as for total phenolic compounds, reaching 366% and 180% respectively compared to control. The time of 36 hours of exposure of plants to salicylic acid resulted in the largest mean both betacyanin content of, as for total phenolic compounds. Now for total flavonoids, the highest exposure times (36 and 48-h) the elicitors had the lowest averages, differing from control and of 12-h of exposure. The extracts of leaf of apaga-fogo presented higher antioxidant capacity when subjected to 36 and 48-h of exposure to salicylic acid. Based on the results we can conclude that the use of elicitors in the culture medium is unfavorable to the growth of plants of A. tenella in vitro, however increases of form significantly the production of compounds of interest in this study, well as the antioxidant capacity of the plant extracts. / Alternanthera tenella Colla, (Amaranthaceae), popularmente conhecida como apaga-fogo, apresenta compostos bioativos como betacianina e compostos fenólicos, sendo utilizada na medicina popular como diurético, anti-inflamatório e antitérmico. O uso de elicitores aliados à cultura de tecidos é uma alternativa para promover alterações no metabolismo secundário das plantas, pois pode estimular a produção de compostos biologicamente ativos, porém pode tornar se prejudicial ao crescimento da planta. O presente trabalho teve como objetivo avaliar a influência do ácido salicílico sobre o crescimento de plantas de A. tenella cultivadas in vitro e verificar seu potencial para otimizar a produção de compostos fenólicos, flavonoides e betacianinas, bem como verificar a capacidade antioxidante dos extratos das plantas. Para isso, em um primeiro experimento, segmentos nodais foram inoculados em meio MS com diferentes concentrações de ácido salicílico (0, 100, 200, 300 e 400 μM) onde permaneceram por 35 dias. As plantas foram avaliadas quanto ao crescimento e às concentrações de betacianinas e compostos fenólicos totais quando expostas as diferentes concentrações do elicitor. Separadamente, em um segundo experimento, as plantas cresceram no substrato vermiculita acrescido de meio MS líquido e após 35 dias foram expostas a concentração de 400 μM de ácido salicílico por 0, 12, 36 e 48-h. Após esse período foram quantificados os compostos bioativos (betacianina, fenóis totais e flavonoides totais) e verificada a capacidade antioxidante de forma não-enzimática das plantas submetidas aos tratamentos. Em relação às variáveis de crescimento o elicitor mostrou-se visivelmente prejudicial, ocorrendo diminuição das médias já na menor concentração utilizada (100 μM). A concentração de 300 μM de ácido salicílico promoveu o aumento na quantidade de betacianina e compostos fenólicos totais, chegando a 366% e 180%, respectivamente, em relação ao controle. O tempo de 36-h de exposição das plantas ao ácido salicílico resultou nas maiores médias tanto para teor de betacianina, como para compostos fenólicos totais. Já para flavonoides totais, os maiores tempos de exposição (36 e 48-h) ao elicitor apresentaram as menores médias, diferindo do controle e das 12-h de exposição. Os extratos de folhas de apaga-fogo apresentaram maior capacidade antioxidante quando submetidas a 36 e 48-h de exposição ao ácido salicílico. Com base nos resultados pode-se concluir que o uso do elicitor no meio de cultura é desfavorável ao crescimento das plantas de A. tenella in vitro, porém aumenta de forma significativa a produção dos compostos de interesse deste estudo, assim como a capacidade antioxidante dos extratos das plantas.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Phenolic compounds in plants have attracted considerable attention during the last decade due to their contribution to human health in reduce the risk of chronic diseases. The grape is one of the largest sources of these substances and like other plants, the synthesis of these compounds is usually associated with defense responses against stress factors, also called elicitors. More recently, ultrasound has been studied as possible abiotic elicitors of secondary metabolites due to mechanical stress and microstreaming induced by acoustic cavitation. Thus, this work aimed evaluate the application of different power densities (53 and 113 W cm-2) and exposure times (1, 5 and 10 minutes) to ultrasound in Isabel and Cabernet Sauvignon grapes about the effects on total polyphenols of juices made from this two cultivars. The samples were also subjected to the determination of total anthocyanins, tannins, total soluble solids, total acidity, pH and sensory evaluation. The Isabel cultivar showed more significant results in response to ultrasound, an increase of up to 83% in the levels of total polyphenols of juice, as well as significant increase in the content of total anthocyanins and tannins. These results were also accompanied by an increase in total soluble solids and total acidity of the juices. The application of ultrasound emphasized color, odor and taste of the Isabel grape juice, and the sonicated treatments were preferred by tasters. About Cabernet Sauvignon cultivar, it only showed a little variation in the most parameters. Power density and exposure time to ultrasound influenced the most of parameters evaluated in juices regarding to degree of response to sonication. Treatment with 5 and 10 min of exposure to ultrasound and power density of 113 W cm-2, is the combination that proved more effective for the elicitation of phenolic compounds by activation of plant secondary metabolism, improving the quality of the Isabel grape juice. / Compostos fenólicos em plantas têm atraído grande atenção durante a última década devido à sua contribuição para a saúde humana, podendo reduzir os riscos de doenças crônicas. A uva é uma das maiores fontes dessas substâncias e assim como os demais vegetais, tem a síntese desses compostos geralmente associada a respostas de defesa contra fatores de estresse, também chamados elicitores. Mais recentemente, o ultrassom vem sendo estudado como possível elicitor abiótico de metabólitos secundários devido a estresse mecânico e microagitação induzidos por cavitação acústica. Dessa forma, este trabalho foi conduzido com o intuito de avaliar a aplicação de diferentes densidades de potência (53 e 113 W cm-2) e tempos de exposição (1, 5 e 10 minutos) ao ultrassom em uvas Isabel e Cabernet Sauvignon quanto aos efeitos sobre os polifenóis totais dos sucos elaborados com as duas cultivares. As amostras também foram submetidas à determinação de antocianinas totais, taninos, sólidos solúveis totais, acidez total, pH e análise sensorial. A cultivar Isabel mostrou resultados mais significativos em resposta ao ultrassom, com elevação de até 83% nos níveis de polifenóis totais do suco, bem como, importante aumento no conteúdo de antocianinas totais e taninos. Estes resultados também foram acompanhados por um aumento nos sólidos solúveis totais e acidez total dos sucos. A aplicação do ultrassom acentuou cor, odor e sabor dos sucos de uvas Isabel, sendo os tratamentos sonicados os preferidos pelos provadores. Quanto a cultivar Cabernet Sauvignon, esta somente apresentou pequena variação na maioria dos parâmetros analisados. A densidade de potência e tempo de exposição ao ultrassom influenciaram a maioria dos parâmetros avaliados nos sucos quanto ao grau de resposta à sonicação. Tratamentos com 5 e 10 min de exposição ao ultrassom, com densidade de potência de 113 W cm-2 mostraram ser a combinação mais efetiva para a elicitação de compostos fenólicos mediante ativação do metabolismo secundário da planta, melhorando a qualidade dos sucos de uva Isabel.

Conselho Nacional de Desenvolvimento Científico e Tecnológico / The present study aimed to evaluate the propolis ethanolic extract obtained by microwave assisted extraction regarding to its phenolic compounds amount, antioxidant activity and its use in the Tuscany Italian sausage. The amount of total phenolic content and antioxidant activity was evaluated by the DPPH method. After optimization, the selected extraction parameters were used to obtain extracts used in 0, 5%, 1% and 2% (p/v) concentrations in the Tuscan Italian sausage manufacture, wich were analyzed during 56 days. It was analyzed humidity, protein, ash and fat. The sausages were analyzed every seven days for pH, color, TBARS index and microbiological analyses. The sensory analysis was evaluated through the acceptability test with 9 points in the Hedonic Scale and the acceptability index was calculated. The results obtained for centesimal composition are in accordance to the Brazilian law. Regarding color, the product presented a yellowish color, that is, a lighter one. In the final storage period the TBARS value for the sausage with 2% extract was 0,249±0,021 and the control one was 1,641±0,037 MDA/Kg de amostra. The values for Staphylococcus positive coagulase; Salmonella; Total Coliforms and fecal Coliforms were lower than the values limited by law. The acceptability index (IA %) for taste and smell of the products with the highest concentration of propolis was not satisfactory, being lower than 70%. For 0,5% and 1% concentrations the acceptability indexes for color, smell, taste, texture and general appearance were satisfactory, higher than 70%. It is possible to conclude that the extraction with microwaves is efficient and the addition of 0,5%, 1% and 2% propolis extract can be used in the Tuscan Italian sausage manufacture to make its shelf life longer facing the lipid oxidation. / O presente estudo teve por objetivo avaliar o extrato etanólico de própolis obtida por extração assistida por micro-ondas focalizada quanto ao teor de compostos fenólicos, atividade antioxidante e sua aplicação em linguiça toscana. Nos extratos de própolis obtido foram quantificados o conteúdo de fenólicos totais e atividade antioxidante pelo método de DPPH. Após os extratos serem comparados, o que obteve melhor resultado foi utilizado, nas concentrações de 0,5%, 1,0% e 2,0% (p/v) na fabricação de lingüiça toscana sendo posteriormente analisadas sob armazenamento refrigerado a 4ºC durante 56 dias. Foram realizadas análises de umidade, proteínas, cinzas, e gordura. As lingüiças foram analisadas a cada sete dias em relação ao pH, cor, índice de TBARS e análises microbiológicas. A análise sensorial foi avaliada através do teste de aceitabilidade com Escala Hedônica de nove pontos e calculado índice de aceitabilidade. Os resultados obtidos na composição centesimal dos produtos estão de acordo com o exigido pela legislação brasileira. Com relação a cor do produto, este apresentou uma coloração com uma tonalidade mais amarelada ou seja, mais clara. No período final de estocagem o valor de TBARS para lingüiça com 2,0% de extrato foi de 0,249±0,021MDA/Kg de amostra e a controle de 1,641±0,037. Os valores para Staphylococcus coagulase positiva; Samonella; Coliformes totais e Coliformes fecais foram inferiores aos limites estabelecidos pela legislação. Os índices de aceitabilidade (IA%) para os atributos sabor e odor do produto com maior concentração de própolis foi insatisfatório, sendo abaixo de 70%. Já nas concentrações de 0,5% e 1,0% os índices de aceitabilidade para os atributos cor, odor, sabor, textura e aparência global foram satisfatórios, sendo acima de 70%. Conclui-se que a extração através do micro-ondas é eficiente e que a adição de 0,5%, 1,0% e 2,0% de extrato de própolis pode ser utilizada na elaboração de lingüiça toscana visando prolongar a vida de prateleira frente a oxidação lipídica.

La vigne a une importance économique majeure au niveau mondial. La plupart des vignobles sont greffés et comportent une variété (cépage, Vitis vinifera) greffée sur des porte-greffes de Vitis sauvages (hybrides de V. berlandieri, V. riparia et V. rupestris). Le potentiel qualitatif du raisin à la vendange dépend d’un équilibre subtil entre teneur en sucres, acidité, concentration en polyphénols et en précurseurs d’arômes. Les mécanismes contrôlant l’équilibre sucres/acides/polyphénols sont influencés par différentes contraintes abiotiques auxquelles la vigne est soumise, en particulier l’azote, en interaction avec le matériel végétal (cépage et porte-greffe). Des travaux antérieurs suggèrent par ailleurs qu’une partie des effets de la contrainte hydrique sont en fait liés à une carence azotée entraînée indirectement par la diminution d’absorption d’eau. Le système racinaire (porte-greffe) joue un rôle important dans l’absorption, la réduction, le transport et le stockage de l’azote, et dans l’équilibre hydrique de la plante. Dans ce contexte, nous avons étudié les mécanismes impliqués dans la régulation de la synthèse des flavonoïdes des baies en réponse à la nutrition azotée, à la nature du porte-greffe et du greffon. Des plants des cépages Cabernet Sauvignon et Pinot Noir cultivés soit en pots en conditions semi-contrôlées soit au vignoble ont été soumis à des niveaux d’alimentation azotée différents. Les analyses agronomiques ont permis de confirmer une augmentation des teneurs en azote des différents organes de la plante (limbes, pétioles et baies) ainsi qu’une augmentation de la surface foliaire et de la masse des bois de taille sous l’effet du traitement azoté. Les analyses métabolomiques des pellicules de baies révèlent une accumulation de métabolites secondaires dont la nature diffère en fonction des différentes combinaisons porte-greffe/greffon. De plus, une augmentation de la synthèse d’anthocyanes et des flavonols a été observée dans les pellicules de baies en réponse à la diminution de la nutrition azotée. L’apport azoté se traduit aussi par une augmentation du degré de polymérisation moyen des tannins, alors que les teneurs en flavan-3-ols et procyanidines des pépins et des pellicules ne sont pas affectées par les différents apports azotés. Les analyses transcriptomiques globales (génome complet) et ciblées (qPCR) ont mis en évidence des modifications des transcrits de gènes liés au métabolisme des flavonoïdes en réponse à la nutrition azotée. La variation de l’apport azoté influence également l’expression des gènes régulateurs positifs (facteurs de transcription de type MYB) et négatifs (protéine de type Lateral organ Boundary Domain (LBD)) des gènes de la biosynthèse des flavonoïdes chez la vigne. / Grapevine has a major economic importance worldwide. Most vineyards are grafted and include a variety (Vitis vinifera) grafted over a wild Vitis rootstock (hybrids of V. berlandieri, riparia and rupestris). Grape berry quality at harvest depends on a subtle balance between acidity and the concentrations of sugars, polyphenols and precursors of aroma compounds. The mechanisms controlling the balance of sugars/acids/polyphenols are influenced by the abiotic environment, in particular nitrogen supply, and interact with the genotypes of both the scion variety and the rootstock. Previous work suggests that some of the effects of water stress are in fact linked to a nitrogen deficiency driven indirectly by the reduction of water absorption. The root system (i.e rootstock) plays an important role in the uptake, reduction, transport and storage of nitrogen, and the water balance of the plant. In this context, we studied the mechanisms involved in the regulation of the synthesis of flavonoids in berries in response to nitrogen nutrition with different scion/rootstock combinations. Two varieties (Cabernet Sauvignon and Pinot Noir) were subjected to different nitrogen supplies in two experimental systems, in pots under semi-controlled conditions and in a vineyard. Agronomic analysis confirmed that high nitrogen supply increased the nitrogen content of different organs (leaf blades, petioles and berries) as well as leaf surface area and cane pruning weight. Metabolomic analyses of berry skins revealed an accumulation of secondary metabolites whose nature depended on the different rootstock/scion combinations studied. In addition, an increase in the synthesis of anthocyanins and flavonols was observed in the berry skins in response to the decrease in nitrogen nutrition. High nitrogen supply also increased the average degree of polymerization of tannins, while the contents of flavan-3-ols and procyanidins in the seeds and skins of the berries were not affected. Global transcriptome (using RNA sequencing) and targeted (qPCR) analyses showed changes in the abundance of transcripts of genes related to the metabolism of flavonoids in response to nitrogen status. Nitrogen supply also influenced the transcript amounts of positive (MYB) and negative (Lateral Boundary Organ Domain) transcription factors controlling of the biosynthesis of flavonoids.

Les pommes sont considérées comme bénéfiques pour la santé, de part leur teneur en polyphénols qui leur confèrent l’étiquette "d’aliment santé". Les procédés de transformation agroalimentaire des fruits et légumes génèrent des coproduits qui sont considérés comme sans valeur par les industriels. Or, au regard des volumes générés, ce sont des sources abondantes de nutriments.Une voie possible de valorisation de ces coproduits est la réintroduction dans des produits alimentaires en tant qu’ingrédient, de façon à proposer des aliments enrichis en molécules fonctionnelles. Pour ce faire, deux voies ont été explorées: l’utilisation de poudres d’épidermes de pommes pour stabiliser une émulsion et l’extraction des polyphénols au moyen de la technologie à base de CO2 supercritique. Ce travail examine également des solutions pour remédier au problème de brunissement enzymatique, comme le traitement thermique et l’addition de poudres de fruits naturellement riches en agents antibrunissement.Les phases de préparation des pommes sont de réelles étapes de transformation qu’il faut optimiser et maîtriser afin de préserver toutes les propriétés antioxydantes des coproduits végétaux. Nos travaux ont également permis de mettre en évidence des propriétés plus fonctionnelles des pelures de pommes, comme agent stabilisant d’émulsions. L’impact des paramètres d’extraction a été mis en évidence par l’exploration de plusieurs conditions telles que la masse chargée, le broyage, le protocole ainsi que la température et la composition du fluide extractant. / Apples are considered beneficial to health, because of their content of phenolic compounds that confer the label "health food". Fruits and vegetables food processes generate byproducts that are considered worthless by industrial. However, given the volumes generated, they are rich sources of nutrients.One possible way of valorization of these byproducts is the reintroduction of food as ingredient, to propose fortified foods with functional molecules. To do this, two approaches have been explored: the use of apple peels powders to stabilize an emulsion and the extraction of phenolic compounds by supercritical CO2+ethanol. This work also discusses solutions to remedy the enzymatic browning, such as heat treatment and the addition of naturally rich fruit powders as anti-browning agents.Apples preparation phases are real transformation steps which it’s necessary to optimize and master to preserve all the antioxidant properties of fruits and vegetables byproducts. Our works also highlight more functional properties of apple peels as a stabilizing emulsions agent. The impact of extraction parameters was demonstrated by the exploration of several conditions such as the loaded weight, the grinding, the protocol as well as the temperature and composition of supercritical fluids.

Les composés phénoliques sont des métabolites secondaires chez les végétaux, qui possèdent un large éventail de propriétés biologiques et fonctionnelles, par exemple antioxydantes, antimicrobiennes. Comme ils sont très répandus dans le règne végétal, les sous-produits issus de la transformation végétale représentent une source intéressante et peu coûteuse des polyphénols qui pourraient être utilisés comme additifs naturels, inoffensifs et multifonctionnels, avec une valeur de marché ajoutée. Toutefois, leur polarité limite leur utilisation dans des systèmes émulsionnés complexes, tels que ceux rencontrés en formulations cosmétiques, alimentaires, ou pharmaceutiques. Sur cette base, la lipophilisation s’est avérée être une stratégie efficace pour améliorer le caractère hydrophobe de ces composés bioactifs et, par conséquent, leurs propriétés fonctionnelles.Le principal objectif de ce travail a été d'ajouter de la valeur à des sources végétales riches en polyphénols par l'extraction, caractérisation et enfin, par lipophilisation des principaux polyphénols issus de ces sources. Dans ce contexte, Hibiscus sabdariffa L. ou roselle, cultivée dans le monde entier et considérée comme une importante source d’anthocyanes, avec des propriétés colorantes uniques, ainsi que des propriétés antioxydantes et antimicrobiennes, a été utilisée pour l'extraction assistée par ultrasons, suivi d’une seule étape de fractionnement par chromatographie en phase liquide à basse pression, pour isoler ces principaux anthocyanes. Les composés ainsi obtenus ont été utilisés pour évaluer leur possibilité à être lipophilisés en utilisant différentes longueurs de chaînes alkyles. Egalement, les propriétés colorantes des anthocyanes issues d’hibiscus ont été calculées par la méthode de pH jumps. Ainsi, les constantes de pseudo équilibre (pK'a 2,9), d’acidité (pKa 3,7) et d’hydratation (pK'h 3,0) de la delphinidine-3-O-sambubioside, ont été estimées.La delphinidine-3-O-sambubioside et la cyanidine-3-O-sambubioside ont été les principaux anthocyanes obtenus à partir de l’extrait d'hibiscus. Ces composés ont été lipophilisés avec succès pour la première fois, par voie chimique. Les produits ont été caractérisés par des techniques de chromatographie et spectrométrie. L'acide protocatéchique (PA), qui peut être extrait de la fleur d'hibiscus, a été utilisé comme molécule modèle, et il a été lipophilisé par voie chimique avec une série homologue d'alcools. Afin d'évaluer l'impact de la lipophilisation sur leurs propriétés fonctionnelles, leur activités antioxydantes (DPPH, ORAC, CAT), et antifongique (méthode de microdilution) ont été évaluées avant et après lipophilisation. De manière générale, les molécules estérifiées avec des alcools de chaîne courte et moyenne (de C1 à C6) ont montré des activités améliorées par rapport au composé non estérifié.En conclusion, cette étude a montré que la lipophilisation, en utilisant une longueur de chaîne optimale, pourrait avoir un impact positif sur les propriétés antioxydantes et antifongiques des composés phénoliques. Ces travaux de thèse ont également démontré que cette stratégie peut être appliquée à différentes familles de composés phénoliques, tels que les anthocyanes et les acides phénoliques, pour l’obtention des molécules pouvant être potentiellement utilisées comme additifs dans des produits de formulation. / Phenolic compounds are a group of secondary plant metabolites with a broad range of inherent biological and functional properties e.g. antioxidants and antimicrobial agents, all bundled within the phenol functional group. Since they are one of the most widely distributed molecules in the vegetal kingdom, by-products issued from the vegetal processing represent an interesting and inexpensive source of polyphenols that could be used as natural, innocuous and multifunctional ingredients, with an added market value. However, the polarity of these phenolic compounds limits their use in complex emulsified systems, such as food, pharmaceutical or cosmetic formulations. Based on this, lipophilization has showed to be an efficient strategy to enhance the hydrophobicity of these bioactive compounds and, consequently, their functional properties. The aim of this work was to add value to polyphenol-rich plant sources by the extraction, characterization and finally, lipophilization of their predominant polyphenols. In this context, Hibiscus sabdariffa L. or roselle flower, an inexpensive, worldwide produced source of anthocyanins, with unique color, and, antioxidant and antimicrobial properties, was used for ultrasound-assisted extraction and single-step fractionation by low-pressure liquid chromatography, to isolate its main anthocyanins. The obtained compounds were used to evaluate their suitability for lipophilization with different chain length alkyl donors. Moreover, the color properties of hibiscus anthocyanins were studied by the pH jumps method, which allowed the calculation of the pseudo equilibrium (pK'a 2.9), acidity (pKa 3.7) and hydration constants (pK'h 3.0).The obtained hibiscus anthocyanins (delphinidin-sambubioside and cyanidin-sambubioside) were successfully chemically lipophilized. The products were characterized by means of LC/ESI-MS. Moreover, protocatechuic acid (PA), which can be extracted from hibiscus flowers, was used as model molecule, and it was chemically lipophilized with a homologous series of alcohols. In order to evaluate the impact of the lipophilization on their functional properties, protocatechuates were assessed for their antioxidant (DPPH, ORAC, CAT), and antifungal (microdilution method) activity, before and after chemical lipophilization. In general, protocatechuates with short to medium alkyl chain lengths (from C1 to C6) showed improved activities compared to the parent compound. In conclusion, this study showed that, lipophilization using the optimal chain length could have a positive impact in the functional properties, e.g. antioxidant and antifungal activities, of phenolic compounds. Here was also demonstrated that this strategy can be applied to different families of phenolic compounds, such as anthocyanins and phenolic acids, representing an alternative to synthesize molecules that might have potential use as additives in formulated products.

Le réchauffement climatique planétaire annoncé ne sera pas sans conséquence sur le métabolisme de la baie et en particulier sur la teneur en composés phénoliques. Les objectifs de cette thèse visent à mieux comprendre les processus de régulation associant le microclimat des baies et la synthèse des composés phénoliques. Par des approches moléculaires et biochimiques, ce travail a permis de mieux décrire les réponses spécifiques des baies de raisin (cultivar Cabernet-Sauvignon) aux facteurs rayonnement et température.L'analyse du transcriptome des baies exposées soit à un stress thermique soit à un stress lumineux a mis en lumière deux processus, une réponse rapide ayant lieu dès les premières heures de traitement et une réponse apparaissant au bout de plusieurs jours d’exposition aux stress. Cette étude a également permis de valider le système expérimental de découplage des effets du rayonnement et de la température.L'analyse des profils d’expression d’une vingtaine de gènes intervenant dans la voie de biosynthèse des flavonoïdes révèle des différences dans la réponse transcriptionnelle des gènes en fonction du stress et du stade développement auquel il a été appliqué. Néanmoins cette réponse n'est pas ou peu corrélée avec les variations des teneurs en anthocyanes et flavonols observées. Les teneurs en anthocyanes sont fortement réduites par l’action de la chaleur alors que les teneurs de certains flavonols augmentent sous l’influence du rayonnement. Au niveau des acides présents dans la pulpe des baies, l’acide malique voit sa teneur réduite sous l’effet de la chaleur ainsi que d’une forte intensité lumineuse. Les analyses montrent un impact important du stress thermique sur la teneur de la phénylalanine, de la tyrosine et de la lysine dans la pellicule.Parallèlement, l'initiation d’une étude du métabolome des pellicules de baies de raisin a été entreprise par UPLC-ESI-LTQ-Orbitrap™ et a permis d'acquérir des bases solides pour optimiser le protocole utilisé en vue d'analyses futures. Cette étude permet de dresser une liste préliminaire de métabolites d’intérêts.Enfin, le gène VvGOLS1 (Galactinol synthase 1) voit son expression stimulée dans les baies exposées au stress thermique, ce qui se traduit par une accumulation de galactinol, précurseur de la voie des RFOs. Un rôle de molécule «signal» est envisagé pour le galactinol. Un régulateur de la transcription de VvGOLS1 a également été identifié par des expériences d'expression transitoire en protoplastes. Il s'agit du facteur de transcription VvHsfA2, dont l'expression est également stimulée par le stress thermique. Dans ce contexte, la caractérisation des gènes de la famille des Hsfs (Heat Stress Factors) a été initée. / Global warming will affect berry metabolism, and especially phenylpropanoïd contents. This PhD work aimed to acquire a better understanding on the cellular processus linking the microclimate and the phenolic synthesis. By molecular and biochemical approaches, we extended this study to detail specific responses taking place in berries under heat and light stress.Transcriptomic analysis of heat-stressed and light-stressed berries showed the existence of two processes that occur in exposed berries. The first one triggers a rapid and transient expression of genes within the first hours of treatment. The second one mobilizes a set of genes showing increase in their expression after several days of stress exposure. Furthermore, this study validated the experimental set used to discriminate the effects of light and temperature, respectively.Expression analysis of 20 genes involved in the flavonoid biosynthetic pathway revealed strong differences among the transcriptional responses, depending on the nature of stress and the developmental stage of the berry. However, expression patterns of genes involved in the biosynthesis of flavonoid could not fully explain the changes in anthocyanin and flavonol contents. This suggests that additional regulation processes such as post-traductional modifications of enzymes or metabolite degradation might take place in berries under abiotic stress. Anthocyanin content decreases under heat stress whereas flavonol content increases under high light. Malic acid increases in berry exposed to heat stress and high light. Moreover, heat-stressed berries showed an accumulation of phenylalanine, tyrosine and lysine in skin but not in pulp.In parallel, a metabolomic analysis was initiated on stress exposed berry skins by using UPLC-ESI-LTQ-Orbitrap™ technology. The first experiments revealed contrasted metabolite contents in berries according to the stress applied, and highlighted several metabolites of interest. The preliminary assays will help optimize this powerful tool for futures analysis.Finally, expression of VvGOLS1 (Galactinol synthase 1) was strongly induced in grape berries exposed to heat stress, in good agreement with the observed galactinol accumulation. Role of galactinol as a signaling molecule is discussed. Transient expression experiments revealed that VvGOLS1 expression is regulated at the transcriptional level through VvHsfA2 action. VvHsfA2 expression is also stimulated under heat stress. In this context, characterization of the grapevine heat stress factors was initiated.