Farkašovský M.: Biological activity of plant metabolites XXIX. Alkaloids of selected cultivars of Narcissus triandrus L. and their influence on acetylcholinesterase and butyrylcholinesterase. Diploma thesis. Charles University in Prague, Faculty of Pharmacy in Hradec Králové, Department of pharmaceutical botany and ecology, Hradec Králové 2015, 70 pages Screening of seven bulb samples of Narcissus genus was performed. It included cultivars Narcissus triandrus cv. Hawera, Narcissus triandrus cv. Ice Wings, Narcissus triandrus cv. Stint, Narcissus triandrus cv. Tresamble, Narcissus cyclamineus cv. February Gold, Narcissus cyclamineus cv. Greenlet and Narcissus cyclamineus cv. Itzim. Primary extract was prepared by boiling crushed bulbs in ethanol 95% and then it was condensated. After extraction in diethyl ether and ethyl acetate was pure alkaloid extract dried with air flow in water bath. Isolated alkaloid extracts were tested for their inhibition activity on acetylcholinesterase and butyrylcholinesterase. Also GC-MS analysis was provided to identify alkaloids. These alkaloids were identified with GC-MS analysis - epinorgalanthamine, galanthamine, galanthine, haemanthamine, hippeastrine, homolycorine, cherylline, incartine, lycoramine, lycoramine-acetate, lycorine, narwedine, neruscine, sanguinine...

Les Polluants Organiques Persistants (POPs), largement présents dans l'environnement via les stations d'épuration des eaux usées (STEP) et l'activité anthropique de l'Homme constituent une source de contamination du milieu aquatique. L'évaluation du risque environnemental de ces composés implique leur analyse chimique quantitative et spécifique et l'estimation de leur origine vis-à-vis des zones contaminées. Cette étude consiste en une analyse qualitative et quantitative, à l'échelle internationale, pour une trentaine de composés organiques représentatifs en termes d'utilisation et pertinents en termes d'impact environnemental. L'approche mise en œuvre intègre une étape de détection et une étape d'évaluation de la toxicité des composés ciblés afin d'estimer leur niveau de risque écotoxicologique. La détection de ces composés dans les matrices sédiments a nécessité la mise au point d'une méthode d'extraction Soxhlet et le développement d'une méthode de détection par GC-MS. La validation de cette méthode a permis de mener à bien une campagne d'échantillonnage de 54 stations répartis sur 4 bassins versants (l'Arc, l'Huveaune, la Durance, l'étang de Berre et le Oued Medjerda), afin d'évaluer le niveau de contamination par les POPs. Ce travail se propose d'effectuer un échantillonnage orienté (avant et après des ouvrages majeurs tels que STEP, Industries …) de sédiments des différents cours d'eaux de la zone Méditerranée et d'étudier à la fois les taux de contamination ainsi que l'origine des POPs. Sur le plan régional et international, ce sujet répond à une demande sociétale forte. / Persistent Organic Pollutants (POPs), largely present in the environment via sewage wastewater (WWTP) and anthropogenic activities are a source of contamination of the aquatic environment. The environmental risk assessment of these compounds involves their specific and quantitative chemical analysis and estimation of their origin next to the contaminated areas. This study was a qualitative and quantitative analysis, internationally, for about thirty organic compounds representative in terms of use and relevant in terms of environmental impact. The approach used incorporates a detection step and a step for evaluating the toxicity of the compounds targeted to estimate their level of ecological risk. The detection of these compounds in the sediment matrix required the development of a Soxhlet extraction method and the development of a method of detection by GC-MS. Validation of this method allowed to carry out a sampling campaign of 54 stations spread over four watersheds (Arc, Huveaune, the Durance, the Etang de Berre and Oued Medjerda), to assess the level of contamination by POPs. This work intends to carry out a guided sampling (before and after major works such as STEP, industries ...) sediments of different rivers in the Mediterranean area and study both infection rates and the origin of POPs. At the regional and international levels, the subject responds to a strong societal demand. Indeed, it is essential to characterize the quality of streams and to estimate their contamination. This work has allowed a consistent inventory level of contamination. The PAH analysis is done by gas chromatography-mass spectrometry (GC / MS).

Prostanoids are oxygenated derivatives of arachidonic acid with a wide range of physiological effects in vertebrates including modulation of inflammation and innate immune responses. Nonsteroidal anti-inflammatory drugs (NSAIDs) act through inhibition of cyclooxygenase (COX) conversion of arachidonic acid to prostanoids. In order to better understand the potential of environmental NSAIDS for interruption of normal levels COX products in fishes, we developed an LC/MS/MS-based approach for tissue analysis of 7 prostanoids. Initial studies examining muscle, gut and gill demonstrated that prostaglandin E2 (PGE2) was the most abundant of the measured prostanoids in all tissues and that gill tissue had the highest and most consistent concentrations of PGE2. After short-term 48-h laboratory exposures to concentrations of 5, 25, 50 and 100 ppb ibuprofen, 50.0ppb and 100.0 ppb exposure concentrations resulted in significant reduction of gill tissue PGE2 concentration by approximately 30% and 80% respectively. The lower exposures did not result in significant reductions when compared to unexposed controls. Measured tissue concentrations of ibuprofen indicated that this NSAID had little potential for bioaccumulation (BCF 1.3) and the IC50 of ibuprofen for inhibition of PGE2 production in gill tissue was calculated to be 0.4 µM. Short-term laboratory exposure to ibuprofen did not result in significant alteration of concentrations of PGE2 at environmentally relevant concentrations.

O consumo de cocaína e crack no Brasil é um problema de saúde pública, ainda mais grave quando realizado por gestantes, que colocam em risco a sua vida e a do feto. A identificação dessas substâncias através de cromatografia gasosa acoplada à espectrometria de massas e utilizando como matriz biológica o mecônio, é uma técnica eficiente para detecção da exposição fetal à cocaína. O mecônio apresenta algumas vantagens em relação às outras matrizes biológicas como uma ampla janela de detecção dos analitos e de fácil coleta por ser não-invasiva. Diante disso, o objetivo do presente estudo foi desenvolver e validar uma metodologia de preparo do mecônio para identificação de cocaína, benzoilecgonina, cocaetileno e o éster metilanidroecgonina em seus extratos, usando a cromatografia gasosa acoplada a espectrometria de massas (CG-EM). Os analitos foram inicialmente extraídos com metanol, sendo posteriormente, purificados usando cartuchos de extração em fase sólida do tipo Bond Elut Certify I. A determinação desses analitos foi realizada usando um CG-EM do tipo íon trap, no modo full scan de detecção. O método foi validado, segundo critérios estabelecidos pela ANVISA, na faixa de linearidade de 20 a 1000 ng/g para a cocaína e o cocaetileno; 40 a 1500 ng/g para a benzoilecgonina e 60 a 1500 ng/g para o éster metilanidroecgonina, usando 0,5 g de mecônio por análise. A resposta do detector apresentou-se linear na faixa estudada e o limite de detecção encontrado foi de 10ng/g para a cocaína; 20ng/g para o cocaetileno; 30ng/g para a benzoilecgonina e 40 ng/g para o éster metilanidroecgonina. O coeficiente de variação intra-ensaio variou de 3,01% a 10,15% e o inter-ensaio variou entre 5,31% a 11,12%; a exatidão variou entre 91,47% e 105,31%. A recuperação encontrada foi superior a 56,30%. A especificidade foi determinada para os seguintes interferentes: AAS (ácido acetilsalicílico), alprazolam, anfetamina, cafeína, dipirona, efedrina, fenilefrina, fluoxetina, metoclopramida, nicotina, sulfato ferroso e THC (tetraidrocanabinol). Após o término da validação, o método foi aplicado em amostras de mecônio coletadas de recém-nascidos no Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto (HCFMRP USP). A coleta destas amostras foi realizada após a autorização das mães em participarem da pesquisa, através da assinatura do Termo de Consentimento Livre e Esclarecido, e após preencherem um questionário sobre consumo de drogas. O método desenvolvido e validado mostrou ser eficiente na identificação de cocaína, metabólitos e derivados em mecônio. / Cocaine and crack use is an important public health problem in Brazil, even though when this is made by pregnants, risking the babies lives besides their own. The identification of such substances through gas chromatograph-mass spectrometry using meconium as biological matrix is highly efficient on detecting fetal exposure to cocaine. The meconium presents some advantages in comparison with other matrices such as analite wide window detection and collection facilities by non-invasive methods. The purpose of this study was to develop and validate a method for meconium sample preparation for a gas chromatography-mass spectrometry (GC-MS) confirmation of meconium extracts for cocaine, benzoylecgonine, cocaethylene and anhydroecgonine methyl ester. The analytes were initially extracted from the matrix by methanol. Then, a solid-phase extraction with Bond Elut Certify I cartridges was applied. Analytes were determined in a GC-MS ion trap, full scan mode. The method was validated in the range of 20 -1000 ng/g for cocaine and cocaethylene; 40 -1500 ng/g for benzoylecgonine and 60 -1500 ng/g for anhydroecgonine methyl ester, using 0.5 g of meconium per assay. The detector response was linear in the studied range and limit of detection were found to be 10ng/g to cocaine; 20ng/g to cocaethylene; 30ng/g to benzoylecgonine and 40 ng/g to anhydroecgonine methyl ester. Intra-batch coefficients of variation oscillated between 3.01% and 10.15% and inter-batch oscillated between 5.31% and 11.12%; accuracy were in range 91.47% - 105.31%. The recoveries were higher than 56.30%. Selectiviy was determined for these interferents: AAS (acetylsalicylic acid), alprazolam, amphetamine, caffeine, dipyrone, ephedrine, phenylephrine, fluoxetine, metoclopramide, nicotine, iron sulfate and THC (tetrahydrocanabynol). Finally, the method was applied to analysis of meconium collected from newborns in the Hospital das Clínicas da Faculdade de Medicina de Ribeirão Preto (HCFMRP USP). The sample collections were made after mothers´s authorization, and after they signed a term and answered a self-report about use of drugs. The developed and validated method was efficient on identification of cocaine, metabolites and derivatives in meconium.

Soil organic matter (SOM) plays an important role in the global carbon cycle. Therefore, it is important to understand the stability of SOM, which is related to several processes. Its intrinsic properties may be related to its stability, for example black carbon is considered to be relatively resistant to degradation. In most soils, the dark horizons coincide with the superficial layers or horizons due to the greater accumulation of organic matter, but in the southern states of Brazil, the presence of soils with dark subsurface horizons is frequently observed. The dark subsurface horizon of these soils are similar to a sombric horizon. Aspects about its origin, formation and preservation have not yet been fully elucidated. The objective of this work is to understand the formation of \'sombric\' horizons in soils of the region from Tijucas do Sul (Paraná, Brazil). Five soil profiles were described and collected, from which three originate from a toposequence and contained a \'sombric\' horizon (P1-P3), a reference soil that is representative of the area (P5) and an intermediate soil (P4) that showed morphology between the reference soil and the soils with a \'sombric\' horizon. To this end SOM is studied for its molecular composition by the pyrolysis technique coupled to gas chromatography and mass spectrometry (pyrolysis-GC-MS). In addition, 13C isotopic composition (δ13C) and phytolytic composition were studied in order to understand paleoclimatic conditions. These results will be related to past environmental conditions using 14C dating techniques, and supported by classical soil analysis. The samples of the horizons were submitted to the SOM chemical fractionation, generating two fractions: extractable fraction with NaOH (EXT) and residue (RES). The morphology of the profiles showed an intense biological activity in A horizons and a wide distribution of microfragments of charcoals. The lateral continuity of \'sombric\' horizons in toposequence soils was also observed, which differentiated them (P1-P3) from buried A horizons. The distribution of SOM in the fractions studied was the same for the five profiles studied: EXT> RES. Products related to wildfires such as polyaromatics (PAHs; BC) were found in all profiles, but in greater relative abundance in the \'sombric\' horizons, indicating a higher incidence of fire during the formation of these horizons and these compounds can be related to the maintenance of dark color of the \'sombric\' horizons. In relation to the paleoclimatic conditions it was observed that the dark subsurface horizons were developed during the Mid-Holocene under vegetation composed mainly by C4 grasses with shrubs, evidencing a drier climate corresponding to a higher fire incidence. / A matéria orgânica do solo (MOS) desempenha um papel importante no ciclo global do carbono. Portanto, é importante entender a estabilidade da MOS, que está relacionada a vários processos. As suas propriedades intrínsecas podem estar relacionadas com a sua estabilidade, por exemplo, o \"black carbon\" é considerado relativamente resistente à degradação. Na maioria dos solos, os horizontes escuros coincidem com as camadas ou horizontes superficiais devido ao maior acúmulo de matéria orgânica, mas nos estados do sul do Brasil, a presença de solos com horizontes subsuperficiais escurecidos é frequentemente observada. O horizonte subsuperficial escurecido destes solos assemelha-se a um horizonte sômbrico. Aspectos sobre sua origem, formação e preservação ainda não foram totalmente elucidados. O objetivo deste trabalho é compreender a formação de horizontes \'sômbricos\' em solos da região de Tijucas do Sul (Paraná, Brasil). Foram descritos e coletados cinco perfis de solo, dos quais três estão localizados em uma topossequência e continham um horizonte similar ao sômbrico (P1-P3), um solo de referência representativo da área (P5) e um solo intermediário (P4) que apresentou uma morfologia entre o solo de referência e os solos com horizonte \'sômbrico\'. Para este fim, a MOS foi estudada pela sua composição molecular através da técnica de pirólise acoplada à cromatografia gasosa e espectrometria de massa (pirólise-GC-MS). Além disso, estudou-se a composição isotópica 13C (δ 13C) e a composição fitolítica, a fim de compreender as condições paleoclimáticas que foram relacionados com as condições ambientais passadas usando técnicas de datação com 14C, e suportados pelas análises clássicas de solo. As amostras dos horizontes foram submetidas ao fracionamento químico MOS, gerando duas frações: fração extraível com NaOH (EXT) e resíduo (RES). A morfologia dos perfis mostrou uma intensa atividade biológica nos horizontes A e uma ampla distribuição de microfragmentos de carvão. Observouse também a continuidade lateral de horizontes \'sômbricos\' em solos da topossequência (P1-P3), diferenciando-os dos horizontes A enterrados. A distribuição da MOS nas frações estudadas pela pirólise foi a mesma para os cinco perfis: EXT> RES. Os produtos relacionados a incêndios florestais como os poliaromáticos (PAHs, BC) foram encontrados em todos os perfis, mas em maior abundância relativa nos horizontes sômbricos, indicando uma maior incidência de incêndio durante a formação destes horizontes. Os PAHs podem estar relacionados com a manutenção da cor escura dos horizontes \'sômbricos\'. Em relação às condições paleoclimáticas observou-se que os horizontes subsuperficiais escurecidos foram desenvolvidos durante o Holoceno Médio sob vegetação composta principalmente por gramíneas C4 com arbustos, evidenciando um clima mais seco correspondente a maior incidência de incêndio.

The revision of the CEN/TR 15522-2:2012 methodology contains new biomarkers to facilitate forensic fingerprinting of the variety of oil types that can be a part of different crimes and the purpose of this project is to validate the biomarkers of the new methodology. Biomarkers were validated by examining corresponding diagnostic ratios compatibility with the internationally used sampling cloth, discriminating power, correlation and simulated weathering sensibility through GC-SIM-MS analysis followed by statistical evaluation with t-tests, diagnostic power, Pearson correlation matrices and MS-PW plots respectively. Results based on most of the diagnostic ratios showed good compatibility with the internationally used sampling cloth, expected patterns of biodegradation and photo-oxidation except for observed photo-oxidation of hydro PAHs and that normative ratios and informative ratios with high diagnostic powers, but with strong correlations for some of the tested ratios, could be identified in diesel oils. Due to delimitations however such as the limited number of oils with similar origins that were analyzed the results should be regarded as guidelines that can be expanded.

Neste trabalho, o método de extração em fase sólida (SPE) com análises por cromatografia gasosa acoplada à espectrometria de massas (GC-MS) foi desenvolvido para a determinação de levoglucosano em amostras de água de chuva da região de Ribeirão Preto-SP, Brasil. O monossacarídeo levoglucosano foi proposto como um marcador específico para queima de biomassa, uma vez que o mesmo é liberado durante a pirólise da celulose em temperaturas acima de 300ºC. Após a otimização das variáveis SPE, as melhores condições dentre as avaliadas foram as seguintes: cartuchos SPE recheados com a fase extratora quitosana (1,0 g), volume de amostra igual a 5,0 mL, vazão de 1,0 mL min-1, solvente de eluição: metanol (volume igual a 2,5 mL), sem a adição de sal. O método SPE/GC-MS para a determinação de levoglucosano em amostras de chuva apresentou faixa linear no intervalo de concentração de 0,05 µg mL-1 a 0,50 µg mL-1, r 0,9967, exatidão de 95 a 110% e precisão inter ensaios com coeficientes de variação (CV) menores que 15%, exceto para o limite de quantificação (0,05 µg mL-1) o qual apresentou CV inferior a 20%. Segundo as determinações (SPE/GC-MS) de levoglucosano em amostras de água de chuva coletadas no período 03/08/2011 à 09/04/2012, as concentrações médias de levoglucosano na safra de cana-de-açúcar são aproximadamente quatro vezes superior ao período de entressafra. Este fato é justificado pela elevada taxa de queima da palha da cana-de-açúcar na região de Ribeirão Preto-SP no período safra, e consequentemente, maior emissão de material particulado para a atmosfera incluindo levoglucosano, COD, entre outros. / In this work, the method of solid phase extraction (SPE) with gas chromatography - mass spectrometry (GC-MS) analysis was developed for the determination of levoglucosan in rainwater samples collected in the region of Ribeirão Preto-SP, Brazil. The monosaccharide levoglucosan (1,6-Anhydro--D- Glucopyranose) was proposed as a specific marker for biomass burning, since it is released during the pyrolysis of cellulose at temperatures above 300 ° C. After optimization of the SPE variables, the best conditions were found to be as follows: SPE cartridges filled with the extraction phase chitosan (1,0 g), sample volume equal to 5,0 mL, flow rate of 1,0 mL.min-1 and elution with methanol (volume of 2,5 mL) without added salt. The use of the method SPE/GC-MS for to determination of levoglucosan in rainwater samples was in the range of concentration going from 0,05 to 0,50 g.mL-1, r 0,9967. The accuracy was 95-110%, determination of the inter assay precision gare coefficients of variation (CV) less than 15%, except for the limit of quantification (0,05 g.mL-1), for which CV was less than 20%. According to SPE/GC-MS determinations of levoglucosan in rainwater samples collected between 03/08/2011 and 09/04/2012, the average concentrations of this compound found during the harvest of sugar cane were approximately four times those obtained during the off-season. This fact is explained by the high rate of sugar cane burning in Ribeirão Preto-SP during harvest, which culminates in larger emissions of particulate matter to the atmosphere, including levoglucosano and COD, among others.

Made available in DSpace on 2017-07-24T19:37:57Z (GMT). No. of bitstreams: 1 JULIANE MARIA BER.pdf: 4116262 bytes, checksum: 58a85b36bbcbe8e5b731b5c3d9014a71 (MD5) Previous issue date: 2007-12-14 / The aim of the present work was to carry out an ethnopharmacological survey of the vegetal species used by the inhabitants of the municipality of Céu Azul (located in the extreme west of the State of Paraná, Brazil). A specific questionnaire was used to get the referring data to the uses and ways of preparation of the species used as medicinal. During the interviews, the plant material (exsiccate) was collected in duplicate with the aid of the informers for posterior identification and deposit in the Museu Botânico Municipal de Curitiba. The uses of 47 species pertaining to 12 botanical families were surveyed in this field research. One of the main objectives of the present work was to evaluate the possible use of forest medicinal species in Céu Azul, especially because an extensive area of the municipality belongs to the National Park of the Iguaçu. However, th 47 plants cited by the population can be divided in a group with 42 exotic species and another one with only 6 Brazilian plants, and none of which is being removed from the reserve. The data base of The International Plant Names Index was used to prevent inappropriate botanical names and to verify the current use of synonyms. A survey of the scientific information on the chemical composition and bioactivity was carried out for each cited species. The main source of scientific information for this work was the data base of the Institute of Scientific Information and the Chemical Abstracts on-line. In general, the Céu Azul population makes use of medicinal plants sufficiently known in the scientific sense, but there is also the use of little studied species. For four of the cited species (Alternanthera bettzickiana, Alternanthera dentata, Plectranthus neochilus, Tradescantia pallida), we could not find any scientific reference on its bioactivity. The results of this survey have been concomitantly interpreted as indicative for the election of one of these species, sufficiently known for the Céu Azul population, for deepening of study in the chemistry area. The chemical composition of the essential oil of Plectranthus neochilus was determined using gas chromatography techniques coupled to mass spectrometry. The main components were the monoterpenes α-pinene (9.7), thujene (9.0), terpin-4-ol (4.8) and sabinene (4.7); the sesquiterpenes caryophyllene (19.9) and caryophyllene oxide (15.5); and the diterpene methyl communate (4.4). The presence of pinene and caryophyllene as principal components of the essential oil could be related to the traditional use of Plectranthus against stomach ache. Keywords: medicinal plants, Céu Azul, ethnopharmacology, Plectranthus neochilus, essential oil, GC-MS. / O presente trabalho teve por objetivo realizar um levantamento etnofarmacológico de espécies vegetais utilizadas pelos moradores do município de Céu Azul (localizado no extremo oeste do Paraná). Utilizou-se um questionário específico para obter os dados referentes aos usos e modos de preparo das espécies utilizadas como medicinais. Durante as entrevistas, o material vegetal (exsicata) foi coletado em duplicata com o auxílio dos informantes para posterior identificação e depósito no Museu Botânico Municipal de Curitiba. Na pesquisa de campo foi constatada a utilização de 47 espécies pertencentes a 12 famílias botânicas. Um dos principais propósitos do presente levantamento foi avaliar o possível uso de espécies medicinais florestais no município de Céu Azul, especialmente porque uma extensa área do município pertence ao Parque Nacional do Iguaçu. No entanto, as 47 plantas citadas pela população ceuazulense podem ser divididas em um grupo com 42 espécies exóticas e outro com apenas 6 plantas brasileiras, nenhuma das quais é retirada da reserva. Utilizou-se o banco de dados do The International Plant Names Index para evitar incorreções nos binômios botânicos e verificar o uso atual de sinônimos. Para cada espécie citada, foi realizado um levantamento das informações científicas sobre a composição química e a bioatividade. As principais fontes de informações científicas foram o banco de dados do Institute for Scientific Information e o Chemical Abstracts on-line. Em geral, a população de Céu Azul faz uso de plantas medicinais bastante conhecidas no meio científico, mas há também a utilização de espécies pouco estudadas. Para quatro das espécies utilizadas (Alternanthera bettzickiana, Alternanthera dentata, Plectranthus neochilus, Tradescantia pallida), não foram encontradas referências científicas sobre suas bioatividades. Desta forma, os resultados deste levantamento foram concomitantemente interpretados como indicativos para a seleção de uma destas espécies, bastante conhecida e utilizada pela população de Céu Azul, para aprofundamento de estudo, Plectranthus neochilus. Determinou-se a composição química do óleo essencial das folhas desta espécie vegetal por técnicas de cromatografia gasosa acoplada à espectrometria de massas. Os principais componentes encontrados foram os monoterpenos pineno (9,7), tujeno (9,0), terpinen-4-ol (4,8) e sabineno (4,7); os sesquiterpenos cariofileno (19,9) e óxido de cariofileno (15,5); e o diterpeno comunato de metila (4,4). A ocorrência de pineno e cariofileno como principais componentes do óleo essencial pode estar relacionada ao uso tradicional desta espécie vegetal contra dores estomacais.

Made available in DSpace on 2017-07-24T19:38:01Z (GMT). No. of bitstreams: 1 Elementos pre-textuais.pdf: 567197 bytes, checksum: b26ed97be7d0d26d6ba01f298cc6bef8 (MD5) Previous issue date: 2009-06-29 / This study aimed to chemically analyze extracts of propolis samples and leaves of plant species of the Baccharis genus in the region of Ponta Grossa - PR, to classify the regional propolis compared to the green propolis (PG12). Baccharis dracunculifolia, the source of green propolis, may present metabolic changes related to environmental factors. To compare with published data, the essential oils obtained by hydrodistillation from the leaves of female (0.81%) and male (0.85%) specimens of B. dracunculifolia harvested in the dry season were analyzed by GC-MS. The two samples showed a low monoterpenes/sesquiterpenes ratio (0.85/96.88% ♀; 2.35/ 95.25% ♂). The monoterpenes b-pinene, limonene and terpinen-4-ol were identified. The main sesquiterpenes hydrocarbons are b-caryophyllene (♀ 7.88%; ♂ 8.88%), b-humulene (♀ 4.61%; ♂ 2.18%), germacrene D (♀ 5.53%; ♂ 4.04%) trans-b- guaiene (♀ 7.98%; ♂ 6.06%) and cadinene (♀ 7.34%; ♂ 5.48%). The oxygenated sesquiterpenes (E)-nerolidol (♀ 24.69%; ♂ 17.16%) and spathulenol (♀ 14.62%; ♂ 21.39%) were found as the major constituents. The high yield, the low monoterpenes / sesquiterpenes ratio and the high proportions of oxygenated sesquiterpenes are the main characteristics of the essential oil of B. dracunculifolia from this region. The leaves of three species of Baccharis (B. dracunculifolia, B. semiserrata and B. uncinella) were extracted using a Soxhlet apparatus with the sequence of solvents chloroform, ethyl acetate and methanol, and the extracts were analyzed by TLC, UV and HPLC. Free phenolic acids were identified: caffeic acid (Bd ♀ and Bd ♂), p-coumaric acid (Bd ♀), ferulic acid (Bd ♀, ♂ Bd, Bu and Bs) and the flavonoid rutin (Bd ♀, Bd ♂, Bu). Thus, it is demonstrated for the first time, the co-occurrence of polar phenolic substances in these three plant species. The ethanol extracts obtained from 13 samples of propolis were analyzed by TLC-HR and UV-VIS, allowing the classification of the regional propolis in three groups. The most characteristic sample of each type (ADI, WS- 3, RS-2) was chosen for analysis by HPLC in comparison with green propolis PG12. Fresh sprouts of male and female specimens of B. dracunculifolia were extracted by rinsing with methanol and the extracts were analyzed by HPLC. All extracts of propolis and B. dracunculifolia showed artepillin C ensuring that this plant is the source of regional propolis. The caffeic acid, ferulic acid and p-coumaric acid were identified in the four samples of propolis. Apigenin is present only in the regional propolis samples, between 2.4% and 3.9%, while crisin was identified only in the PG12, but as a main component (18.1%). An unidentified flavonoid (lma x 365 nm, 42,96 min) is the main substance present in the three samples of regional propolis and also the extract obtained by the rinsing of the plant sprouts. All the ethanol extracts of regional propolis showed excellent levels of antioxidant activity when tested against the radical DPPH, several of them getting to be more effective than the PG12. A comparative analysis of antioxidant power and chemical composition of the samples indicates that the unidentified flavonoid is responsible for the strong antioxidant activity of ADI and WS-2 compared to RS-2 and PG12. / Neste trabalho, foram analisados extratos de amostras de própolis e de folhas de espécies vegetais do gênero Baccharis da região de Ponta Grossa – PR, visando classificar a própolis regional em comparação com a própolis verde (PG12). Baccharis dracunculifolia, a fonte da própolis verde, pode apresentar variações metabólicas relacionadas a fatores ambientais. Para comparar dados publicados sobre esta planta com dados obtidos do vegetal coletado nesta região, foram analisados por CG-EM os óleos essenciais obtidos por hidrodestilação das folhas de espécimes feminino (0,81%) e masculino (0,85%) de B. dracunculifolia colhidos na estação seca. As duas amostras apresentaram baixa razão monoterpenos/sesquiterpenos (0,85/96,88% ♀; 2,35/95,25% ♂). Foram identificados os monoterpenos b-pineno, limoneno e terpinen-4-ol. Os sesquiterpenos hidrocarbonetos principais foram: b-cariofileno (♀ 7,88%;♂ 8,88%), a-humuleno (♀ 4,61%; ♂ 2,18%), germacreno D (♀ 5,53%; ♂ 4,04%), trans-b- guaieno (♀ 7,98%; ♂ 6,06%) e d-cadineno (♀ 7,34%; ♂5,48%). Os sesquiterpenos oxigenados (E)-nerolidol (♀ 24,69%; ♂ 17,16%) e espatulenol (♀ 14,62%; ♂ 21,39%) se encontraram como constituintes majoritários. O alto rendimento, a baixa relação monoterpenos/sesquiterpenos e as altas proporções de sesquiterpenos oxigenados são as principais características do óleo essencial de B. racunculifolia desta região. As folhas de três espécies de Baccharis (B. dracunculifolia, B. semiserrata e B. uncinella) foram extraídas em aparelho Soxhlet com a sequência de solventes clorofórmio, acetato de etila e metanol, analisando-se os extratos por CCD, UV e CLAE. Foram dentificados os ácidos fenólicos livres: ácido caféico (Bd ♀ e Bd ♂), ácido p-cumárico (Bd ♀), ácido ferúlico (Bd ♀, Bd ♂, Bu e Bs) e o flavonóide rutina (Bd ♀, Bd ♂, Bu). Assim, demostrou-se, pela primeira vez, a co-ocorrência de substâncias fenólicas polares nestas três espécies vegetais. Os extratos etanólicos obtidos de 13 amostras de própolis foram analisados por CCD-AR e UV-VIS, permitindo classificar as própolis regionais em três grupos. Foi escolhida a amostra mais característica de cada grupo (ADI, WS-3, RS-2) para análise por CLAE em comparação com a própolis verde PG12. Brotos frescos dos espécimes masculino e feminino de B. dracunculifolia foram extraídos por enxágüe com metanol e também submetidos à análise por CLAE. Todos os extratos, de própolis e de B. dracunculifolia, apresentaram artepilina C garantindo que esta é a fonte vegetal da própolis regional. Os ácidos caféico, ferúlico e pcumárico foram identificados nas quatro amostras de própolis. Apigenina estaria presente somente nas própolis regionais, entre 2,4% e 3,9%, enquanto a crisina foi identificada somente na PG12, mas como componente principal (18,1%). Um flavonoide (lmáx 365 nm, 42,96 min) é a substância principal das três amostras de própolis regionais e também do extrato obtido por enxágue dos brotos da planta, mas não foi identificado. Todos os extratos etanólicos de própolis regionais apresentaram excelentes níveis de atividade antioxidante quando testados frente ao radical DPPH, vários deles chegando a ser mais efetivos que o da PG12. Uma análise comparativa do poder antioxidante e da composição química das amostras permitem concluir que o flavonóide não identificado é o responsável pela fortíssima atividade antioxidante de ADI e WS-2 em comparação com RS-2 e PG12.

Les exsudats des plantes sont des substances naturelles complexes. Ils sont utilisés par plusieurs populations d’Afrique et trouvent de nombreuses applications : en thérapeutique, en pharmacie galénique, en parfumerie, etc. Au Congo, une grande majorité de ces substances est encore mal connue. Ce travail de thèse a consisté à inventorier les plantes à exsudats du Congo et à mettre au point une méthodologie d’analyse physico-chimique permettant la caractérisation des exsudats d’arbres d’origine botanique certifiée.La méthodologie d’inventaire adoptée nous a permis de recenser 135 plantes à exsudats appartenant à 87 genres. Il en ressort de cette étude que les exsudats des plantes inventoriées ont multiples usages (glu, insectifuge, allume-feu, antidouleur, etc.).La stratégie mise en place pour leur caractérisation fait appel à plusieurs techniques analytiques (SPIR, SMIR, CPG-SM, GCxGC-MS) et de traitement d’échantillon (Hs-SPME, hydrodistillation, extraction au Soxhlet, silylation, méthylation, pyrolyse). Dans le cadre de ce travail deux exsudats ont été caractérisés. L’exsudat de D. edulis (G. Don) K.J. Lam qui est constitué de monoterpènes hydrocarbonés et de triterpènes, dont les majeurs sont : 3-épi-α et -β-amyrine. L’exsudat de G. demeusei (Harms) J. Léonard dont la fraction volatile est constituée essentiellement de sesquiterpènes et la fraction peu-volatile d’acides diterpéniques de type labdane (acide éperuique, cativique, copalique, ozique, etc.).Pour exploiter l’efficacité de séparation élevée de la GCxGC-MS, nous nous sommes confronté aux problèmes de la non-répétabilité des cartes chromatographiques obtenues en 2D. Nous avons alors mis en place une méthodologie permettant de vérifier l’épaisseur de film et l’homogénéité des dépôts de phases stationnaires afin de sélectionner des colonnes courtes identiques pour la GCxGC. Cependant, les résultats préliminaires ont montré que même avec des colonnes de deuxième dimension identiques, on pouvait avoir des différences au niveau de carte 2D. / Plant exudates are natural complex substances. They are used by several populations in Africa and have many applications: in therapeutics, in galenic pharmacy, in perfumery, etc. In Congo, a great majority of these substances are not yet well known. This thesis work aimed to make an inventory of plant exudates from Congo and to develop a methodology for performing physico-chemical analysis which may enable characterization of exudates from botanical certified trees.Through the methodology that we developed, 135 plant exudates belonging 87 genera were registered. It was observed in this study that these exudates are used for many purposes (glue, insect repellent, making fire, anti-pain, etc).The strategy implemented for characterizing the exudates included several analytical (SNIR, SMIR, GC-MS, GCxGC-MS) and sample processing (Hs-SPME, hydrodistillation, extraction using the Soxhlet, silylation, methylation pyrolyse) techniques. With regard to this work, two exudates were characterized. The exudates from D. edulis (G. Don) K.J. Lam which contained monoterpene hydrocarbons and triterpenes, with the major constituents being: 3-epi-α and -β-amyrine. The exudates from G. Demeusei (Harms) J. Léonard was found to have the sesquiterpenes as major constituents of the volatile fraction and the labdane-type diterpenes (eperuique acide, cativique acid, copalique acid, ozique acid, etc) as major constituents of the poor-volatile fraction.To explore the efficacy of elevated separation of the GCxGC-MS, we faced some problems related to the non-repeatability of chromatographic cards obtained in 2D. Therefore, we put in place a methodology which enables us to verify the film thickness and homogeneity of layers of stationary phases in order to select identical short columns for GCxGC. However, the preliminary results have shown that even using second identical dimension columns, we could observed differences in 2D cards