This diploma thesis deals with possibilities of using of aronia fruit pomace. The theoretical part is focused on botanical characteristics of aronia fruit, description of HPLC and anthocyanins dyes. The effects of various extraction condition on content of some polyphenolic compounds (gallic and vanilla acids, katechin and rutin) in aronia berry pomace are investigated in the experimental part. The mentioned compounds were detected with HPLC. Thanks to this method the content of carbohydrates was also detected. A quantity of anthocyanins dyes was determined by pH differential method.

In this paper we present the inhibitory effect of a variety of structurally modulated/modified polyphenolic compounds on purified F 1 or membrane bound F 1F o Escherichia coli ATP synthase. Structural modulation of polyphenols with two phenolic rings inhibited ATP synthase essentially completely; one or three ringed polyphenols individually or fused together inhibited partially. We found that the position of hydroxyl and nitro groups plays critical role in the degree of binding and inhibition of ATPase activity. The extended positioning of hydroxyl groups on imino diphenolic compounds diminished the inhibition and abridged position enhanced the inhibition potency. This was contrary to the effect by simple single ringed phenolic compounds where extended positioning of hydroxyl group was found to be effective for inhibition. Also, introduction of nitro group augmented the inhibition on molar scale in comparison to the inhibition by resveratrol but addition of phosphate group did not. Similarly, aromatic diol or triol with rigid or planar ring structure and no free rotation poorly inhibited the ATPase activity. The inhibition was identical in both F 1F o membrane preparations as well as in isolated purified F 1 and was reversible in all cases. Growth assays suggested that modulated compounds used in this study inhibited F 1-ATPase as well as ATP synthesis nearly equally.

<p>U okviru disertacije ispitani su i upoređeni pokazatelji kvaliteta integralnog p&scaron;eničnog i integralnog heljdinog bra&scaron;na dobijenog mlevenjem netretiranih (netretirano integralno heljdino bra&scaron;no) i autoklaviranih zrna heljde (autoklavirano integralno heljdino bra&scaron;no). Nakon ispitivanja nutritivnih i funkcionalnih svojstava pomenutih bra&scaron;na, kreirane su i proizvedene integralne taljatele sa različitim sadržajem integralnog heljdinog bra&scaron;na (10&ndash;30%).<br />Pokazatelji senzorskog, nutritivnog i funkcionalnog kvaliteta proizvedenih taljatela sa integralnim heljdinim bra&scaron;nima ispitani su u poređenju sa kontrolnim uzorkom taljatela od integralnog p&scaron;eničnog bra&scaron;na.<br />U ispitivanim uzorcima bra&scaron;na i taljatela određeni su osnovni hemijski sastav, sadržaj mineralnih materija, sadržaj ukupnih rastvorljivih polifenola, utvrđen je kvalitativni i kvantitativni sastav polifenolnih jedinjenja, ispitana je antiradikalska aktivnost na DPPH˙ i određen je sadržaj fitinske kiseline i 5-hidroksimetilfurfural.<br />Pored analize nutritivnih i funkcionalnih svojstava, na taljatelama su sprovedena ispitivanja fizičkih osobina (dimenzije) i svojstva taljatela pri kuvanju. Instrumentalno su izmereni boja i teksturna svojstva nekuvanih i kuvanih taljatela. Senzorska ocena taljatela sprovedena je uz primenu panela utreniranih ocenjivača, metodom bodovanja i panela potro&scaron;ača, testom dopadljivosti.<br />Dobijeni rezultati ukazuju da se supstitucijom dela integralnog p&scaron;eničnog bra&scaron;na integralnim heljdinim bra&scaron;nom (autoklaviranim ili netretiranim) postiže unapređenje nutritivnog i funkcionalnog profila uzoraka obogaćenih taljatela, uz manje ili vi&scaron;e izraženo naru&scaron;avanje senzorskog profila, u zavisnosti od stepena supstitucije i vrste integralnog heljdinog bra&scaron;na.</p> / <p>This study investigated quality parameters of wholegrain wheat flour and wholegrain buckwheat flour from non-treated buckwheat grains (non-treated wholegrain buckwheat flour) and autoclaved buckwheat grains (autoclaved wholegrain buckwheat flour). After investigation of nutritional and functional properties of aforementioned flours, new wholegrain tagliatelle formulations, with different wholegrain buckwheat flour content (10&ndash;30%) have been developed.<br />Sensory, nutritional and functional quality parameters of buckwheat containing tagliatelle samples were analysed and compared with control tagliatelle sample from wholegrain wheat flour.<br />Proximate composition, content of minerals, content of total polyphenols and quantitative and qualitative composition of polyphenols were investigated in both, flour samples and produced uncooked and cooked tagliatelle samples. Antiradical activity on DPPH radicals, and content of less favourable compounds, phytic acid and HMF, have been tested, as well.<br />Tagliatelle physical properties, dimensions, cooking properties, colour and textural properties were determined by instrumental and sensory methods. Tagliatelle acceptability was evaluated by the consumers. The obtained results indicate that wholegrain buckwheat flour (autoclaved or non-treated) incorporation into tagliatelle formulation led to an improvement of nutritional and functional profiles of tagliatelle samples, followed by greater or lesser deterioaration of sensory profile, depending on substitution level and type of wholegrain buckwheat flour.</p>

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / Antioxidants are compounds known as free radicals react with and/or reactive oxygen species in order to idle them, preventing the oxidative damage. Many studies have pointed to the imbalance between oxidants and antioxidants as the cause of many diseases and because of the great importance given to this issue, there is a strong demand for studies involving the identification and quantification of compounds with antioxidant activity. This work was then identify and quantify the polyphenolic compounds full of grains of Brazilian cultivars of barley, grown in the municipality of Ibiaçá/RS, in the agricultural year of 2005 and 2006, from the Research Center of Embrapa/Wheat, Passo Fundo/RS and assess the weather conditions (average temperature, precipitation index and insolation) between the time of planting and harvesting barley in the quantification of phenolic compounds between the different seasons. The samples were characterized chemically related to the presence of polyphenols, using the method of separation based on the system of high performance liquid chromatography in reversed phase (RP-HPLC) with UV-VIS detection to 254 nm. We identified the phenolic compounds rutin, caffeic acid, ferulic acid, quercitrin and myricetin, with rutin and caffeic acid the most abundant among cultivars. Alongside was a quantification of phenolic compounds by the Folin-Ciocalteau technique of aiming at a comparison with the chromatographic technique. The techniques have proved satisfactory for the purposes of identification and quantification of differentiation in barley samples analyzed. There was a chemical differentiation of the varieties of barley in relation to polyphenolic compounds identified and quantified and between varieties of different years of cultivation. In the assessment of climatic factors, it was observed that a lower average temperature, a higher rainfall and less sunshine received by barley between planting and harvest seasons, reflected in an increase of total phenols using the Folin-Ciocalteu and quantification of HPLC flavonoid rutin. / Os antioxidantes são compostos conhecidos por reagirem com radicais livres e/ou espécies reativas de oxigênio, de forma a inativá-los, prevenindo os danos oxidativos. Muitos estudos têm apontado para o desequilíbrio entre oxidantes e antioxidantes como o causador de muitas patologias, e devido à grande importância dada a este tema, existe uma forte demanda de estudos envolvendo a identificação e quantificação dos compostos com atividade antioxidante. O objetivo deste trabalho foi então quantificar e identificar os compostos polifenólicos de grãos integrais de cultivares brasileiras de cevada, cultivadas no município de Ibiaçá/RS, no ano agrícola de 2005 e 2006, provenientes do Centro de Pesquisa da Embrapa/Trigo, Passo Fundo/RS, e avaliar as condições climáticas (temperatura média, índice pluviométrico e insolação) entre a época de plantio e colheita da cevada na quantificação dos fenólicos totais entre as diferentes safras. As amostras foram caracterizadas quimicamente com relação à presença de polifenóis, empregando-se o método de separação baseado no sistema de cromatografia líquida de alta eficiência em fase reversa (RP-HPLC) com detecção UV-VIS a 254nm. Foram identificados os compostos fenólicos rutina, ácido caféico, ácido ferúlico, quercitrina e miricetina, sendo a rutina e o ácido caféico os mais abundantes entre as cultivares. Paralelamente foi realizada uma quantificação dos fenólicos totais pela técnica de Folin-Ciocalteu, visando uma comparação com a técnica cromatográfica. As técnicas mostraram-se satisfatórias para os objetivos de quantificação e identificação na diferenciação das amostras de cevada analisadas. Observou-se a diferenciação química das variedades de cevada com relação aos compostos polifenólicos identificados e quantificados e entre variedades de diferentes anos de cultivo. Na avaliação dos fatores climáticos, foi observado que uma menor temperatura média, um maior índice pluviométrico e uma menor insolação recebida pela cevada entre as épocas de plantio e colheita, refletiram em um aumento dos fenóis totais pelo método de Folin-Ciocalteu e na quantificação do flavonóide rutina por HPLC.

Au cours de ce travail de thèse nous avons étudié l’influence des apports en azote nitrique sur le contenu en polyphénols chez la tomate. Plusieurs dispositifs de culture hors sols ont été utilisés (hydroponie, laine de roche, NFT). Nous avons quantifié les principaux composés phénoliques de la tomate, à savoir : l’acide chlorogénique, la rutine, le kaempférol rutinoside dans les parties végétatives de la plante (limbe, tige et racine), ainsi que des dérivés de l’acide caféique et la naringénine chalcone dans les fruits. Ces composés ont été analysés par chromatographie liquide à haute performance. Nous avons observé que les limbes étaient le compartiment le plus sensible aux conditions de nutrition. Nous y avons observé une multiplication par deux des concentrations en polyphénols lors d’une diminution des apports en azote de 15 à 0.05mM NO3-. Nous avons mis en évidence, en établissant des courbes de réponses, que ces augmentations s’effectuaient de façon nettement plus importante dans des conditions de nutrition azotée limitante pour la croissance de la plante. Dix à 20 jours de carence ou de limitation semblent nécessaires pour obtenir une modification du contenu en polyphénols à l’échelle de la plante. Dans les fruits les concentrations en polyphénols ont été modifiées par le niveau de fertilisation azotée, mais nous n’avons pas mis en évidence d’augmentations très significatives. L’impact d’autres facteurs, comme les conditions climatiques (lumière et température) a également été pris en compte. Nos résultats semblent indiquer que les conditions climatiques régissent le contenu en polyphénols de manière plus importante que la nutrition azotée / During my PhD we studied the effects of nitrate supply on tomato polyphenolics content. Several cultural systems were used (hydroponic, rockwool culture, NFT). We quantified the main tomato phenolic compounds: chlorogenic acid, rutine, kaempferol rutinoside in vegetative parts (leaf, stem, root), together with some caffeic acid derivates and naringenine chalcone in fruits. These compounds were analyses by High Performance Liquid Chromatography. We noticed that leaves were the more responsive compartment of the plant, to the nutrition conditions. We observed a two-fold increase in polyphenolics concentrations when nitrate supply decrease from 15 to 0.05mM. We found, by drawing response curves, that this increase was more important when nitrate supply limited plant growth. Ten to 20 days seem necessary to observe a modification, at the plant level, of polyphenolics content. In tomato fruits, polyphenolics concentrations were modified by the nitrate supply, but we do not observed very significant increases. The effects of other environmental factors, such as climate conditions (light, temperature) were studied. Our results seem to indicate that climate is more important than nitrogen nutrition for the determination of the polyphenolic compounds concentrations

La Valle d'Aosta, pur non avendo grandi superfici destinate alla viticoltura, è caratterizzata dalla presenza di un ricco patrimonio ampelografico di cui fanno parte dodici varietà autoctone a bacca rossa: Bonda, Cornalin, Crovassa, Fumin, Mayolet, Ner d'Ala, Petit rouge, Premetta, Roussin, Roussin de Morgex, Vien de Nus e Vuillermin. Malgrado i numerosi vitigni menzionati, nella produzione dei vini a D.O.C troviamo in prevalenza il Petit rouge e poche altre varietà che, nell'insieme, rappresentano circa il 34% della produzione totale. Al fine di implementare la presenza di prodotti tipici che esprimano i caratteri distintivi della zona di provenienza sono state approfondite le conoscenze dei vitigni autoctoni per evidenziarne le attitudini e le potenzialità per la produzione di vini rossi di qualità. Allo stesso scopo sono stati valutati gli effetti dell'applicazione di alcune tecniche di vinificazione alternative sull'espressione dei caratteri qualitativi del Petit rouge, le cui uve sono da tempo vinificate in purezza o in assemblaggi nella produzione di vini a D.O.C. / Aosta Valley has a little viticulture surface and it is characterised by a rich ampelographic collection including the following twelve red grape autochthonous varieties: Bonda, Cornalin, Crovassa, Fumin, Mayolet, Ner d'Ala, Petit rouge, Premetta, Roussin, Roussin de Morgex, Vien de Nus e Vuillermin. Only the 34% of the D.O.C. wine production is represented by some of the above mentioned cultivars, in particular the Petit Rouge, despite of the huge number of cultivars. The Petit rouge is used in purity or in miscellaneous in the D.O.C. wine production. In order to implement the presence of typical products, this study was carried out to evaluate the aptitude of Aosta Valley ancient grapes to produce quality red wines. Moreover to improve quality of the most diffused Aosta Valley red wine the effects of some alternative winemaking were evaluated on the Petit rouge characters.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / This paper describes the development of methodology for capillary electrophoresis with UV detection for the determination of 14 polyphenolic antioxidants in extracts of Connarus perrottetii var. angustifolius Radlk, a native plant of the Amazon forest. The method using capillary zone electrophoresis (CZE) allows simultaneous determination of 3-acetyilcoumarine, resveratrol, 6-hydroxycoumarin, catechin, rutin, ferulic acid, quercitrin, kaempferol, fisetin, myricetin, quercetin, caffeic acid, gallic acid and 4 hydroxicinnamic acid in the optimized conditions: borate buffer 20 mmol L-1 (pH 9.2) as work electrolyte containing 15% methanol (v /v), -20 kV potential separation, temperature 25 °C; hydrodynamic injection gravity 20 cm for 60 s. The method was validated on parameters of linearity, limit of detection, quantification limit, precision and accuracy, and was used in the analysis of aqueous infusion and ethanol, butanol and ethyl acetate plant fractions. The method was capable of identifying the antioxidants present in samples with high selectivity and sensitivity. In infusion and in ethanol and butanol fractions was found the presence of catechin and rutin as major compounds, which makes up about 0.5% of the total mass of medicinal plant. The obtained dry extracts (ethanol and butanol) allowed the concentration of rutin and catechin around 20 times compared to the dry plant. In addition, tests were performed to measure the antioxidant activity of the extracts against DPPH radical, superoxide anion radical, hydroxyl and peroxyl. A sample of aqueous infusion of 10% showed the highest activity against most free radicals. It corroborates the results of characterization by capillary electrophoresis, where catechin and rutin were determined in relatively high concentrations in the aqueous infusion. But all treatments of the samples showed good antiradicalar behavior with the methodologies applied. Against DPPH, for example, the ethyl acetate fraction showed a potential of 94,43%. Against superoxide and hydroxyl radicals, the butanol fraction showed the best performance: 64,23% and 74,97%, respectively. / Este trabalho descreve o desenvolvimento de metodologia em eletroforese capilar com detecção UV para a determinação de 14 antioxidantes polifenólicos em extratos de Connarus perrottetii var. angustifolius Radlk, planta nativa da Amazônia brasileira. O método empregando eletroforese capilar de zona (CZE) permite a determinação simultânea de 3-acetilcumarina, resveratrol, 6-hidroxicumarina, catequina, rutina, ácido ferúlico, quercitrina, canferol, fisetina, miricetina, quercetina, ácido cafeico, ácido gálico e ácido 4-hidroxicinâmco nas condições otimizadas: eletrólito de trabalho tampão borato 20 mmol L-1 (pH 9,2) contendo metanol 15% (v/v), potencial de separação -20 Kv, temperatura 25 °C; injeção hidrodinâmica por gravidade em 20 cm durante 60 s. O método foi validado nos parâmetros de linearidade, limite de detecção, limite de quantificação, precisão e exatidão e foi aplicado na análise da infusão aquosa e frações etanólica, butanólica e acetato de etila da planta. O método foi capaz de identificar os antioxidantes presentes nas amostras com alta seletividade e sensibilidade. Na infusão e nas frações etanólica e butanólica foi encontrada a presença de catequina e rutina como compostos majoritários, os quais compõe cerca de 0,5% da massa total de planta medicinal. Os extratos secos obtidos (etanólico e butanólico) possibilitaram a concentração de rutina e catequina em torno de 20 vezes em relação à planta seca. Além disso, foram realizados testes para mensurar a atividade antioxidante dos extratos frente aos radicais DPPH, ânion radical superóxido, hidroxila e peroxila. A amostra de infusão aquosa a 10% foi a que apresentou maior atividade frente à maioria dos radicais livres. Esse resultado corrobora os resultados de caracterização por eletroforese capilar, onde catequina e rutina foram determinadas em concentrações relativamente altas na infusão aquosa. Porém, todos os tratamentos das amostras apresentaram bom comportamento anti-radicalar com as metodologias aplicadas. Frente ao radical DPPH, por exemplo, a fração acetato de etila demonstrou um potencial de 94,43%. Frente aos radicais superóxido e hidroxila, a fração butanólica apresentou o melhor desempenho: 64,23% e 74,97%, respectivamente.

Corynebacterium diphtheriae, Pseudomonas aeruginosa, Ricinus communis, Shigella dysentariae, and Vibrio cholerae produce AB toxins which share the same basic structural characteristics: a catalytic A subunit attached to a cell-binding B subunit. All AB toxins have cytosolic targets despite an initial extracellular location. AB toxins use different methods to reach the cytosol and have different effects on the target cell. Broad-spectrum inhibitors against these toxins are therefore hard to develop because they use different surface receptors, entry mechanisms, enzyme activities, and cytosolic targets. We have found that grape seed extract provides resistance to five different AB toxins: diphtheria toxin (DT), P. aeruginosa exotoxin A (ETA), ricin, Shiga toxin, and cholera toxin (CT). To identify individual compounds in grape seed extract that are capable of inhibiting the activities of these AB toxins, we screened twenty common phenolic compounds of grape seed extract for anti-toxin properties. Three compounds inhibited DT, four inhibited ETA, one inhibited ricin, and twelve inhibited CT. Additional studies were performed to determine the mechanism of inhibition against CT. Two compounds inhibited CT binding to the cell surface and even stripped bound CT off the plasma membrane of a target cell. Two other compounds inhibited the enzymatic activity of CT. We have thus identified individual toxin inhibitors from grape seed extract and some of their mechanisms of inhibition against CT. This work will help to formulate a defined mixture of phenolic compounds that could potentially be used as a therapeutic against a broad range of AB toxins.