Magister Scientiae - MSc / The overall aims of this project were to assess the pharmaceutical quality and consistency of commercially available solid dose Sutherlandia frutescens containing products (viz. tablets & capsules) by exploring the use of monitoring the pharmaceutical presentation, flavonoid profile and antioxidant activity levels and to develop/or adapt methods and specifications that may be used for the quality control of such products.Stability tests were conducted on all of the selected SCP. The products were stored under elevated temperatures and environmental humidity conditions and total phenol, antioxidant and chromatographic analysis was conducted on these samples. Samples of each of the SCP were hydrolyzed using HCL and then analyzed using HPLC to test the stability of the flavonoids present in each product. The SCP investigated in this study physically appeared to be of quite good “pharmaceutical” quality, but generally lacked information on the date of manufacture and lacked package inserts, or when these were present they contained insufficient information. Based on the results obtained, it is recommended that, the manufacturers of SCP pay more attention to the information provided on the package inserts and the storage conditions for their products. Further the levels of antioxidant activity, total phenols and flavonoid (sutherlandins A to D) be used as specifications to control the quality of commercially available solid dose Sutherlandia frutescens containing preparations on an individual basis. / South Africa

The determination of anthocyanins in red grapes and wines has been of increasing interest in the last years, as they play an important role in colour quality of red wines revealing also many human health beneficial effects. They contribute e.g. to the reduction of coronary heart disease, but exhibit also antimutagenic, anti-inflammatory, anti-carcinogenic and antioxidant properties. In this doctoral thesis, the complex study of grape skin alcoholic and water extracts, prepared from Alibernet and St. Laurent wine grape varieties is presented. Extracts were prepared from three different amounts (0,5; 1,0 and 1,5 g) of lyophilized grape skin powder using the Pressurized fluid extraction (PFE) and the Pressurized Hot Water Extraction (PHWE) at different temperatures ranging from 40 up to 120°C and pressure of 15 MPa. Methanol, ethanol and water were used as a solvents. Total phenolic compound content (TPC) of individual extracts was determined using the Folin-Ciocalteau assay and their tristimulus color values (CIE Lab) were estimated, using the UV-VIS spectrophotometer. The identification and quantification of anthocyanins by high-performance liquid chromatography with diode array detection (HPLC-DAD) was performed. In addition, pH values of all extracts were also measured using the combinated glass electrode. Antioxidant activity of extracts was tested by EPR spectroscopy in Fenton system (H2O2/Fe2+) generating reactive radicals (•OH, O2-•, •CH3) followed by spin trapping technique, using 5,5-dimethylpyrroline-N-oxide (DMPO) as spin trap. In addition, radical scavenging activity of extracts was assessed applying 2,2-diphenyl-1-picrylhydrazyl (•DPPH) free radical and 2,2'-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) cation radical (ABTS•+) assays. All the experimental data were processed with principal component analysis (PCA) and canonical discriminant analysis (CDA) to specify the optimum extraction conditions for extract preparation from the perspective of the potential further application of the extracts as food supplements or food colour enhancers. The results indicated that grape skins of both varieties are a promising source of anthocyanins with prospective application in food industry.

The work was devoted to the technology of flavored beers and consisted of two parts. In the first one, we dealt with their basic division, commonly used raw materials, the description of individual technological steps and the difference between the production of ordinary and flavored beers. They have also been divided based on their origin and assigned to the typical geographical areas in which they are produced. In the second - experimental part - nine micro samples were created and by help of FermentoFlash destillation, and spectrophotometer the basic parameters were determined. Such as the original extract, weight, volume, alcohol and color of wort. Also, the total polyphenol content and antioxidant capacity were determined using DPPH and FRAP methods. A sensory evaluation was made in order to detect sensory differences depending on the phase in which the fruit was added. The results were subjected to statistical research and reviewed and compared the results published by other authors.

The ethanol extract of A. phylicoides was investigated for its antioxidant activity using the DPPH scavenging method. The extract showed good antioxidant results with a EC50 value of 10.64 ± 0.0842 µ/ml. The extract was also tested for antibacterial activity against microorganisms (Staphylococcus aureus, Enterococus faecalis, Bacillus cereus, Bacillus subtilis, Bacillus pumilus, Pseudomonas aeruginosa, Escherichia coli, Klebsiella pneumonia) commonly known to pose a threat in the wellbeing of man. All tested microorganisms were significantly inhibited by the extract with the MIC values ranging from 3.13 µg/ml to 6.25 µg/ml. Folin-Ciocalteu’s reagent method was used to determine total phenolic content of dried and freshly prepared crude extract of A. phylicoides. Higher total phenolic content (28.28 ± 0.019 mg GAC/100g) and antioxidant activity (EC50, 10.64 ± 0.084 µg/ml) was observed in the dried extract compared to the fresh extract with a TPC value of 23.04 ± 0.003 mg GAC/100g and EC50 of 13.97 ± 0.066 µg/ml. Bioassay-guided fractionation of ethanolic extract from aerial parts of Athrixia phylicoides using silica and sephadex column chromatography led to the isolation of four known flavanoids, 5-hydroxy-6,7,8,3’,4’,5’-hexamethoxyflavon-3-ol (1), 3-0- demethyldigicitrin (2), 5,6,7,8,3’,4’-hexamethoxyflavone (3) and Quecertin (4). Due to the low yield, no further tests were done on compound 3. A DPPH-scavenging assay was performed to evaluate the antioxidant activity of the isolated compounds. All the tested compounds showed potent antioxidant activity with EC50 values ranging from 1.27 to 3.41 µg/ml. Compound 4 showed a higher antioxidant activity (EC50, 1.27 µg/ml) than vitamin C (EC50, 2.66 µg/ml) used as a control. The MIC values of the isolated compounds against tested microorganisms varied from 20 to more than 40 µg/ml. All the tested compounds showed no activity against S. aureus, B. pumilus, K. pneumonia and P. aeruginosa at the highest concentration tested (40 µg/ml). These compounds together with the extract were further analyzed by XTT assay on Vero cells. The extract showed a low toxicity effect on the cells at lower concentrations exhibiting EC50 value of 107.8 ± 0.129 µg/ml. Compound 4 showed minimal toxicity effect on the cells with a EC50 value of 81.38±0.331 µg/ml, compared to Compound 1 and 2 which exhibited EC50 values of 27.91 ± 0.181 µg/ml and 28.92 ± 0.118 µg/ml respectively. The results obtained from this study provide a clear rationale for the medicinal uses of Athrixia phylicoides. / Dissertation (MSc)--University of Pretoria, 2010. / Plant Science / unrestricted

L'huile de Sésame a un grand intérêt économique et industriel et est utilisée comme ressource biologique riche en protéines, en antioxydants et comme substrat de bioprocédés pour la production d'enzymes, d'antibiotiques ou de biopesticides. Nous avons préparé, par transestérification de l'huile via un ester méthylique, un mélange d'esters de l'acide vanillique (VNA) et obtenu un produit plus hydrosoluble VNA-2, qui a ensuite été encapsulé dans des billes de pectinate de calcium ou de zinc, visant à une libération contrôlée dans le colon. La dissolution des billes a été étudiée aux pH de l'estomac et du colon, et en milieu enzymatique. L'activité antioxydante du principe actif libéré a été évaluée in vitro par des tests DPPH et TRAP et des tests in vivo sont en cours sur le lapin.L'huile de Tamanu, huile végétale non comestible extraite des graines de Calophyllum inophyllum, est utilisée en médecine polynésienne traditionnelle et comme matière première en cosmétique; elle possède des propriétés thérapeutiques: effets antimicrobien, anti-inflammatoire, analgésique, anti-tumoral, cytoprotecteur anti-UV. Sa capacité antioxydante, celle de ses fractions résineuses et de coumarines isolées ont été évaluées par des tests basés sur la réduction de radicaux libres stables (DPPH) ou le piégeage de radicaux libres dérivés de l'oxygène impliqués dans le stress oxydant (TRAP). Les résultats montrent que la résine de Tamanu contient les principales fractions antioxydantes. La molécule la plus active au regard des deux tests est l'Inocalophylline B. Les inophyllums C et E présentent une activité antioxydante modérée vis à vis du test DPPH bien que ne portant pas de fonction hydroxyle / Sesame oil is of great economic and industrial interest. It is used as a protein or antioxidant rich biological resource and as a substrate for bioprocesses for enzymes, antibiotics or biopesticides production. We esterified sesame oil with methanol and then transesterified the resulting mixture with vanillic acid (VNA) in order to get a more water soluble mixture of vanillic esters of its fatty acids (VNA-2). VNA-2 was encapsulated in Ca or Zn pectine beads with the objective of a controlled release in the colon. Beads dissolution was followed in acidic (stomach) and basic (colon) media as well as in enzymatic medium. The antioxidant activity of the liberated substrate was evaluated by in vitro DPPH and TRAP tests and compared to that of the non-encapsulated product. In vivo experiments of drug release in the rabbit are in progress.Tamanu oil is a non-edible vegetable oil extracted from the seeds of Calophyllum inophyllum, widely used in traditional Polynesian medicine and as raw material in cosmetics. It exhibits various therapeutic properties: antimicrobial and anti-inflammatory, analgesic and anti-tumor, cytoprotective anti-UV. Antioxidant capacity of Tamanu oil, its resinous extracts and coumarins isolated from the latter were evaluated using standard tests, based on the reduction of a stable free radical (DPPH) or the scavenging of oxygen-derived radicals involved in oxidative stress. The results show that the resin contains Tamanu oil major antioxidant fractions. The more active molecule in both tests is inocalophylline B. The inophyllums C and E show a moderate antioxidant activity with respect to the DPPH test, while not bearing an hydroxyl group.

Dans l’optique de comprendre et maitriser le vieillissement des vins, en particulier des vins blancs, il est nécessaire d’approfondir nos connaissances sur les mécanismes physico-chimiques d’oxydation liés aux processus d’oxygénation. Pour cela, il est important d’avoir des outils permettant la quantification de la capacité antioxydante des vins blancs, et l’identification des composés impliqués dans celle-ci, afin de pouvoir anticiper l’aptitude au vieillissement d’un vin.Lors de cette étude, des analyses de la capacité antioxydante par DPPH et Résonnance Paramagnétique Electronique (RPE) ont été réalisées sur un grand nombre de vins en cours d’élevage et issus de plusieurs millésimes. En parallèle des analyses métabolomiques, principalement par Chromatographie Liquide couplée à la Spectrométrie de Masse (UPLC-Q-TOF-MS) mais également par Spectrométrie de Masse à Résonance Cyclotronique des Ions et à Transformée de Fourier (FT-ICR-MS) ont été réalisées.L’optimisation de la méthode DPPH, pour l’analyse des vins blancs a révélé une réactivité importante des composés soufrés, avec des capacités antioxydantes similaires à celles des composés phénoliques. La comparaison de cette méthode optimisée à la méthode de référence du laboratoire (RPE) sur plus de 106 vins, a montré la complémentarité de l’information apportée par ces deux différentes méthodes de mesure de capacité antioxydante. Le traitement statistique des corrélations avec les analyses métabolomiques réalisées sur 287 vins a permis de révéler une liste de 380 marqueurs moléculaires associés à la capacité antioxydante des vins. Une méthode alternative de mesure du potentiel antioxydant des vins blancs a également été développée, qui repose sur l’identification de composés nucléophiles naturellement présents et susceptibles de piéger les quinones formées au cours de mécanismes d’oxydation. Outre des composés soufrés, nous avons montré que différents peptides ayant des propriétés antioxydantes présentent de telles propriétés nucléophiles. Dans un second temps, l’étude de différentes pratiques œnologiques (élevage, hyperoxygénation ou sulfitage) a permis de déterminer leurs impacts sur la capacité antioxydante des vins mais également sur les marqueurs moléculaires associés. Il apparait ainsi une augmentation significative de la capacité antioxydante des vins au cours de l’élevage. De plus, cette augmentation de la capacité antioxydante est associée à une consommation de peptides en début d’élevage, ainsi qu’à l’apparition de nouveaux composés dans les vins et ce indépendamment du millésime. Les composés qui apparaissent semblent être majoritairement des composés phénoliques provenant potentiellement du bois ou de l’autolyse des levures. Les vins issus de moûts protégés par l’ajout précoce de sulfites ont une capacité antioxydante plus élevée par rapport aux vins issus de moûts hyperoxygénés. De plus, la protection du moût à un impact important sur la composante moléculaire soufrée retrouvée dans les vins. Ainsi, il a été observé une quantité beaucoup plus faible de composés soufrés (peptides ou non) dans les vins issus de moûts hyperoxygénés montrant donc une consommation de ces composés dans les mécanismes d’oxydation du vin. En résumé, ces résultats révèlent pour la première fois l’importance des composés soufrés dans les mécanismes de protection des vins blancs de Bourgogne vis-à-vis de l’oxydation, par leur capacité de piégeur de radicaux mais également de piégeur de quinones. / In order to understand and control the aging of wines, particularly white wines, it is necessary to deepen our knowledge about the physico-chemical mechanisms of oxidation related to oxygenation processes. For this, it is important to have tools to quantify the antioxidant capacity of white wines, and to identify the compoiunds involved, in order to anticipate the aging ability of a wine.In this study, analyzes of the antioxidant capacity by DPPH and Electron Paramagnetic Resonance (EPR) were carried out on a large number of wines during aging and from several vintages, in parallel with metabolomic analyzes, mainly carried out by Liquid Chromatography Coupled to Mass Spectrometry (UPLC-Q-TOF-MS) but also by Fourier Transform Ion Cyclotron Resonance Mass Spectrometry (FT-ICR-MS).Optimization of the DPPH method for the analysis of white wines, showed a high reactivity of sulfur compounds with similar antioxidant capacities to those of phenolic compounds. The comparison of this optimized method with the laboratory reference method (EPR) on more than 106 wines, showed the complementarity of the information provided by these two different methods of measuring an antioxidant capacity.Combining antioxidant capacity measurements and metabolomic analyzes, it was possible to determine a list of molecular markers related to the antioxidant capacity of white wines. During this study, a focus on nucleophilic compounds present in wines has also been realized, these compounds being able to react with the quinones formed during oxidation and thus to play a role in the oxidation mechanisms of white wines. Besides sulfur compounds, we showed that several peptides with antioxidant properties could exhibit such nucleophilic behavior. In a second step, the study of different oenological practices (aging, hyperoxygenation or adding SO2 to must) allowed to determine their impacts on the antioxidant capacity of wines but also on the associated molecular markers. It thus appears a significant increase in the antioxidant capacity of the wines during aging. In addition, this increase in antioxidant capacity is associated with a consumption of peptides at the beginning of aging, but also with the appearance of compounds in wines, regardless of the vintage. The compounds that appear are potentially phenolic compounds which can come from wood or lees autolysis. Wines from protected musts with sulfites addition, have a higher antioxidant capacity compared to wines from hyperoxygenated musts. In addition, the protection of musts has a significant impact on the sulfur component found in wines. Thus, a much smaller amount of sulfur compounds (peptides or not) has been observed in wines derived from hyperoxygenated musts, showing a consumption of these compounds in the oxidation mechanisms. Overall, these results reveal for the first time the importance of sulfur compounds in the mechanisms of protection of white wines from Burgundy against oxidation, by radical scavenging capacity and quinone trapping.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The Rhamnaceae family is composed of some 58 genera and about 900 species. Scutia buxifolia Reissek species, popularly known as coronilha, is a local plant from South America, with a dispersion that comprise Rio Grande do Sul state in Brazil, Argentina and Uruguay. The stem bark and leaves decoction are popularly used as cardiotonic, antihypertensive and diuretic. This work aims to isolate and identify flavonoids and triterpenes present in Scutia buxifolia and analyze ability to capture free radicals. Stem bark and leaves of S. buxifolia were collected in October 2007 in Dom Pedrito-RS (coordinates 30º 59'09''S and 54º27'44''W). The material is deposited in the herbarium of the Department of Biology UFSM cataloged under the registration number SMBD 10919. The dried plant material was ground and macerated using ethanol: water (70:30, v/v) as solvent. The crude extract was fractionated with solvents of increasing polarity (CH2Cl2, AcOEt, n-BuOH). In AcOEt fraction of the leaves was characterized quercetin, quercetin 3-0-rhamnosíde, quercetin 3-0-β-Dglucopyranoside and rutin, and the CH2Cl2 fraction of stem bark the steroids β-sitosterol and stigmasterol, and the triterpene lupeol. The compounds were analyzed by 1H-NMR and 13CNMR and spectroscopic data were compared with those obtained from the literature. The compounds were quantified by HPLC in the fraction used for insolation. The crude extract and fractions of S. buxifolia were investigated for lipid peroxidation, antioxidant activity and total content of phenols. The total phenolic content ranged from 141.09 ± 0.71 to 323.47 ± 2.62 mg/g for stem bark and 72.09 ± 0.50 to 383.34 ± 2.31 mg/g for leaves of S. buxifolia. Crude extract and fractions caused a sharp fall in TBARS production with IC50 from 2.93 ± 2.17 to 40.46 ± 2.51 μg/mL for the leaves and 0.66 ± 0.17 to 27.3 ± 1.23 μg/mL for the stem bark. The order of capture of free radicals was: AcOEt > n-BuOH > CH2Cl2 > crude extract, for the two parts of the plant. / A família Rhamnaceae é composta por cerca de 58 gêneros e aproximadamente 900 espécies. A espécie Scutia buxifolia Reissek, é conhecida popularmente como coronilha, é uma planta nativa da América do Sul, com uma dispersão que compõe o estado do Rio Grande do Sul no Brasil, Argentina e Uruguai. A decocção das cascas do tronco e folhas é utilizada popularmente como cardiotônica, hipotensora e diurética. O presente trabalho objetivou isolar e identificar flavonóides e triterpenos presentes em S. buxifolia bem como analisar a capacidade de capturar radicais livres. As cascas do tronco e as folhas de S. buxifolia foram coletadas em outubro de 2007 no município de Dom Pedrito RS. O material está depositado no herbário do Departamento de Biologia da UFSM catalogado sob o número de registro SMBD 10919. O material vegetal foi seco, moído e macerado utilizando como solvente etanol:água (70:30, v/v). Fez-se fracionamento do extrato bruto com solventes orgânicos de polaridades crescentes (CH2Cl2, AcOEt, n-BuOH). Na fração AcOEt das folhas caracterizou-se os flavonóides quercetina, quercetina 3-0-rhamnosídeo, quercetina 3-0-β-D-glicopiranosídeo e rutina. Da fração CH2Cl2 das cascas do tronco, os esteróides β-sitosterol e estigmasterol, e o triterpeno lupeol. Os compostos isolados foram analisados por 1H-RMN e 13C-RMN e seus dados espectroscópicos foram comparados com os obtidos da literatura. Os compostos isolados foram quantificados por CLAE. O extrato bruto e as frações de S. buxifolia foram investigadas quanto a peroxidação lipídica, atividade antioxidante e conteúdo de fenóis totais. O conteúdo de fenólicos totais variou de 141,09 ± 0,71 a 323,47 ± 2,62 mg/g para as cascas do tronco e de 72,09 ± 0,50 a 383,34 ± 2,31 mg/g para as folhas de S. buxifolia. O extrato bruto e as frações provocaram uma queda acentuada na produção de TBARS com IC50 de 2,93 ± 2,17 a 40,46 ± 2,51 μg/mL para as folhas e 0,66 ± 0,17 a 27,3 ± 1,23 μg/mL para a cascas do tronco. Para o ensaio do DPPH a ordem de captação de radicais livres foi: AcOEt > n-BuOH > CH2Cl2 > extrato bruto, para as duas partes da planta.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O camu-camuzeiro (Myrciaria dubia (Kunth) Mc Vaugh) é uma espécie frutífera pertencente à família Myrtaceae. Possui potencial econômico, por se tratar do fruto com maior quantidade de vitamina C, chegando a atingir 6.112 mg de ácido ascórbico por 100 g-1 de polpa. Objetivou-se com o presente trabalho caracterizar os frutos de camu-camu detectando-se o melhor ponto de colheita para o fruto, assim como testar tecnologias pós-colheita e de armazenamento em sua conservação. Foram realizados dois experimentos. O primeiro, objetivou a determinação do ponto de colheita onde os frutos foram colhidos em três diferentes estádios de maturação (imaturo, semi-maturo e maturo) e armazenados por oito dias. No segundo, foram testadas embalagens e temperaturas de armazenamento constituindo os seguintes tratamentos: T1 (sem embalagem a temperatura ambiente, 22 C); T2 (sem embalagem a 15 C); T3 (sem embalagem a 20 C); T4 (PET a temperatura ambiente, 22 C); T5 (PET a 15 C); T6 (PET a 20 C); T7 (PVC a temperatura ambiente, 22 C); T8 (PVC a 15 C); e T9 (PVC a 20 C). Os delineamentos utilizados foram inteiramente casualizados em arranjo fatorial. Os frutos foram analisados diariamente quanto a: perda de massa fresca, pH, teores de sólidos solúveis (SS), acidez titulável (AT), ácido ascórbico (AA), vitamina C total, carotenoides, antocianinas, clorofilas A e B, flavonoides e compostos fenólicos totais, assim como a atividade antioxidante (FRAP e DPPH) e o índice de maturação (SS/AT). No estádio semi-maturo houve uma maior conservação dos atributos de qualidade (SS, AT e menor perda de massa), assim como do teor de ácido ascórbico e da atividade antioxidante (FRAP). Os pigmentos carotenoides, flavonoides e antocianinas, assim como o teor de vitamina C apresentaram maiores conteúdos nos frutos colhidos maturos, sendo esse estádio considerado o indicado para a extração desses biocompostos funcionais. Esse ponto de colheita também correspondeu ao maior conteúdo médio de fenólicos totais e da atividade antioxidante (DPPH). Assim o melhor ponto de colheita para a extração de pigmentos e biocompostos antioxidantes do camu-camu é o estádio maturo. Quando a intenção for obter maior vida de prateleira, o melhor ponto de colheita é o semimaturo, por conservar os atributos qualitativos por mais tempo. No armazenamento pós-colheita do camu-camu concluiu-se que a melhor conservação dos atributos qualitativos e a melhor conservação dos pigmentos e biocompostos antioxidantes ocorreu quando os frutos foram armazenados a 15 C em conjunto com a bandeja de poliestireno expandido recoberta com filme de PVC. / The camu-camu tree (Myrciaria dubia (Kunth) Mc Vaugh) is fruit-bearing tree belonging to the family Myrtaceae. It possesses economic potential for being concerned with the fruit of greatest amount of vitamin C, coming to reach 6.112 mg of ascorbic acid per 100 g-1 of pulp. It was aimed through the present wok to characterize the camu-camu fruit by detecting the best harvest date for the fruit as well as to test post-harvest and storage technologies in their conservation. Two experiments were conducted. The first one aimed at the determination of the harvest date where the fruits were collected at three different ripening stages (unripe, semiripe and ripe) and stored for eight days. In the second one, packages and storage temperatures constituting the following treatments were tested: T1 (with no package at room temperature, 22 C); T2 (with no package at 15 C); T3 (with no package at 20 C); T4 (PET at room temperature, 22 C); T5 (PET at 15 C); T6 (PET at 20 C); T7 (PVC at room temperature, 22 C); T8 (PVC at 15 C); and T9 (PVC at 20 C). The designs utilized were completely randomized in factorial arrangement. The fruit were analyzed daily as to: the loss of fresh mass, pH, contents of soluble solids (SS), titrable acidity (TA), ascorbic acid (AA), total vitamin C, carotenoids, anthocyanins, chlorophylls A e B, flavonoids and total phenolic compounds as well as the antioxidant activity (FRAP and DPPH) and the maturation index (SS/TA). At the semiripe stage, there was a greater conservation of the quality attributes (SS, TA and less mass loss) as well as of the ascorbic acid content and of the antioxidant activity (FRAP). The pigments carotenoides, flavonoids and anthocyanins as well as the content of vitamin C presented higher amounts in the fruit collected matures; that stage being regarded as the appropriate for the extraction of those functional biocompounds. That harvest date also corresponded to the highest average content of total phenolics and of the antioxidant activity (DPPH). So, the best harvest date for the extraction of both pigments and antioxidant biocompounds of camu-camu is the ripe stage. When the intention is obtaining longest shelf-life, the best harvest date is the semi-ripe, for conserving the qualitative attributes for longer. In the post-harvest storage of the camu-camu, it follows that the best conservation of the qualitative attributes and the best conservation of the pigments and antioxidant biocompounds occurred when the fruit were stored at 15 C jointly with the expanded polystyrene tray covered with PVC film.

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.

Ce travail de recherche est centré sur la valorisation de deux Euphorbiacées : Ricinus communis et Jatropha curcas. La première est une espèce autochtone connue comme plante dont l’huile des graines est utilisée pour ses vertus cosmétiques quant à la deuxième, c’est une espèce allochtone récemment introduite à titre expérimental en Tunisie et connue comme plante bioénergetique. Pour le ricin huit populations Tunisiennes ont été étudiées: Riadh Andalous, Nefza, Béja, Nabeul, Hammamet, Bouficha, Khanguet Hajej et Aouled Amer. Quant au jatropha, le matériel végétal est récolté dans la station expérimentale de Nabeul (Tunisie). Il s’agit de huit populations qui proviennent d’Arusha en Tanzanie, de Mozambique, de Suriname et de Brésil à partir de cinq provenances à savoir : Paranà, Norte de Minas, Mato Grosso, Regiao sudeste et Vale do Jequitinhonha. Le travail comprend une première partie consacrée à une étude bibliographique. Une deuxième partie est consacrée aux matériels et méthodes utilisés et une dernière partie qui montre l’ensemble des résultats obtenues. Ainsi, les résultats montrent que les extraits des feuilles des populations des deux espèces étudiées sont plus riches en composés phénoliques que les extraits des racines. L’étude phytochimique a montré que le ricin contient surtout de l’acide gentisique. Quant au jatropha, il contient surtout de l’épicatechine et de la naringine. L’étude chimique des huiles fixes des deux espèces montrent que l’huile de ricin renferme essentiellement d’acide ricinoléique. Quant à l’huile de jatropha, elle contient deux acides gras en proportions majeures qui sont l’acide oléique et l’acide linoléique. L’évaluation des activités anti-oxydantes des extraits des deux espèces indique une corrélation positive entre ces activités et les teneurs en composés phénoliques. Par ailleurs, l’étude de l’activité anti-acétylcholinestérase des extraits testés montrent que ces extraits sont des inhibiteurs de l’AChE plus puissants que la galanthamine utilisée comme contrôle positif. Notre étude a confirmé que les différents extraits de ricin et de jatropha, autres que les huiles fixes, peuvent donc être exploités pour d’autres activités biologiques, parmi les quelles l’action inhibitrice de l’AChE une des principales cibles des traitements contre la maladie l’Alzheimer et le piégeage des radicaux libres, en raison de leur richesse en composés phénoliques / The aim of this study is to promote two Euphorbiaceae plants the Ricinus communis and the Jatropha curcas, the first one is known for its oil used in the cosmetic products, whereas the second one is known especially for its seeds used in the production of biodiesel. For the castor plant, eight Tunisian populations are studied: Riadh Andalous, Nefza, Beja, Nabeul, Hammamet, Bouficha, Khanguet Hajej and Aouled Amer. For the jatropha, recently introduced in Tunisia, the plant material is collected from the Nabeul station (Tunisia). Eight populations coming from Arusha in Tanzania, Mozambique, Suriname and Brazil from regions of Paraná, Minas Norte, Mato Grosso, Regiao sudeste and Vale do Jequitinhonha are studied. In first, a bibliographic study is made. In second, the used materials and methods are cited. Fanilly, all the results are mentioned. The study shows that the phenolic compounds are higher in the leaves extracts than in the roots extracts for both species. The phytochemical study shows that the gentisic acid is the major phenolic compound identified in the castor plant extracts. In contrast, the epicatechin and the naringin are the most important phenolic compounds identified in the jatropha extracts. The GC-MS analysis reveals that the castor oil contains mainly the ricinoleic acid. For the jatropha oil, two major fatty acids are identified: the oleic and the linoleic acids. Results of the antioxidant properties of leaves and roots of both species indicate a positive correlation between the leaves and the roots activities and their contents of phenolic compounds. Furthermore, the anti-acetylcholinesterase activity of the tested extracts shows for the first time that some tested extracts are more active than the galantamine used as a positive control. Our study confirmed that, in addition to their oils, different extracts of the castor plant and the jatropha can be used for biological activities such as the scavenging free radicals and the inhibitory action of AChE enzyme, which is a major target for treatments against the Alzheimer's disease due to their high levels of phenolic compounds. Owing to the activities of the leaves and the roots extracts confirmed by this study, the agricultural exploitation of the castor plant and the jatropha can be economically more profitable