Amorphous solid dispersion (ASD) is a popular method to increase drug solubility and consequently poor drug bioavailability. Cellulose ω-carboxyesters were designed and synthesized specifically for ASD preparations in Edgar lab that can meet the ASD expectations such as high Tg, recrystallization prevention and pH-triggered release due to the free -COOH groups. Rifampicin (Rif), Ritonavir (Rit), Efavirenz (Efa), Etravirine (Etra) and Quercetin (Que) cellulose ester ASDs were investigated in order to increase drug solubility, prevent release at low pH and controlled release of the drug at small intestine pH that can improve drug bioavailability, decrease needed drug content and medication price to make it affordable in third world countries, and extent pill efficiency period to improve patient quality of life and adherence to the treatment schedule. The studies were compared with cellulose based commercial polymers to prove the impact of the investigation and potential for the application. Furthermore, the in vitro results obtained were further supported by in vivo studies to prove the significant increase in bioavailability and show the extended release. The need of new cellulose derivatives for ASD applications extended the research area, the design and synthesis of a new class of polymers, alkyl cellulose ω-carboxyesters for ASD formulations investigated and the efficiency of the polymers were summarized to show that they have the anticipated properties. The polymers were synthesized by the reaction of commercial cellulose alkyl ethers with benzyl ester protected, monofunctional hydrocarbon chain acid chlorides, followed by removal of protecting group using palladium hydroxide catalyzed hydrogenolysis to form the alkyl cellulose wcarboxyalkanoate. Having been tested for ASD preparation, it was proven that the polymers were efficient in maintaining the drug in amorphous solid state, release the drug at neutral pH and prevent the recrystallization for hours, as predicted. / Ph. D.

Yes / Chronic obstructive pulmonary disease (COPD) in humans, describes a group of lung conditions characterised by airflow limitation that is poorly reversible. The airflow limitation usually progresses slowly and is related to an abnormal inflammatory response of the lung to toxic particles. COPD is characterised by oxidative stress and an increased risk of lung carcinoma. The 2-amino-3-methylimidazo [4,5-f]quinoline (IQ) is one of a number of mutagenic/carcinogenic heterocyclic amines found mainly in well-cooked meats which are thus part of the regular diet. Antioxidants are very important in order to protect the cells against oxidative damage. The aim of the present study was to assess the effects of IQ on the level of DNA damage and susceptibility to a potent mutagen in peripheral blood cells of COPD patients. DNA damage and the frequency of micronuclei (MNi) were evaluated using the Comet and micronucleus assays, respectively. Differential expressions of both mRNA and protein of the endogenous antioxidant enzyme catalase were evaluated with quantitative polymerase chain reaction (qPCR) and Western blot analysis, respectively. Furthermore, the effect of bulk and nano forms of quercetin and their combination with IQ were examined. Results of the present study clearly demonstrated that MNi frequency in the peripheral blood lymphocytes exhibited a positive correlation with the DNA damage as evident from the different Comet assay parameters. Increase of the endogenous antioxidant catalase also showed there was a stimulation of this enzyme system by IQ. Whereas, the endogenous antioxidant quercetin significantly reduced oxidative stress in COPD patients and healthy individuals. / Libyan Government

Dans le diabète de type 2 établi, l'hyperglycémie chronique, un taux élevé d'acides gras libres et l'inflammation induisent un stress oxydatif (SO) au niveau de la cellule beta. Le SO, qui apparaît dès le stade de pré-diabète, peut induire un dysfonctionnement précoce de cette cellule. Ainsi, la protection de la cellule β par des molécules anti-oxydantes pourrait ralentir la progression du pré-diabète au diabète.La quercétine, un flavonoïde, a présenté des propriétés antidiabétiques dans plusieurs études in vivo. Cependant, très peu de données traitent de son mécanisme d'action directement au niveau de la cellule beta. Dans ce contexte, nous avons étudié les effets de la quercétine au niveau de la cellule beta dans des conditions physiologiques et des conditions de SO.Nos résultats montrent qu'en présence de concentrations stimulantes de sécrétagogue, la quercétine potentialise la sécrétion d'insuline par un mécanisme impliquant l'augmentation de calcium intracellulaire et la potentialisation de ERK1/2 via l'activation des voies de la PKA et de la CaMK II. De plus, la quercétine protège la cellule beta du SO en sur-activant ERK1/2. Le resvératrol et la NAC, deux antioxydants de référence, sont inactifs dans ces conditions expérimentales.En absence de concentrations stimulantes de sécrétagogue, la quercétine induit une sécrétion d'insuline modérée en augmentant le calcium intracellulaire suite à une activation directe des CaV de type L. Dans ces conditions, l'activation de ERK1/2 induite par la quercétine, qui est indépendante de l'activation des voies de la PKA et de la CaMK II, ne serait pas impliquée dans le mécanisme sécrétoire. Nos résultats indiquent que le mécanisme d'action de la quercétine au niveau de la cellule β ne repose pas uniquement sur ses capacités anti-oxydantes mais fait intervenir des cibles pharmacologiques et la régulation de voies de signalisation intracellulaires. / In type 2 diabetes, chronic hyperglycaemia, elevated free fatty acids and inflammation induce oxidative stress (OS) in pancreatic β cell. SO, which appears at the stage of pre-diabetes, may induce early dysfunction of this cell. Thus, the β cell protection by antioxidant molecules could slow the progression of pre-diabetes to diabetes.Quercetin, a flavonoid, has shown antidiabetic properties in several in vivo studies. However, very few data address its mechanism of action directly at the β cell. In this context, we studied the effects of quercetin at the β cell under physiological conditions and conditions of OS.Our results show that in the presence of stimulating concentrations of secretagogue, quercetin potentiates insulin secretion by a mechanism involving increased intracellular calcium and potentiation of ERK1 / 2 via activation of the PKA and the CaMK II pathways. In addition, quercetin protects beta cell from OS via a suractivation of ERK1/2. Resveratrol and NAC, two antioxidants of reference are inactive under these experimental conditions.In the absence of stimulating concentration of secretagogue, quercetin induced moderate insulin secretion by increasing the intracellular calcium via a direct activation of L-type CaV Under these conditions, the activation of ERK1/2 induced by quercetin, which is independent of the activation pathways of PKA and CaMK II to, would not be involved in the secretory mechanism.Our results indicate that the mechanism of action of quercetin at the β cell not only based on its antioxidant capacity but involves pharmacological targets and the regulation of intracellular signaling pathways.

La quercétine (QCT) est un composé polyphénolique d'origine végétale connu pour ses activités antioxydantes et ses effets bénéfiques sur la santé. Sa solubilité, sa stabilité, sa biodisponibilité et ses activités biologiques peuvent être améliorées par une acylation sélective de ses groupements hydroxylés. Ce travail vise à étudier la possibilité d'une acétylation enzymatique de la QCT par la lipase B de Candida antarctica (CALB), la lipase la plus exploitée industriellement pour des estérifications régio- et énantiosélectives. Dans une perspective d'ingénierie rationnelle de l'enzyme, une démarche de modélisation moléculaire est mise en oeuvre pour mieux comprendre les interactions qui régissent le positionnement et l'orientation du substrat dans le site actif de la lipase. Dans une première partie expérimentale, l'absence d'activité d'acétylation de la CALB envers la QCT, en présence d'un excès d'acétate de vinyle, a été confirmé. Dans une seconde partie, cette inactivité de la CALB a été expliquée à l'aide de simulations de docking et de dynamique moléculaire. Elle résulte d'une orientation inappropriée du donneur d'acyle liée à la sérine catalytique et d'une proximité insuffisante des hydroxyles de la QCT vis-à-vis des résidus catalytiques. L'éloignement de la QCT de la triade catalytique est due à la rigidité de la molécule, l'étroitesse du site actif ainsi qu'à des interactions hydrophobes et électrostatiques entre le substrat et les résidus de la cavité. En revanche, cette approche de simulation moléculaire prédit un bon positionnement des deux substrats dans le site actif de la lipase de Pseudomonas cepacia (PCL), laquelle est capable d'acétyler la QCT. Dans une troisième partie, l'influence de mutations de deux résidus impliqués dans les liaisons de stabilisation hydrophobe de la QCT dans la CALB a été investiguée par simulation. La substitution d'isoleucines par des valines et des alanines conduit à une augmentation du volume de la poche catalytique et une mobilité accrue de la QCT. Mais ces mutations sont insuffisantes pour permettre un positionnement adéquat de l'acétate et de la QCT par rapport à la triade catalytique. La dernière partie focalise sur les interactions électrostatiques entre la QCT et le site actif de CALB. Les orientations du substrat dans la cavité suite à une méthylation ou une acétylation des groupements hydroxyles de la QCT sont précisées / Quercetin (QCT) is a plant-produced polyphenolic compound well-known for its antioxidant activities and beneficial health effects. Its solubility, stability, bioavailability and biological activities may be improved by a selective acylation of its hydroxyl groups. This work aims at studying the possibility of QCT enzymatic acetylation by Candida antarctica lipase B (CALB), the most industrially exploited lipase for regio- and enantioselective esterifications. In prospect of the rational enzyme design, a molecular modeling approach was implemented to understand the interactions that govern the substrate positioning and orientation in the lipase's active site. In a first experimental part, the absence of CALB acetylation activity towards quercetin in excess of vinyl acetate was confirmed. In a second part, this inactivity of CALB was explained by means of docking and molecular dynamics simulations. This results from an inappropriate positioning of the acyl donor linked to the catalytic serine and from an insufficient proximity of QCT hydroxyls vis-à-vis catalytic residues. The distance of QCT from the catalytic triad is due to its rigidity and to the narrow active site as well as to hydrophobic and electrostatic interactions between the substrate and the cavity residues. On the contrary, this molecular simulation approach predicts an appropriate positioning of both substrates in the active site of Pseudomonas cepacia lipase (PCL), which can perform QCT acetylation. In a third part, the impact of mutations of two residues implicated in the stabilization of QCT by hydrophobic interactions in CALB was investigated through simulations. The substitution of isoleucines by alanines and valines led to an increase in the catalytic pocket volume which intensified the mobility of QCT. However, these mutations are insufficient to allow an appropriate positioning of acetate and QCT in relation to the catalytic triad. The last part of this work focuses on the electrostatic interactions between QCT and CALB's active site. The substrate orientation in the cavity following methylation or acetylation of QCT's hydroxyl groups was clarified

Natural flavonoids and flavonolignans feature beneficial properties for living organisms such as antioxidant and hepatoprotective effects, anticancer, chemoprotective, dermatoprotective and hypocholesterolemic activities. Their metabolism in mammals is complex, the exact structure of their metabolites still remains partly unclear and the standards are usually not commercially available. Hence, this project focused on the preparation of potential and defined biotransformation Phase II sulfated metabolites of silymarin flavonolignans: silybin, 2,3-dehydrosilybin, isosilybin, silychristin, silydianin and flavonoids quercetin, taxifolin, rutin and isoquercitrin. Pure sulfated derivatives were prepared using aryl sulfotransferase from Desulfitobacterium hafniense and aryl sulfotransferase from rat liver. Using heterologously expressed PAPS (3'-phosphoadenosine-5'-phosophosulfate) - independent arylsulfotransferase from Desulfitobacterium hafniense and cheap p-nitrophenyl sulfate as sulfate donor, sulfated flavonolignans and flavonoids were obtained in high yields. Silymarin flavonolignans afforded exclusively monosulfates at the position C-20 (C-19 in the case of silychristin), except 2,3-dehydrosilybin that yielded also the 7,20-O-disulfated derivative. Isoquercitrin and rutin were selectively sulfated...

Made available in DSpace on 2015-05-14T12:59:23Z (GMT). No. of bitstreams: 1 arquivototal.pdf: 4714773 bytes, checksum: e1d2845582482ef401e124b0aa366365 (MD5) Previous issue date: 2008-08-19 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior - CAPES / Leishmaniases are a complex of infectious parasitic diseases caused by species of the Leishmania genus. These diseases comprise a large spectrum of manifestations ranging from localized self-healing cutaneous lesions to fatal visceral infections. In Brazil, the visceral leishmaniasis is caused by Leishmania chagasi which affects ca. 2 million people every year with estimated 90% of cases occurring in Northeastern. Current treatments of leishmaniasis are based on first line pentavalent antimonials or other drugs like amphotericin B and pentamidine. Toxicity of those drugs, their high sideeffects, besides the high cost of treatment, difficulty for administering them and the surge of resistance are their great drawbacks. These aspects have stimulated the search for new leishmanicide agents, like the isolation and identification of natural compounds which could provide new therapeutic models for the treatment of leishmaniases. It was aimed in the present work a comparative evaluation of potentially antileishmania natural isolate compounds and their action on promastigote and amastigote forms of Leishmania chagasi by observing the cycle of this parasite in vitro. Assays were carried out with usnic acid, coumarin, quercetin, and reserpine which showed to have significant antileishmanial activity on promastigote forms of Leishmania chagasi, presenting IC50 equal to 0.0417; 1.07; 0.271 and 1.7 mM, respectively. It was possible in our experimental conditions of standardized cultivation of parasites, to establish their life cycle in vitro, by observing their metamorphosis from promastigote to mastigote. The in vitro life cycle was characterized by parasite cultivations with establishment of: 1) lag (initial), log or exponential and stationary stages of promastigotes; 2) transformation of promastigotes to intracellular amastigotes by infection of murine macrophages; this step favoured the promastigotes infectivity; and 3) amastigote isolation and transformation to promastigotes completing the cycle. Furthermore, it was also possible to establish the in vitro transformation of promastigotes in axenic amastigotes of Leishmania chagasi and its use for the evaluation of antileishmanial activity. In this case, the natural compound usnic acid exhibited antileishmanial activity against axenic amastigote forms with IC50 of 1,16 mM. It was not observed any similarity of IC50 on antileishmanial activity of usnic acid and pentamidine between the axenic amastigote and promastigote forms. This demonstrates that it is important to characterize the action of compound on each life form of the parasites. It is concluded that all assayed compounds carried out in this work had antileishmanial activity on promastigote forms, mainly, and the effect of concentration was different between promastigote, of lower IC50 values, and axenic amastigotes forms, of greater IC50 values. / As leishmanioses constituem-se por um complexo de doenças infectoparasitárias, causadas por parasitas do gênero Leishmania, e apresentam um espectro de sintomas variando de simples lesões cutâneas de cicatrização espontânea a lesões viscerais letais. A leishmaniose visceral, causada pela espécie Leishmania chagasi, no Brasil infecta cerca de duas mil pessoas por ano, sendo cerca de 90% dos casos no Nordeste. Os tratamentos atuais são baseados em compostos antimôniais pentavalentes, ou na utilização de outras drogas como a anfotericina B e a pentamidina. A toxicidade desses agentes, com efeitos colaterais graves, o alto custo dos tratamentos, as dificuldades de administração e o surgimento de resistência são grandes desvantagens, tornando essencial a busca por novos agentes leishmanicidas, como a identificação de compostos naturais isolados, que podem fornecer novos modelos terapêuticos no tratamento das leishmanioses. A presente pesquisa teve como objetivo avaliar comparativamente o potencial antileishmania de compostos naturais isolados sobre as formas promastigotas e amastigotas de Leishmania chagasi, estabelecendo o ciclo do parasito in vitro. Os compostos naturais ensaiados, ácido úsnico, cumarina, quercetina e reserpina apresentaram significativa atividade antileishmania sobre as formas promastigotas de Leishmania chagasi, com IC50 igual a 0,0417; 1,07; 0,271 e 1,7 mM respectivamente. Nas condições de cultivo padronizadas no presente estudo foi possível estabelecer o ciclo de vida in vitro dos parasitos, com passagem destes pelas duas principais formas de vida: promastigotas e amastigotas. O ciclo in vitro foi caracterizado pelo cultivo das promastigotas com estabelecimento: 1) das fases: lag (inicial), log ou exponencial e estacionária dos parasitos; 2) transformação das formas promastigotas em amastigotas intracelulares, pela infecção de macrófagos murinos, o que favoreceu a infectividade das promastigotas; e 3) pelo isolamento das formas amastigotas e transformação destas em promastigotas novamente, completando o ciclo. Além disto, foi também possível estabelecer a transformação in vitro das promastigotas em amastigotas axênicas de L. chagasi e o cultivo das formas axênicas para ensaios de ação antileishmania sobre essa forma. Nesse caso, o composto natural isolado ácido úsnico apresentou atividade antileishmania sobre amastigotas axênicas de L. chagasi, com IC50 igual a 1,16 mM. Não foi observada similaridade da IC50 na atividade antileishmania do ácido úsnico e da pentamidina entre as formas amastigotas axênicas e promastigotas, o que demonstra a importância de caracterizar a ação dos compostos sobre cada forma de vida dos parasitos. Conclui-se que os compostos isolados têm atividade antileishmania sobre as formas promastigotas e o ácido úsnico sobre as formas amastigotas axênicas de L. chagasi.

Submitted by Socorro Pontes (socorrop@ufersa.edu.br) on 2017-03-02T14:51:05Z No. of bitstreams: 1 ChristianeMCR_TESE.pdf: 1824459 bytes, checksum: 8d67db5817b6096ad2d44e2a3043a10d (MD5) / Approved for entry into archive by Vanessa Christiane (referencia@ufersa.edu.br) on 2017-03-21T15:02:11Z (GMT) No. of bitstreams: 1 ChristianeMCR_TESE.pdf: 1824459 bytes, checksum: 8d67db5817b6096ad2d44e2a3043a10d (MD5) / Made available in DSpace on 2017-03-21T15:02:28Z (GMT). No. of bitstreams: 1 ChristianeMCR_TESE.pdf: 1824459 bytes, checksum: 8d67db5817b6096ad2d44e2a3043a10d (MD5) Previous issue date: 2016-03-03 / Emepa - Empresa Estadual de Pesquisa Agropecuária / The social and economic importance of yellow mombin is due to the marketing of its raw fruits and derivatives. The non-existence of commercial clones and the high plant size are the main obstacles to its rational cultivation being necessary to make the cloning a viable instrument through the interspecific rootstocks in order to downsize the plants heights. Studies done with yellow mombin grafted onto rootstocks of yellow mombin and hog-plum did not markedly downsize the plant height but these clones were not evaluated in other enviroments. The yellow mombin shows potential for remedies. In the pharmaceutical industry it is used as phytotherapics against the Herpes virus. In works done with this species some bioactive compounds were found in the leaves such as flavonoids and tannins. Although this potential, some researches on chemical characterization and genetic diversity among genotypes are incipient. The objective of this study was to evaluate the vegetative growth of yellow mombin clones grafted onto interspecific rootstocks in different environments and the genetic diversity as to bioactive compounds among accessions of the Emepa-PB´s germplasm bank. In order to evaluate the clones development two trials with a completely randomized block design were performed. The trial 1 was conducted in a 3 x 4 x 3 factorial design with four repetitions, totaling 36 treatments, being three different interspecific rootstocks with yellow mombin (S. mombin), hog-plum (S. tuberosa) and coarse mombin (S. venulosa), four graftings of yellow mombin clones (Itaitinga, Gereau, Lagoa Redonda and Genipabu) and three different places (João Pessoa, PB, Ipanguaçu, RN and Itabuna, BA). The trial 2 was conducted in a 4 x 3 factorial design, with four repetitions, totaling 12 treatments. Three graftings of yellow mombin (Emepa 8.2, Emepa 8.6 and Emepa 20) obtained from the grafting between yellow mombin clones, grafted onto yellow mombin, plus one genotype of ungrafted yellow mombin, were used in the same places. The evaluated variables for the two trials at the 8, 39 and 46 months old were plant height, stem circumference, crown spread, crown format, number of branches and percentage of dead plants. The conclusions are that the environment, the grafting and the rootstocks did not influence the size and vigor of the clones allowing the identification of the morphological characteristics variations in each environment; that in Trial 1, for the three enviroments, the graftings of yellow mombin onto yellow mombin, hog-plum and coarse mombin rootstocks did not form clones with ideal sizes so that they cannot be recommended for commercial plantation; that in Trial 2, in the three enviroments, the Emepa 8.2 showed smaller size, bigger crown spread, major number of branches, 100% crown in a spread format and it can be recommended for new studies and later multiplication for commercial plantation. Leaves and barks of 27 yellow mombin accessions of the Emepa´s germplasm bank, in João Pessoa, PB, were used. The extractions were performed with methanol and the identification and quantification of bioactive compounds were checked through the High-Performance Liquid Chromatography (HPLC). For the grouping of accessions were used the Principal Components Analysis (PCA) and the UPGMA method. The standardized Euclidean distancewas used as a measure of genetic diversity. Rutin, quercetin, ellagic acid and geraniin compounds were quantified in leaves and ellagic acid and geraniin compounds were quantified inbarks. The highest amounts were found in leaves. Seven groups were formed. The accession group A3 was the most divergent with high amounts of rutin, quercetin and ellagic acid in leaves. The conclusion is that there is variability among accessions and it must be preserved and exploited through the genetic improvement programs / A importância socioeconômica da cajazeira é devido à comercialização dos frutos e do consumo de seus derivados. A inexistência de clones comerciais e o porte alto da planta constituem-se em entraves ao seu cultivo racional, necessitando viabilizar a clonagem com porta-enxertos interspecíficos visando reduzir o porte das plantas. Estudos realizados com a cajazeira enxertada na própria espécie e em umbuzeiro não demonstraram a diminuição significativa do porte da planta. No entanto, esses clones não foram avaliados em outros ambientes. A cajazeira apresenta potencial para medicamentos. Na indústria farmacêutica é utilizada como fitoterápico contra o vírus do Herpes. Em trabalhos realizados por outros pesquisadores, foram identificados compostos bioativos nas folhas tais como flavonoides e taninos. Apesar desse potencial, as pesquisas sobre a caracterização química e a divergência genética entre os genótipos são incipientes. O objetivo deste trabalho foi avaliar o desenvolvimento de clones de cajazeira enxertados sobre porta-enxertos interespecíficos em diferentes ambientes e a diversidade genética quanto a compostos bioativos entre acessos do banco de germoplasma da Emepa-PB. Para avaliação do desenvolvimento dos clones, foram realizados dois ensaios em delineamento inteiramente casualizado. O Ensaio 1 foi conduzido em esquema fatorial 3 x 4 x 3, com quatro repetições, totalizando 36 tratamentos, sendo três porta-enxertos interespecíficos com cajazeira (S. mombin), umbuzeiro (S. tuberosa) e cajazeira-grande (S. venulosa), quatro enxertos de clones de cajazeira (Itaitinga, Gereau, Lagoa Redonda e Genipabu), e três locais (João Pessoa, PB, Ipanguaçu, RN e Itabuna, BA). O Ensaio 2 foi conduzido em esquema fatorial 4 x 3, com quatro repetições, totalizando 12 tratamentos. Foram utilizados três exertos de acessos de cajazeira (Emepa 8.2, Emepa 8.6 e Emepa 20) enxertados sobre cajazeira, mais um pé-franco de cajazeira, nos mesmos locais. As variáveis avaliadas, aos 8, 39 e 46 meses de idade, para os Ensaios 1 e 2, foram altura de planta, perímetro de caule, largura e formato de copa, número de ramos e mortalidade de plantas. Concluiu-se que o ambiente, os enxertos e os porta-enxertos de espécies de Spondias influenciaram no desenvolvimento dos clones, permitindo identificar variabilidade nas características morfológicas em cada ambiente; no Ensaio 1, nos três ambientes, a enxertia da cajazeira sobre porta-enxertos de cajazeira, cajazeira-grande e umbuzeiro não formaram clones com porte ideal para recomendação comercial; no Ensaio 2, nos três ambientes, o clone Emepa 8.2 apresentou menor porte, maior largura de copa, maior número de ramos e 100% de formato de copa esgalhada, podendo ser recomendado para novos estudos e posterior multiplicação para exploração comercial. Para avaliação da diversidade genética, foram utilizadas folhas e cascas de cajazeira de 27 acessos do banco de germoplasma da Emepa, em João Pessoa, PB. As extrações foram realizadas com metanol e a quantificação foi determinada por Cromatografia Líquida de Alta Eficiência (CLAE). Para o agrupamento dos acessos utilizou-se a Análise de Componentes Principais (ACP) e o método aglomerativo UPGMA. Foi utilizada a distância euclidiana média padronizada como medida de dissimilaridade genética. Foram quantificados nas folhas os compostos rutina, quercetina, ácido elágico e geraniina e foram quantificados nas cascas os compostos ácido elágico e geraniina. Os maiores valores foram obtidos nas folhas. Foram formados 7 grupos. O acesso A3 foi o mais divergente com valores elevados de rutina, quercetina e geraniina nas folhas. Concluiu-se que há variabilidade entre os acessos e que esta deve ser preservada e explorada em programas de melhoramento genético / 2017-02-24

This work concerns the amount of certain phenols in some of the representatives of families Alliaceae, Lamiaceae and Apiaceae. Phenols in plants are widely represented and highly concentrated. Flavonoids are one of the smallest, yet quite significant classes of phenols. Ingestion of food containing flavonoids can prevent certain diseases such as Arteriosclerosis. cardiovascular and tumor diseases. This work focuses, for their special biological effects, on five flavonoids: Kaempferol, Quercetin, Myricetin, Apigenin and Luteolin. For determining the content of phenols a method of High-performance liquid chromatography (HPLC) was used. Three representative of the family Alliaceae, two representatives of the family Lamiaceae and three representatives of the family Apiaceae were analysed. Only edible parts of the plants were used for the analysis and the amount of phenols was compared in certain plants planted in beddings to the plants planted in greenhouses. At first, the qualitative representantion of phenols was ascertained by the HPLC method. The result is chromatographic profiles, which were then used in calculating the amount of particular phenols. Then the total amounts of kaempferol, quercetin, myricetin, apigenin and luteolin were found out thanks to HPLC method. The biggest amount of total kaempferol was determined in petroselium hortense planted in beddings (588 mg/kg of fresh sample) and in a greenhouse (340 mg/kg of fresh sample). The biggest amount of quercetin was determined in red onion planted in a greenhouse (773 mg/kg of fresh sample) and in ocimum basilicum planted in beddings (535 mg/kg of fresh sample). The biggest amount of apigenin was determined in petroselium hortense planted in a greenhouse (1790 mg/kg of fresh sample) and in petroselium chrispum planted in beddings (3690 mg/kg of fresh sample).

The work has been inquired into the problem of content determination of phenolic substances in medicinal plants traditionally used in Czech Republic. Phenolic substances belong to a group of natural compounds, which are purely plant origin. Flavonoids are a part of this extensive group of compounds. As for flavonoids, most attention is paid to quercetin and rutin. It is caused by their easy availability and very significant biological activity. These compounds embody a lot of positive biological effects. They have expressive antioxidant properties, inhibit lipid peroxidation, scavenge free oxygen radicals and bond into the chelates they inactivate some prooxidant metal ions. The latest researches have shown that thanks to their properties natural flavonoids can occurence of chronical diseases, such as arterosklerosis, cardiovascular or tumor disorder. Flavonoids are exploited both in traditional and modern medicine. Same other studies have evidenced that valuable sources of these biologically effective substances are traditionally used medicinal plants. Content of phenolic substances was determined by method of micellar electrokinetic capillary chromatography (MECC) and liquid chromatography (HPLC) in collection of 8 medicinal plants usually used in Czech Republic. For analysis was used freeze-dried and dried plant material. In freeze-dried material was found the highest content of total quercetin in Filipendula ulmaria L. (14200 mg/kg of dry weight) and in Betula pendula Roth. (11800 mg/kg of dry weight). The highest content of rutin contained Sambucus nigra L. (17700 mg/kg of dry weight). Similar values was measured in dried plant material. The content of total quercetin and rutin during drying was unchanged.

The thesis deals with measuring the content of chosen phenolic substances in some specimen of the genera Chenopodium L. and Atriplex L. Two independent analytical methods were used to determine the content of phenolic substances, namely the Micellar Electrokinetic Capillary Chromatography (MECC) method and the High - Performance Liquid Chromatography (HPLC) method. Two cultured species of the genera Spinacia and Atriplex, three freely growing specimen of the genus Chenopodium and three freely growing species of the genus Atriplex were analysed. The analysis concerned the leaves and the inflorescence of these species.The total content of quercetin and rutin was determined by the MECC method. The highest total content of quercetin was found in the leaves of the Garden Orache (4240 mg/kg of dry matter), the lowest total content of quercetin was found in the inflorescence of the Atriplex prostrata DC. (19.6 mg/kg of dry matter). Rutin was only found in four samples, the rest of the samples contained rutin in quantities below the limit of quantification. The highest content of rutin was found in the leaves of the Lamb´s Quarters (868 mg/kg of dry matter).