A película do amendoim apresenta coloração vermelha intensa e é rica em compostos fenólicos, todavia o extrato líquido é instável e de difícil comercialização. Os objetivos deste trabalho foram extrair os pigmentos da película do amendoim, estudar o processo de secagem por atomização deste extrato, caracterizar os pós obtidos, bem como sua propriedade antioxidante, antimicrobiana e estabilidade durante o armazenamento, sempre comparando com o controle. Quanto a caracterização do extrato líquido foi analisado fenólicos totais, pH, umidade, cinzas, proteínas, lipídeos, açúcares, aflatoxina e acidez. Os extratos foram então misturados com agente carreador nas concentrações de 10, 20 e 30%, estes foram atomizados em spray dryer com temperaturas do ar de entrada de 130, 150 e 170ºC. Para obtenção da amostra controle, parte do extrato foi liofilizado sem a presença de agente carreador. Os pós obtidos foram caracterizados quanto à umidade, atividade de água, higroscopicidade, morfologia, tamanho das partículas, cor, teor de fenólicos totais, solubilidade, estabilidade, propriedade antimicrobiana e antioxidante pelos métodos: ORAC, DPPH e HPLC-ABTS on line. Na caracterização do extrato líquido foram obtidos os seguintes resultados: 0,24% de proteína, 2,31% de açúcares, 0,09% de cinzas, 1,41% de lipídeos, 0,91% de acidez titulável, 42,88 mg de ác. gálico eq./g de extrato (fenólicos totais), pH de 5,30, 90,93% de umidade e 1,81 ng aflatoxina/mL extrato (aflatoxina). Na caracterização dos pós, a umidade foi menor para pós secos a 170ºC e para concentração de 10% de maltodextrina e foi sempre menor que o controle. A higroscopicidade foi menor quanto maior a concentração de maltodextrina, pois esta tem baixa higroscopicidade. Temperaturas mais altas geraram pós mais higroscópicos, pois esses tinham menor umidade. Com relação à solubilidade, os valores variaram de 85,60 a 91,91%, obtendo-se maiores valores para pós com maiores concentrações de maltodextrina, pois esta apresenta elevados valores de solubilidade. O controle apresentou solubilidade mais baixa de 77,62%. Já os parâmetros de cor tiveram influência apenas da concentração de maltodextrina, sendo que as amostras com menor concentração apresentaram cor mais acentuada, o que era de se esperar visto que a maltodextrina apresenta coloração branca. Portanto, o controle sempre apresentou a coloração mais intensa frente aos pós atomizados. A temperatura de 170ºC originou pós de superfícies mais lisas, apresentando assim melhor escoamento. Com o estudo de estabilidade, verificou-se que as amostras atomizadas tiveram menor perda de cor em relação à liofilizada, e as amostras com maior concentração de maltodextrina (30%) preservaram melhor os fenólicos totais, destacando-se a temperatura de 150 ºC. Em relação à atividade antioxidante, o tratamento T5 se destacou frente aos outros, apresentando o melhor valor de atividade antioxidante por todos os métodos estudados. O extrato da película de amendoim em pó também apresentou atividade antimicrobiana contra as bactérias Gram-positivas Staphylococcus aureus e Listeria monocytogenes, apresentando ainda capacidade bactericida para Staphylococcus aureus. Frente a esse estudo, tem-se na película de amendoim um potencial aditivo natural, podendo ser utilizado como pigmento em pó que apresenta excelente estabilidade, baixa higroscopicidade e alta solubilidade; além de apresentar atividades biológicas como capacidade antioxidante e antimicrobiana. / The peanut skin presents intense red color and is rich in phenolic compounds, however the liquid extract is unstable and difficult to commercialize. The aim of this project was to extract the pigments of peanut skin, study the process of spray drying this extract, characterize the powders as well as its antioxidant properties, antimicrobial properties and storage stability, when compared to the control. The extract was characterized measuring the phenolic compounds, pH, moisture, ash, protein, lipids, sugars, aflatoxin and acidity. The extracts were then mixed with the carrier agent at concentrations of 10, 20 and 30%, these were atomized in a spray dryer at inlet air temperatures of 130, 150 and 170 º C. To obtain a control sample, part of the extract was freeze-dried without the presence of maltodextrin. The powders were characterized for moisture, water activity, hygroscopicity, morphology, particle size, color, total phenolic content, solubility, stability, antimicrobial and antioxidant properties by the methods: ORAC, DPPH and ABTS HPLC-online. In the analysis of the liquid extract, the following results were obtained: 0.24% protein, 2.31% sugar, 0.09% ash, 1.41% lipid, 0.91% titratable acidity, 42.88 mg of Galic acid eq. / g extract (phenolic), pH of 5.30, 90.93% moisture and 1.81 ng aflatoxin / mL extract (aflatoxin). In the analysis of the powders, moisture was lower for powders dried at 170ºC and with 10% maltodextrin concentration and was always lower than the control. The hygroscopicity is lower as higher the concentration of maltodextrin, since the last has low hygroscopicity. Higher temperatures generated more hygroscopic powders, because they reduced moisture. In concern to solubility, the values ranged from 85.60 to 91.91%; higher values were obtained for powders with higher concentrations of maltodextrin, since it has high solubility values. The control showed lower solubility of 77.62%. The color parameters were influenced only by the concentration of maltodextrin; the samples with lower concentrations showed more pronounced color, what was to be expected since maltodextrin is white. Therefore, controls have always presented with more intense color than the atomized powders. The temperature of 170 ºC originated powders with smoother surfaces, thus resulting in better flow. In the evaluation of stability, it was found that the atomized sample suffered lower color loss compared to the freeze dried sample, and samples with higher concentrations of maltodextrin (30%) had better preservation of total phenolics, especially at a temperature of 150ºC. In concern to antioxidant activity, the T5 treatment stood out compared to the others, showing the best values of antioxidant activity in all methods studied. The extract of peanut skin in powder also showed antimicrobial activity against Gram-positive bacteria Staphylococcus aureus and Listeria monocytogenes, with bactericidal properties against Staphylococcus aureus. This study clearly showed that peanut skin is a potential natural additive product and may be used as a powdered pigment which has excellent stability, low hygroscopicity and high solubility, besides having biological activities, such as antioxidant and antimicrobial capabilities.

O Brasil possui uma imensa diversidade biológica, na qual muitos compostos bioativos podem ser encontrados e utilizados em beneficio à sociedade. No entanto, processos de degradação do ambiente e introdução de espécies exóticas têm contribuído ao conhecimento e uso limitadosde muitas plantas nativas, sendopequena a quantidade de estudos sobre sua composição química e potencial biológico. A prevenção de doenças crônicas constantemente vem sendo associada à atividade antioxidante de metabólitos secundários dos vegetais, principalmente os fenólicos. Assim, o objetivo deste trabalho foi avaliar a atividade antioxidante e composição fenólica de cinco espécies frutíferas nativas do Brasil (G. brasiliensis, E. leitonii, E. involucrata, E. brasiliensis e E. myrcianthes). Os métodos capacidade de redução do Folin-Ciocalteau, autoxidação do sistema ?-caroteno/ácido linoleico, capacidade de absorção de radicais oxigênio (ORAC), sequestro dos radicais livres DPPH e ânion superóxido foram aplicados na determinação da atividade antioxidante dos extratos etanólicos de folha, casca, semente e polpa das espécies selecionadas. As amostras demonstraram significativa atividade antioxidante e, em alguns casos, superior as frutas comumente consumidas pela população brasileira. Em geral, folhas apresentaram as maiores atividades, mas o destaque foi a semente de E. leitonii que exibiu os melhores resultados em quatro dos cinco métodos utilizados: 120,67 mg AG.g-1 na redução do reagente Folin; 7,08 Gmol Trolox.g-1 no ?-caroteno; EC50 de 0,26 mg.mL-1 e 0,07 mg.mL-1 no sequestro do ânion superóxido e DPPH, respectivamente; 514,75 Gmol Trolox.g-1 no ORAC para o qual a folha de E. involucrata obteve o maior valor (1393,3 Gmol Trolox.g-1). Os extratos das espécies nativas também demonstram efeito antioxidante contra radicais de relevância biológica, como peroxila e superóxido. Por meio de CG-EM e CLAE acoplado a arranjo de fotodiodos, os principais compostos fenólicos encontrados nos extratos vegetais foram catequina, epicatequina e ácido gálico. Este trabalho demonstrou o grande potencial antioxidante das frutíferas nativas brasileiras, evidenciando assim possível efeito positivo em sistemas biológicos. / Brazil has a great biodiversity, in which many bioactive compounds can be found and used to benefit the society. However, environmental degradation processes and introduction of exotic species have contributed to limited use and knowledge of many native plants, reflecting in few studies about chemical composition and biological potential. The prevention of chronic diseases has been constantly associated with the antioxidant activity of plants secondary metabolites, mainly the phenolics. The objective of this study was to evaluate the antioxidant activity and the phenolic composition of five Brazilian native fruitful species (G. brasiliensis, E. leitonii, E. involucrata, E. brasiliensis e E. myrcianthes). The methods of Folin-Ciocalteau reducing capacity, co-oxidation of ?-carotene/linoleic acid system, oxygen radical absorbance capacity (ORAC), DPPH and superoxide free radical scavenging were used to determine the antioxidant activity of ethanolic extracts of leaf, peel, seed and pulp of the selected species. The samples showed significant antioxidant activity and, in some cases, it was superior to fruits commonly consumed by Brazilian population. In general, leaves presented the highest activities, but the seed of E. Leitonii stood out exhibiting the best results in four of the five methods: 120.67 mg GA.g-1 in the Folin reducing; 7.08 Gmol Trolox.g-1 in the ?-carotene; EC50 of 0.26 mg.mL-1 and 0.07 mg.mL-1 in the superoxide and DPPH scavenging, respectively; 514.75 Gmol Trolox.g-1 in the ORAC, for which the E. Involucrata leaf had the highest value (1393.3 GmolTrolox.g-1). The extracts of native species also demonstrate antioxidant effect against radicals of biological relevance, such as peroxyl and superoxide. By GC-MS and HPLC coupled to a photodiode array, the major phenolic compounds found in the plant extracts were catechin, epicatechin and gallic acid. In this study, Brazilian native fruitful presented high antioxidant potential, showing a possible positive effect on biological systems.

Lifestyle diseases represent a large burden on developed societies and account for much morbidity worldwide. Research has shown that eating a diet rich in fruit and vegetables helps to ameliorate and prevent some of these diseases. Antioxidants found in fruits and vegetables may provide a substantial benefit in reducing disease incidence. This thesis examines the antioxidant properties of resveratrol, piceatannol, and pterostilbene, and the ability of Burkitt's Lymphoma (Raji) cells to uptake these three antioxidants. It also studies the effect of the antioxidants in protecting against DNA damage and their role in DNA repair following oxygen radical exposure in Raji cells. The Oxygen Radical Absorbance Capacity (ORAC) assay was used to measure overall antioxidant contribution as well as the ability of Raji cells to uptake antioxidant following exposure to 2,2’-Azobis(2-methyl-propionamide) dihydrochloride (AAPH). The single cell gel electrophoresis (Comet) assay was used to assess DNA damage and DNA repair rates of cells. Results showed that Raji cells, following oxygen radical exposure, significantly uptake pterostilbene (p < 0.0001), but not piceatannol or resveratrol. Piceatannol provided protection against hydrogen peroxide induced DNA damage, but pterostilbene and resveratrol increased DNA damage following hydrogen peroxide treatment. None of the compounds showed any effect on DNA repair. Overall, this study indicates there is merit for further research into the bioactive roles, including antioxidant capacity, of all three compounds. Such research may provide evidence for the more widespread use of these and other food based compounds for preventing lifestyle diseases.

Roasting is an important processing step for developing cocoa flavor, color, and aroma. Cocoa beans contain polyphenolic compounds, which can be desirable antioxidants. Oxygen Radical Absorbance Capacity (ORAC) values can be used as an indicator of health benefits of antioxidants in foods. ORAC values measure total antioxidant capacity of different foods by measuring antioxidant scavenging activity against peroxyl radical induced by 2,2’-azobis (2-amidinopropane) dihydrochloride (AAPH). This measurement of total antioxidant capacity gives a complete assessment during which the inhibition time and inhibition degree are measured as the reaction comes to a completion. ORAC values were determined as Trolox Equivalents (TE). Cocoa beans were also measured for antioxidants using Total Phenolics assay and DPPH assay and measured as gallic acid equivalents (GAE). Cocoa beans from the Ivory Coast were roasted at varying times (10-40 minutes) and temperatures (100ºC -190ºC). It was determined that cocoa beans from the Ivory Coast, roasted at 130ºC for 30 minutes resulted in an ORAC value of 522,789 μmol (micromoles) TE/g, GAE value of 2.46 mg/L as determined by Total Phenolics, and GAE value of 1.48 mg/L as determined by DPPH assay. The analyzed values tended to decrease at the highest temperatures and times of roasting. Cocoa beans from different countries were roasted at 130ºC for 30 minutes and antioxidants were analyzed. It was determined that Dominican Republic and Ecuador had the highest TE values (487,913 and 463958 μmol (micromoles) TE/g respectively). GAE differed and Total Phenolic assay found Haiti had the highest GAE (3.26 mg/L) and DPPH assay found Ivory Coast and Dominican Republic had the highest (0.623 and 0.610 mg/L respectively). If an acceptable flavor, color, and aroma of cocoa can be developed at a roasting temperature closer to 130ºC than to 160ºC, then a greater antioxidant content should occur in dry cocoa powder.

The use of biomarkers is considered a promising alternative, or complement, to traditional ecotoxicological assays. Toxic effects are often initially manifested at the molecular or biochemical level, biomarkers are therefore used as sensitive indicators of toxic exposure. Ideally, biomarkers would also indicate reduced fitness and possible later effects at the individual or population levels. However, implementing biomarkers in ecotoxicology is challenging and few biomarkers have an established connection to reduced individual fitness. The aim of this thesis was to increase the value and improve the interpretation of biomarker responses in ecotoxicological studies by examining the impact of confounding factors and the relationship between oxidative biomarkers and reproductive effects in crustaceans. The sensitivity of biomarkers was confirmed in paper I as toxic effects of pharmaceuticals with conserved drug target orthologs were observed at the molecular and biochemical levels both earlier and at lower concentrations than effects on mortality and reproduction. No toxic effects were observed for the pharmaceutical without identified drug target orthologs, thus stressing the importance of considering toxic mechanisms and being aware of the most likely target when evaluating toxic effects also in non-target species. Many xenobiotics and environmental stressors interfere with oxidative processes, making oxidative biomarkers interesting to study in ecotoxicology and stress ecology. Still, feeding rate was identified as a confounding factor for antioxidant capacity (assayed as oxygen radical absorbance capacity, ORAC) and lipid peroxidation in ecotoxicological studies (paper II). However, ORAC normalized to protein was independent of altered feeding rates, hence it can be applied as a suitable exposure biomarker without considering alterations and effects of feeding rate. The connection between reproduction and oxidative stress is dual, as reproduction both can be inhibited by oxidative stress and induce pro-oxidative processes. Further, a positive association was found between ORAC and the occurrence of embryo aberrations in the benthic amphipod Monoporeia affinis (paper III). An association between antioxidant defense and reproduction was also observed for Daphnia magna (paper IV). Threshold values for identification of exposed individuals and prediction of possible later reproductive effects were established for ORAC. This thesis has contributed to diminishing some of the knowledge gaps limiting the use of oxidative biomarkers in ecotoxicology, by contributing to increased understanding of how oxidative biomarkers relate to important life-traits. Moreover, ORAC has been identified as a suitable biomarker of not only exposure, but also reproductive effects. Future research should continue to establish connections between biomarker responses and effects at higher levels, and focus on providing defined threshold values to enable predictions about later effects. / <p>At the time of the doctoral defense, the following papers were unpublished and had a status as follows: Paper 3: Manuscript. Paper 4: Manuscript.</p>

Orientador: Mário Roberto Maróstica Júnior / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Engenharia de Alimentos / Made available in DSpace on 2018-08-16T13:28:38Z (GMT). No. of bitstreams: 1 Leite_AliceVieira_M.pdf: 971111 bytes, checksum: 075888bf78f40ee6ae18cdf0e3fb131a (MD5) Previous issue date: 2010 / Resumo: O objetivo do presente estudo foi avaliar a produção de casca de jabuticaba em pó, sua composição de antocianinas e efeito antioxidante em animais de laboratório. Muitas frutas tropicais são ricas em antocianinas e existem poucos dados disponíveis sobre a caracterização e quantificação destas antocianinas. A identificação e quantificação de antocianinas de casca de jabuticaba liofilizada foram realizadas por HPLC-PDA e HPLC-ESI-MS/MS e revelaram a presença de dois compostos, delfinidina-3-glicosídeo e cianidina-3- glicosídeo (635,3 e 1964 mg/100 de peso seco), respectivamente. Para as análises do potencial antioxidante foram utilizados os ensaios de ORAC, DPPH e ABTS. Para estes ensaios foram encontrados os seguintes valores, respectivamente, 25514,25±3037,24 YMTEAC.g-1, 45,38 Yg.ml-1, 9457,74±54,29 YMTEAC.g-1. Foram quantificados também os fenólicos totais por Folin-Ciocalteau, e encontrado 556,28±20,37 gGAEb.kg-1 .Neste estudo, a casca de jabuticaba liofilizada mostrou uma grande capacidade antioxidante comprovada por todos os métodos antioxidantes analisados, pela quantidade de compostos fenólicos encontrada e principalmente pela concentração de cianidina-3-glicosídeo e delfinidina-3- glicosídeo. Diante dos dados descritos acima, o efeito antioxidante da ingestão de pó casca de jabuticaba liofilizada no plasma de ratos foi investigado em dois experimentos. No primeiro foi realizado um teste de cinética de absorção, no qual trinta e cinco ratos machos, divididos em 5 grupos, receberam por gavagem 7 mg de antocianinas/ 100 g de peso. A coleta de sangue foi realizada nos tempos 0, 30, 60 e 120 minutos. O potencial antioxidante foi quantificado no plasma por ORAC e TEAC. Neste experimento não ocorreu diferença significativa no potencial antioxidante plasmático quando comparados os diversos tempos. No segundo experimento, 40 ratos machos foram divididos em 4 grupos que consumiram, respectivamente, 0, 1, 2 e 4% de pó de casca de jabuticaba liofilizada acrescido a dieta. Foi observado um aumento no potencial antioxidante plasmático (1.7 vezes pelo método de TEAC e 1.3 vezes por ORAC), porém a concentração de 4% não apresentou efeito antioxidante. A partir dos resultados do potencial antioxidante in vivo, e diante do fato de que a ingestão de uma dieta rica em antioxidantes e com pouca quantidade de gorduras pode reduzir o risco de obesidade e resistência à insulina, investigou-se o efeito da casca de jabuticaba liofilizada sobre as concentrações plasmáticas de glicose, frações lipídicas, aspartato aminotransferase e alanina aminotrasferase em ratos Wistar machos adultos. Os animais foram distribuídos nos seguintes grupos: G0, o grupo controle e G1, G2 e G4, os grupos que receberam 1, 2 e 4%, respectivamente, do pó de casca de jabuticaba liofilizada adicionado à dieta AIN-93M. A suplementação reduziu as concentrações de glicose nos grupos G1 e G2, porém no grupo G3 esse efeito não foi encontrado. O colesterol total sofreu redução para os animais tratados com 1% de cascas de jabuticabas em relação ao grupo controle, mas os animais tratados com 2 e 4% de cascas de jabuticabas sofreram aumento no colesterol total e da fração LDL-colesterol, porém, o valor para colesterol total apresenta-se dentro da faixa normal para ratos Wistar. Os triacilgliceróis e a fração HDL-colesterol não sofreram alterações significativas. Os resultados indicam um possível benefício da suplementação da casca de jabuticaba na redução da glicemia plasmática, dos níveis de LDL e colesterol total e a possível existência de uma dose recomendada / Abstract: The objective of this study was to evaluate the antioxidant effects of anthocyanin and the antioxidant effects of peel jabuticaba powder in laboratory animals. Many tropical fruits are rich in anthocyanins, but there are few data available on the characterization and quantification of these anthocyanins. The identification and quantification of anthocyanins from bark jabuticaba was performed by HPLC-PDA and LC-ESI-MS/MS Q-TOF. The analysis revealed the presence of two compounds, delphinidin-3-glucoside and cyanidin-3-glucoside (635, 3 and 1964 mg/100 dry weight, respectively). ORAC, DPPH and ABTS methods were used for antioxidant analysis and the results were, 25514,24±3037,24 µMTEAC.g-1, 45.38 Yg.ml-1, 9457,74±54,29 µMTEAC.g-1, respectively. The total phenolics (Folin-Ciocalteau method) revealed that the jabuticaba powder contained 556,28±20,37 gGAEb.kg-1. In this study, freezedried peel jabuticaba showed great antioxidant proven by all methods antioxidants tested. The effects of the ingestion of jabuticaba peel in the antioxidant capacity of plasma of rats was investigated in two 'in vivo¿ experiments. In the first test, the absorption kinetics were evaluated. The male rats were divided into 5 groups received, by gavage, 7 mg anthocyanins / 100 g of body weight. Blood collection was performed at 0, 30, 60 and 120 minutes. The antioxidant potential was quantified in plasma by ORAC and TEAC. In this experiment there was no significant difference in plasma antioxidant potential when comparing the different times. In the second experiment, 40 rats were divided into 4 groups that consumed, respectively, 0, 1, 2 and 4% powdered peel jabuticaba lyophilized plus diet. We observed an increase in plasma antioxidant potential (1.7 times by the method of TEAC and ORAC by 1.3 4 times), but the concentration of 4% had no antioxidant effect. We investigated the effect of freeze-dried peel jabuticaba on plasma glucose, lipid fractions, aminotransferase and aspartate aminotransferase alanian in adult male Wistar rats. The animals assigned to groups G0, the control group, G1, G2 and G4 groups receiving 1, 2 and 4%, respectively, of freeze-dried peel jabuticaba added to the AIN-93M diet. Supplementation reduced the concentration of glucose in G1 and G2, but in G4 this effect was not found. Total cholesterol was reduced in animals treated with 1% bark jabuticabas in the control group, but animals treated with 2 and 4% experienced an increase in total cholesterol and LDL cholesterol, but the value for cholesterol total amount is within the normal range for rats. Triglycerides and HDL-cholesterol did not change significantly. The results indicate the possible benefit of supplementing with bark jabuticaba in reducing plasma glucose, LDL cholesterol and total cholesterol and the possible existence of a dose / Mestrado / Nutrição Experimental e Aplicada à Tecnologia de Alimentos / Mestre em Alimentos e Nutrição

O arroz (Oryza sativa L.) é consumido predominantemente na forma polida, porém há uma tendência na procura por grãos integrais não pigmentados e pigmentados, como o preto e o vermelho, e também pelo arroz selvagem, que pertence ao gênero Zizania. Além da composição nutricional e propriedades sensoriais distintas, os tipos pigmentados possuem compostos bioativos, que atribuem cor aos grãos e que têm sido relacionados a efeitos benéficos à saúde. Contudo, uma comparação em termos nutricionais e de bioativos entre esses grãos ainda inexiste. Este projeto visou comparar a composição química, incluindo os compostos fenólicos totais (CFT), os polifenóis majoritários e a capacidade antioxidante de três amostras de arroz-preto produzido no Estado de São Paulo, duas de arroz-preto produzido no Estado do Rio Grande do Sul e 11 de arroz-preto produzido no Estado de Santa Catarina, 9 de arroz-vermelho, produzido nos estados mencionados e, 6 amostras de arroz selvagem, importadas do Canadá e comercializadas em São Paulo; todas as amostras foram produzidas e/ou comercializadas no período de 2009 a 2011. A comparação das variáveis foi feita pelo uso de análise estatística uni- e multivariada. Fez parte do escopo do projeto avaliar o efeito do cozimento sobre a estabilidade dos CFT e a atividade antioxidante. Também foi conduzida uma comparação preliminar dos teores de fitoquímicos lipossolúveis, &#947;-orizanol, tocoferóis e tocotrienóis, entre 7 amostras de arroz-preto e 4 de arroz-vermelho, todas da safra de 2013. Com base nas análises químicas e na análise estatística multivariada, foi possível agrupar os vários tipos de arroz em quatro grupos, significativamente diferentes entre si: arroz selvagem, arroz-preto de grãos longos, arroz-preto de grãos médios e arroz-vermelho. O grupo que mais se diferenciou dos demais foi o arroz selvagem, por apresentar os maiores teores proteicos (12,9 g/100g) e de ácido &#945;-linolênico (0,12 g/100g) e os menores teores de lipídeos (0,9 g/100g), de compostos fenólicos e atividade antioxidante, entre os grupos estudados. Outro grupo com características distintas foi o arroz-preto de grãos longos que se destacou principalmente pelo elevado teor de compostos fenólicos, representados pelas antocianinas, e elevada atividade antioxidante. A cianidina-3-O-glicosídeo foi identificada por CLAE-DAD-MS/MS, como sendo a antocianina majoritária. Os teores de proteínas e de lipídeos, da ordem de 9,8 e 3,6 g/100g, respectivamente, foram maiores do que dos outros dois grupos estudados. O arroz-preto de grãos médios e o vermelho foram similares em termos de composição de nutrientes e CFT. Os teores proteicos em ambos os grupos foi, em média, de 8,8 g/100g. Porém, enquanto no arroz-preto predominam antocianinas, a coloração do arroz-vermelho é conferida por proantocianidinas, com diferenças na sua atividade antioxidante. O arroz-preto de grãos médios apresentou atividade antioxidante mais do que o dobro do arroz-vermelho, no entanto próxima ao do arroz-preto de grãos longos. Este resultado indica que o elevado teor de antocianinas, independente do formato do grão, é responsável pela alta capacidade antioxidante no arroz-preto. A análise multivariada demonstrou que o formato do grão é determinante na diferenciação dos dois grupos de arroz-preto em termos de composição de nutrientes, como proteínas e lipídeos, mas não de fitoquímicos e de atividade antioxidante. O método ORAC foi mais apropriado para avaliar a atividade antioxidante do arroz-preto, por ser mais sensível à presença de antocianinas do que o método de DPPH&#183;. Nos outros dois grupos, vermelho e selvagem o método de DPPH&#183; também pode ser utilizado. O cozimento do arroz provocou perda significativa nos teores de CFT e na atividade antioxidante do arroz-preto e vermelho. No arroz-preto, a perda de CFT foi de 26%, em média, enquanto a perda das antocianinas foi de 50%. A explicação pode ser que durante o cozimento uma parte das antocianinas seja convertida em ácido protocatecóico, que é dosado como CFT. No arroz-vermelho, a redução de CFT foi de 60%, possivelmente devido a uma insolubilização de parte das proantocianidinas. No arroz selvagem, o cozimento não causou perdas significativas. A capacidade antioxidante dos vários grupos de arroz após o cozimento depende do método empregado, sendo fortemente correlacionada com os teores de CFT remanescentes. Assim, o arroz-preto, mesmo depois de cozido, apresentou a maior capacidade antioxidante, seguido do arroz-vermelho e do selvagem. Numa avaliação inicial, os teores dos fitoquímicos lipossolúveis, &#947;-orizanol e homólogos da vitamina E, foram semelhantes em arroz-preto e vermelho, indicando que independem da coloração do pericarpo. / Rice (Oryza sativa L.) is mostly consumed in its milled form; however there is an increasing demand for whole non-pigmented and pigmented rice, such as black, red, and wild rice, which the latter belongs to the genus Zizania. Pigmented rice has particular nutritional composition and sensory characteristics, and in addition high amounts of phenolic compounds, which not only confer color but also has been linked to beneficial effects on human health. To date, little is known about the nutritional and bioactive contents of these grains. The aim of this study was to compare the chemical composition, including the total phenolic compounds (TPC), the majoritarian polyphenols and the antioxidant radical efficiency of the following accessions: three black rice from the state of São Paulo, two black rice from the state of Rio Grande do Sul, eleven black rice genotypes from Santa Catarina state, nine red rice from those states and six wild rice, imported from Canada and marketed in São Paulo. All samples were cultivated and/or marketed from 2009 to 2011. Data were evaluated by uni- and multivariate statistical analysis. The effect of cooking on the stability of TPC and antioxidant capacity was also evaluated. In addition, a preliminary comparison of &#947;- oryzanol, tocopherols and tocotrienols was carried out between two groups: seven black rice and four red rice samples, all of them cultivated in 2013. Based on chemical results and multivariate statistical analysis it was possible to cluster the various types of rice in four groups, significantly different among themselves: wild rice, black long grain rice, black medium grain rice, and red rice. Wild rice was the most dissimilar group due to its highest contents of protein (12.9 g/100g) and &#945;-linolenic acid (0.12 g/100g), and the lowest amounts of lipids (0.9 g/100g), TPC, and antioxidant capacity. Black long grain rice was characterized mainly by its high contents of TPC, especially anthocyanins and by its elevated antioxidant capacity. Cyanidin-3-O-glycoside was identified by HPLC-DAD-MS/MS as being the main anthocyanin. The protein and lipid mean contents in these groups were respectively, 9.8 and 3.6 g/100g and were higher than the amounts in the other next two types of rice. The black medium grain rice and red rice were similar in terms of nutrient composition and TPC. The average amount of protein in both groups was about 8.8 g/100g. However, in black rice prevails anthocyanins, while in red rice coloration is provided by proanthocyanidins, which results in differences in antioxidant activity. Medium and long black grain rice showed a 2-fold higher antioxidant activity than red rice. These findings indicate that the high content of anthocyanins, independent of the grain shape, is responsible for the high antioxidant capacity in black rice. The multivariate analysis demonstrated that the grain shape is fundamental to differentiate black rice in terms of nutrient composition, like protein and lipids, but not in relation to amounts of phytochemicals and antioxidant capacity. ORAC was more suitable than DPPH&#183; methodology to evaluate the antioxidant activity of black rice, due to its high correlation to anthocyanin contents. Conversely, DPPH&#183; can be a consistent method to evaluate antioxidant capacity of red and wild rice. Cooking resulted in significant loss on TPC contents and on the antioxidant capacity of black and red rice. In black rice, 26% of TPC was reduced on average, while the loss of anthocyanins was 50%. The reason may be that during cooking, part of the anthocyanins is converted into protocatechuic acid, which is quantified as TPC. The reduction in TPC in red rice was 60%, possibly due to a partial insolubilization of proanthocyanidins. In wild rice, cooking caused no significant loss of TPC. The antioxidant capacity of different types of cooked rice was dependent on the method used, being strongly correlated with the remaining levels of TPC. Thus, black rice even after cooking showed the highest antioxidant capacity, followed by red and wild rice. From a preliminary evaluation, the contents of lipophilic phytochemicals, &#947-orizanol and vitamin E homologues were similar in black and red rice, which indicates that the contents of these compounds are not dependent of the pericarp color.

Les stilbènes sont des composés phénoliques issus du métabolisme secondairevégétal, dont la distribution au sein du règne végétal est limitée aux espèces qui ont acquis aucours de l’évolution la capacité à synthétiser ces molécules. Leurs impacts et leurs activitésbiologiques tels que les effets neuroprotecteurs, anticancérigènes, antioxydants ont déjàconcerné plusieurs sujets d’étude. C’est dans ce contexte que l’objectif de notre travail a prisnaissance. Dans un premier temps, nous avons isolé et identifié ces molécules chez quelquesespèces de la famille des Cypéracées. Cette étude phytochimique a été réalisée en utilisant unensemble de stratégies analytiques et préparatives faisant appel à la CLHP analytique etpréparative et la CPC (Chromatographie de Partage Centrifuge) pour l’obtention desmolécules pures et à la LC-Masse et la RMN pour l’identification des composées isolés. Dansun second temps, nous avons étudié les activités biologiques in vitro de ces produits telles queles activités antioxydantes par trois méthodes (ORAC, DPPH et MCA) et l’effet de cesstilbènes sur la cytotoxicité neuronale induite par le peptide β-amyloïde avec des cellulesPC12. Nous avons isolé une nouvelle molécule, le carexinol A, pour la première fois, qui amontré une forte activité anti-amyloïde. La dernière partie de cette thèse fait état de l’analyseet du dosage des stilbènes dans des vins tunisiens dont le vin Sidi Zahia qui a donné lesmeilleurs résultats. / Stilbenes are phenolic compounds of plant secondary metabolism, whosedistribution within the plant kingdom is limited to species that have acquired during theevolution the ability to synthesize these molecules. Their impacts and their biologicalactivities such as neuroprotective, anticarcinogenic, antioxidant effects have already touchedseveral topics. It is in this context that the purpose of our work arose. First, we have isolatedand identified these molecules in some species of the sedge family. Phytochemical studieswere performed using a set of analytical and preparative strategies by means of analytical andpreparative HPLC and CPC (Centrifugal Partition Chromatography) for obtaining puremolecules and LC-Mass and NMR for identification of compound isolated. In a second step,we investigated the in vitro biological properties of these products such as antioxidantactivities by three methods (ORAC, DPPH and MCA) and their effect on neuronalcytotoxicity induced by β-amyloid peptide with PC12 cells. We isolated a new molecule, forthe first time, carexinol A, that showed strong anti-amyloid activity. The last part of this thesisrefers to the analysis and determination of stilbenes in Tunisian wines including Sidi Zahiawine that gave the best results.

O arroz (Oryza sativa L.) é consumido predominantemente na forma polida, porém há uma tendência na procura por grãos integrais não pigmentados e pigmentados, como o preto e o vermelho, e também pelo arroz selvagem, que pertence ao gênero Zizania. Além da composição nutricional e propriedades sensoriais distintas, os tipos pigmentados possuem compostos bioativos, que atribuem cor aos grãos e que têm sido relacionados a efeitos benéficos à saúde. Contudo, uma comparação em termos nutricionais e de bioativos entre esses grãos ainda inexiste. Este projeto visou comparar a composição química, incluindo os compostos fenólicos totais (CFT), os polifenóis majoritários e a capacidade antioxidante de três amostras de arroz-preto produzido no Estado de São Paulo, duas de arroz-preto produzido no Estado do Rio Grande do Sul e 11 de arroz-preto produzido no Estado de Santa Catarina, 9 de arroz-vermelho, produzido nos estados mencionados e, 6 amostras de arroz selvagem, importadas do Canadá e comercializadas em São Paulo; todas as amostras foram produzidas e/ou comercializadas no período de 2009 a 2011. A comparação das variáveis foi feita pelo uso de análise estatística uni- e multivariada. Fez parte do escopo do projeto avaliar o efeito do cozimento sobre a estabilidade dos CFT e a atividade antioxidante. Também foi conduzida uma comparação preliminar dos teores de fitoquímicos lipossolúveis, &#947;-orizanol, tocoferóis e tocotrienóis, entre 7 amostras de arroz-preto e 4 de arroz-vermelho, todas da safra de 2013. Com base nas análises químicas e na análise estatística multivariada, foi possível agrupar os vários tipos de arroz em quatro grupos, significativamente diferentes entre si: arroz selvagem, arroz-preto de grãos longos, arroz-preto de grãos médios e arroz-vermelho. O grupo que mais se diferenciou dos demais foi o arroz selvagem, por apresentar os maiores teores proteicos (12,9 g/100g) e de ácido &#945;-linolênico (0,12 g/100g) e os menores teores de lipídeos (0,9 g/100g), de compostos fenólicos e atividade antioxidante, entre os grupos estudados. Outro grupo com características distintas foi o arroz-preto de grãos longos que se destacou principalmente pelo elevado teor de compostos fenólicos, representados pelas antocianinas, e elevada atividade antioxidante. A cianidina-3-O-glicosídeo foi identificada por CLAE-DAD-MS/MS, como sendo a antocianina majoritária. Os teores de proteínas e de lipídeos, da ordem de 9,8 e 3,6 g/100g, respectivamente, foram maiores do que dos outros dois grupos estudados. O arroz-preto de grãos médios e o vermelho foram similares em termos de composição de nutrientes e CFT. Os teores proteicos em ambos os grupos foi, em média, de 8,8 g/100g. Porém, enquanto no arroz-preto predominam antocianinas, a coloração do arroz-vermelho é conferida por proantocianidinas, com diferenças na sua atividade antioxidante. O arroz-preto de grãos médios apresentou atividade antioxidante mais do que o dobro do arroz-vermelho, no entanto próxima ao do arroz-preto de grãos longos. Este resultado indica que o elevado teor de antocianinas, independente do formato do grão, é responsável pela alta capacidade antioxidante no arroz-preto. A análise multivariada demonstrou que o formato do grão é determinante na diferenciação dos dois grupos de arroz-preto em termos de composição de nutrientes, como proteínas e lipídeos, mas não de fitoquímicos e de atividade antioxidante. O método ORAC foi mais apropriado para avaliar a atividade antioxidante do arroz-preto, por ser mais sensível à presença de antocianinas do que o método de DPPH&#183;. Nos outros dois grupos, vermelho e selvagem o método de DPPH&#183; também pode ser utilizado. O cozimento do arroz provocou perda significativa nos teores de CFT e na atividade antioxidante do arroz-preto e vermelho. No arroz-preto, a perda de CFT foi de 26%, em média, enquanto a perda das antocianinas foi de 50%. A explicação pode ser que durante o cozimento uma parte das antocianinas seja convertida em ácido protocatecóico, que é dosado como CFT. No arroz-vermelho, a redução de CFT foi de 60%, possivelmente devido a uma insolubilização de parte das proantocianidinas. No arroz selvagem, o cozimento não causou perdas significativas. A capacidade antioxidante dos vários grupos de arroz após o cozimento depende do método empregado, sendo fortemente correlacionada com os teores de CFT remanescentes. Assim, o arroz-preto, mesmo depois de cozido, apresentou a maior capacidade antioxidante, seguido do arroz-vermelho e do selvagem. Numa avaliação inicial, os teores dos fitoquímicos lipossolúveis, &#947;-orizanol e homólogos da vitamina E, foram semelhantes em arroz-preto e vermelho, indicando que independem da coloração do pericarpo. / Rice (Oryza sativa L.) is mostly consumed in its milled form; however there is an increasing demand for whole non-pigmented and pigmented rice, such as black, red, and wild rice, which the latter belongs to the genus Zizania. Pigmented rice has particular nutritional composition and sensory characteristics, and in addition high amounts of phenolic compounds, which not only confer color but also has been linked to beneficial effects on human health. To date, little is known about the nutritional and bioactive contents of these grains. The aim of this study was to compare the chemical composition, including the total phenolic compounds (TPC), the majoritarian polyphenols and the antioxidant radical efficiency of the following accessions: three black rice from the state of São Paulo, two black rice from the state of Rio Grande do Sul, eleven black rice genotypes from Santa Catarina state, nine red rice from those states and six wild rice, imported from Canada and marketed in São Paulo. All samples were cultivated and/or marketed from 2009 to 2011. Data were evaluated by uni- and multivariate statistical analysis. The effect of cooking on the stability of TPC and antioxidant capacity was also evaluated. In addition, a preliminary comparison of &#947;- oryzanol, tocopherols and tocotrienols was carried out between two groups: seven black rice and four red rice samples, all of them cultivated in 2013. Based on chemical results and multivariate statistical analysis it was possible to cluster the various types of rice in four groups, significantly different among themselves: wild rice, black long grain rice, black medium grain rice, and red rice. Wild rice was the most dissimilar group due to its highest contents of protein (12.9 g/100g) and &#945;-linolenic acid (0.12 g/100g), and the lowest amounts of lipids (0.9 g/100g), TPC, and antioxidant capacity. Black long grain rice was characterized mainly by its high contents of TPC, especially anthocyanins and by its elevated antioxidant capacity. Cyanidin-3-O-glycoside was identified by HPLC-DAD-MS/MS as being the main anthocyanin. The protein and lipid mean contents in these groups were respectively, 9.8 and 3.6 g/100g and were higher than the amounts in the other next two types of rice. The black medium grain rice and red rice were similar in terms of nutrient composition and TPC. The average amount of protein in both groups was about 8.8 g/100g. However, in black rice prevails anthocyanins, while in red rice coloration is provided by proanthocyanidins, which results in differences in antioxidant activity. Medium and long black grain rice showed a 2-fold higher antioxidant activity than red rice. These findings indicate that the high content of anthocyanins, independent of the grain shape, is responsible for the high antioxidant capacity in black rice. The multivariate analysis demonstrated that the grain shape is fundamental to differentiate black rice in terms of nutrient composition, like protein and lipids, but not in relation to amounts of phytochemicals and antioxidant capacity. ORAC was more suitable than DPPH&#183; methodology to evaluate the antioxidant activity of black rice, due to its high correlation to anthocyanin contents. Conversely, DPPH&#183; can be a consistent method to evaluate antioxidant capacity of red and wild rice. Cooking resulted in significant loss on TPC contents and on the antioxidant capacity of black and red rice. In black rice, 26% of TPC was reduced on average, while the loss of anthocyanins was 50%. The reason may be that during cooking, part of the anthocyanins is converted into protocatechuic acid, which is quantified as TPC. The reduction in TPC in red rice was 60%, possibly due to a partial insolubilization of proanthocyanidins. In wild rice, cooking caused no significant loss of TPC. The antioxidant capacity of different types of cooked rice was dependent on the method used, being strongly correlated with the remaining levels of TPC. Thus, black rice even after cooking showed the highest antioxidant capacity, followed by red and wild rice. From a preliminary evaluation, the contents of lipophilic phytochemicals, &#947-orizanol and vitamin E homologues were similar in black and red rice, which indicates that the contents of these compounds are not dependent of the pericarp color.

Regular physical activity (PA) leads to an increased quality of life by means of certain physiological adaptations. Regular PA is beneficial to the human body and its functionality, including the physiological, biochemical and even psychological modalities. During PA an increased burden is placed on all physiological mechanisms due to the increased energy demand, resulting in an adaptation of the physiological systems. Currently the biochemical mechanisms by which these adaptations occur are not well understood or defined. During the flow of electrons through the electron transport chain in the mitochondria free radicals and reactive oxygen species (ROS) are produced. PA results in increased ROS production. The relationship of different exercise intensities and ROS production with resulting DNA damage is unclear. These free radicals and ROS disturb the pro-oxidant anti-oxidant balance resulting in oxidative stress. When this balance is disturbed oxidative stress could lead to potential oxidative damage, Oxidative damage occurs in lipid, protein and nucleic acid macromolecules. ROS can attack DNA bases or deoxyribose residues to produce damaged bases and/or single and double strand breaks. When the DNA is regarded and the damages are replicated it could cause mutations or apoptosis, affecting the cell function and physiology. The purpose of this study was to investigate the influence of different aerobic intensities on oxidative DNA damage and repair in middle aged men by means of the Comet assay. Five PA males and five physically inactive males were assigned to an experimental and control group respectively. The subjects did not differ significantly at baseline. The VO2-max of each subject was determined at baseline. Subjects were then randomly assigned to 60, 70, 80 and 90% of individual baseline VO2-max intensities for an acute exercise intervention of 30 minutes on a bicycle ergometer. Blood sampling was done at baseline, post-exercise and 24 hours post-exercise for oxygen radical absorbance capacity (ORAC) and hydroperoxide analysis (dROM). Peripheral blood was obtained for DNA damage testing by means of Comet analysis at baseline, post-exercise, 5, 15, 30 minutes, and also 6, 12, 24, 48 and 72 hours after exercise. The results obtained indicated that subjects who regularly participate in PA had an increased baseline reading of ORAC and dROM values. ORAC levels after each acute exercise session increased, with the highest increase in the control group, with a decrease in the direction of baseline readings 24 hours post exercise. A biphasic damage-repair cycle over the 72 hour period was observed with the Comet analysis. The most damaged cells occur directly after acute exercise. The highest incidence of DNA damage over a 72 hour period was observed at 70% VO2-max, with the least amount of damage after 90% VO2-max. In conclusion the study indicates stress proteins or other kinds of physiological reaction to minimize the damaging effect of oxidative stress, is in place to restore the cell's homeostasis. Thus PA results in the development of oxidative DNA damage. To minimize DNA damage the optimal intensity for acute physical exercise is between 70-80% VO2-max. At higher intensities the release of stress proteins are initiated to buffer the damaging effect of oxidative stress and to restore homeostasis. / Thesis (M.Sc. (Human Movement Science))--North-West University, Potchefstroom Campus, 2007.