Dans cette étude, nous nous sommes intéressés aux propriétés physiques et chimiques de six accessions de Mammea americana afin de pouvoir identifier les accessions les plus prometteuses pour la vente en frais ou la transformation. Nous avons également cherché à évaluer l’activité antioxydante de la pulpe, identifier et quantifier ses composés phénoliques et optimiser leur extraction à l’aide de la technique assistée par ultrasons.Nos résultats montrent que les accessions étudiées présentent des caractéristiques physiques, physico-chimiques et fonctionnelles significativement différentes. Pavé 11, Lézarde et Ti Jacques sont intéressants pour la vente en frais, en raison de leurs fruits sucrés avec une teneur élevée en caroténoïdes et composés phénoliques totaux. Sonson, Pavé 11 et Lézarde présentent une adaptabilité à la transformation. La composition polyphénolique de la pulpe déterminée par HPLC-DAD et UPLC-MS, a mis en évidence la présence d’acides phénoliques, de tanins condensés, de flavonols et flavanols dans nos échantillons. D’autre part, les tests d’activité antioxydante (DPPH et ORAC) révèlent que Ti jacques est l’extrait le plus actif. Un plan d’expérimentation a été mis en œuvre afin d’optimiser l’extraction des polyphénols à l’aide d’une technique d’extraction assistée par ultrasons. Les résultats montrent que l’extrait obtenu est riche en polyphénols et contient les mêmes teneurs en acides phénoliques et flavonols comparé à celui obtenu par la méthode conventionnelle. De plus, l’extrait obtenu à l’aide d’un solvant « vert » possède de bonnes propriétés organoleptiques. / Our work focuses on the physical and chemical properties of six mamey apple cultivars in order to select elite cultivars suitable for food processing or as table fruit. The antioxidant activity of the fruit pulp, the identification and quantification of the polyphenols responsible for it, and ultrasound assisted extraction method were also investigated.According to our results, the postharvest routes for every cultivar could be different. Pavé 11, Lézarde and Ti Jacques were found to be good for consumption, giving sweeter fruits with high total phenolic and carotenoid contents. Sonson, pavé 11 and Lézarde had suitable characteristics for the manufacturing of mamey products. The polyphenolic composition of the pulp determined by HPLC-DAD and UPLC-MS showed the presence of phenolic acids, condensed tannins, flavonols and flavanols. The results of the antioxidant test (DPPH and ORAC) point out that the most antioxidant cultivar was Ti Jacques. The design and optimization of the ultrasound assisted extraction method has done for polyphenols extraction. The results showed that the polyphenols rich extract contains the same content of phenolic acids and flavonols in comparison to the conventional method. Additionally, the dry extract obtained with a “green” solvent, had good organoleptic properties.

Ces travaux ont pour but d’identifier des métabolites dans des teintures-mères (TMs) utilisées en homéopathie et en phytothérapie pour mieux contrôler la qualité des TMs et de mieux comprendre leur mode d’action sur la santé humaine. Nous avons étudié, par une approche de métabolomique globale, 19 TMs et un produit leader : le L52, fabriqués par les Laboratoires Lehning. Premièrement, nous avons utilisé des approches non-ciblées en construisant des banques de données GC-MS et UPLC-MS/MS, afin d’identifier un maximum de molécules dans chaque extrait. Puis, nous avons fractionné certaines TMs et purifié des métabolites inconnus pour une identification fine en HRMS. Nous avons identifié de nombreuses molécules dans chaque TM, montrant que ces dernières sont très riches en molécules pouvant être utilisées pour le contrôle-qualité des TMs et valorisées pour la santé humaine. / Tinctures defined as hydro-alcoholic extracts have been used from centuries in homeopathy and phytotherapy, but their chemical compositions remain still unknown. During my PhD, metabolomics analyses of nineteen tinctures and one leader product, L52, made by Laboratoires Lehning, were conducted using untargeted metabolomic approach. We build GC-MS and UPLC-MS/MS databases to identify a large amount of metabolites. Then, we used semi-preparative HPLC with both UV and mass detection to isolate some compounds from tinctures. We used UPLC-HRMS to obtain chemical formula, a prerequisite for metabolites identification. Finally, we identified a broad range of different metabolites in each tincture, highlighting the metabolic complexity of the TMs. These molecules can now be used for quality-control and valued for a better understanding of these products on human health.

Les coumarines et furocoumarines sont des phytoalexines synthétisées par certaines familles de plantes (ex : Rutacées dont font partie les agrumes), pour se défendre contre les bioagresseurs. Les furocoumarines peuvent être toxiques pour l’homme, lorsqu’elles sont combinées à certains médicaments : c’est l’effet pomelo. Aujourd’hui, la plupart des cytochromes P450 impliqués dans la synthèse des furocoumarines chez les Apiacées, ont déjà été caractérisés. En revanche, malgré l’importance économique des agrumes, nous en savons très peu sur la voie de biosynthèse des coumarines et furocoumarines chez ces plantes. Dans ce travail, nous avons créé, optimisé et validé une méthode d’analyse en chromatographie liquide à ultra haute performance couplée à un spectromètre de masse (UPLC-MS), pour identifier et quantifier 28 coumarines et furocoumarines dans la peau et la pulpe d’agrumes. Cette méthode nous a permis de chémotyper 62 variétés d’agrumes, distinguées par leur faible ou forte capacité de production de ces composés. En parallèle, un travail de bioinformatique sur des banques publiques d’ADN génomique d’agrumes, a permis d’identifier sept gènes présentant de fortes homologies de séquences avec ceux intervenant dans la synthèse des furocoumarines chez Pastinaca sativa (CYP71) et chez Arabidopsis thaliana (CYP82). Une analyse quantitative de leur niveau d’expression chez des agrumes, a montré que quatre d’entre eux étaient plus fortement exprimés chez les fruits fortement producteurs de coumarines et furocoumarines. Le clonage de ces gènes et leur expression hétérologue chez la levure, a révélé la fonction de CYP82D64 de pomelo et de Combava, qui hydroxyle la xanthotoxine pour donner la 5-hydroxy-xanthotoxine. La synthèse des coumarines et furocoumarines chez les agrumes, ainsi mieux appréhendée, nous a permis de proposer un schéma de sélection variétale visant à abaisser les taux de ces composés chez les Citrus. Nous avons aussi montré l’évolution convergente des CYP71 et CYP82 dans leur synthèse chez les Apiacées et les Rutacées respectivement. La découverte du premier cytochrome P450 de Citrus intervenant dans la production de ces composés, ouvre de nouvelles perspectives quant à l’élucidation de leur voie de biosynthèse chez les agrumes / Coumarins and furanocoumarins are phytoalexines synthesized by some plant families (e.g. Rutaceæ that include citrus), to defend themselves against bioaggressors. Furanocoumarins can be toxic for humans, when combined with some drugs: this is the grapefruit juice effect. Nowadays, most of the cytochrome P450s involved in the furanocoumarin synthesis in Apiaceæ, have already been characterized. However, despite the economical importance of citrus, a little is known about the coumarins and furanocoumarins pathway in these plants. In this work, we created, optimized and validated an analytical method by ultra high performance liquid chromatography coupled with mass spectrometry (UPLC-MS), to identify and quantitate 28 coumarins and furanocoumarins in citrus peel and pulp. This method allowed us to chemotype 62 citrus varieties, distinguished by their low or high capacity to produce these compounds. In parallel, a bioinformatic work on public banks of genomic DNA from citrus, allowed to identify seven genes with high sequence homologies with those involved in the synthesis of furanocoumarins in Pastinaca sativa (CYP71) and in Arabidopsis thaliana (CYP82). A quantitative analysis of their expression level in citrus showed that four of them were more expressed in high coumarins and furanocoumarins producing fruits. The cloning of these genes and their heterologous expression in yeast, revealed the function of grapefruit and Combava CYP82D64, which catalyzes the hydroxylation of xanthotoxin in 5-hydroxy-xanthotoxin. The synthesis of coumarins and furanocoumarins in citrus, then better apprehended, allowed us to propose a breeding scheme aiming at decreasing the levels of these compounds in Citrus. We also showed the convergent evolution of CYP71 and CYP82 in their synthesis in Apiaceæ and in Rutaceæ respectively. The discovery of the first cytochrome P450 from Citrus involved in the production of these compounds, opens up new prospects for the elucidation of their biosynthetic pathway in citrus

Afin d’étudier la santé d’un écosystème marin et le potentiel toxique d’une contamination telle que celle liée à la présence d’hydrocarbures aromatiques polycycliques (HAP), il est nécessaire, outre de connaître les niveaux de contamination du milieu, de pouvoir accéder à la fraction toxique à laquelle les organismes aquatiques ont été exposés et de connaître les effets toxiques des contaminants incriminés. L’exposition et la contamination des organismes aquatiques aux HAP ont généralement été évaluées par le dosage des HAP bioaccumulés dans les tissus. Or, cette approche est critiquable si l'on tient compte des capacités de biotransformation des organismes, notamment des vertébrés, et des propriétés toxiques des produits de transformation formés. Dans ce contexte, l’objectif de cette thèse est d’étudier les phénomènes de bioccumulation et de biotransformation des HAP chez les organismes marins via l’étude des métabolites de HAP. Un effort de validation de biomarqueurs pertinents pour évaluer la génotoxicité des HAP en lien avec la contamination chimique des tissus et la production de métabolites est nécessaire. Des méthodes de dosage des métabolites de HAP dans les matrices biologiques ont tout d’abord été mises au point. Ces outils analytiques sensibles, innovants et performants ont ensuite été appliqués lors d’expositions de poissons à des HAP via différentes voies de contamination en milieu contrôlé. Ils ont permis une meilleure connaissance des phénomènes de biotransformation des HAP. Enfin, des études de terrain ont été réalisées, notamment dans le cadre de l’étude de la contamination de la Baie de Seine, montrant l’applicabilité du dosage des métabolites de HAP dans l’évaluation de l’exposition des organismes aux HAP en milieu naturel. Dans le cadre d’une approche intégrée chimie/biologie, ces travaux ont permis d’apporter une contribution dans le transfert méthodologique des biomarqueurs de génotoxicité des HAP pour des applications en surveillance de l’Atlantique Nord et notamment dans la Manche. / In order to study the health of a marine ecosystem and the toxic potential of a contamination such as that related to the presence of polycyclic aromatic hydrocarbons (PAHs), it is necessary, in addition to the determination of environmental contamination levels, to have access to the fraction for aquatic organisms have been exposed to and to identify the toxic effects of the contaminants. The exposure and contamination of aquatic organisms to PAHs have generally been evaluated by the quantification of bioaccumulated PAHs in tissues. However, this approach is criticable when taking into account the biotransformation capabilities of organisms such as vertebrates and the toxic properties of biotransformation products. In this way, the aim of this study is to study PAH bioaccumulation and biotransformation phenomena through the PAH metabolites study. An effort for the validation of relevant biomarkers to evaluate the link between the genotoxicity of PAHs, PAHs body burden and PAH metabolites production, is necessary. Analytical techniques to quantify PAH metabolites in biological matrices have first been set up. Then, these sensible, innovating and powerful analytical tools have been applied to the study of fish exposures to PAHs through differents contamination sources in controlled conditions. This allowed to have a better understanding of PAH biotransformation phenomena. Finally, field studies have been led, notably to study the contamination of the Seine bay, demonstrating the applicability of the quantification of PAH metabolites to evaluate the exposure and the contamination of organisms to PAHs in natural environment. Within the framework of an integrated approach chemistry/biology, this work led to a contribution in the methodological transfer of biomarkers of PAH genotoxicity

Submitted by Sandro Camargo (sandro.camargo@unipampa.edu.br) on 2015-05-08T02:43:19Z No. of bitstreams: 1 127110045.pdf: 1527824 bytes, checksum: 960729a0a23069d5d40ec997cc3034b8 (MD5) / Made available in DSpace on 2015-05-08T02:43:19Z (GMT). No. of bitstreams: 1 127110045.pdf: 1527824 bytes, checksum: 960729a0a23069d5d40ec997cc3034b8 (MD5) Previous issue date: 2014-07-17 / O gênero Cuphea, popularmente conhecido no Brasil por “sete-sangrias”, tem seu uso medicinal reconhecido devido aos efeitos diurético, hipotensor e cardioprotetor. No sul do Brasil, em região característica do bioma Pampa, foi encontrada a espécie Cuphea glutinosa Cham. & Schltdl. Embora o uso popular, esta espécie é pouco descrita na literatura. O presente trabalho tem como objetivos o estudo da composição química dos extratos de C. glutinosa e a avaliação das atividades antifúngica e anti-hipertensiva. O material vegetal foi coletado na cidade de Uruguaiana (RS, Brasil), identificado e depositado em herbário. Após secagem e trituração do material vegetal, foram obtidos os extratos hidroetanólicos através de maceração exaustiva com etanol 40% (v/v) para folhas e etanol 70% (v/v) para raízes. Para a infusão, utilizou-se água a 80oC. As análises cromatográficas foram realizadas em equipamento cromatógrafo a líquido Prominence Shimadzu, em técnica por CLAE e CLUE. Utilizou-se sistema de fase reversa, eluição por gradiente com fase móvel composta por acetonitrila:metanol (4:1) e ácido fórmico 0,1% pH 3,0, coluna C18 analítica e fast, e detecção por UV-DAD e ESI-MS. Os teores de polifenóis totais e de flavonóides foram determinados por método colorimétrico, seguindo metodologia padronizada. A atividade antifúngica in vitro foi realizada utilizando o método de microdiluição em caldo, determinando-se a CIM, in-vitro, contra diferentes isolados clínicos. Para avaliação do potencial anti-hipertensivo in vivo, foram realizadas medições da pressão sanguínea pelo método de monitoramento hemodinâmico invasivo, através da inserção de cateter na artéria carótida. Os resultados de teor de fenólicos totais indicaram predominância destes componentes em extratos obtidos de folhas e por maceração, conforme os valores obtidos: 1,8501 mg EAG/mL (folhas) e 0,8467 mg EAG/mL (raízes) para infusão, e 3,7284 mgEAG/mL (folhas) e 2,6266 mg EAG/mL (raízes) para maceração. Quanto ao teor de flavonóides, os resultados quantitativos foram: 7,0959 mg/g (folhas) e 0,5664 mg/g (raízes) para a infusão, e 7,9511 mg/g (folhas) e 0,5994 mg/g (raízes) para maceração. Na análise cromatográfica, os extratos obtidos das folhas de C. glutinosa apresentaram picos cromatográficos bem separados, em perfil reprodutível. Na determinação por CLUE-MS, os dados de íon molecular e fragmentos de massa indicaram a composição predominante em flavonóides, sugerindo-se os componentes quercetina-3-O-glicosídeo, quercetina-3- arabinosídeo, quercetina-3-glicuronídeo, isoramnetina e quercetina-5-O-β-glicopiranosídeo. Para o potencial antifúngico, os extratos das folhas e raízes apresentaram atividade in vitro contra Candida parapsilosis, Candida tropicalis e Trichosporon asahii, com CIM variando na faixa de 1,9-62,5 μg/mL. Nos testes hemodinâmicos realizados, os extratos das folhas não apresentaram efeito significativo sobre a pressão arterial. A identificação dos componentes em C. glutinosa, derivados de quercetina, torna promissora novas investigações a fim de aprofundar o conhecimento a respeito desta espécie, em especial na busca de respostas para a relatada ação anti-hipertensiva. / The Cuphea genus, popularly known in Brazil as "sete-sangrias", is used traditionally due the diuretic, hypotensive and cardioprotective effects. In southern Brazil, in characteristic region of Pampa biome, it was found the species Cuphea glutinosa Cham. & Schltdl. Although used popularly, this species is few reported in the literature. The present work aimed to study the chemical composition of extracts from C. glutinosa and to evaluate the antifungal and anti -hypertensive activities. The plant material was collected in the city of Uruguaiana (RS, Brazil), identified and deposited in a herbarium. After dryness and milling, the hydroethanolic extracts were obtained through exhaustive maceration using ethanol 40% (v/v) for leaves and ethanol 70% (v/v) for roots. The infusions were prepared using water at 80 °C. The chromatographic analyses were performed in liquid chromatography Prominence Shimadzu, for HPLC and UPLC assays. The method was conducted using reverse phase system, gradient elution with mobile phase composed by acetonitrile:methanol (4:1) and formic acid 0.1% pH 3.0, C18 analytical and fast column, and detection by UV-DAD and MS. The polyphenols and flavonoids contents were determined by colorimetric method. The in vitro antifungal activity was conducted by using the broth microdilution method, determining the MIC against different clinical isolates. For evaluation of in vivo anti-hypertensive potential, the blood pressure was measured by the method of invasive hemodynamic monitoring, through of insertion the catheter into the carotid artery. The results of phenolic content indicated the high concentration of these compounds in leaves extracts obtained by maceration: 1.8501 mgEAG/mL (leaves) and 0.8467 mgEAG/mL (roots) for infusion, and 3.7284 mgEAG/mL (leaves) and 2.6266 mgEAG/mL (roots) for maceration. For flavonoids, the contents were: 7.0959 mg/g (leaves) and 0.5664 mg/g (roots) for infusion, and 7.9511 mg/g (leaves) and 0.5994mg/g (roots) for maceration. In the chromatographic analyses, the leaf extracts from C. glutinosa presented chromatographic peaks well separated and reproducible. In the determination by UPLC-MS, the molecular ion and mass fragments indicated the predominant composition in flavonoids, suggesting the compounds quercetin-3- O-glucoside, quercetin-3-arabinoside, quercetin-3-glucuronide, isorhamnetin and quercetin-5- O-β-glucopiranoside. For the antifungal potential, the leaf and roots extracts presented activity against Candida parapsilosis, Candida tropicalis e Trichosporon asahii, with MIC values ranging 1,9-62,5 μg/mL. In the hemodynamic tests performed, the leaves extracts did not present significant effect in the arterial pressure, although a tendency in pressure reduction could be observed. The identification of quercetin derivatives in C. glutinosa becomes promisor further investigations about this species, mainly in respect to the anti-hypertensive action.

The hip-flask defence is a common claim in drunkdrinking cases. In Sweden and Norway two different models are used to determinethese cases. In Sweden one blood and two urine samples taken 60 minutes apartare used for analysis. In Norway two blood samples taken 30 minutes apart areused. Sweden focuses on the rise or fall of alcohol concentration in urine(UAC), and the ratio between UAC and blood alcohol concentrations (BAC). Norwayfocuses on the rise or fall of the alcohol metabolite ethylglucuronide (EtG) and the ratio between BAC and EtG. The aim of this study wasto test the models for multiple intakes and with different alcoholic beverages.Thirtyfive participants ingested two doses, first0.51 g/kg of beer and later either 0.25, 0.51 or 0.85 g/kg of beer, wine orvodka. Blood and urine samples were obtained before and after alcoholingestion. Alcohol was measured by GC-HS, and the alcohol metabolite byUPLC-MS/MS. The results showed that there are kineticdifferences between single and repeated intakes, that there are no significantdifferences in kinetics from different alcoholic beverages and thatthe Norwegian model appears to be the stronger one in hip-flask determination.

<p>Entomophagy, which is the habit of eating insects, has become relevant in the past few years as it could potentially help reduce current and future food insecurity, due to the highly nutritious and sustainable characteristics of edible insects. In addition to the nutritional content of insects, research on the potential bioactive components of insect extracts has also gained popularity. In this study, extracts from house cricket (<i>Acheta domesticus</i>) from two farms and their corresponding feeds were obtained using a microwave-assisted extraction. Further phenolic characterization led to the identification and quantification of 4-hydroxybenzoic acid, <i>p</i>-coumaric acid, ferulic acid and syringic acid as major phenolic compounds in both <i>A. domesticus </i>extracts as well as both feed extracts. Additionally, <i>in vitro</i> antioxidant activity was evaluated using 2,2-diphenyl-1-picrylhydrazyl radical cation (DPPH) and 2,2’-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical assays. In both <i>in vitro</i> assays, <i>A. domesticus</i> extracts showed higher antioxidant activity compared to the feeds. Antibacterial activity against <i>E. coli</i> and <i>L. innocua</i> was also evaluated using the microwell method. The <i>A. domesticus </i>extracts showed a selective inhibition (p<0.05) towards the gram-positive bacteria <i>L. innocua </i>between a period of 4 to 8 h. This inhibition is thought to have occurred as a result of the presence of phenolic acids and antimicrobial peptides, while the feed extracts did not exhibit any inhibitory activity towards any of the bacteria. The finding of the same phenolic acids in <i>A. domesticus</i> and their corresponding feed could imply the capacity of <i>A. domesticus</i> to absorb and sequester dietary phenolics that may provide additional health benefits when the insect is consumed, unveiling new benefits of entomophagy. </p>

Per- and polyfluorinated substances (PFAS) are used in the production of ski wax to reduce the friction between the snow and the ski. In this occupational study of ski wax technicians’ exposure to PFAS and particulate aerosol we have collected whole blood (wb) (n =94), air (n =84) and aerosol (n =159) samples at World Cup events from 2007-2011. We have analysed the blood, air and aerosol with respect to 13 perfluorocarboxylic acids (PFCAs), 4 perfluorosulfonic acids (PFSAs), 3 fluorotelomer alcohols (FTOHs), 3 fluorotelomer acids (FTCAs) and 3 unsaturated fluorotelomer acids (FTUCAs). Further, we assessed the exposure to 3 particulate aerosol fractions (inhalable, respirable and total aerosol) in air. In comparison to a general population, several of the PFCA blood levels are elevated in the technicians’, primarily  erfluorooctanoate (PFOA) and perfluorononate (PFNA) with concentrations up to 628 and 163 ng/mL wb, respectively. Further,  we detected FTUCAs and FTCAs in the blood, suggesting biotransformation of FTOHs to PFCAs. The metabolites 5:3 and 7:3 FTCA were detected in all blood samples at levels up to 6.1 and 3.9 ng/mL wb. Levels of perfluorohexadecanoic acid PFHxDA) and perfluorooctadecanoic acid (PFOcDA) were detected in the technician’s blood at mean concentration up to 4.22 ng/mL wb and 4.25 ng/mL wb. The FTOH levels in air of the wax cabin during work ranged up to 997 000 ng/m3 (average=114 000 ng/m3 ) and PFOA up to 4 890 ng/m3 (average= 526 ng/m3 . FTOHs were not detected in aerosols but PFOA showed average levels of 12 000 ng/m3 (range=1 230- 46 900 ng/m3 ). The occupational exposure limit (OEL) of 2 mg/m3 was exceeded in 37% of the personal measurements with aerosol  concentrations up to 15 mg/m3 . Keywords : Perfluorinated, polyfluorinated, FIS, occupational exposure, ski wax,  iotransformation, metabolism, fluorotelomer alcohol, fluorotelomer acid, aerosol, dust, UPLC/MS-MS, GC/MS-MS

Diploma work focuses on the determination of selected representatives of perfluoroalkyl substances in complex matrices, particularly, in sewage sludge and feed. In case of sewage sludge, samples were extracted into methanol and three extractions techniques were compared: accelerated Soxhlet, pressurized liquid extraction and Powley method. Powley method showed sufficient efficiency, the lowest matrix effect and minimal background. Perfluorooctane sulfonate was present in all tested samples of sewage sludge (0.74–38.02 ng.g-1). Other detected compounds were perfluorodecanoic acid and perfluorooctanoic acid. Three extraction techniques were tested on feed samples (QuEChERS, fast methanol extraction and Powley method into acetonitrile). Powley method provided the cleanest extracts and showed simultaneously the highest recovery of native perfluoroalkyl substances and the lowest matrix effects. Perfluorooctane sulfonate and perfluorooctanoic acid were rarely present in samples of complete and supplemental feeds, while other perfluoroalkyl substances were found at levels below limit of quantification or they were not detected at all. Perfluorooctane sulfonate (0.100–2.768 ng.g-1), perfluoroundecanoic acid and perfluorotridecanoic acid were determined in all fish meal samples.

La consommation des composés pharmaceutiques est en constante croissance à travers le monde et leur utilisation peut entraîner une accumulation dans l’environnement d’où leur désignation à titre de contaminants émergents. De nombreux groupes de recherche se penchent donc sur les questions de leurs effets sur des organismes vivants, sur la santé de certains écosystèmes ou encore sur l’efficacité de différents types de traitement des eaux usées. Or, peu de données sont disponibles quant aux concentrations présentes dans différentes matrices telles que les eaux usées ou l’eau de surface dans lesquelles elles sont déversées ou encore l’impact que peuvent avoir les hôpitaux sur l’occurrence des composés pharmaceutiques. La recherche dans ce domaine peut s’avérer un outil clé dans l’établissement de normes et de limites pour différents contaminants puisque celles-ci sont encore marginales dans la plupart des pays du monde, surtout au niveau des médicaments. Dans le cadre de cette thèse, différentes méthodes analytiques ont été développées pour faire l’analyse de classes ciblées de médicaments par chromatographie liquide couplée à la spectrométrie de masse afin de quantifier ceux-ci dans diverses matrices environnementales. Un des objectifs du présent ouvrage est donc de détailler les travaux de développement et de validation de méthodes analytiques robustes. Une première méthode d’extraction en phase solide en ligne avec la chromatographie liquide ultra-haute performance couplée à la spectrométrie de masse en tandem (SPE en ligne UPLC-MS/MS) a été développée afin de faire l’analyse de huit différentes classes de composés pharmaceutiques susceptibles d’être retrouvés dans les eaux usées. Afin de déterminer la distribution et l’adsorption des composés ciblés sur la phase solide de différentes matrices, une méthode d’extraction et de purification a également été optimisée de telle sorte que les extraits de matière particulaire et de sédiments soient compatibles avec cette méthode analytique. Ces méthodes ont été appliquées sur de nombreuses eaux usées d’hôpitaux et d’usine de traitement des eaux au Québec (Canada). Les produits de chimiothérapies présentent un défi analytique supplémentaire puisqu’ils sont polaires comparativement à la plupart des autres classes de composés pharmaceutiques. Il y a donc peu d’études sur la présence et le comportement de ces contaminants bien que ceux-ci présentent un risque écotoxicologique potentiellement important. Un deuxième objectif de cette thèse a donc été de comparer différentes alternatives pour la préconcentration et la séparation chromatographique, notamment la chromatographie à interaction hydrophile (HILIC). Celle-ci a mené à la validation de deux méthodes analytiques dont la sensibilité est de l’ordre des ng/L pour ces composés dans des eaux usées brutes. Ces méthodes de SPE en ligne UPLC-MS/MS ont été appliquées à de nombreux échantillons d’effluents d’hôpitaux ainsi qu’à des usines de traitement des eaux usées au Québec (Canada). D’autre part, la méthode SPE en ligne UPLC-MS/MS pour l’analyse des multiples classes de composés pharmaceutiques a été validée pour l’eau de surface enfin d’atteindre un dernier objectif de cet ouvrage qui consiste à évaluer différentes sources de contamination de cette matrice à proximité de zones densément peuplées. Durant de larges campagnes d’échantillonnage sur le fleuve Saint-Laurent, échelonnées sur une période de cinq ans et ayant couvert une zone géographique de près de 700 km entre le Lac Ontario et l’estuaire, plus de 400 échantillons ont été prélevés et analysés. Parmi ceux-ci figurent des échantillons provenant de 56 rivières tributaires au fleuve prélevés afin de déterminer l’impact de celles-ci en termes de pollution en composés pharmaceutiques et de les comparer avec des points de rejet d’eau usée de villes telles que Montréal ou Québec. Enfin, ces mêmes méthodes ont été appliquées lors de projets en collaboration avec des chercheurs se penchant sur les effets écotoxicologiques des composés pharmaceutiques sur des organismes aquatiques dans des matrices d’eau de surface ainsi que pour de développement de technologies alternatives pour le traitement des eaux usées. / A constant increase in pharmaceutical compounds worldwide can accentuate problems related to these pollutants of emerging concern in environmental matrices. Therefore, many researchers are studying their effects on living organisms, the health of impacted ecosystems, and the efficiency of different types of wastewater treatment systems. However, there is a lack of available data on the occurrence and concentrations of pharmaceuticals in different matrices, such as wastewaters or receiving surface waters, and the relative importance of hospital effluents on the load of these contaminants. Research in this field can be a key tool on different drugs since only a few countries have implemented norms and guidance, especially for medications. As part of this thesis, different analytical methods have been developed for the analysis of selected drugs, and liquid chromatography coupled with mass spectrometry was used to quantify multi-class pharmaceuticals occurring in various environmental matrices. One objective of this manuscript is to develop and validate robust analytical methods. A first extraction method on-line with liquid chromatography coupled to tandem mass spectrometry (on-line SPE UPLC-MS/MS) was developed to analyze eight different pharmaceutical classes susceptible to be found in wastewaters. With the aim of evaluating the partitioning and adsorption of target compounds onto a solid phase, an extraction and purification method was optimized to ensure that particulate matter and sediment extracts were compatible with the analytical method. These optimized methods were then applied to several hospital effluents and wastewater treatment plants samples in the province of Quebec (Canada). Chemotherapy agents present an additional analytical challenge since these drugs are relatively polar compared with most of other pharmaceutical classes. This explains the low number of publications relating to their occurrence and behaviour even though they present a high ecotoxicological hazard. A second objective of this thesis was to compare different potential alternatives for enrichment and chromatographic separation of these pollutants, notably the use of hydrophilic interaction chromatography (HILIC). This work led to the validation of two analytical methods for which the sensitivity reached ng/L levels for these compounds in raw sewage. These methods were successfully applied to samples from numerous hospital effluents and wastewater treatment plants from the province of Quebec (Canada). Also, a multi-class pharmaceutical method was validated for surface water in order to achieve the last objective of this thesis which consisted in the determination of the different contamination sources of this matrix near densely populated areas. During major sampling campaigns on the Saint-Lawrence River, spanning over five years and covering a geographical area of approx. 700 km between Lake Ontario and the estuary, a total of more than 400 samples were collected and analyzed. Among those, 56 different rivers tributary to the Saint-Lawrence were sampled to establish the impact they may have on pharmaceutical pollution. Data were compared with wastewater rejection points from cities, such as Montreal and Quebec. Finally, these methods were applied in the context of collaboration projects with researchers focusing on measuring ecotoxicological effects of pharmaceutical compounds on living organisms in surface water matrices and on the development of alternative wastewater treatment technologies.