Qualitative and quantitative analysis of anthocyanins and other phenolic compounds from a purple corn extract was performed. The purple corn extract had cyanidin-3-glucoside, pelargonidin-3-glucoside, peonidin-3-glucoside and its respective acylated anthocyanin-glucosides. Cyadinin-3glucoside was the main constituent (44.4 ?? 4.7%) followed by the acylated cyanidin-3-glucoside (26.9 ?? 8.0%). Other phenolic compounds present in the purple corn corresponded to protocatechuic acid, vanillic acid, and p-coumaric acid. In addition, quercetin derivatives, a hesperitin derivative and pcoumaric and ferulic acid derivatives were found. Fractionation of phenolic compounds yielded two main fractions, an anthocyanin-rich water fraction (WF) and an ethyl acetate fraction (EAF). Evaluation of antimutagenic activity in both fractions revealed higher antimutagenic activity in the ethyl acetate fraction compared to the anthocyanin-rich fraction. On the other hand, antioxidant activity of the anthocyanin-rich fraction was higher compared to the ethyl acetate fraction. Further fractionation of the anthocyanin-rich fraction in a Toyopearl HW40 gel permeation column yielded five sub-fractions which showed no difference in antimutagenic activity except for the water sub-fraction WF-V. All the sub-fractions were active as antimutagens and antioxidants. Further fractionation of the ethyl acetate fraction yielded four sub-fractions that showed to be active as antimutagens and antioxidants. Ethyl acetate sub-fraction EAF-IV was the most active as an antimutagen. HPLC-DAD characterization of that sub-fraction revealed mainly the presence of a quercetin derivative with UV-visible spectral characteristics similar to rutin but with a little longer retention time. The mechanism of antimutagenic action by the phenolic compounds present either in the anthocyanin-rich fraction or the ethyl acetate fraction and sub-fraction EAFIV seems to be a contribution of a direct action on the enzymes involved in the activation of the mutagen and to the scavenging activity of the mutagen nucleophiles, as demonstrated by our assays.

Made available in DSpace on 2014-06-11T19:27:56Z (GMT). No. of bitstreams: 0 Previous issue date: 2008-05-29Bitstream added on 2014-06-13T20:36:36Z : No. of bitstreams: 1 almeida_dc_me_botfm.pdf: 330108 bytes, checksum: 81beba7a4d6ff8045b1bfcf3c2d5173d (MD5) / Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Considerando que as estratégias para a prevenção de doenças relacionadas a danos no DNA, ou que envolvam a eliminação da exposição humana a fatores de risco nem sempre são possíveis, a identificação de agentes quimioprotetores torna-se uma alternativa relevante. Nesse contexto, a própolis, por seu amplo espectro de ação biológica e devido a sua complexa composição química, fácil obtenção e importância econômica para o país, é um composto que vem sendo bastante estudado, especialmente com relação a sua atividade terapêutica na saúde humana. Assim sendo, o presente estudo objetivou avaliar a ação protetora do extrato aquoso de própolis verde (12,5, 25, 50, 100 e 200 μg/ml) e duas de suas frações (F4 e F5; 25, 50 e 100 μg/ml) sobre danos quimicamente induzidos no DNA de células das linhagens CHO (células de ovário de hamster chinês) e HepG2 (células de hepatoblastoma humano), sob três protocolos de tratamento. Como agentes indutores de danos genéticos foram utilizados o peróxido de hidrogênio (H2O2), a 4-n-óxido-nitroquinolina (4NQO), o metil metano sulfonato (MMS) e a n-dietilnitrosamina. Os dados mostraram que tanto o extrato aquoso da própolis quanto suas frações foram eficientes na proteção contra danos genotóxicos, nas duas linhagens celulares e nos diferentes protocolos de tratamento, quando avaliados pelo teste do cometa. No entanto, quando a própolis foi testada pelo teste do micronúcleo, sua atividade antimutagênica foi menos evidente, sendo detectada apenas em situações específicas. Concluindo, os resultados demonstraram que tanto o extrato aquoso da própolis como suas frações foram capazes de reduzir os danos quimicamente induzidos no DNA, por mecanismos antioxidantes e de modulação de sistema enzimático de metabolismo de drogas. / Since it is not always possible to reduce human exposure to mutagens, attempts have been directed to identify potential antimutagens and anticarcinogens for protecting human population against environmental diseases. Propolis has been used in folk medicine since ancient times, and its antimicrobial, antiparasitic, antiviral, antiinflammatory, antitumoral and antioxidant properties have been described. In view of the great therapeutic interest on this bee product, this study was designed to evaluate the antigenotoxic and antimutagenic effect of an aqueous extract of propolis (AEP) and two of its fractions (F4 and F5) on Chinese Hamster Ovary (CHO) and Human Hepatoblastoma (HepG2) cell lines by using comet assay and cytokinesis-block micronucleus assay (CBMN). Chemicals with different mechanisms of mutagenicity (hydrogen peroxide, 4-nitroquinoline-1-oxide, methyl methanesulphonate (MMS), and or n-nitrosodiethylamine) were used in order to better understand propolis chemopreventive action. Data showed that both AEP and the fractions reduced genotoxicity as depicted by the comet assay. However, lower protective effect was detected in the micronucleus assay. In conclusion, the results confirmed propolis as an effective chemopreventive agent, mainly on primary-induced DNA damage. This effect might be attributed to its antioxidant activity and also to its capability in modulate drug metabolizing enzymes.

Orientador: Daisy Maria Fávero Salvatori / Banca: José Maurício Sforcin / Banca: Verônica Pimenta Vicentini / Resumo: Considerando que as estratégias para a prevenção de doenças relacionadas a danos no DNA, ou que envolvam a eliminação da exposição humana a fatores de risco nem sempre são possíveis, a identificação de agentes quimioprotetores torna-se uma alternativa relevante. Nesse contexto, a própolis, por seu amplo espectro de ação biológica e devido a sua complexa composição química, fácil obtenção e importância econômica para o país, é um composto que vem sendo bastante estudado, especialmente com relação a sua atividade terapêutica na saúde humana. Assim sendo, o presente estudo objetivou avaliar a ação protetora do extrato aquoso de própolis verde (12,5, 25, 50, 100 e 200 μg/ml) e duas de suas frações (F4 e F5; 25, 50 e 100 μg/ml) sobre danos quimicamente induzidos no DNA de células das linhagens CHO (células de ovário de hamster chinês) e HepG2 (células de hepatoblastoma humano), sob três protocolos de tratamento. Como agentes indutores de danos genéticos foram utilizados o peróxido de hidrogênio (H2O2), a 4-n-óxido-nitroquinolina (4NQO), o metil metano sulfonato (MMS) e a n-dietilnitrosamina. Os dados mostraram que tanto o extrato aquoso da própolis quanto suas frações foram eficientes na proteção contra danos genotóxicos, nas duas linhagens celulares e nos diferentes protocolos de tratamento, quando avaliados pelo teste do cometa. No entanto, quando a própolis foi testada pelo teste do micronúcleo, sua atividade antimutagênica foi menos evidente, sendo detectada apenas em situações específicas. Concluindo, os resultados demonstraram que tanto o extrato aquoso da própolis como suas frações foram capazes de reduzir os danos quimicamente induzidos no DNA, por mecanismos antioxidantes e de modulação de sistema enzimático de metabolismo de drogas. / Abstract: Since it is not always possible to reduce human exposure to mutagens, attempts have been directed to identify potential antimutagens and anticarcinogens for protecting human population against environmental diseases. Propolis has been used in folk medicine since ancient times, and its antimicrobial, antiparasitic, antiviral, antiinflammatory, antitumoral and antioxidant properties have been described. In view of the great therapeutic interest on this bee product, this study was designed to evaluate the antigenotoxic and antimutagenic effect of an aqueous extract of propolis (AEP) and two of its fractions (F4 and F5) on Chinese Hamster Ovary (CHO) and Human Hepatoblastoma (HepG2) cell lines by using comet assay and cytokinesis-block micronucleus assay (CBMN). Chemicals with different mechanisms of mutagenicity (hydrogen peroxide, 4-nitroquinoline-1-oxide, methyl methanesulphonate (MMS), and or n-nitrosodiethylamine) were used in order to better understand propolis chemopreventive action. Data showed that both AEP and the fractions reduced genotoxicity as depicted by the comet assay. However, lower protective effect was detected in the micronucleus assay. In conclusion, the results confirmed propolis as an effective chemopreventive agent, mainly on primary-induced DNA damage. This effect might be attributed to its antioxidant activity and also to its capability in modulate drug metabolizing enzymes. / Mestre

Presented diploma work is focused on study of antioxidants in different kinds of dried fruits. Analyses of ascorbate, tocopherols, carotenoids and flavonoids were performed using RP-HPLC with spectrophotometric detection. Analysis of dried fruits showed high level of vitamins and phenolocs mainly in berries. High level of carotenoids was observed in dried apricots and plums. Further, antioxidant activity of dried fruit extracts was tested by ABTS method. High antioxidant activity was found mainly in dried apples, cranberries and blueberries. The biological test with yeast Sascharomyces cerevisiae D7 was used for the analysis of antimutagenic efects of dried fruits. High antimutagenic activity exhibited dried cranberries and blueberries. Most of tested dried fruits with high antimutagenic effect exhibited also high antioxidant activity as well as high content of some antioxidants. No direct correlation was found among these parameters. Last part of this work was focused on study of influence of drying on antioxidant content in two types of apples. Drying at mild controlled conditions exhibited no significant negative effect on active substance content; in some samples their concentration was observed.

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No. of bitstreams: 6 Dissertação - Camila Regina do Vale - 2012.pdf: 3235850 bytes, checksum: 687e2cdf37b9bac962f29a62c285b353 (MD5) Dissertação - Camila Regina do Vale - pt 1.pdf: 149467 bytes, checksum: 7f1815d7489ba931bb9892b778028608 (MD5) Dissertação - Camila Regina do Vale - pt 2.pdf: 213195 bytes, checksum: d971df42608e298f9a5b9ef182198144 (MD5) Dissertação - Camila Regina do Vale - pt 3.pdf: 135423 bytes, checksum: 05f3ace3a177dce48ae73b89c7dcafa5 (MD5) Dissertação - Camila Regina do Vale - pt 4.pdf: 209379 bytes, checksum: 0d8927034cc8da946756c3ffc2fd1539 (MD5) license_rdf: 23148 bytes, checksum: 9da0b6dfac957114c6a7714714b86306 (MD5) Previous issue date: 2012-10-23 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / Hymenaea courbaril L. (Family Fabaceae), popularly known in Brazil as jatobá, is a tropical species that occurs in semi-deciduous forest of the South America. The species has been used in Brazil for culinary purposes and in folk medicine to treat arthritis, gastric dysfunction, inflammation and respiratory diseases. Due to the spreading use of this plant as a therapeutic resource and food, the present study aimed to evaluate the toxic, genotoxic, recombinogenic and antigenotoxic effects of Hymenaea courbaril sap (Hycs) using the mouse bone marrow micronucleus test and somatic mutation and recombination test (SMART) in Drosophila melanogaster. To evaluate the clastogenic and aneugenic activities by micronucleos test the animals were treated with 3 concentrations of Hycs (5, 10 and 15 mL/kg body weight). To evaluate the anti-clastogenic and anti-aneugenic activities, the animals were simultaneously treated with Hycs and mitomycin C (4mg/kg body weight). To evaluate the mutagenic and recombinogenic activities by SMART, three-day-old larvae derived from standard (ST) and high bioactivation (HB) crosses were treated with 3 doses of Hycs (0.3, 1.5 or 3 mL) for approximately 48 hours. To evaluate the antimutagenic and antirecombinogenic activities, larvae derived from both crosses were cotreated with 3 doses of Hycs (0.3, 1.5 or 3 mL) and doxorubicin (0.125 mg/mL). Our results in the mouse bone marrow micronucleus test showed that SHyc exhibited no cytotoxic, clastogenic and/or aneugenic effects, but showed anticytotoxic, anti-clastogenic and /or anti-aneugenic activities in mouse bone marrow. The results for the SMART test showed no mutagenic and recombinagenic effects, but antimutagenic and anti-recombinogenic activities were found in both crosses in somatic cells of Drosophila melanogaster. / Hymenaea courbaril L. (família Fabaceae), popularmente conhecida no Brasil como jatobá, é uma espécie tropical que ocorre na floresta semi-decídua da América do Sul. A espécie tem sido utilizada no Brasil para fins culinários e na medicina popular para tratar artrite, disfunção gástrica, inflamação e doenças respiratórias. Devido ao uso generalizado dessa planta como um recurso terapêutico e alimentar, o presente estudo teve como objetivo avaliar os efeitos tóxicos, genotóxicos, recombinogênicos e antigenotóxicos da seiva de Hymenaea courbaril (SHyc), usando o teste do micronúcleo em medula óssea de camundongo e o teste de recombinação e mutação somática (SMART) em Drosophila melanogaster. Para avaliar a ação clastogênica e aneugênica pelo ensaio do micronúcleo, os animais foram tratados com 3 concentrações de SHyc (5, 10 e 15 mL/kg de peso corporal). Para avaliar a atividade anticlastogênica e antianeugênica , os animais foram tratados simultaneamente com SHyc e mitomicina C (4mg/kg de peso corporal). Para avaliar as atividades mutagênica e recombinagênica pelo teste SMART, larvas de terceiro estágio provenientes dos cruzamentos padrão (ST) e alta bioativação (HB) foram tratadas com 3 doses de SHyc (0,3; 1,5 ou 3 mL), por aproximadamente 48 horas. Para avaliar a atividade antimutagênica e anti-recombinogênica, larvas provenientes de ambos os cruzamentos foram co-tratadas com 3 doses de SHyc (0,3, 1,5 ou 3 mL) e doxorubicina (0,125 mg/mL). Nossos resultados para o teste do micronúcleo em medula óssea de camundongos mostraram que SHyc não apresentou efeitos citotóxicos, clastogênicos e/ou aneugênicos , mas apresentou atividade ancitotóxica, anticlastogênica e/ou antianeugênica em medula óssea de camundongos. Os resultados para o teste SMART/asa não demonstraram efeitos mutagênicos e recombinagênicos, mas a acão antimutagênica e anti-recombinogênica foi evidenciado em células somáticas de Drosophila melanogaster.

Chan Wai-Lung, William. / Thesis submitted in: December 2000. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2001. / Includes bibliographical references (leaves 124-131). / Abstracts in English and Chinese. / Abstract --- p.i / Abstract (Chinese Version) --- p.iii / Acknowledgements --- p.v / Table of Contents --- p.vi / List of Tables --- p.ix / List of Figures --- p.xii / List of Abbreviations --- p.xvi / Chapter 1 --- Introduction --- p.1 / Literature Review --- p.4 / Chapter 1.1 --- A Brief Introduction of Cancer --- p.4 / Chapter 1.2 --- Natural Products as a Drug --- p.5 / Chapter 1.2.1 --- Development of terrestrial plants as a drug --- p.6 / Chapter 1.2.1.1 --- Anticancer drugs from terrestrial plants and Chinese medicinal herbs --- p.7 / Chapter 1.2.2 --- Development of marine organisms as a drug --- p.8 / Chapter 1.2.2.1 --- Anticancer drugs from marine organisms --- p.9 / Chapter 1.3 --- Anticlastogenic Study - an Anticancer Study --- p.10 / Chapter 1.3.1 --- Anticlastogenesis mechanisms study --- p.11 / Chapter 1.3.2 --- In vivo anticlastogenic study --- p.13 / Chapter 1.4 --- Anticlastogenic Study of Chinese Medicinal Herbs and Marine Algae --- p.17 / Chapter 1.4.1 --- Selection of nine Chinese medicinal herbs and three marine algae for anticlastogenic screening --- p.18 / Chapter 1.5 --- Methods of Investigation --- p.20 / Chapter 1.5.1 --- Extraction methods --- p.20 / Chapter 1.5.2 --- Single cell gel electrophoresis (Comet assay) --- p.21 / Chapter 2 --- Materials and Methods --- p.27 / Chapter 2.1 --- Materials --- p.27 / Chapter 2.1.1 --- Chinese medicinal herbs --- p.27 / Chapter 2.1.2 --- Marine algae --- p.27 / Chapter 2.1.3 --- Animals --- p.27 / Chapter 2.1.4 --- Chemicals and solutions --- p.28 / Chapter 2.2 --- Methods --- p.31 / Chapter 2.2.1 --- Crude extraction of natural products --- p.31 / Chapter 2.2.1.1 --- Water extraction of Chinese herbs --- p.31 / Chapter 2.2.1.2 --- Water extraction of marine algae --- p.31 / Chapter 2.2.2 --- Test for the effective dosage of clastogen ethyl methanesulfonate (EMS) to BALB/c mice --- p.31 / Chapter 2.2.2.1 --- In vitro test --- p.32 / Chapter 2.2.2.2 --- In vivo test --- p.32 / Chapter 2.2.3 --- Anticlastogenic bioassays --- p.33 / Chapter 2.2.3.1 --- In vitro anticlastogenic screening --- p.33 / Chapter 2.2.3.2 --- In vitro anticlastogenic mechanisms investigation --- p.33 / Chapter 2.2.3.3 --- In vivo anticlastogenic screening --- p.34 / Chapter 2.2.3.4 --- Different in vivo anticlastogenic treatment schedules --- p.35 / Chapter 2.2.4 --- Single cell gel electrophoresis assay (Comet assay) --- p.36 / Chapter 2.2.5 --- White blood cell viability determination --- p.37 / Chapter 2.2.6 --- Statistical analysis --- p.38 / Chapter 3 --- Results --- p.40 / Chapter 3.1 --- Extraction amount of different natural products and cell viability checking --- p.40 / Chapter 3.1.1 --- Chinese medicinal herbs --- p.40 / Chapter 3.1.2 --- Seaweeds --- p.40 / Chapter 3.1.3 --- Cell viability --- p.42 / Chapter 3.2 --- Effective dosage of clastogen EMS to BALB/c mice peripheral white blood cells --- p.42 / Chapter 3.2.1 --- In vitro --- p.42 / Chapter 3.2.2 --- In vivo --- p.42 / Chapter 3.3 --- In vitro anticlastogenic screen test and mechanisms investigation --- p.44 / Chapter 3.3.1 --- In vitro anticlastogenic screen test --- p.44 / Chapter 3.3.1.1 --- Chinese herbs --- p.44 / Chapter 3.3.1.2 --- Seaweeds --- p.53 / Chapter 3.3.2 --- In vitro anticlastogenic mechanisms investigation --- p.55 / Chapter 3.3.2.1 --- H. dilatata --- p.56 / Chapter 3.3.2.2 --- S. angustifolium --- p.56 / Chapter 3.3.2.3 --- S. siliquastrum --- p.63 / Chapter 3.4 --- In vivo anticlastogenic screen test and mechanisms investigation --- p.66 / Chapter 3.4.1 --- In vivo anticlastogenic screen test --- p.66 / Chapter 3.4.1.1 --- Chinese herbs --- p.66 / Chapter 3.4.1.2 --- Seaweeds --- p.73 / Chapter 3.4.2 --- Different treatment methods in in vivo anticlastogenic test --- p.86 / Chapter 3.4.2.1 --- Simultaneous application method --- p.86 / Chapter 3.4.2.2 --- Pre-drug treatment method --- p.91 / Chapter 3.4.2.3 --- Post drug treatment method --- p.91 / Chapter 4 --- Discussion --- p.94 / Chapter 4.1 --- Cell viability and water extracts in Chinese medicinal herbs and marine algae --- p.94 / Chapter 4.2 --- Clastogenic effect of EMS to pWBCs of BALB/c mice --- p.94 / Chapter 4.3 --- In vitro anticlastogenic screen test of nine water extracts of Chinese medicinal herbs and three water extracts of marine algae --- p.99 / Chapter 4.4 --- In vitro anticlastogenic mechanisms investigation of three \03 marine algae extracts --- p.103 / Chapter 4.5 --- In vivo anticlastogenic screen test of Chinese herbs extracts and seaweeds extracts --- p.108 / Chapter 4.6 --- Different administration methods in in vivo anticlastogenic test --- p.115 / Chapter 4.6.1 --- Intraperitoneal route of administration --- p.115 / Chapter 4.6.2 --- In vivo pre- and post-treatment methods --- p.116 / Chapter 5 --- Summary and Conclusion --- p.120 / References --- p.124

Le cancer est une maladie multifactorielle dont la première étape est souvent une mutation. Il est envisageable de prévenir l'apparition de cette maladie en limitant l'apparition de mutations. Le benzo(a)pyrène et le 1-nitropyrène sont deux mutagènes et cancérogènes très répandus dans notre environnement. Ces deux composés entrainent une réponse inflammatoire chez l'homme qui à son tour induit un stress oxydant aboutissant à des mutations. L'objectif de cette étude est de rechercher l'activité antimutagène de sept plantes médicinales et de deux molécules naturelles anti-inflammatoires, avec un intérêt plus développé pour l'Harpagophytum procumbens (HP) et son principal iridoïde : l'harpagoside. Elle consiste également à vérifier si l'activité anti-inflammatoire peut être reliée à une activité antimutagène. L'activité antimutagène a été étudiée au niveau des mutations chromosomiques à l'aide du test des micronoyaux sur lymphocytes humains, et au niveau des mutations ponctuelles à l'aide du test d'Ames. Tous les extraits HP, à l'exception de l'extrait méthanolique, possèdent une activité antimutagène importante dans le test des micronoyaux, mais aucun dans le test d'Ames. Pour les six autres plantes anti-inflammatoires, plus de la moitié des extraits possèdent une activité antimutagène. Nous avons montré que l'activité antioxydante n'est pas ou peu impliquée dans l'activité antimutagène. Il est probable que d'autres mécanismes d'action soient impliqués tels que l'inhibition de l'inflammation (NF-ĸB). C'est la première fois que ce type d'études est réalisé sur des plantes possédant une activité anti-inflammatoire et plus particulièrement sur l' Harpagophytum procumbens. / Cancer is a multifactorial disease in which the first step is often a mutation in the genome. It is then possible to prevent the onset of the disease by limiting the occurrence of mutations. Benzo (a) pyrene (BaP) and 1-nitropyrene (1-NPY) are two widespread mutagens and carcinogens in our environment. These two compounds produce an inflammatory response in humans which in turn induces oxidative stress leading to gene mutations. The objective of this study was to investigate the antimutagenic activity of seven medicinal plants and two natural anti-inflammatory molecules, especially in Harpagophytum procumbens (HP) with its major iridoid: harpagoside. However, also to check whether the anti-inflammatory activity may be related to antimutagenic activity. The antimutagenic activity was investigated with chromosomal mutations using the in vitro cytokinesis-block micronucleus assay in primary cultures of human lymphocytes, and at the point mutations using the Ames test. All HP extracts, except for the methanol extract, showed a significant anti-mutagenic activity in the micronucleus test. No antimutagenic activity could be detected by the Ames assay. For the six other anti-inflammatory plants, more than half of the extracts possessed an antimutagenic activity. We have shown that the antioxidant property was not responsible for the antimutagenic activity. Thus, it is likely that other mechanisms of action are involved, such as anti-adduct mechanism, inhibition of metabolism or inhibition of inflammation (NF-ĸB). This is the first report of antimutagenic properties of anti-inflammatory plants and more particularly of the Harpagophytum procumbens.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / In Brazil, the economic potential of the germplasm of medicinal species is a treasure to be preserved and utilized, increasingly becoming a form of alternative therapy available for the population to use. However, the indiscriminate and uncontrolled use can, cause more harm than benefits to health and it is important to understand these plants, from cellular levels and the action on living organisms. The medicinal species Echinodorus grandiflorus and Sagittaria montevidensis belong to the family Alismataceae, known respectively as leather hat and false leather hat in Brazil. For E. grandiflorus the attributed medicinal potentials are depurative, diuretic, anti-inflammatory, antiophidic and anti-rheumatic action, were leaves are the main part of the plant used, either as a tea or infusion. S. montevidensis has no known medicinal properties and having a wide range of leaf morphologies, leading to population confused with leather hat . The present study evaluated the genotoxic and antiproliferative extracts of the dried leaves of E. grandiflorus and S. montevidensis using the cell cycle of Allium cepa, and performing analysis by high performance liquid chromatography (HPLC-DAD) for the quantification of the extracts of the compounds. The test system is well suited for the analysis of genotoxicity, being validated by the International Programme on Chemical Safety (IPCS, WHO) and can alert on the possible damages that can be caused to organisms exposed to the substances in the plants. For the test, plant material was collected in the field in Rio Grande do Sul, in the municipalities of: Santa Maria, São Sepé, Silveira Martins and Pinhal Grande and in the commercial form of E. grandiflorus, separated into two experimental groups. The extracts were prepared, by infusing dry leaves for 15 min., at two concentrations, 6g.L-1 e 24g.L-1, and two controls, water (negative control) and glyphosate 15% (positive control). The chromatographic analysis (HPLC-DAD), revealed the presence of flavonoids and phenolic acids for both species, higher concentrations of caffeic acid and flavonoid glycoside in E. grandiflorus and phenol glycoside in S. montevidensis. This study showed that extracts of E. grandiflorus and S. montevidensis show genotoxic potential and when used in high concentrations, antiproliferative potential. / No Brasil, o potencial econômico de germoplasma de espécies medicinais é uma riqueza a ser preservada e utilizada, tornando-se cada vez mais uma forma de terapia alternativa acessível para o uso da população. No entanto, o uso indiscriminado e sem controle pode causar mais danos à saúde do que benefícios, sendo importante o conhecimento dessas plantas, desde os níveis celulares bem como ação sobre os organismos vivos. As espécies medicinais Echinodorus grandiflorus e Sagittaria montevidensis pertencem à família Alismataceae, conhecidas respectivamente como chapéu-de-couro e falso chapéu-de-couro. Para E. grandiflorus é atribuído potencial medicinal como ações depurativa, diurética, antiinflamatória, antiofídica e antirreumática, sendo as folhas a principal parte utilizada da planta, seja na forma de chá ou infusão. S. montevidensis não apresenta suas propriedades medicinais conhecidas e tendo uma grande variedade morfológica das folhas, isso leva a população a confundi-la com chapéu-de-couro . O presente estudo teve o objetivo de avaliar o potencial genotóxico e antiproliferativo de extratos das folhas secas de E. grandiflorus e S. montevidensis, sobre o ciclo celular de Allium cepa, bem como realizar análise por cromatografia líquida de alta eficiência (CLAE-DAD) para quantificação dos compostos dos extratos. O sistema teste A. cepa é bem aceito para a análise da genotoxicidade, sendo validado pelo Programa Internacional de Segurança Química (IPCS, OMS), podendo vir a alertar sobre possíveis danos que substâncias podem causar aos organismos a elas expostos. Para o teste foi coletado material vegetativo no habitat natural no Rio Grande do Sul, nos munícipios de: Santa Maria, São Sepé, Silveira Martins e Pinhal Grande e na forma comercializada de E. grandiflorus, separados em 2 grupos experimentais. Os extratos preparados, por infusão de 15 min. das folhas secas, em duas concentrações 6g. L-1 e 24g.L-1, para cada espécie e 2 controles, água (controle negativo) e o glifosato 15% (controle positivo). A análise cromatográfica (HPLC-DAD) revelou a presença de flavonóides e ácidos fenólicos para ambas as espécies. Sendo, maiores teores de ácido cafeico e flavonóide glicosídeo em E. grandiflorus e glicosídeo fenol em S. montevidensis. Este estudo mostrou que os extratos de E. grandiflorus e S. montevidensis apresentam potencial genotóxico e quando usadas em elevadas concentrações, potencial antiproliferativo.

This diploma project deals with antioxidant and antimutagenic properties of selected herbal and fruit teas commonly used in Czech population. Influence of different tea packaging (bag teas and loose leaf teas) on bioactive compound content was compared. Further, effect of long-term storage in common household conditions was studied. Antioxidant properties of teas were characterized using some group parameters - total antioxidant activity ("Randox Total Antioxidant Status Kit"), total phenolics and total flavonoids - as well as some individual representatives of low molecular weight antioxidants. Higher antixidant content was found in herbal teas than in fruit teas. Comparing bag teas with loose leaf teas higher antioxidant activity was shown in loose leaf teas. Individual antioxidants were analyzed using HPLC method with spectrophotometric detection and verified by on-line LC/MS. IN all tea samples catechins - catechin, epicatechin, epicatechin gallate and other flavonoids - rutin, morin, quercetin, kaempferol, myricetin and luteolin were determined. In most of teas high level of catechin and rutin was detected. The highest level of flavonoids was determined in herbal poured teas. Ascorbic acid content was also determined by HPLC method. Higher vitamine C level was found in most of fruit teas and in rose hip tea. Antimutagenicity of tea extracts was tested by in vitro test using Saccharomyces cerevisiae D7 yeast. High antimutagenic activity showed mainly nettle tea, tutsan tea and most of fruit teas. During long-term storage (1 year, 20°C, darkness) a significant decrease of all analyzed antioxidant parameters was followed. Higher lost of antioxidants was found in fruit teas when compared with the herbal ones.

The presented doctoral thesis is focused on the study of the biological effects of active compounds found in cereals, on the development of methods of analysis these effects, on the determination of the content of active substances and characterization of the relationship between composition and biological effects of cereals and cereal products. For the analysis several kinds of raw cereal samples (flakes, flour, germ, bran), flavored extruded cereal products and also samples of paddy and husked rice were chosen. To major types of analysed active compounds belong mainly phenolic compounds in the form of glycosides and aglycones, and also saccharides. Group parameters such as total polyphenols, flavonoids and total and reducing saccharides were determined spectrophotometrically, individual phenolics and saccharides were determined by high performance liquid chromatography (HPLC). TEAC, DPPH and CLAMS methods were used to determine the antioxidant activity. Indirect methods of determination of substances with antioxidant effect were used as well. Results of the total antioxidant activity were compared with values of antimutagenic/genotoxic activity obtained by several microbial test systems. Antimutagenic effect was expressed as a percentage of inhibition of effect of standard mutagen and could be considered as a potential preventive effect of cereals to DNA arising primarily by free radicals effect. The highest values of group and individual phenolics, antioxidant and antimutagenic activity were found in germs, bran, in buckwheat products and in coloured and raw rice. In flavoured cereal products addition of chocolate or fruit positively influences content of active phenolic substances as well as sugars, antixidant and antimutagenic activity. In a representative sample of Czech population, questionnairy study was performed to monitor interest in cereals and consumer preferences. The most of consumers consider cereals with chocolate flavour as less healthy than confirmed results of laboratory analyses. In this study some new food products were developed. Several types of model cereal products containing plant (fruit, vegetables) extracts were proposed. Extracts were added to cereals in freeze-dried and encapsulated form. The highest positive effect exhibited addition of local forrest fruit extract. Within preparation of encapsulated extracts several methods of preparing lipid or saccharides particles were also tested. The encapsulation efficiency of the methods and stability and size of particles were analysed. Optimal type of fortified cereal foods could be suggested based on the acquired results. Selected cereals were used as alternative carbon substrates (processed or raw) for the cultivation of microorganisms to produce enriched biomass usable in the feed industry. We can conclude that cereals in raw as well as processed form belong to universal foods and rich sources of biologically active substances. They can be processed by many ways. They can be used for direct consumption, as a part of new products and also undirectly as a substrate for feedstock.