This thesis deals with the preparation of the combined alcoholic beverages based on wine and fruit juices. The fruit juices were obtained from the selected fruits as honeysuckle, mulberry, dogwood, rose hips, elderberry, buckthorn and aronia. Some chemical properties e. g. total fenolics, total anthocyanins, vitamin C and sacharides were determined for characterization of the mentioned juices. As well as chemical parameter of juices, the same chemical parameters were determined on wine. There were prepared three sets of combined beverages from wine and juices. The two best evaluated beverages were chosen on the basis of their sensory evaluation. In the end, the same chemical properties were determined in both particular juices and resulting beverages.

This Diploma thesis focuses on the study of penetration of selected phenolic substances contained in extract from black elder (Sambucus nigra) via modelling systems represented by two types of synthetic membranes and a real system represented by a biological membrane. As a biological membrane, we have chosen skin from pig´s earlobe. Experiments of transdermal absorption and absorption through synthetic membranes were conducted through vertical (Franz´s) cells. Selected substances contained in the extract from black elder were rutin and chlorogenic acid. The main focus of this work was to determine how readily, or if at all, these two selected phenolic substances penetrate the skin and selected synthetic membranes. The percentage of absorbed amount of the substance was determined by HPLC analysis.

No description available.

I det här projektet extraherades värdefulla komponenter från äpple-pomace som är en restprodukt från cidertillverkning. Restprodukten var tillhandahållen av en svensk cidertillverkare. Extraktionen fokuserades mot pektin och fenolföreningar som uppvisar antioxiderande aktivitet. Extraktionsprocessen designades ur ett miljövänligt perspektiv, inga skadliga kemikalier användes. Fenolära substanser extraherades initialt från pomacen med en vattenlösning innehållande 50% etanol. Därefter studerades kompositionen och den antioxiderande aktiviteten i dessa extrakt. Fokuseringen i projektet låg främst på pektin. Pektin extraherades med subkritiskt vatten vid tre olika pH (3,5 och 7) samt vid två skilda temperaturer (120°C och 140°C) i antingen 5, 10 och 15 minuters sekvenser. Pektinet karakteriserades med avseende på extraktionsutvinningsgrad, sockerkomposition, molekylvikt och antioxiderande aktivitet. Resultaten jämfördes för att undersöka effekten av pH, temperatur och extraktionstid. Gelningsegenskapen hos olika pektin studerades som ett bevis på koncept i ett empiriskt experiment där hög-viskösa vätskor bestående av 5% pektin och 60% sackaros framställdes. Extraherade fenolföreningar har potential att fungera som naturligt utvunna antioxidanter i kosmetiska produkter och pektin kan tänkas användas som reologimodifierare i vattenbaserade formuleringar med låga pH-värden utan behov av kemiska modifieringar. / In this project, apple pomace from a Swedish cider making factory as a by-product was used as a raw material to extract valuable compounds. The extraction was focused on pectin and phenolic compounds with antioxidant activity. For the extraction procedures environmentally friendly processes were chosen without using any harsh chemicals. Phenolic compounds wereinitially extracted from the pomace using 50% aqueous ethanol and then the composition, total phenolic content and antioxidant activity were studied in these extracts. The main focus was on pectins, which were extracted by subcritical water at three different pH conditions (pH 3, 5 and 7) and two different temperatures (120°C and 140°C) in 5-, 10- and 15 minute sequences. Then the pectins were characterized in terms of extraction yield, sugar composition, molecular weight and antioxidant activity and the results were compared in terms of the effect of pH, temperatur eand extraction time. The gelling properties of the different pectins were also studied as a proof of concept in an empirical experiment, where highly viscous liquids were obtained at 5% pectinand 60% sucrose content. In summary, the extracted phenolic compounds have potential to function as naturally derived antioxidants in cosmetics and the pectin may be used as a rheology modifier in water-based formulations of low pH without any additional chemical modifications.

No description available.

<p>U okviru disertacije su razvijeni postupci za iskori&scaron;ćenje sporednog produkta prerade paradajza (tropa) kao polazne sirovine za proizvodnju prehrambenih proizvoda sa dodatom vredno&scaron;ću. Sprovedena istraživanja kretala su se u dva smera: proizvodnja lipofilnih i hidrofilnih ekstrakata od liofilizovanog tropa paradajza, primenom rastvarača koji su dozvoljeni za upotrebu u prehrambenoj industriji (n-heksana i 95% etanola) i njihova karakterizacija u pogledu sadržaja i sastava fitohemikalija i biolo&scaron;ke aktivnosti in vitro; razvoj formulacija i tehnolo&scaron;kih postupaka za kreiranje gotovih proizvoda sa povećanim sadržajem prehrambenih vlakana i njihova karakterizacija u smislu nutritivnih, reolo&scaron;kih i senzorskih svojstava. Proizvedena su tri proizvoda sa povećanim sadržajem prehrambenih vlakana: kečap od svežeg tropa paradajza, kečap od liofilizovanog tropa paradajza u prahu i marmelada od liofilizovanog tropa paradajza u prahu.<br />Proizvedeni lipofilni i hidrofilni ekstrakti pokazali su sposobnost redukcije jona gvožđa, kao i antiradikalsku aktivnost na DPPH, azot (II)-oksid i superoksid-anjon radikale. Karotenoidi detektovani u lipofilnim ekstraktima (likopen i &beta;-karoten) povezani su sa vi&scaron;om antiradikalskom aktivno&scaron;ću na superoksid-anjon radikal, dok su fenolna jedinjenja detektovana u hidrofilnim ekstraktima (hidroksicinamati, flavonoli i dihidrohalkoni) bila efikasnija u redukciji jona gvožđa. Etanolni ekstrakt tropa paradajza pokazao je inhibitornu aktivnost na angiotenzin-konvertujući enzim pri koncentraciji ekstrakta od 10 mg/ml.<br />Kečap proizveden od svežeg tropa paradajza, kečap proizveden od liofilizata tropa paradajza i marmelada proizvedena od liofilizata tropa paradajza su bili okarakterisani povećanim sadržajem prehrambenih vlakana (preko 3 g na 100 g proizvoda), na osnovu kog mogu da se deklari&scaron;u kao izvor vlakana. Marmelada je proizvedena od liofilizovanog tropa paradajza bez dodatka hidrokoloida, sa 20% manjim sadržajem &scaron;ećera u odnosu na komercijalne proizvode.<br />Reolo&scaron;ka svojstva kečapa proizvedenog od svežeg tropa paradajza su prevashodno zavisila od koncentracije nerastvorljivih čestica. Rezultati senzorske ocene sprovedene uz primenu panela treniranih ocenjivača, ukazuju da su miris na paradajz i aroma paradajza bili najvažniji za jasno razlikovanje kečapa proizvedenog od svežeg tropa paradajza od komercijalnih proizvoda.<br />Kečap i marmelada proizvedeni od liofilizovanog tropa paradajza se, kao reolo&scaron;ki sistemi, mogu opisati modifikovanom frakcionom Kelvin-Voigt-ovom jednačinom kao viskoelastična tela. Ispitivanje model sistema sastavljenih od različitih odnosa liofilizata tropa paradajza i vode tretiranih na temperaturi od 60&deg;C i 100&deg;C ukazalo je da su vrednosti modula elastičnosti (G&rsquo;) zavisile od koncentracije liofilizata tropa i da su G&rsquo; vrednosti niže na 100&deg;C u odnosu na 60&deg;C. Test dopadljivosti kečapa i marmelade od liofilizovanog tropa paradajza, sproveden primenom hedonske skale od 1 do 7, ukazuje da su oba proizvoda prihvaćena od strane potro&scaron;ača (prosečna ocena za ukupnu dopadljivost veća od 4).</p> / <p>This dissertation describes processes for utilization of tomato processing byproduct (tomato pomace) as raw material for value-added food products. Research was conducted in two directions: one is production of lipophilic and hydrophilic tomato pomace extracts using solvents that are allowed for use in the food industry (n-hexane and 95% ethanol) and their characterization in terms of content and composition of phytochemicals and their in vitro biological activity; the other one is development of formulations and technological processes for the creation of tomato pomace based products with increased content of dietary fiber and their characterization in terms of nutritional, rheological and sensory properties. Three products with increased dietary fiber content were produced: ketchup was produced from fresh tomato pomace, and ketchup and jam were produced from lyophilized tomato pomace powder.<br />Produced lipophilic and hydrophilic extracts showed ferric ion reducing ability, as well as DPPH, nitric oxide and superoxide anion radical scavenging activity. Carotenoids detected in lipophilic extracts (lycopene and &beta;-carotene) were correlated with higher superoxide anion radical scavenging activity, while phenolic compounds detected in hydrophilic extracts (hydroxycinnamates, flavonols and dihydrochalcones) were more effective in ferric ion reduction.<br />Ketchup produced from fresh tomato pomace, ketchup produced from lyophilized tomato pomace, and jam produced from lyophilized tomato pomace were characterized by increased content of dietary fiber (more than 3 g per 100 g of product). Jam was produced from the lyophilized tomato pomace without addition of hydrocolloids, with 20% lower sugar content in comparison with commercial products. Rheological properties of ketchup produced from fresh tomato pomace primarily depended on the insoluble particles content. The results of sensory analysis indicate that tomato odor and flavor were the most important for clear separation of ketchup produced from fresh tomato pomace from commercial products.<br />Ketchup and jam produced from lyophilized tomato pomace could be described, as rheological systems, by the application of modified fractional Kelvin-Voigt model as viscoelastic solids. The examination of model systems comprised of different lyophilized tomato pomace:water ratios heat treated at 60&deg;C and 100&deg;C indicated that storage modulus (G&rsquo;) values depended on the concentration of lyophilized tomato pomace and that G&rsquo; values were lower at 100&deg;C than at 60&deg;C. Preference test for ketchup and jam, conducted by application of 7-point hedonic scale, indicated that both products were accepted by the consumers (average score for overall acceptability higher than 4).</p>

L’industrie libanaise du raisin et de la viniculture acquit dans les quelques années passées une renommée internationale du fait de la bonne qualité du vin produit. Il a été démontré que les constituants suspectés d’être responsable des caractéristiques biologiques du raisin sont les composés phénoliques. Ces derniers sont des métabolites secondaires des plantes, impliquées dans la protection vis-à-vis des conditions extrêmes environnementales et dans la défense contre les parasites. Ces composés contribuent aussi au goût, à l’astringence et à la couleur des produits issus du raisin. De plus, ils exhibent un pouvoir antioxydant, de ce fait un large spectre d’effets pharmacologiques leur sont attribués i.e. des propriétés anti-inflammatoire, anti-carcinogène et antiallergique. Ils peuvent aussi réduire le risque des maladies cardio-vasculaires. La production des composés phénoliques dans les fruits peut être affectée par le microclimat et les caractéristiques du terroir.En premier lieu, nous avons optimisé l’extraction des composés phénoliques totaux et des anthocyanes du raisin cultivé au Liban. Un modèle statistique basé sur la méthodologie de la surface de réponses a été utilisé pour étudier les effets de paramètres tels que le temps, la température, le choix et ratio du solvant organique dans le mélange sur la procédure d’extraction et la determination des conditions optimales. La Chromatographie Liquide à Haute Performance ou HPLC a aussi été utilisée pour suivre l’évolution de quelques composés phénoliques en fonction de la modification des paramètres d’extraction.En second lieu, afin d’étudier l’impact des composés phénoliques sur la santé humaine, l’effet de l’extrait de raisin obtenu à paritir du cépage Cabernet Sauvignon (Vitis Vinifera L. Cv. Cabernet Sauvignon) sur les cellules d’adénocarcinome colorectal – HT-29 a été déterminé.La viabilité, la prolifération, l’apoptose et le cycle cellulaire ainsi que le potentiel de membrane mitochondrial en présence de l’extrait de raisin, ont été mesurés à l’aide des techniques de microscopie, de cytométrie en flux et d’histochimie.La technologie des puces à ADN a été utilisée pour détecter des changements au niveau de l’expression des gènes dans les cellules HT-29 et pour révéler les voies de signalisation impliquées dans la réponse cellulaire aux composés phénoliques. Cette étude couvre différents aspects de la biochimie des composés phénoliques et aide à mieux comprendre leur impact sur la santé humaine, surtout au niveau cellulaire. De plus, ce projet de thèse ajoute à la valeur de l’industrie libanaise du raisin et de ses produits en proposant des conditions optimales pour extraire, analyser les composés phénoliques et montrer leurs qualités à l’échelle cellulaire. / Due to wine high quality, the Lebanese grape industry and wineries became in the past few years a business of worldwide trust. The components proved to be responsible of the grapes biological characteristics are the phenolic compounds. These are secondary plant metabolites especially implicated in plant protection against environmental extremes and defense against parasites. They as well contribute to flavor, astringency and color characteristics of grape products. Moreover, they are powerful antioxidants and exhibit a wide range of biochemical and pharmacological effects, including anti-inflammatory, anti-carcinogenic and anti-allergic properties. They can also reduce the risk of coronary heart diseases. The phenolic compounds production in fruits may be affected by microclimate and soil characteristics.Our study aimed first of all to optimize the extraction of total phenolic compounds and antocyanins from Lebanese planted grapes.Statistical experimental design based on the response surface methodology was employed to investigate the effects of parameters like time, temperature, organic solvent / water ratio on the extraction procedure, and the determination of optimum conditions.HPLC (High Performance Liquid chromatography) was also used to monitor the evolution of some phenolic compounds when modifying the extraction parameters.Second of all, to study the impact of phenolic compounds on human health, the effect of the extract obtained from Cabernet Sauvigon (Vitis vinifera L. Cv. Cabernet Sauvignon) grapes was determined on a colorectal adenocarcinoma cell line; HT-29. Cell viability/proliferation, apoptosis, cell cycle and mitochondrial membrane potential in presence of the extracts, were measured using microscopy, flow cytometry and fluorescent staining.The DNA chips technology was used to detect changes at the gene expression level of the cell line used in this study after incubation with grape extracts, and to reveal certain pathways implicated in cellular response to phenolic compounds.This study covers different aspects in the biochemistry of phenolic compounds and helps in a better comprehension of their impact on human health at the cellular level. This also provides a better value to the Lebanese grape industry and products by proposing optimal conditions to extract, monitor the phenolic compounds and by showing their roles on the cellular level

A geração de resíduos sólidos pelas atividades agroindustriais tem criado a demanda por um reaproveitamento tecnológico desses materiais. Assim, o objetivo deste trabalho foi avaliar o potencial bioativo e tecnológico de resíduos agroindustriais, como fontes naturais de compostos fenólicos com atividade antioxidante. Foram analisados resíduos agroindustriais vinícolas, de indústrias produtoras de polpas congeladas de frutas (açaí, cajá, cupuaçu e graviola) e provenientes do beneficiamento de café e de laranja. Inicialmente, foi realizado um estudo para a determinação das condições ótimas de extração, empregando planejamento experimental multivariado com delineamento composto central rotacional, cujos resultados foram avaliados empregando a técnica de superfície de resposta. Na sequência, foram feitos a triagem dos resíduos, baseada na atividade antioxidante, e a caracterização fenólica dos extratos hidroalcoólicos obtidos dos resíduos agroindustriais. De acordo com os resultados de atividade antioxidante, engaço de uva da variedade Chenin Blanc (EC) e semente de açaí (SA) foram os resíduos selecionados, os quais seguiram para as etapas de concentração e fracionamento bioguiado de sua(s) molécula(s) bioativa(s), as quais foram posteriormente identificadas por UHPLC-ESI-LTQ-MS. Extratos brutos e concentrados foram avaliados in vitro quanto à capacidade de desativação de espécies reativas de oxigênio (radicais peroxila, ânion superóxido e ácido hipocloroso) e então, aplicados em óleo de soja, emulsão e suspensão de lipossomos, a fim de se avaliar a efetividade desses extratos como antioxidante natural em matrizes lipídicas. Concentrações intermediárias de etanol (40-60%) e alta temperatura (96°C), exceto para semente de açaí (25°C), foram as condições ótimas para a extração de antioxidantes dos resíduos agroindustriais. Epicatequina, ácido gálico, catequina e procianidina B1 foram os compostos de maior ocorrência, quando avaliados pela técnica de HPLC-DAD. O EC apresentou a maior atividade antioxidante global e SA a maior atividade entre os resíduos de polpas de frutas, laranja e café. A concentração dos extratos brutos de EC e SA, pela resina Amberlite XAD&reg;-2, produziu aumento significativo da atividade antioxidante. Além disso, extratos brutos e concentrados apresentaram atividade antiproliferativa e anti-inflamatória. Os extratos concentrados foram fracionados por meio de Sephadex LH-20, a partir da qual foi possível identificar quatro frações de maior bioatividade para o EC e três para o SA. Procianidina B1, catequina, epicatequina e resveratrol foram identificados no extrato concentrado e frações de EC. Dezoito procianidinas poliméricas, catequina, epicatequina foram os principais compostos identificados em SA, por meio de UHPLC-ESI-LTQ-MS. Resveratrol também foi encontrado em SA pela primeira vez. Quando avaliados em óleo de soja, EC e SA demonstraram atividade pro-oxidante. Contudo, elevada atividade antioxidante foi verificada quando essas amostras foram aplicadas em sistemas lipídicos coloidais, pois retardaram o consumo de oxigênio em uma emulsão óleo/água e o período de indução na produção de dienos conjugados em uma suspensão de lipossomos. Portanto, os resíduos agroindustriais EC e SA possuem potencial tecnológico de reaproveitamento industrial podendo ser considerados possíveis matérias-primas para a obtenção de extratos ricos em antioxidantes ou pela extração de antioxidantes naturais de uso pelas indústrias farmacêutica e/ou de alimentos. / The solid waste generation by agroindustrial activities have created the demand for a technology reuse of these materials. The objective of this study was to evaluate the bioactive and technological potential of agroindustrial residues as sources of phenolic compounds with antioxidant activity. Agroindustrial residues from wineries, from industries producing frozen fruit pulps (acai, caja, cupuaçu and soursop) and from the processing of coffee and orange, were analyzed. Initially, a study using a multivariate experimental planning with central composite rotatable design was performed, whose results were evaluated by response surface technique. After, it was made the screening based on the antioxidant activity and the phenolic characterization of hydroalcoholic extracts of optimized agroindustrial residues. According to the results obtained for the antioxidant activity the residues grape stem from the variety Chenin Blanc (EC) and açaí seed (SA) were selected and followed to the steps of concentration and bioguided fractionation of their(s) molecule(s) bioactive(s), which were subsequently identified by UHPLC-ESI-LTQ-MS. Crude and concentrated extracts were evaluated in vitro for the deactivation capability of reactive oxygen species (peroxyl radicals, superoxide and hypochlorous acid) and then applied to soybean oil, emulsion and liposome suspension in order to evaluate the effectiveness these extracts as natural antioxidant in lipid matrices. Intermediate concentrations of ethanol (40-60%) and high temperature (96 °C), except for acai seed (25 °C) were the optimal conditions for the extraction of antioxidants from agroindustrial residues. Epicatechin, gallic acid, catechin and procyanidin B1 compounds were the most frequent molecules, when assessed by HPLC-DAD. The EC had the highest overall antioxidant activity and SA the greatest activity between residues of fruit pulp, orange and coffee. Concentration of the crude extracts of EC and SA, by Amberlite XAD&reg;-2 resin, produced a significant increase in antioxidant activity. Furthermore, crude and concentrated extracts showed antiproliferative and anti-inflammatory activity. The concentrated extracts were fractionated by Sephadex LH-20, from which it was possible to identify four fractions of greater bioactivity for the EC and three for SA. Procyanidin B1, catechin, epicatechin and resveratrol were identified in the concentrated extract and EC fractions. Eighteen polymeric procyanidins, catechin, epicatechin were the major compounds identified in SA by UHPLC-ESI-LTQ-MS. Resveratrol has also been found for the first time in SA. When evaluated in soybean oil, EC and SA demonstrated pro-oxidant activity. However, high antioxidant activity was observed when these samples were evaluated on lipid colloidal systems, for delayed oxygen consumption in an emulsion oil/water and the induction period in the production of conjugated dienes in a liposome suspension. Therefore, the agroindustrial residues EC and SA have technological potential for industrial reuse and, thus, can be considered as raw material for obtaining antioxidant-rich extracts or by extraction of natural antioxidants useful for pharmaceutical and/or food industries.

O Brasil apresenta grande potencial para a exploração da apicultura, dado ao seu vasto território e flora diversificada, o que permite diferentes variedades de méis com propriedades únicas. O estado do Paraná é um dos maiores produtores de méis do país e o investimento em produção que atenda aos mercados mais exigentes estimulou a produção do mel orgânico. O conhecimento desde a antiguidade sobre os efeitos benéficos à saúde pelo mel vem estimulando a pesquisa deste alimento nobre. Assim, este trabalho teve por objetivo estudar méis orgânicos brasileiros certificados (MO) para a caracterização do perfil fenólico, volátil, além da avaliação da capacidade de sequestro das espécies reativas de oxigênio e nitrogênio. Os méis foram coletados nos apiários de apicultores com certificação orgânica de dois municípios do sul do Paraná, General Carneiro e Turvo-PR. Nos ensaios foram utilizados extratos fenólicos dos méis, obtidos por meio da utilização da resina Amberlite® XAD®2, bem como méis brutos in natura. Os extratos apresentaram conteúdo de compostos fenólicos significativo, sendo o melato (MO5), de General Carneiro, o de maior teor (117,68± 4,40 mg EAG/g). Para as análises de sequestro das espécies reativas de oxigênio e nitrogênio, os extratos fenólicos foram sempre superiores aos méis brutos in natura. Os extratos fenólicos, de maneira geral, apresentaram alta capacidade de sequestro para o radical peroxila (ROOo), ácido hipocloroso (HOCl) e óxido nítrico (NOo). Em relação aos melatos, o extrato MO7 apresentou alta capacidade para o sequestro do HOCl (EC50= 4,83 ± 0,13 ?g/mL), enquanto que o MO5 foi melhor para o sequestro do NOo (EC50=2,16 ± 0,18 ?g/mL). Pelo método HPLC-ABTS on-line foi possível identificar e quantificar a contribuição para a atividade antioxidante do ácido ferúlico no extrato (MO1) e do flavonoide kanferol na amostra (MO4). O ácido ascórbico foi identificado e quantificado por HPLC somente nos melatos (MO3, MO5 e MO7). Pela técnica de LC-MS/MS foram identificados a presença dos seguintes compostos fenólicos: ácido caféico, rutina e hesperidina em todos os extratos. A análise de compostos voláteis por SPME-CG/EM mostrou a presença de dois compostos, encontrados apenas nos melatos, que foram o terpineno-4-ol, que possui ação antifúngica, antiparasitológica e anti-inflamatória; e o 3,4-dimetil-1-deceno, podendo assim serem utilizados como marcadores químicos destes méis. O conhecimento da composição química destes méis, bem como a composição fenólica bioativa, contribui para o fornecimento de antioxidantes naturais para a dieta, atenuando assim os efeitos negativos dos radicais livres / Brazil has a great potential to explore beekeeping due to its vast territory and diversified flora, what allows different varieties of honeys with unique characteristics. Parana state is one of the largest honey producers and the investment in production that meets the most demanding markets stimulated the organic honey production. The knowledge since early in history regarding the beneficial health effects promoted by honey is stimulating the scientific research of this noble food. Thus, this paper aimed to study certified Brazilian organic honeys (MO) in order to determine the phenolic and volatile profiles, and also the evaluation of radical scavenging capacity against Nitrogen and Oxygen Reactive Species (RNS and ROS, respectively). The honeys were collected from apiaries from beekeepers with the organic certification from two municipalities of southern Parana, General Carneiro and Turvo, PR. In the essays, phenolic extracts were obtained from honeys by using Amberlite® XAD®2 resin, as well as crude in natura honeys. The extracts showed a significant content in phenolic compounds, with honeydew (MO5), from General Carneiro, showing the highest content (117,68 ± 4,40 mg AGE/g). For the analyzes to determine the radical scavenging capacity against RNS and ROS, the phenolic extracts always showed up superior results in comparison to crude in natura honeys. Phenolic extracts showed, in general, great capacity to scavenge peroxyl radical (ROOo), hypochlorous acid (HOCl) and nitric oxide (NOo). In relation to honeydews MO7 extract showed the highest capacity to scavenge HOCl (IC50= 4,83 ± 0,13 ?g/mL) while MO5 was the sample with better capacity to scavenge NOo (IC50=2,16 ± 0,18 ?g/mL). By using HPLC-ABTS on-line method it was possible to identify and to quantify the ferulic acid in MO1 extract, a compound with an important contribution to the antioxidant activity of this sample, as well as the flavonoid kaempferol in MO4 sample. Ascorbic acid was identified and quantified by HPLC only in the honeydew samples (MO3, MO5 and MO7). The analyzes developed by LC-MS/MS techniques indicated the presence of the phenolic compounds caffeic acid, rutin and hesperidin in all the extracts. The analysis of volatile substances developed by SPME-GC/MS promoted the identification of two compounds found only in the honeydew samples. The compounds were the terpinen-4-ol, which has antifungal, antiparasitological and anti-inflammatory activities; and 3,4-dimethyl-1-decene. Both compounds can be used as chemical markers of these honeys. The knowledge of the chemical composition of the studied honeys, as well as their bioactive phenolic composition, contributes to supply natural antioxidants to human diet, thus attenuating the negative effects of free radicals

Esta tese apresenta o desenvolvimento de metodologias analíticas em sistemas de solução em fluxo com detecção amperométrica e espectrofotométrica explorando a utilização de tecidos vegetais como fonte enzimática para a biocatálise de reações. Desempenhos satisfatórios foram obtidos com a utilização do mesocarpo fibroso do coco (Cocus nucifera, L.) e com a casca e/ou polpa de frutos da palmeira leque (Latania sp), fontes de polifenol oxidase. Uma metodologia, simples e rápida, para a determinação dos parâmetros bioquímicos da polifenol oxidase foi desenvolvida ao longo do presente trabalho com a enzima naturalmente imobilizada no tecido do coco, pela eliminação das etapas de extração e de purificação e associando-se análise em fluxo e espectrofotometria. Parâmetros cinéticos importantes como o efeito do pH, a constante de Michaelis-Menten (Km), a velocidade máxima (Vmax),a energia de ativação (Ea) e os parâmetros térmicos (valores D, z e Q10)foram determinados utilizando como substrato o catecol. A atividade relativa junto a diferentes substratos e o efeito de inibidores foram também avaliados. Aplicações envolvendo estes tecidos vegetais em reatores empacotados (associados com FIA) ou incorporados em eletrodos de pasta de carbono (medidas realizadas em batelada) para a determinação de produtos fenólicos como catecol, fenol e dopamina, inibidores da atividade enzimática como o sulfito, bem como para a quantificação do conteúdo de água, demonstram a sua versatilidade. Os produtos fenólicos foram quantificados com boa sensibilidade (na faixa de &#181;mol L-1) pela redução amperométrica das respectivas quinonas, produzidas pela oxidação enzimática. Aplicações com amostras reais, dentre as quais água de rio e resíduos de uma fábrica de papel puderam ser implementadas. A determinação do conteúdo de água em meio aquo-restrito explorou a estimulação, pela água, da atividade catalitica da polifenoI oxidase naturalmente imobilizada no mesocarpo fibroso do coco na presença de catecol. Esta propriedade foi utilizada para a determinação rápida e reprodutível do conteúdo de água em amostras comerciais de álcool, utilizando acetonitrila como carregador. A quantificação de sulfito baseou-se no seu efeito inibidor sobre a polifenol oxidase naturalmente imobilizada em relação à catálise oxidativa do catecol. Diversos interferentes comumente presentes em amostras de vinho foram também investigados. Durante este trabalho, foi desenvolvida uma simples e interessante maneira de conservar o mesocarpo fibroso do coco como fonte de polifenol oxidase naturalmente imobilizada. O tecido desta fruta foi desidratado e moído. O pó deste tecido, estocado a mais de dois anos à temperatura ambiente, ainda apresenta boa atividade enzimática. / The development of analytical methodologies exploring plant tissues as enzymatic source for biocatalysis of many reactions is presented in this thesis. Amperometry and spectrophotometry was associated with flow analysis for analytical purposes and for biochemical characterization of naturally immobilized polyphenol oxidase. Good performance was achieved with tissues from the fruits of two palm trees: the fibrous mesocarp of green coconut fruits, Cocus nucifera, L., and the skin and the pulp of green fruits from Latania sp. Both tissues are very effective in the biotransformation of o-diphenols to the corresponding o-quinones. A simple, fast, and new methodology for the determination of the biochemical parameters of the poyphenol oxidase naturally immobilized on the fibrous tissue was developed (eliminating the extraction and purification of enzymes) by association of flow injection analysis and spectrophotometry. For coconut tissue, cinetic parameters like pH effect, Michaelis-Menten constant (Km) and maximum rate (Vmax), activation energy (Ea) and thermal parameters (D, z and Q10values) on catechol biotransformation were determined. Also the response for several substrates as too for various inhibitors was explored. Amperometric quantification of phenolic compounds was made using the vegetal tissue in form of packed reactors (associated with FIA) or incorporated in carbon paste electrodes (measurements in batch). Very good response for catechol, phenol and dopamine was registered for this compounds, with very high sensitivity (&#181;mol L-1 range). Applicability to real samples as for river water and for a paper plant waste water was verified. The same amperometry-FIA system was employed for enzymology in non-aqueous medium. The activity of the polyphenol oxidase contained in coconut tissues are strongly dependent of water. The strategy involves the stimulation by the content of water on the biocatalytic activity of the enzyme in the presence of catechol substrate. This strategy was used for the determination of water content in alcohol samples, in medium of dry acetonitrile. The inhibition of the enzymatic activity produced by many compounds can be explored for an indirect quantification of the inhibitor. A flow injection amperometric procedure was developed for the determination of sulphite ion based in its inhibitory effect on the activity of polyphenol oxidase on the oxidation of catechol. The effect on the bioreactor performance of potential interferents commonly present in wine samples were also investigated. During this work, it was developed a simple and interesting way to preserve coconut tissue. The mesocarp of this fruit can be dried and grounded. These tissues still with very good activity after more than two years in \"shelf temperature\".