Tesi sul tema "SPE-HPLC-MS/MS"

Alguns compostos orgânicos como interferentes endócrinos e ativos farmacêuticos estão sendo encontrados em águas superficiais e vêm chamando a atenção de orgãos públicos e da comunidade científica. Dentre esses compostos, foram enfatizados nesse trabalho, o hormônio sintético 17α-etinilestradiol, o surfactante (4-nonilfenol), bisfenol A, o antibiótico β-lactâmico amoxicilina e a cafeína como traçador de atividade antrópica. Estes compostos são continuamente introduzidos em corpos d\'água pelo lançamento de esgoto in natura o que afeta a qualidade da água, a saúde dos ecossistemas e impacta o suprimento de água potável. Considerando a baixa taxa de remoção desses contaminantes pelo tratamento de água atualmente utilizado no Brasil e o estado de degradação dos mananciais de água para abastecimento público, esse trabalho apresenta o desenvolvimento de metodologia analítica para determinação dos compostos citados e a avaliação da eficiência de remoção quando o tratamento convencional de água é aplicado. A determinação foi baseada em métodos já estabelecidos e envolveram as etapas de limpeza e concentração de amostra usando extração em fase sólida (SPE) e análise por cromatografia a líquido de alta eficiência (HPLC) com detector de arranjo de diodos (DAD) e espectrometria de massas. As amostras do Reservatório do Guarapiranga foram utilizadas para ensaios em escala laboratorial que simulavam o tratamento convencional de água em planta real. Os resultados mostraram a adequação dos métodos utilizados para a separação e determinação desses compostos em amostras ambientais. Além disso, apontaram para a necessidade de se avaliar a utilização de metodologias não convencionais que otimizem a remoção pelo tratamento de água atualmente utilizado.
Some organic compounds, such as endocrine disruptors and pharmaceuticals, are being found in the surface water, and are concerning both the public organs and the scientific community. Among these compounds, the synthetic hormone 17α-ethinylestradiol, the surfactant (4-nonylphenol), bisphenol-A, the β-lactam antibiotic amoxicillin and the marker of human activity caffeine were highlighted in the present work. These compounds are continuously drained to the water bodies by sewers, affecting not only the water quality itself, but also the ecosystems, and impacting the drinking water availability. Currently in Brazil, the removal rate of these contaminants is low, whereas the water sources for public supply are highly degraded. In this context, the present work develops an analytical method to detect the aforementioned compounds, and to evaluate the efficiency of its removal by conventional water treatments. Detection was based on pre-established methods and steps involved clean up and samples concentration by solid phase extraction (SPE), high performance liquid chromatography with diode array detection (DAD), and mass spectrometry. Recovery tests were done using environmental samples from Guarapiranga Reservoir (São Paulo, SP). This samples were further used in laboratorial tests, simulating real scale water conventional treatments. Current results revealed the adequacy of the chosen methods in the separation and determination of the abovementioned compounds from natural samples. Also, they highlight the need for the evaluation of non-conventional methodologies that optimize removal by current water treatment.

De nombreux composés d’origine anthropique sont détectés dans l’environnement, particulièrement dans les eaux de surface utilisées comme ressource. Parmi ces molécules, les pesticides, bien qu’ils soient parfois présents à de très faibles concentrations, peuvent causer des dommages sanitaires ou environnementaux. La préparation de l’échantillon et la méthode de détection sont les deux points critiques de la qualité du dosage de tels substances dans les eaux. L’extraction demeure notamment une étape fastidieuse qui génère des problèmes de récupération. Pour les réduire, une technique de chromatographie liquide avec préconcentration en ligne (SPE-LC) couplée à un spectromètre de masse de type piège ionique linéaire (LIT) a été développée et validée en matrice eau propre (type eau d’Evian) et en eau de rivière (type rivière Loire - Nantes). Les critères de performances retenus étaient les suivants : exactitude, reproductibilité, sélectivité et sensibilité afin d’atteindre aisément des limites de quantification (LQ) au niveau 10 ng/L, pour un grand nombre de molécules. Près de 210 pesticides sont analysés dans le même run chromatographique en mode ElectroSpray positif. 2 ml d’échantillon d’eau sont percolés à travers une cartouche appropriée. Les LQ atteintes dans les deux matrices sont, pour la plupart des molécules inférieures ou égales à 10 ng/L. Les résultats montrent que la SPE-LC couplée à la spectrométrie de masse en tandem (MS/MS) est une technique adaptée, entièrement automatisable, pour réaliser une analyse multi-résidus de routine dans ces types d’eaux. La dégradation des molécules a par la suite été étudiée par la recherche de quelques métabolites, et aussi par la modification des conditions de stockage, montrant ainsi l’importance d’évaluer la stabilité des composés en milieu aqueux (hydrolyse, photolyse). L’étude s’est ensuite focalisée sur la qualité d’identification des composés par l’utilisation du mode d’acquisition MRM-EPI (Full scan MS/MS) disponible sur ce type de spectromètre. Ce mode de balayage se révèle comme un outil de choix pour assurer l’identification en dosage multi-résidus. Enfin, le thème d’effet matrice a été largement abordé, car la technique développée donnait lieu à des suppressions de signal non négligeables pour un certain nombre de composés (exaltations de signal plus rarement). L’origine de ces effets matrice a été évaluée par des essais d’introduction post-colonne, montrant que l’ionisation par Electrospray est un point critique. Il a fallu apporter des solutions à la maîtrise et à la réduction de ces effets incommodants pour la restitution juste d’une concentration d’un analyte présent dans un échantillon d’eau. L’étalonnage interne s’est montré efficace et ainsi a été choisi, par application ou non d’un étalon interne adapté pour une molécule précise
To analyze swiftly and reliably numerous pesticides in waters, an on-line solid phase extraction liquid chromatography technique (on-line SPE-HPLC) coupled with triple quadripole linear ion trap mass spectrometry (Q-Trap MS/MS) procedure was developed for a large range of chemical families with detection limits at the 10 ng/L level. In this study, triazines, phenylureas, organophosphorus compounds and others (near than 210 compounds) were tested in the same ESI positive chromatographic run (less than 25 min). 2 ml of water sample (filtered if necessary) were injected into the SPE-HPLC system through a preconcentration cartridge. Analytes were eluted by a suitable mobile phase towards an analytical column (monolithic stationary phase), then detected and quantified by the mass spectrometer (MS/MS). The LOQ were achieved under the 10 ng/L level for the most of compounds in the both matrices (tap water and river water). Degradation of molecules was studied by research of few metabolites, and also by modification of storage conditions, showing the importance to evaluate stability of compounds in aqueous environment (hydrolysis, photolysis). The study was then focused on the identification by using the MRM-EPI scan mode (Full scan MS/MS), available on this kind of mass spectrometer. Results show that on-line-SPE-LC technique, coupled with hybrid triple quadripole linear ion trap mass spectrometry is a suitable way to determine multiple organic residues in waters. Finally, matrix effects theme was largely approached, since developed technique gave place non negligible signal suppressions for few compounds (signal enhancement more rarely). Matrix effects origin was evaluated by post-column introductions tests, showing that Electrospray ionization is a critical point. It was necessary to bring solutions of the control and the reduction of these annoying effects in the just restoration of an analyte concentration. The internal calibration showed effective and so was chosen, by application or not of an internal standard adapted for a particular molecule

L'analyse de la composition chimique de deux orchidées de la sous-tribu des Aeridinae, Aerides rosea Lodd. ex. Lindl. & Paxton et Acampe rigida (Buch.-Ham. ex Sm.) P. F. Hunt, a été menée en ayant recours à des techniques de couplages et grâce à une stratégie de déréplication. Dix dérivés phénanthréniques ont ainsi été identifiés dans les tiges d'A. rosea, dont deux nouvellement décrits. Quatre stilbénoïdes, trois dérivés d'acides phénoliques et quatre esters d'acide cinnamique ont été identifiés dans les tiges d'A. rigida. Nous avons également effectué des dosages de traceurs dans différents échantillons de deux représentants de la tribu des Vandeae : Vanda teres (Roxb.) Lindl. et Vanda coerulea Griff. ex. Lindl., afin de mettre en évidence d'éventuelles variations de composition en fonction des facteurs environnementaux et du stade de croissance végétative. Enfin, nous avons mis au point des conditions d'isolement préparatif par chromatographie de partage centrifuge afin de purifier des marqueurs biologiques préalablement identifiés dans les tiges de Vanda teres : trois glucospyranosyloxybenzyl - malates ainsi que leur précurseur biosynthétique.

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Universidade Federal de Sao Carlos
The growth of industrial deployment and urbanization play a major role in the degradation of global environmental and resource depletion mainly in countries in development such as Brazil. The problems regarding water pollution in Latin America have been well-documented, and there is no evidence of substantive effort to change the situation. There are a large number of organic pollutants that leads several damages to the ecosystem, threatening human health and are flushed to wastewater treatment plants (WWTPs). The WWTPs are responsible for the removal of natural and anthropogenic pollutants from the sewage and, for this, play an important role in protection of human health and the environment. Due to the varying operational conditions, process and quality, the wastewater treatments need to be monitored continuously to ensure a reliable and efficient operation. Therefore, the NMR analyses are widely used in the study of different components of wastewater such as complex organic matter (fulvic and humic acids), sludge and organic contaminants. In the present work, we employed two methods to study wastewater samples: non-targeted 1D (13C and 1H) and 2D NMR spectroscopy analyses to characterize the largest possible number of compounds in urban wastewater; and analysis by HPLC-(UV/MS)-SPE-ASS-NMR to detect and elucidate the non-specific recalcitrant in treated wastewater, sparing the common use of standards. In the non-targeted 1D NMR analysis, we employed also a method to study wastewater samples by chemometric and quantitative analyses to monitor the seasonal variations and identify anomalous discharges in urban wastewater system. The set of data were constituted of several compounds on which the concentration ranges considerably with treatment and seasonality. An anomalous discharge, the influence of storm water on the wastewater composition and recalcitrant compounds (LAS surfactant homologues) in the effluent were further identified. The seasonal variations and abnormality in the composition of organic compounds in sewage indicated that the employed procedure can be useful to identify the pollution source, to follow up the efficiency of WWTPs plants to design preventive actions in order to protect equipment and preserve the environment.
Atualmente, o crescimento industrial, a urbanização e as atividades rurais, como a agricultura, são um dos responsáveis pela deterioração do meio ambiente, principalmente nos países em desenvolvimento, como o Brasil. Problemas relacionados com a poluição das águas fluviais na América Latina têm sido bem documentados, porém não há evidências de esforços substanciais para alterar essa situação. Existem vários poluentes orgânicos presentes no esgoto tratado pelas Estações de Tratamento de Esgoto (ETEs) que podem causar danos aos ecossistemas e, com isso, ameaçar a saúde humana. As ETEs são responsáveis pela remoção da maioria dos poluentes antropogênicos ou naturais presentes no esgoto sanitário doméstico, e por isso, desempenham um papel fundamental na preservação do meio ambiente e consequentemente da saúde pública. Devido às variações das condições operacionais, processos e qualidade dos descartes, o tratamento do esgoto sanitário doméstico precisa ser monitorado continuamente para assegurar eficiência operacional. Para o estudo de contaminações ambientais, a RMN tem-se configurado como uma valiosa ferramenta, uma vez que ela tem contribuído significativamente no estudo de componentes químicos presentes nas águas e no lodo de esgoto; nas análises de interações com as substâncias húmicas e fúlvicas, além de contaminações ambientais por fármacos, pesticidas, surfactantes e seus produtos de degradação, dentre outros. No presente trabalho, foram propostos dois métodos para estudar as águas oriundas do esgoto anterior e posterior ao tratamento ocorrido na ETE do município de São Carlos-SP: análises de compostos orgânicos não específicos por meio da espectroscopia de RMN 1D (1H e 13C) e 2D (COSY, HSQC e HMBC), com o intuito de caracterizar o maior número de substâncias dissolvidas nas matrizes; análises desenvolvidas através do sistema hifenado HPLC-(UV/MS)-SPE-ASS-NMR, para detectar e elucidar os compostos não específicos recalcitrantes, em baixas concentrações e sem o uso de padrões. Ademais, foram empregadas análises quimiométricas e quantitativas (RMNq) no primeiro método (compostos orgânicos não específicos) para, desta forma, monitorar variações sazonais e identificar descargas anômalas no sistema comum de escoamento do esgoto sanitário doméstico. Portanto, em ambas as matrizes de esgoto (não tratado e tratado pela ETE) foram caracterizados 50 compostos orgânicos, em que as concentrações variaram consideravelmente com o tratamento da ETE e de acordo com a sazonalidade da coleta. Também foram verificados uma descarga anormal no sistema comum de esgoto e influências das águas fluviais sobre a composição das águas de esgoto, além de serem completamente elucidados compostos homólogos recalcitrantes pertencentes a classe dos surfactantes LAS (alquilbenzeno sulfonados de cadeia linear) no efluente. As variações sazonais e anormalidade na composição química no esgoto indicaram que o procedimento empregado pode ser especialmente útil para identificar fontes poluidoras e acompanhar a eficiência do tratamento das ETEs para que, desta forma, sejam tomadas ações preventivas a fim de proteger todo sistema de tratamento e equipamentos e, com isso, minimizar os danos causados ao meio ambiente.

This diploma thesis deals with the determination of macrolide antibiotics in wastewater, especially with erythromycin, clarithromycin and roxithromycin. In this time are these pharmaceuticals prescribed quite frequently. Solid phase extraction (SPE) was used for the isolation and the purification of selected analytes from an aqueous matrix; as the suitable procedure was found the using Oasis HLB cartridges. High performance liquid chromatography with mass spectrometry detection (HPLC-MS) was optimized for its analysis of selected pharmaceuticals. The optimized method was used for the determination of pharmaceuticals in real water samples, which was taken at the inflow and the outflow of the urban wastewater treatment plant in Brno-Modřice.

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Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Atualmente, cerca de 18 compostos são utilizados como biocidas de reforço (metálicos) em tintas anti-incrustantes, como por exemplo Piritionato de Zinco (Zn(PT)2), Zineb e Ziram. É relevante o desenvolvimento de um método analítico para determinação das concentrações ambientais de biocidas metálicos. O presente estudo teve como objetivos i)desenvolver uma metodologia de extração em fase sólida (SPE) e cromatografia líquida de alta eficiência acoplada (HPLC) ao espectrômetro de massas com plasma acoplado indutivamente (ICP-MS) para determinação de Zn(PT)2, Zineb e Ziram e ii) quantificar, in lab, a fração lábil total do Piritionato de Zinco, Zineb e Ziram e in situ, a fração lábil total do zinco, utilizando a técnica de difusão em filmes finos por gradiente de concentração (DGT) em solução padrão e água estuarina, respectivamente. Na técnica de SPE foi utilizado o sorbente de sílica funcionalizado com fenil apresentando excelente retenção para Zn(PT)2, Zineb e Ziram (94 ± 0,1%, 85 ± 0,04% e 93 ± 0,1%, respectivamente) e recuperações entre 85% e 110%. Na determinação dos biocidas de zinco utilizando o acoplamento HPLC-VGroove-ICP-MS com diluição pós coluna cromatográfica, a fase móvel composta por metanol e 0,006 mol L-1 de acetato de amônio (50:50, v v-1) apresentou o melhor desempenho na separação do Zn(PT)2, Zineb, Ziram. A curva analítica obtida para o Zn(PT)2 apresentou coeficiente de correlação, LD e LQ satisfatórios para os isótopos 64Zn (0,98, 0,575 mg L-1, 1,916 mg L-1), 66Zn(0,99, 0,480 mg L-1 , 1,600 mg L-1), 68Zn(0,98, 0,602 mg L-1, 2,007 mg L-1). Na técnica DGT foi utilizado o agente ligante resina Chelex® 100 para avaliar a labilidade do Zn(PT)2, Zineb e Ziram em água estuarina. Os resultados demonstraram que a fração lábil total do Zn ficou em torno de 100% para o Zineb (111%) e Ziram (109%), ou seja, estes biocidas formaram espécies totalmente lábeis na amostra de água estuarina e para Zn(PT)2 foi de 75% indicando espécies parcialmente lábeis. Os resultados obtidos na técnica SPE, no acoplamento HPLC-VGroove-ICP-MS e DGT demonstraram que os métodos apresentam desempenho satisfatório para a determinação de Piritionato de Zinco, Zineb e Ziram.
About 18 compounds are used as booster biocides (metal) in antifouling paints such as Zinc Pyrithione (Zn(PT)2), Zineb and Ziram. It is important to develop an analytical method for determining of the environmental concentrations of zinc biocides. The present study had as objectives i) to develop a solid phase extraction (SPE) and high performance liquid chromatography (HPLC) coupled to the inductively coupled plasma mass spectrometer (ICP-MS) for the determination of Zn(PT)2, Zineb and Ziram and ii) quantify in lab the total labile fraction of Zn(PT)2, Zineb and Ziram and in situ the total labile fraction of zinc using the diffusive gradient in thin films (DGT) technique in standard solution and estuarine water, respectively. In the SPE technique, the silica sorbent functionalized with phenyl presented excellent retention for Zn(PT)2, Zineb and Ziram (94 ± 0.1%, 85 ± 0.04% and 93 ± 0.1%, respectively) and recoveries between 85% and 110%. In the determination of zinc using the HPLC-VGroove-ICP-MS coupling with post-column chromatographic dilution, the mobile phase composed of methanol and 0.006 mol L-1 of ammonium acetate (50:50, v v-1) of presented the best performance in the separation of Zn(PT)2, Zineb and Ziram. The analytical curve obtained for Zn(PT)2 presented satisfactory correlation coefficient, LD and LQ for the isotopes 64Zn (0.98, 0.575 mg L -1, 1.916 mg L-1), 66Zn (0.99, 0.480 mg L-1, 1.600 mg L-1), 68Zn (0.98, 0.602 mg L-1, 2.007 mg L-1). In the DGT technique, the Chelex® 100 resin binder was used to evaluate the lability of the zinc biocides Zn(PT)2, Zineb and Ziram in estuarine water. The results showed that the total labile fraction of Zn was around 100% for Zineb (111%) and Ziram (109%), that is, these biocides formed totally labile species in the estuarine water sample and for Zn(PT)2 was 75% indicating partially labile species. The results obtained in the SPE technique in the HPLC-VGroove-ICP-MS coupling and DGT demonstrated that the methods present satisfactory performance for the determination of Zn(PT)2, Zineb and Ziram.
CNPq: 164326/2015.
FAPESP: 2015/03397-4.

Orientador: Amauri Antonio Menegário
Banca: Didier Gastmans
Banca: Lauren Nozomi Marques Yabuki
Banca: Luciana Polese
Banca: Anne Hélène Fostier
Resumo: Atualmente, cerca de 18 compostos são utilizados como biocidas de reforço (metálicos) em tintas anti-incrustantes, como por exemplo Piritionato de Zinco (Zn(PT)2), Zineb e Ziram. É relevante o desenvolvimento de um método analítico para determinação das concentrações ambientais de biocidas metálicos. O presente estudo teve como objetivos i)desenvolver uma metodologia de extração em fase sólida (SPE) e cromatografia líquida de alta eficiência acoplada (HPLC) ao espectrômetro de massas com plasma acoplado indutivamente (ICP-MS) para determinação de Zn(PT)2, Zineb e Ziram e ii) quantificar, in lab, a fração lábil total do Piritionato de Zinco, Zineb e Ziram e in situ, a fração lábil total do zinco, utilizando a técnica de difusão em filmes finos por gradiente de concentração (DGT) em solução padrão e água estuarina, respectivamente. Na técnica de SPE foi utilizado o sorbente de sílica funcionalizado com fenil apresentando excelente retenção para Zn(PT)2, Zineb e Ziram (94 ± 0,1%, 85 ± 0,04% e 93 ± 0,1%, respectivamente) e recuperações entre 85% e 110%. Na determinação dos biocidas de zinco utilizando o acoplamento HPLC-VGroove-ICP-MS com diluição pós coluna cromatográfica, a fase móvel composta por metanol e 0,006 mol L-1 de acetato de amônio (50:50, v v-1) apresentou o melhor desempenho na separação do Zn(PT)2, Zineb, Ziram. A curva analítica obtida para o Zn(PT)2 apresentou coeficiente de correlação, LD e LQ satisfatórios para os isótopos 64Zn (0,98, 0,575 mg L-1, 1,916 mg L-1... (Resumo completo, clicar acesso eletrônico abaixo)
Abstract: About 18 compounds are used as booster biocides (metal) in antifouling paints such as Zinc Pyrithione (Zn(PT)2), Zineb and Ziram. It is important to develop an analytical method for determining of the environmental concentrations of zinc biocides. The present study had as objectives i) to develop a solid phase extraction (SPE) and high performance liquid chromatography (HPLC) coupled to the inductively coupled plasma mass spectrometer (ICP-MS) for the determination of Zn(PT)2, Zineb and Ziram and ii) quantify in lab the total labile fraction of Zn(PT)2, Zineb and Ziram and in situ the total labile fraction of zinc using the diffusive gradient in thin films (DGT) technique in standard solution and estuarine water, respectively. In the SPE technique, the silica sorbent functionalized with phenyl presented excellent retention for Zn(PT)2, Zineb and Ziram (94 ± 0.1%, 85 ± 0.04% and 93 ± 0.1%, respectively) and recoveries between 85% and 110%. In the determination of zinc using the HPLC-VGroove-ICP-MS coupling with post-column chromatographic dilution, the mobile phase composed of methanol and 0.006 mol L-1 of ammonium acetate (50:50, v v-1) of presented the best performance in the separation of Zn(PT)2, Zineb and Ziram. The analytical curve obtained for Zn(PT)2 presented satisfactory correlation coefficient, LD and LQ for the isotopes 64Zn (0.98, 0.575 mg L -1, 1.916 mg L-1), 66Zn (0.99, 0.480 mg L-1, 1.600 mg L-1), 68Zn (0.98, 0.602 mg L-1, 2.007 mg L-1). In the DGT technique, t... (Complete abstract click electronic access below)
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Les globules rouges suscitent un intéret clinique particulier en raison de leur importance biologique. Il a été récemment démontré que certaines pathologies érythrocytaires pouvaient être associées à une composition lipidique anormale. Parmi ces pathologies figure la maladie de Gaucher, un trouble lysosomal rare causé par une déficience en β-glucocérébrosidase, enzyme dont l'absence entraîne une accumulation excessive de certains sphingolipides dans l'organisme. Cette surcharge lipidique altère les propriétés morphologiques et rhéologiques des globules rouges, provoquant des anomalies hématologiques et vasculaires.Dans cette thèse, nous avons étudié le lien entre la composition lipidique des globules rouges et son implication dans la maladie de Gaucher. En particulier, nous nous sommes concentrés sur deux catégories de lipides : les sphingolipides et les phospholipides, qui jouent un rôle clé dans les altérations érythrocytaires observées. Deux approches complémentaires ont été utilisées : la première utilise la spectrométrie de masse dans le but d'identifier de nouveaux biomarqueurs. La seconde exploite des techniques d'imagerie infrarouge à différentes échelles pour étudier la répartition des lipides dans les globules rouges.Nous avons mis au point une méthode rapide de préparation d'échantillons en ligne avec la chromatographie liquide couplée à la spectrométrie de masse (SPE-HPLC-MS/MS), pour doser 30 sphingolipides et phospholipides en moins de 15 min. Par la suite cette méthode a été appliquée pour quantifier les 30 espèces lipidiques dans le plasma et le culot globulaire de sujets contrôles et de patients Gaucher. Parmi ces espèces lipidiques, certaines étaient surexprimées et n'étaient pas associées à la maladie de Gaucher, ouvrant de nouvelles perspectives pour le diagnostic et le suivi thérapeutique.A l'issue de ce dosage nous nous sommes intéressés à l'exploration du lipidome du globule rouge sain pour pouvoir le comparer à celui du globule rouge Gaucher. Pour cela une approche semi-ciblée a été menée en utilisant un analyseur triple quadripolaire. Ceci nous a permis d'identifier plus de 266 espèces moléculaires réparties sur 12 classes lipidiques différentes.Ces résultats ont contribué à une meilleure connaissance du lipidome du globule rouge sain et sont prometteurs pour l'identification de nouveaux biomarqueurs et la compréhension de diverses pathologies affectant le lipidome érythrocytaire.En complément de cette approche, nous avons étudié la morphologie et la composition chimique des globules rouges en utilisant des techniques d'imagerie vibrationnelle infrarouge à différentes résolutions : à l'échelle microscopique avec la microspectroscopie infrarouge à transformée de Fourier (μ-FTIR), à l'échelle submicronique avec l'imagerie optique thermique (O-PTIR) et à l'échelle nanométrique grâce à l'AFM-IR.Cette étude a été réalisée en trois étapes principales : (1) l'élaboration d'un protocole de préparation permettant la fixation des globules rouges, (2) l'évaluation de leur stabilité chimique et morphologique, et enfin (3) une analyse multi-échelles comparant des globules rouges sains à ceux affectés par la maladie de Gaucher.Grâce à notre protocole de fixation, nous avons pu préserver les globules rouges sans altération pendant 10 jours, ce qui nous a permis de mener à bien l'analyse multi-échelle. L'AFM-IR s'est révélée être la seule technique capable de mettre en évidence des différences significatives entre les globules rouges sains et Gaucher. L'AFM-IR a permis de visualiser les différentes morphologies des cellules Gaucher et de comparer la distribution des lipides à travers des cartographies chimiques, révélant des zones d'intérêt au niveau de la membrane érythrocytaire
Red blood cells attract particular clinical interest due to their biological significance. It has recently been demonstrated that certain erythrocyte pathologies may be associated with abnormal lipid composition. Among these pathologies is Gaucher's disease, a rare lysosomal disorder caused by a deficiency in β-glucocerebrosidase, an enzyme whose absence leads to excessive accumulation of certain sphingolipids in the body. This lipid overload alters the morphological and rheological properties of red blood cells, causing hematological and vascular abnormalities.In this thesis, we studied the link between the lipid composition of red blood cells and its implication in Gaucher's disease. In particular, we focused on two categories of lipids: sphingolipids and phospholipids, which play a key role in the erythrocyte alterations observed. Two complementary approaches were used: the first uses mass spectrometry with the aim of identifying new biomarkers. The second uses infrared imaging techniques at different scales to study the distribution of lipids in red blood cells.We developed a rapid sample preparation method compatible with liquid chromatography coupled to mass spectrometry (SPE-HPLC-MS/MS) to quantify 30 sphingolipids and phospholipids in less than 15 minutes. This method was then applied to quantify the 30 lipid species in the plasma and red blood cell pellet of control subjects and Gaucher patients Among these lipid species, some were overexpressed and were not associatedAfter this quantification, we focused on exploring the lipidome of healthy red blood cells and comparing it to that of Gaucher red blood cells. To achieve this, a semi-targeted approach was conducted using a triple-quadrupole analyzer, which allowed us to identify more than 266 molecular species spread across 12 different lipid classes.These results have contributed to a better understanding of the lipidome of healthy red blood cells and are promising for the identification of new biomarkers and the diagnosis of various pathologies affecting the erythrocyte lipidome.As a complement to this approach, we focused on the morphological and chemical study of red blood cells using vibrational infrared imaging techniques at different resolutions: microscopic with Fourier-transform infrared spectroscopy (FTIR), submicron with optical photothermal infrared imaging (O-PTIR), and nanometric with atomic force microscopy (AFM-IR).Our study was structured around three steps: (1) developing a preparation protocol allowing the fixation of red blood cells, (2) evaluating their chemical and morphological stability, and finally (3) conducting a multi-scale analysis comparing healthy red blood cells with those affected by Gaucher's disease.Due to our fixation protocol, we were able to preserve red blood cells without degradation for 10 days, which enabled us to successfully conduct the multi-scale analysis. AFM-IR proved to be the only technique capable of highlighting significant differences between healthy and Gaucher red blood cells. AFM-IR allowed us to visualize the distinct morphologies of Gaucher cells and to compare lipid distribution through chemical mapping, revealing areas of interesting in the erythrocyte membrane

L'évaluation de l'inertie des matières plastiques au contact de denrées alimentaires est régie par le règlement européen N°10/2011. Au Liban, l'absence de législations relatives à l'usage des matières en plastiques a conduit à des emballages non contrôlés au niveau de la production et du stockage, ce qui peut induire un risque sanitaire pour le consommateur. L'objectif de ce travail a été d'apprécier les phénomènes de migration des contaminants du PET vers l'eau. Pour cela, l'analyse de certains migrants potentiels a été initiée tels que des COV et des SV afin de vérifier la conformité de leur teneur par rapport aux LMA dans les règlementations internationales. Le dosage des aldéhydes dans l'eau a été effectué par dérivation avec la 2,4-DNPH, suivie d'une étape de préconcentration sur une cartouche en C18, puis séparation et quantification par HPLC-UV. L'analyse des composés SV a été réalisée par SPME-GC-FID sur une fibre en PDMS/DVB. L'analyse des BTEX et du styrène a été réalisée par HS-Trap-GC-FID. Ces méthodes ont été validées conformément à la norme NF T 90-210 avec des LQ inférieures aux LMA. L'analyse des COV dans l'eau embouteillée et dans le PET a été réalisée par HSTrap- GC-MS. La présence de FA, d'AA, de DEP et des BTEX dans l'eau embouteillée en PET a été confirmée. Il a été mis en évidence que la température et l'exposition solaire influencent la migration des aldéhydes. Les concentrations d'AA trouvées après incubation à 40ºC ne sont pas négligeables et peuvent modifier les propriétés organoleptiques de l'eau. L'analyse du PET montre la présence des hydrocarbures, des aldéhydes, des alcools, et des cétones
The inertness’ value of plastics that are on contact with foodstuffs is governed by the European rule N°10/2011. In Lebanon, the lack of the Rules and Regulations of plastics’ usage led for a non-controlled packing in production as in storage, which will cause a sanitary risk to the consumer and to the population. The purpose of this research was to appreciate the migration’s phenomena of contaminants from the PET to the water. For this, there was an analysis of potential migrants such as VOC and SV to verify their tenor’s conformity relative to the MCL in international regulations. The aldehydes’ dosage in the water was performed by the derivation with the 2,4-DNPH, followed by a concentration on a C18 cartridge, then separation and quantification was done by HPLC-UV. The SV compounds’ analysis was realized by SPME-GC-FID method on a PDMS / DVB fiber. The BTEX and the styrene’s analysis was done by HS-trap-GC-FID method. These methods were validated in accordance with standard NF T 90-210 with a LQ less than the MCL. The volatile compounds in the bottled water and in the PET were analyzed by HS-trap-GC-MS method. The presence of FA, AA, DEP and BTEX in the bottled water with the PET was confirmed. It was highlighted that temperature and sun exposition influence the aldehydes’ migration. The acetaldehyde’s concentration after incubation at 40ºC wasn’t negligible and can modify the water’s organoleptic properties. The PET’s analyze reveals the presence of hydrocarbons, aldehydes, alcohols and ketones

The theoretical part of this diploma thesis consists of current matters regarding the presence of perfluorinated organic compounds in living environment elements. The attention has been focused on perfluoroktanoic acid (PFOA), (PFOS), and (FOSA). Physical-chemical characteristics of these organic pollutants and their toxicological and environmental aspects have been described. Possible ways of the pollutants analytical determination have been specified. The experimental part has explored effects of extraction techniques (sonication, pressurized solvent extraction, solid-phase extraction) used for PFAS separation from seats of fire soil sample. Identification of the analytes have been achieved by liquid chromatography/mass spektrometry.

Pharmaceuticals are biological active compounds with different functional groups, physico-chemical and biological properties. These chemical compounds are called as “new contaminants” which cumulate in various environmental components. These contaminants input to environment from industrial processes, hospitals and health care institutions or household sources and these pollutants can to negatively interact with environmental components. Pharmaceuticals are separated according to structure and their effects to live organisms. On the basis of drug consumption in Czech Republic the non-steroidal anti-inflammatory drugs are the most using pharmaceuticals, because these drugs can be obtain without prescriptions. The choosing analgesics for study were monitored in waste water from waste waters treatment plant (WWTP Brno – Modřice) and in surface water from two sampling sites of river Kretinka. Pharmaceuticals were monitored in surface waters in Czech Republic, in Scotland (river Thurso) and in Taiwan (river Erren, Agongdian, Yanshuei and canal of Tainan) Solid phase extraction (SPE) is used for extraction of study pharmaceutical from water system. The high performance liquid chromatography (HPLC) with diode array detection or mass spectrometry was used for determination of drugs in surface waters from Czech Republic. The water samples from Scotland and Taiwan were analysed and the high performance liquid chromatography with mass spectrometry (HPLC/MS) was used for determination of pharmaceuticals. The method of HPLC/MS enables the determination of study compounds in the very low range of concentrations (ng ? l-1). All monitored pharmaceuticals were identified and quantified in water samples from river Kretinka in Czech Republic, river Thurso (Scotland) and rivers Erren, Agongdian, Yanshuei and canal of Tainan in Taiwan and the obtained data were compared.

Les Hydrocarbures Aromatiques Polycycliques (HAP) sont principalement émis par les activités humaines et représentent l'un des principaux groupes de polluants dans l'air, les sols, les eaux et les aliments. Par conséquent, ils font l'objet d'une surveillance atmosphérique, et alimentaire. Cette étude s'attache au cas particulier de la femme enceinte et son fœtus. En effet, l'exposition pendant la grossesse est à l'origine de malformations fœtales, de prématurité et de troubles pouvant survenir ensuite chez l'enfant mais aussi chez l'adulte. À ce jour, seuls quelques HAP réglementés par l'environnement ont été recherchés et quantifiés dans le sang maternel ou le sang de cordon ombilical. De plus, les données sur les performances analytiques sont souvent manquantes. Ainsi ce projet vise pour la première fois en France à développer et valider des méthodes analytiques et biologiques pour la surveillance de l'exposition aux HAP durant la grossesse. Dans une première partie, la quantification simultanée des 24 HAP réglementés dans les sérums de sang maternel et de sang de cordon ombilical a été développée. Tout d'abord, l'analyse des 24 HAP par chromatographie en phase liquide et par détections UV et fluorescence (HPLC-UV-FD) et par chromatographie en phase gazeuse couplée à la spectrométrie de masse (GC-MS) a été optimisée. Le prétraitement des échantillons de sérum a ensuite été développé. La précipitation des protéines présentes dans le sérum a été optimisée avec un plan d'expérience afin d'interrompre toutes les interactions entre les protéines et les HAP. Un protocole d'extraction sur phase solide (SPE), avec un supportt à base de silice greffée C18, a ensuite été optimisé pour extraire et concentrer les HAP. Pour améliorer les facteurs d'enrichissement, les fractions d'élution de SPE ont ensuite été évaporées et 2 approches ont été évaluées: une évaporation totale ou partielle (méthode "dernière goutte"). Le protocole analytique final impliquant la préparation d'échantillon suivi par l'analyse en HPLC-UV-FD ou GC-MS a été validé avec succès à partir d'un mélange de sérums maternels dopés et un mélange de sérums dopés provenant de sangs de cordons ombilicaux avec l'approche des profils d'exactitude. Ensuite, des échantillons de sérum provenant de sang maternel et de sang de cordon ombilical ont été analysés par HPLC-UV-FD et GC-MS. Enfin, les méthodes analytiques développées précédemment ont été utilisées pour l'étude du transfert placentaire des HAP avec le modèle de perfusion du cotylédons humains ex vivo. Les perfusions ont été réalisées pour la première fois avec un mélange de 14 HAP (concentration totale : 1 microM) et leur transfert placentaire a été démontré. De plus, des cultures cellulaires de trophoblastes humains exposées avec ce mélange de 14 HAP (concentration totale : 1 microM) ont été réalisées pour évaluer l'impact de l'exposition aux HAP sur les fonctions placentaires. Le dysfonctionnement de certaines fonctions placentaires telles que la fusion et la différentiation des trophoblastes, les fonctions endocrines et les fonctions de détoxifications
Polycyclic Aromatic Hydrocarbons (PAHs) are mainly emitted by human activities and are one of the main pollutant groups in air, soils, waters, and food. Therefore, they are subject to atmospheric, environmental and food monitoring. This study focuses on the particular case of exposure of pregnant women and their fetus, which are at risks. Indeed, PAH exposure can lead to fetal malformations, prematurity and even disorders in children and adult. To date, only a few PAHs regulated by US-EPA were searched and quantified in maternal or umbilical cord blood. Moreover, analytical performance data are often lacking. Hence, this study aims for the first time in France, at developing analytical and biological methods for the monitoring of PAH exposure during pregnancy. In a first part, the simultaneous determination of the 24 regulated PAHs in sera from maternal and umbilical cord bloods was developed. First, the analysis of the 24 PAHs by liquid chromatography and UV and fluorescence detections (HPLC-UV-FD) and by gas chromatography hyphenated to mass spectrometry (GC-MS) were optimized. The sample pretreatment of the plasma samples was next developed. The precipitation of the proteins present in the plasma was optimized with a Design of Experiment to disrupt all the interactions between them and the PAHs. A solid phase extraction (SPE) protocol, with a C18-based sorbent, was next optimized to extract and concentrate the PAHs. To improve the enrichment factors, the elution SPE fractions were next evaporated and 2 approaches were evaluated: a total or a partial evaporation ("last drop" method). The final analytical protocol involving the sample pretreatment followed by HPLC-UV-FD or GC-MS was successfully validated with spiked pooled sera from maternal and umbilical cord bloods with the accuracy profile approach. Then, sera samples from maternal and umbilical cord bloods were analyzed by LC/UV-FD and GC/MS. Finally, the previously developed analytical methods were used for a study with the ex vivo human cotyledon perfusion model. Perfusions were carried out for the first time with a mix of 14 US-EPA PAHs and their placental transfer was demonstrated. In addition, human trophoblast cell cultures with 1 µM with 14 US-EPA PAHs were carried out to evaluate the impact of PAH exposure on placental functions. The dysfunction of certain placental functions such as trophoblast fusion and differentiation, endocrine functions and detoxification functions was demonstrated

L’utilisation intensive de certains pesticides et leur relative persistance vont de pair avec la présence de résidus dans l’eau de surface et l’eau potable mais aussi dans les produits agricoles disponibles pour les consommateurs, y compris les denrées alimentaires. À l’heure actuelle, les effets des pesticides sur la vie aquatique et d’autres organismes non ciblés sont relativement bien connus, et la possibilité des effets sur l’être humain fait débat. Des normes de qualité ont été proposées pour l’eau, que ce soit des critères pour l’eau potable ou des critères de protection de la vie aquatique pour l’eau de surface. Des limites maximales de résidus (MRL) de pesticides ont également été établies pour certains produits, notamment les fruits et légumes. Un des défis pour les chercheurs est la mise en œuvre de nouvelles méthodes analytiques sensibles et robustes pour la quantification ultra-trace de ces composés, afin de déterminer si les différents échantillons sont conformes aux directives ou aux MRL. L’analyse des pesticides modérément polaires dans des matrices complexes repose tout d’abord sur la méthode d’extraction. Plusieurs options sont disponibles, telles que l’extraction liquide-liquide ou en phase solide (SPE, Solid Phase Extraction) pour les matrices aqueuses, ou encore dSPE de type QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) pour les matrices solides. Actuellement, la chromatographie en phase liquide couplée à la spectrométrie de masse en tandem représente un choix pertinent pour les analyses ultra-traces, mais sa mise en œuvre peut présenter certains défis. Dans ce contexte, les principaux objectifs de ce travail de recherche sont les suivants : i) proposer des méthodes analytiques rapides, sensibles et robustes pour déterminer des pesticides multi-classes aux niveaux d’exposition que l’on retrouve dans différentes matrices comme l’eau potable, les denrées alimentaires et l’urine comme matrice biologique, et ii) évaluer le lien entre les sources de contamination des divers pesticides et leur mobilité afin de documenter la distribution spatiale et temporelle dans l’eau de surface et l’eau potable au Québec. Pour les échantillons aqueux, une méthode SPE en ligne entièrement automatisée couplée à la chromatographie liquide haute performance et spectrométrie de masse en tandem a été développée. La méthode proposée est rapide (8 min par échantillon) avec des limites de détection comprises entre 0.1 et 5 ng L-1 pour les pesticides de la famille des néonicotinoïdes et l’atrazine. Pour les produits alimentaires tels que les fruits et légumes, l’optimisation d’une méthode de type QuEChERS a été réalisée. La méthode permet d’atteindre des niveaux de détection entre 0.05 ng g-1 et 2 ng g-1 pour une gamme de 22 pesticides couvrant 7 classes différentes, incluant les organophosphorés, les carbamates, les néonicotinoïdes et les triazines, entre autres. La robustesse des diverses méthodes a été démontrée par des expériences de contrôle qualité inter- et intra-journaliers afin de garantir l’exactitude, la précision et l’absence d'effets matriciels pour de longues séquences d’analyse. Les méthodes validées ont été appliquées à des échantillons réels, y compris des échantillons d’eau du robinet couvrant 52 villes de la province du Québec (Canada), 68 échantillons d’eau de surface (fleuve Saint-Laurent et tributaires), et 133 échantillons de laitue, pomme, raisin et tomates achetés sur les marchés locaux. Les résultats indiquent une forte occurrence de l’atrazine, la thiaméthoxame, la clothianidine et l’imidaclopride dans les échantillons d’eau et les quatre produits alimentaires.
The extensive use of certain pesticides and their relative persistence go on par with the presence of residue levels in surface water and drinking water, but also in agricultural products available to consumers (including foodstuffs). There are potential effects on aquatic life and non-target organisms, and the possibility of effects in humans remains a topical issue. Quality standards have been proposed for water, including criteria for drinking water and criteria for the protection of aquatic life (surface water). Maximum residue limits (MRLs) for pesticides have also been established for foodstuff, including fruits and vegetables. One of the challenges for researchers is the implementation of sensitive and robust analytical methods for the ultra-trace quantification of these compounds, with a view to determining whether the samples are compliant with guidelines or MRLs. The analysis of moderately polar pesticides in complex matrices relies notably on the extraction method. Diverse options are available, including liquid-liquid or solid phase extraction (SPE) for aqueous samples, and dSPE approaches such as QuEChERS (Quick, Easy, Cheap, Effective, Rugged and Safe) for solid samples. Liquid chromatography coupled to tandem mass spectrometry is usually selected for separation and detection at the ultra-trace level, but there are some pitfalls. In this context, the main objectives of the present research were as follows: i) to propose fast and robust analytical methods to determine multi-class pesticides at different exposure routes including drinking water and food, and ii) to evaluate the link between the contamination sources of various pesticides and their mobility to document their distribution in surface water and tap water in Quebec. For water samples, a fully automated on-line SPE method coupled to ultra-highperformance liquid chromatography tandem mass spectrometry was developed. The proposed method is rapid (8 min per sample) with detection limits between 0.1 and 5 ng L- 1 for neonicotinoids and atrazine. For food products (fruits and vegetables), a QuEChERS method was investigated. The optimized procedure shows limits of detection between 0.05 ng g-1 and 2 ng g-1 for a total of 22 pesticides encompassing 7 different classes, including organophosphorus compounds, carbamates, neonicotinoids and triazines, among others. The robustness of the various methods has been demonstrated by inter-day and intra-day iv quality control experiments to ensure suitable accuracy, precision, and the absence of matrix effects in long LC-MS batch sequences. The validated methods were applied to real samples, including tap water samples from 52 municipalities in the province of Quebec (Canada), 68 surface water samples from the St. Lawrence River and its main tributaries, and 133 fruits and vegetables samples (lettuce samples, apples, grapes and tomatoes) purchased from local markets. The results indicate a high occurrence of atrazine, thiamethoxam, clothianidin, and imidacloprid in the water samples and the four food products.

Modern separation and spectrometric techniques for biological sample lipidom investigation Due recent progress in field of mass spectrometry the lipidomics, part of metabolomics, is increasing its importance for broad fields of biological study. The aim of this study is to test the lipid extraction techniques and to optimize the preseparation and separation of lipids suitable for mass spectrometry detection. The fragmentation patterns of four, the most abundant lipid classes of glycerolipids (PC, PE, TG, DG), were acquired for the proposed system. These patterns were compared with literature. The most appropriate method for extraction was declared technique according Folch based on methanol and chloroform solution. The preseparation due SPE method is very useful tool for lipid determination. The optimized were focused to reach higher recovery especially in polar lipid fraction. Proposed HPLC system is based on methanol with ammonium buffer, water and isopropanol. The testing was done on three columns with different type of sorbents (Gemini, Syncronis and Kinetex). The separation was evaluated according mass spectrometer response, shape and wide of particular analytes peaks. Composition contains 20% of water was determinate as the best and also the best separation was achieved by Kinetex column. The proposed...

Wprowadzenie do powszechnego użytku katalizatorów samochodowych spowodowało wzrost emisji metali z grupy platynowców (PGE) do środowiska. Podwyższony poziom zawartości platyny, palladu i rodu jest obserwowany szczególnie w okolicach dużych szlaków komunikacyjnych, rzadko jednak przekracza 1 μg g-1. Celem pracy doktorskiej było określenie wpływu, jaki pierwiastki z tej grupy wywierają na środowisko. Aby to było możliwe konieczne jest dysponowanie metodami gwarantującymi zarówno oznaczanie całkowitych zawartości analitów na odpowiednich poziomach stężeń, jak i śledzenie przemian jakim ulegają i identyfikację ich form chemicznych. Aby uzyskane wyniki oznaczeń były wiarygodne należy szczególną uwagę zwrócić na sposób przygotowania badanych materiałów do analizy. Pod względem analitycznym, oznaczanie śladowych ilości platyny, rodu czy palladu w próbkach środowiskowych nie jest zadaniem prostym. Złożoność matrycy próbek oraz niski poziom stężeń sprawiają, że metody oznaczeń muszą charakteryzować się odpowiednio niskimi granicami oznaczalności. Często w procedurze analitycznej wprowadza się dodatkowy etap zatężania analitów i uproszczenia matrycy próbki. W ramach prowadzonych badań zoptymalizowano procedury roztwarzania próbek gleb i piasków kwarcowych z wykorzystaniem mieszaniny kwasów HNO3 i HCl. W oparciu o wyniki uzyskane dla certyfikowanego materiału odniesienia – pyłu drogowego BCR-723 wykazano efektywne przeprowadzenie PGE do roztworu. Do wydzielania analitów z matrycy wykorzystano metodę ekstrakcji do fazy stałej. W trakcie badań zastosowano sorbenty kationo- i anionowymienne oraz eluenty o różnej sile elucyjnej: 0,1 mol L-1 tiomocznik w 0,1 mol L-1 HCl, 2 mol L-1 HCl, 0,025-0,053 mol L-1 bufor amonowy. Opracowane procedury przygotowania próbek zastosowano do monitorowania PGE w próbkach gleb i piasków kwarcowych, pobieranych z poletek monitoringowych rozmieszczonych wzdłuż ciągów komunikacyjnych i eksponowanych na zanieczyszczenia związane z ruchem drogowym. Szczególną uwagę zwrócono na możliwość zastosowania w oznaczeniach metody woltamperometrii inwersyjnej z adsorpcyjnym zatężaniem (AdSV), która, ze względu na niskie granice oznaczalności może być alternatywą do metody spektrometrii mas z plazmą indukcyjnie sprzężoną (ICP MS). Na podstawie uzyskanych wyników wykazano wpływ ruchu drogowego na wzrastające skażenie środowiska oraz wskazano katalizatory samochodowe, jako główne emitery pierwiastków z grupy platynowców. Odkładanie się PGE w glebach sprawia, że mogą być one pobierane i akumulowane przez rośliny, co może powodować zaburzenia ich prawidłowego funkcjonowania. W oparciu o materiał roślinny pozyskany w ramach upraw hydroponicznych określono wpływ platyny, palladu i rodu na rozwój gorczycy białej (Sinapis alba L.). Rośliny narażone na stres związany z obecnością różnych soli i nanocząstek PGE zostały scharakteryzowane pod kątem całkowitej zawartości metali pobranych z pożywki i przetransportowanych do części nadziemnych (ICP MS), obecności nanocząstek w wybranych tkankach (SP ICP MS, TEM), jak i tworzenia kompleksów z ligandami obecnymi lub syntezowanymi de novo w roślinie. Powstałe w tkankach fitochelatyny (PC2, PC3 i PC4) zidentyfikowano metodą wysokosprawnej chromatografii cieczowej z detekcją fluorescencyjną (HPLC FLD) oraz spektrometrią mas z jonizacją przez elektrorozpraszanie (ESI MS). Dodatkowo dzięki wykorzystaniu metody SEC ICP MS, wykryto i rozdzielono grupy związków palladu powstających w komórkach Sinapis alba L. Z kolei zastosowanie metody HILIC ESI MS umożliwiło zidentyfikowanie wybranych połączeń Pd z ligandami organicznymi, jak histydyna i nikotianamina, odpowiedzialnymi za procesy detoksykacji metali.
Introduction of car catalysts for general use caused an increase in emissions of platinum group elements (PGEs) to the environment. The increased level of platinum, palladium and rhodium is observed, especially in the vicinity of main communication routes, but still, rarely exceeds 1 μg g-1. The aim of the doctoral thesis was to determine an impact which PGEs may have on the environment. To make that possible, it's necessary to develop procedures, guaranteeing both, determination of the total content of analytes at appropriate concentration levels, as well as monitoring the changes they undergo and identification of their chemical forms. In order to get the reliable results, particular attention should be paid to the proper way of sample preparation before analysis. The determination of trace amounts of platinum, rhodium or palladium in environmental samples is not a simple task. The complexity of the sample matrix and the low concentration level of analytes cause that the determination methods must have sufficiently low limits of quantification. Often an additional step of analytes pre-concentration and separation from the matrix is introduced in the analytical procedure. As a part of the conducted research, the digestion procedures of soil and quartz sands samples, using the mixture of HNO3 and HCl acids, were optimized. Based on the results obtained for certified reference material (BCR-723; road dust) the possibility of efficient analytes transfers into the solution was proven. To isolate analytes from the matrix the solid phase extraction method was suggested. During the experiments, cation and anion exchange sorbents, as well as the eluents of various elution strength were used: 0.10 mol L-1 thiourea in 0.10 mol L-1 HCl, 2.0 mol L-1 HCl, 0.025-0.053 mol L-1 ammonium buffer. Developed sample preparation procedures were used to monitor PGEs in soil and quartz sands samples received from the monitoring plots distributed along high-ways and exposed to traffic-related contamination. Particular attention was paid to the possibility of using adsorptive stripping voltammetry (AdSV), which, due to low limits of quantification, can be an alternative method to inductively coupled plasma mass spectrometry (ICP MS). Based on the results obtained, the influence of road traffic on the increasing environmental pollution was proven, along with the indication of car catalysts as main emitters of the platinum group elements. The deposition of PGEs in soils makes that they can be taken up and accumulated by plants, which may cause disturbances in their proper functioning. The influence of platinum, palladium and rhodium on the growth of white mustard (Sinapis alba L.) was defined based on the plant material obtained from hydroponic cultivation. The plants, exposed to stress from the presence of various salts and nanoparticles of PGEs, have been characterized for the total content of metals taken from the nutrient solution and transported to aboveground organs (ICP MS), the presence of nanoparticles in selected tissues (SP ICP MS, TEM) and the formation of complexes with ligands present or de novo synthesized in the plant. The phytochelatins (PC2, PC3 and PC4), formed in plant cells, were identified by high-performance liquid chromatography with fluorescence detection (HPLC FLD) and electrospray ionization mass spectrometry (ESI MS). Additionally, thanks to the use of the SEC ICP MS method, some palladium compounds formed in Sinapis alba L. cells were detected and separated. The application of HILIC ESI MS method enabled identification of several Pd complexes with organic ligands, like histidine and nicotianamine, responsible for metal detoxification processes.