Veterinary drugs and growth promoters are a part of many chemical hazards that can be found in the modern food chain, and they pose risks to human health such as antimicrobial resistance. The presence of these drug residues increased with intensive farming practices and is an issue of global concern. Food safety authorities around the world establish control programs by scientifically assessing the risk for each drug and set maximum levels for each drug in various matrices. The testing laboratories and analytical methods to test samples from food and feed chain form the core of such programs and provide evidence for regulatory authorities to take decisions. In the drug residue testing, there is a need for improved methods of testing that can provide high selectivity, high throughput, robustness and affordability. In that sense, one main goal of this thesis was to develop analytical methodologies for various families of veterinary drugs and growth promoters in feed, food and biological samples from food producing animals. First three chapters form the introduction of this thesis. Chapter 1 provides an overview on food safety hazards, food safety authorities, legislations for control of veterinary drugs and growth promoters and the role of analytical laboratories in the control program. Chapter 2 covers sample preparation techniques and the typically used chromatographic (LC) and detection (MS) techniques for drug residue testing. Chapter 3 reviews various applications in literature and focus on trends in developing analytical methodologies for food safety testing which move in the direction of comprehensive analysis of samples owing to very high number of contaminants and increasing number of unknown contaminants. Chapter 4 presents a method developed and validated for analyzing eight sulphonamides in six types of feed based on liquid chromatography (LC) and ultraviolet detection. Chapter 5 presents the results from a systematic study of various hydrophilic interaction chromatography (HILIC) stationary phases for analysis of aminoglycosides and also reports two analytical methods based on HILIC- tandem mass spectrometry (MS/MS) for analysis of ten aminoglycosides in honey and animal kidney samples. This work was carried out in collaboration with laboratory of Agencia Salut Publica de Barcelona (ASPB) and the methods were validated according to Decision 2002/657/EC and implemented for routine analysis of samples in accordance with the requirements of ISO 17025: 2005. The laboratory of ASPB acquired an LC- high resolution mass spectrometry (HRMS) instrument and in order to incorporate the instrument for routine targeted analysis, an exploration of various parameters of the equipment was necessary. In that sense, a systematic study to explore various modes of acquisition has been conducted and a method to analyze nine hormones in urine has been transferred to HRMS and is reported in Chapter 6. Gaining understanding about the functioning of the instrument, an improved method to analyze aminoglycosides based on HRMS was developed and reported in Chapter 5.4. Moreover, the LC- HRMS instrument with quadrupole-orbitrap hybrid analyzer has been incorporated in the laboratory for resolving ambiguous test results. A case study of resolving a false positive result from the analysis of ronidazole in meat was presented in Chapter 7. This case study highlights the pitfalls with low resolution mass spectrometry and existing confirmation criteria for identification with mass spectrometric detection. The HRMS in food safety testing for comprehensive analysis of samples, a non targeted screening workflow using various HRMS data mining and analysis tools have been developed and presented in Chapter 8. At the end of this thesis, conclusions and outlook are presented followed by a list of bibliographic literature. / Los medicamentos veterinarios y promotores del crecimiento forman parte de los muchos peligros químicos que se pueden encontrar en la cadena alimentaria moderna. Estos pueden representar un riesgo para la salud humana puesto que están relacionados con la aparición de la resistencia a los antimicrobianos. La presencia de estos residuos de medicamentos ha aumentado debido a las prácticas de agricultura intensiva y es un tema de preocupación global. Las autoridades de seguridad alimentaria establecen programas de control mediante la evaluación científica del riesgo de cada medicamento, y también establecen los niveles máximos de cada medicamento en diversas matrices. Los laboratorios de ensayo y métodos analíticos para analizar muestras de alimentos y piensos forman el núcleo de este tipo de programas y proporcionan evidencias a las autoridades reguladoras en su toma de decisiones. En el campo de análisis de residuos, existe una necesidad de mejorar los métodos de análisis para que puedan proporcionar una alta selectividad, alto rendimiento, elevada robustez y razonable asequibilidad. En ese sentido, el objetivo principal de esta tesis fue el desarrollo de metodologías analíticas para varias familias de fármacos de uso veterinario y promotores del crecimiento en piensos, alimentos y muestras biológicas de los animales que producen alimentos. Los primeros tres capítulos son la introducción de esta tesis. El Capítulo 4 presenta un método desarrollado y validado para el análisis de ocho sulfonamidas en seis tipos de piensos mediante cromatografía de líquidos (LC) y detección ultravioleta. El Capítulo 5 presenta los resultados de un estudio sistemático de diferentes fases estacionarias de cromatografía de interacción hidrófila (HILIC) para el análisis de aminoglucósidos y también reporta dos métodos analíticos basados en espectrometría de masas en tándem acoplada a HILIC para el análisis de diez aminoglucósidos en muestras de riñón de animales y miel. Un estudio sistemático para explorar distintos modos de adquisición en un instrumento de espectrometría de masas de alta resolución (HRMS, Q-Orbitrap) acoplada a cromatografía líquida y un método para analizar nueve hormonas en orina se detalla en el capítulo 6. Un método mejorado para analizar los aminoglucósidos basado en HRMS fue desarrollado y se presenta en el Capítulo 5.4. Un estudio de un caso práctico sobre la resolución de un resultado falso positivo en análisis de ronidazol en carne se presenta en el Capítulo 7. Se ha desarrollado un workflow de análisis non targeted utilizando diversas herramientas de minería de datos y de análisis, que se ha aplicado a una muestra de sedimento y presentado en el capítulo 8.

Tesi realitzada a l'Institut de Diagnosi Ambiental i Estudis de l'Aigua del Consell Superior d'Investigacions Científiques i al Laboratori Dr. Oliver-Rodés, S.A. / The thesis with title "Identification of plastic material constituents in water and development of migration assays" has as the main objective the identification of the plastic components in the water for human consumption and evaluate the risk according to their migration ability, toxicity and daily intake. To achieve this goal, it was developed analytical methods based on solid phase extraction (SPE) followed by gas chromatography couplet to mass spectrometry (GC-MS) and based on stir bar sorptive extraction (SBSE) followed by gas chromatography coupled to tandem mass spectrometry (GC-MS/MS) for the determination of the plastic components and other environmental contaminants in water springs and bottled waters. Once developed, the presence of plasticizers and herbicides was determined in the springs and fountains of Spain for most of the bottled waters. The same bottled waters were analysed after packaging and after one year stored. Determination of plastic material components and other contaminants was also done in several bottled water sampled in countries around the world that were packaged in plastic containers made of polyethylene terephthalate (PET). In order to determine the migration of plastic components in water, migration assays were performed for the most common plastics used in bottle packaging. Among the analysed plastics, there were polycarbonate (PC) and its possible substitute, TritanTM. Both plastics were evaluated with in vitro and in vivo toxicology tests to determine the endocrine disrupting activity of the compounds identified in the migration assays, as for example bisphenol A (BPA) that was identified in PC samples. Finally, migration tests were performed for domestic water treatment devices to assess the possible changes in the composition of the water according to the migration of contaminants. / La tesi doctoral amb títol "Identification of plastic material constituents in water and development of migration assays" té com objectiu principal identificar la presència de components del plàstic a l’aigua destinada al consum humà i avaluar el seu risc segons la seva capacitat de migració, toxicitat i ingesta diària. Actualment existeix una preocupació creixent pel progressiu augment en la quantitat d’espècies químiques que s’alliberen al medi ambient o són incorporades a la dieta com additius alimentaris o fàrmacs. A nivell mundial es sintetitzen de forma rutinària més de 60.000 espècies químiques i cada any s’afegeixen entre 200 i 1000 espècies noves a la llista. També cal afegir la creixent llista de fàrmacs i cosmètics, en molts casos dissenyats per assemblar-se o contrarestar hormones naturals, anomenats disruptors endocrins. Els disruptors endocrins han generat una alarma global durant les últimes dècades. Es tracta de compostos que afecten al metabolisme hormonal dels éssers vius i es caracteritzen per tenir un efecte biològic a concentracions molt baixes. Els disruptors endocrins es troben al medi ambient, aliments i aigües. I degut a què les aigües minerals estan exposades a aquests compostos ja sigui per contaminació mediambiental o pels mètodes d’envasatge i emmagatzematge, ha augmentat la preocupació socioeconòmica i política per la presència en les aigües de consum. Per aconseguir l'objectiu mencionat anteriorment es van desenvolupar metodologies analítiques basades en l’extracció en fase sòlida (SPE) seguida de cromatografia de gasos acoblada a espectrometria de masses (GC-MS) i extracció per adsorció en barretes agitadores (SBSE) seguida de cromatografia de gasos acoblada a espectrometria de masses en tàndem (GC-MS/MS) per a la determinació de components del plàstic i altres contaminants ambientals en aigües de deus i aigües envasades. Un cop desenvolupades, es va determinar la presència de plastificants i herbicides en els punts d’emergència o captació de deus i brolladors espanyols, en la corresponent aigua envasada i en l’aigua emmagatzemada durant un any per avaluar la migració de contaminants durant el procés d’envasat i emmagatzematge. Pel què fa a l'anàlisi d'aigües envasades també es va determinar la presència de contaminants ambientals i els components del plàstic procedents de la migració dels envasos de polietilè tereftalat (PET) en aigües de vint-i-set països d’arreu del món. Per a poder determinar la migració de components del plàstic a l'aigua es van realitzar estudis de migració forçada en els principals tipus d’envasos comercials d’aigua per a avaluar el tipus i quantitat de compostos que es desprenen dels diferents materials polimèrics. Entre els plàstics que es van analitzar hi havia el policarbonat (PC) i el seu possible substitut, el TritanTM. En ambos plàstics es va avaluar l’activitat de disrupció endocrina del bisfenol A (BPA) i altres components del plàstic presents en els extractes de PC i el compostos migrats del plàstic TritanTM mitjançant assajos de toxicitat in vitro i in vivo. Finalment, també es van realitzar assaigs de migració en aparells domèstics de tractament d’aigua per a avaluar els possibles canvis en la composició de l’aigua en referència als contaminants que puguin migrar.

Las zonas costeras son las regiones más vulnerables de todo el medio marino, especialmente por su colindancia con las zonas de desarrollo humano tierra adentro. La escasez de estudios relacionados con la presencia, distribución e impacto de los contaminantes orgánicos en estas zonas marinas ha sido el detonante para la elección del tema de este trabajo. Para realizar una adecuada vigilancia ambiental del medio marino de compuestos orgánicos emergentes y prioritarios, se desarrollaron 3 metodologías multiresiduales utilizando como técnica de análisis, identificación y cuantificación a la cromatografía de gases acoplada a espectrometría de masas y diferentes técnicas de preconcentración/aislamiento como la extracción en fase sólida (SPE), extracción por adosorción en barras agitadoras (SBSE) y extracción asistida por ultrasonidos (UAE). Las metodologías multiresiduales mostraron un excelente rendimiento (entre 69 y 120%) y una buena sensibilidad (límites de detección entre 0.01 a 38 ng L-1 para aguas de mar) lo cual las hace óptimas para ser utilizadas como herramientas en la vigilancia ambiental de contaminantes orgánicos prioritarios y emergentes. Estas permitieron el análisis de 51 contaminantes orgánicos semivolátiles de las familias de los hidrocarburos policíclicos aromáticos, alquilfenoles, policlorobifenilos, polibromodifenil éteres, pesticidas organoclorados, ftalatos y bisfenol A en aguas de mar, aguas residuales, aguas de río, sedimentos y mejillones. Las técnicas fueron aplicadas para el análisis de muestras recolectadas en la zona del litoral Catalán y costas y estuarios de las provincias de Asturias-Cantabria-País Vasco en España. Adicionalmente, se evaluó la presencia de compuestos perfluorados en la región del litoral Catalán. Los estudios de vigilancia ambiental permitieron identificar 47 contaminantes orgánicos en aguas costeras de las dos regiones de España, siendo las concentraciones detectadas mayores en estuarios y puertos que en costas y bocas de ríos. Las concentraciones detectadas se encontraron por debajo de los Niveles de Calidad Ambiental establecidos por la Directiva 105/2008/CE, excepto en algunos sitios con un mayor impacto industrial, urbano y portuario. Las descargas de efluentes y el transporte de los ríos fueron identificados como las principales fuentes de contaminación de las aguas costeras. Es evidente que existen otras fuentes más, como las actividades marítimas y escorrentías agrícolas, urbanas e industriales. La contaminación descargada a través de los emisarios submarinos de las estaciones depuradoras de aguas residuales (EDAR) se diluye tanto de manera horizontal como vertical y esparcen los contaminantes por las aguas costeras. Además de distribuirse en la columna de agua, los contaminantes orgánicos prioritarios y emergentes estudiados se acumularon en sedimentos, pudiendo ser por tanto disponibles para organismos bentónicos como los mejillones. Se demostró la bioacumulación de contaminantes carcinogénicos y disruptores endocrinos en zonas con elevado impacto antropogénico a través de mejillones trasplantados y expuestos durante 21 días. Esta bioacumulación podría representar un riesgo para otras especies de organismos expuestos a los elevados niveles de contaminantes. Se realizó un análisis de riesgo por medio de cocientes de riesgo o RQs y del denominado procedimiento COMMPS o sistema combinado de fijación de prioridades basado en mediciones y modelos. Se demostró que las zonas potencialmente más afectadas por los contaminantes orgánicos son los puertos y sitios cercanos a las descargas de EDAR y ríos. Gráficamente se apreció la distribución geográfica del riesgo, lo cual permitirá tomar medidas pertinentes para alcanzar una calidad adecuada de las aguas costeras. De esta manera, se identifican las zonas de riesgo y en futuros trabajos será posible ampliar el número de muestras en estas regiones, con la finalidad de determinar si el riesgo se mantiene a través del tiempo. Este trabajo contribuyó a ampliar el conocimiento del comportamiento y efecto de contaminantes orgánicos prioritarios y emergentes en las frágiles zonas costeras. / "Assessment, distribution and impact of priority and emerging organic pollutants in coastal waters" Presented by Juan Ignacio Sánchez Avila Coastal areas are vulnerable marine zones, due to its border with inland human development areas. The scarcity of studies related to the presence, distribution and impact of organic contaminants in these marine areas has been the trigger for choosing the topic of this paper. Three multiresidual methodologies were developed for monitoring 51 emerging and priority pollutants (polycyclic aromatic hydrocarbons, alkylphenols, polychlorinated biphenyls, polybrominated diphenyl ethers, organochlorine pesticides, phthalates and bisphenol A) in sea water, wastewater, river water, sediments and mussels. Gas chromatography coupled to mass spectrometry was used for the analysis of the pollutants and solid phase extraction (SPE) extraction or stir bar sorptive extraction (SBSE) or ultrasonic assisted extraction (UAE) for their preconcentration/ isolation. Methodologies showed excellent performance (between 69 and 120%) and good sensitivity (detection limits 0.01 to 38 ng L-1 for sea water). The techniques were applied to the analysis of samples collected in the Catalan coast and from estuaries and coasts from the Cantabric in Spain. Additionally, the presence of perfluorinated compounds in the Catalan coastal region was assessed. 47 organic pollutants were detected in coastal waters, with major concentrations in estuaries and ports. The concentrations detected were below Environmental Quality Standards established by the Directive 105/2008/CE, except in sampling points with higher industrial, port and urban impact. Effluent discharges and river transport were identified as major sources of pollution of coastal waters. Wastewater treatment plant (WWTP) discharges were diluted both horizontally and vertically and pollutants were spread in coastal waters. Organic pollutants were accumulated in sediments. In transplanted mussels was demonstrated the bioaccumulation of carcinogenic pollutants and endocrine disruptors. Risk analysis was evaluated via “Risk Quotients” and by the COMMPS procedure. The distribution of the risk was also graphically indicated. This work helped to increment the knowledge about the behaviour and effects of priority and emerging organic pollutants in the fragile coastal areas.

Tesi realitzada a l'Institut de Diagnòstic Ambiental i Estudis de l'Aigua (IDAEA-CSIC) / Els compostos orgànics persistents (COPs) són un conjunt de contaminants que comparteixen una sèrie de característiques comuns. Com el seu nom indica, poden romandre en el medi ambient durant llargs períodes de temps, són molt estables i no es degraden amb facilitat. D'altra banda són tòxics i a causa de les seves propietats fisico-químiques es distribueixen per tot el planeta transportant-se a llocs remots allunyats dels focus emissors. A més, en ser lipofílics, tenen una gran capacitat de bioacumulació en els organismes i per tant es biomagnifiquen al llarg de les xarxes tròfiques. La producció de compostos organoclorats durant el darrer segle per part de l'empresa cloro¬àlcali situada a Flix (al sud oest de Catalunya), ha donat lloc a l'acumulació de fangs contaminats a l'embassament d'aquesta població. Tradicionalment, en relació a les concentracions de contaminants orgànics, els organismes han estat tractats de forma parcial en tot aquest tram final del riu Ebre. Es per aquesta raó que en aquesta tesi s'ha realitzat un estudi més complet de la seva distribució i impacte sobre tots els nivells de les xarxes tròfiques fluvials tot començant pels nivells tròfics inferiors i incloent els principals depredadors. Això ha representat l'anàlisi de diversos compostos organoclorats en diferents matrius biòtiques i abiòtiques: els clorobenzens (PeCB i HCB), els policlorobifenils (PCBs), els hexaclorociclohexans (HCHs), el diclorodifeniltricloroetà (DDT) i els seus metabòlits, així com també els policloroestirens (PCEs). Per complementar la recerca s'ha volgut estudiar la presència d'aquests compostos a llocs allunyats de les principals fonts d'emissió. Per fer això, s'han analitzat mostres al llarg de tot el tram baix del riu Ebre, per comprovar si hi havia diferències de concentracions respecte a l'embassament de Flix, diferències en els patrons de distribució de COPs, o en els fenòmens de biomagnificació d'aquests compostos. Així mateix aquesta tesi estudia l'impacte que ha tingut la fàbrica clor-àlcali situada a l'embassament de Flix sobre les xarxes tròfiques del darrer tram del riu Ebre, i com aquest focus de contaminació puntual, ha afectat els llocs situats a distàncies llargues de les emissions i abocaments. Primer s'estudien les concentracions de COPs i les relacions entre els nivells tròfics inferiors de l'embassament de Flix. En segon lloc s'avaluen els nivells de contaminants que tenen els peixos tant a l'embassament com aigües avall. En tercer lloc es determina l'efecte que tenen els compostos organoclorats en els estadis superiors de les xarxes tròfiques fluvials: els ocells. Per finalitzar, es condensa tota la informació relacionada amb la biomagnificació dels contaminants al llarg de la xarxa tròfica dins de l'embassament de Flix. / Persistent organic pollutants (POPs) are a well-known group of contaminants that have in common some characteristics. As its name indicates, they can remain in the environment for long periods of time, they are stable and do not degrade easily. Furthermore, they are toxic and because of their physical and chemical properties they can be distributed throughout the planet reaching remote places far away from pollution sources. In addition, they are lipophilic and with great capacity to bioaccumulate inside the organisms so they are biomagnified along food webs. The production of organochlorine compounds during the last century by the chlor-alkali plant located at Flix (South West Catalonia) has generated an accumulation of polluted muds inside the water reservoir from this locality. Concerning the concentrations of organic pollutants, so far the organisms living in the last stretch of the Ebro River have only been considered partially. This thesis includes a complete study of the distribution and accumulation of these compounds in fluvial food webs starting from the lower trophic levels and including main predators. Different organochlorine compounds have been analyzed in a wide range of biotic and non-biotic matrices: chlorobenzenes (PeCB and HCB), polychlorobiphenyls (PCBs), hexachlorocyclohexanes (HCHs), dichlorodiphenyltrichloroethane (DDT) and its metabolites, as well as the polychlorostyrenes (PCSs). To complement this research, the presence of these compounds has been assessed in sites located far away from main pollution sources. To achieve this, samples from the last stretch of the Ebro River have been analysed in order to assess if there were differences in concentrations in comparison to samples from the Flix water reservoir, differences in distribution patterns of POPs, or differences at the biomagnification processes of these compounds. Firstly it studies concentrations of POPs and the relationships between lower trophic levels from the Flix water reservoir. Secondly it evaluates levels of contaminants from fish inside the Flix water reservoir and downstream. Third, it determines the effect that POPs have at higher trophic levels from fluvial food webs: birds. Finally, it integrates all the information related to the biomagnification of pollutants along the food web within the Flix water reservoir.

In view of the concerns about the presence of pharmaceuticals in the aquatic environment, two main lines of research regarding these substances were followed within this thesis: Their fate was assessed along waste water treatment plants (WWTPs) and Iberian River basins. Their potential ecotoxicological risks to non-target aquatic organisms were evaluated. On this basis, the more specific objectives and the subsequent findings achieved within this thesis are reported as follows: I. The fate and behavior of pharmaceuticals under nitrifying activated sludge treatment in WWTPs and after their release into receiving surface waters (SW) via wastewater (WW) effluent discharge were studied. To that aim, a sensitive analytical protocol based on off-line SPE-LC—MS/MS was developed and validated for the simultaneous determination of diclofenac their metabolites and their transformation products (TPs) in WW. The method was further optimized and validated for additional analysis of sulfamethoxazole and its TPs in WW and SW. The method was applied to the analysis of WW and SW reporting the occurrence of the metabolites and TPs of diclofenac and sulfamethoxazole in the ng L-1 range. Additionally, the microbial mediated biotransformation of diclofenac and other related pharmaceutical structures into nitro and nitroso derivates in the nitrifying activated sludge was investigated. The use of HR-MS/MS allowed the identification of tentative chemical structures of nitrosation/nitration TPs confirmed for all compounds to generate via biotic mechanisms. II. Afterwards, the presence of a list of up to 96 selected pharmaceuticals in SW and sediments from four Iberian River basins characterized by high anthropogenic pressure was assessed. To that, the spatial and temporal distribution of pharmaceuticals were evaluated. pharmaceuticals were widespread and pseudo-persistent micropollutants in the Iberian aquatic environment. Concentration of pharmaceuticals in SW varied from the low to high ng L-1 range; while in sediments they were at the low ng g-1 level. Analgesics/antiinflammatories were the most relevant therapeutic group in SW; while for sediments these were antibiotics. Factors affecting their occurrence (i.e. hydrological conditions and human and animal uses) were also assessed by the application of modeling approaches and statistical tools. Overall, response of pharmaceuticals to river flow was negative, principally due to expected dilution effects. A “plug-flow” model approach was successfully applied to describe the fate of 14 selected pharmaceuticals in terms of attenuation downstream the water course of their main emission source considered as the WWTPs. Significantly positive relationships were found among levels of pharmaceuticals and population density and livestock units in both SW and sediment matrices, thus responding to the anthropic pressures in the catchments. III. Finally, this thesis contributed to the knowledge about the ecotoxicological risk of pharmaceuticals to aquatic ecosystems. To that, individual and combined acute toxicity of pharmaceuticals and other relevant micropollutants to D. magna and V. fischeri were assessed. Individual pharmaceuticals and TPs did not showed acute toxicity to the target aquatic organisms. However PhACs and TPs displayed synergistic effects in mixtures with other toxicants of the same environmental compartments. Additionally, an ecotoxicological risk assessment of 55 pharmaceuticals to D. magna, V. fischeri and fish along four Iberian River basins was conducted. The extensive data presented on the predicted ecotoxicological risk and the computation of pharmaceuticals relative contribution to the whole toxicity of the sample, provided valuable information for further prioritization exercices in the risk assessment of Spanish river basins. Then, the impact of changing pharmaceuticals levels and water flow conditions on the structure and function of river biofilms was studied. PhACs affected the structure and functioning of fluvial biofilms. The biotic response to the two main stressors studied PhACs and hydrology, varied among the different biofilm compartments (photoautotrophs and bacteria). / En vista de las preocupaciones acerca de la presencia de productos farmacéuticos en el medio ambiente acuático, dentro de esta tesis se estudiaron y evaluaron: Su destino a lo largo de las plantas de tratamiento de aguas residuales y de las cuencas de los ríos Ibéricos. Los riesgos ecotoxicológicos que pueden representar para los organismos acuáticos no diana. Sobre esta base, las investigaciones más específicas en esta tesis fueron: I. Se estudió el destino y el comportamiento de los fármacos durante el tratamiento de lodos activados nitrificantes en las plantas de tratamiento de aguas residuales y después de su liberación en las aguas superficiales receptoras a través de los efluentes de aguas residuales. Con ese objetivo, se desarrolló y se validó un protocolo de análisis sensible para la determinación simultánea de diclofenaco sus metabolitos y sus productos de transformación en aguas residuales. Posteriormente, el método fue optimizado y validado para el análisis adicional de sulfametoxazol y sus productos de transformación en las aguas residuales y aguas superficiales. Además, se investigó la biotransformación, mediada por la comunidad microbiana del lodo activado nitrificante, de diclofenaco y otras estructuras farmacéuticas relacionadas a nitro y nitroso derivados. II. Se evaluó la presencia de fármacos en las aguas superficiales y sedimentos de cuatro cuencas hidrográficas Ibéricas caracterizadas por una elevada presión antropogénica. Para ese fin, se evaluó la distribución espacial y temporal de los fármacos. También se evaluaron los factores que influyen en su aparición (es decir, las condiciones hidrológicas y uso humano/animal) mediante la aplicación de métodos de modelización y herramientas estadísticas. III. Finalmente, se contribuyó al conocimiento del riesgo ecotoxicológico de los fármacos en los ecosistemas acuáticos. Para esto, se evaluó la toxicidad aguda individual y combinada de fármacos y otros microcontaminantes a Daphnia magna y Vibrio fischeri. Además, se realizó una evaluación del riesgo ecotoxicológico de los fármacos a D. magna, V. fischeri y peces en las cuencas hidrográficas ibéricas. A continuación, se estudió el impacto de los cambios en los niveles de los fármacos y las condiciones de caudal sobre la estructura y función de los biofilms fluviales.

Tesi realitzada a l'Institut de Diagnòstic Ambiental i Estudis de l'Aigua (IDAEA-CSIC) / Les imatges hiperespectrals són una mesura instrumental singular i de gran interès, ja que proporcionen informació química (espectral) i de distribució espacial (imatge) dels constituents de les mostres. Aquest fet les fa especialment interessants en aplicacions de la indústria farmacèutica, dels camps mediambiental i biomèdic i en la recerca i identificació de materials. L’objectiu d’aquesta tesi ha estat el coneixement de la naturalesa de la mesura de les imatges hiperespectrals amb la finalitat de dissenyar o adaptar eines d’anàlisi de dades més específiques i de proporcionar protocols d’actuació per a la interpretació d’aquest tipus de mesura en funció del tipus de tècnica espectroscòpica utilitzada i del problema químic d’interès. De manera específica, aquest treball s’ha centrat en l’estudi del potencial del mètode de resolució multivariant de corbes per mínims quadrats alternats, MCR-ALS, per a l’anàlisi d’imatges hiperespectrals, que proporciona mapes de distribució i espectres purs dels constituents de les imatges a partir únicament del coneixement de la mesura original. S’ha treballat amb l’anàlisi d’imatges individuals i l’anàlisi conjunta d’imatges obtingudes amb la mateixa tècnica o amb diferents plataformes espectroscòpiques. A partir de l’estudi d’imatges Raman i IR associades a problemes químics de diferents tipologies, s’han proposat protocols d’anàlisi que inclouen el preprocessat de les dades originals, l’obtenció dels mapes de distribució i espectres purs dels constituents de la imatge i el postprocessat dels mapes i espectres resolts per a l’obtenció d’informació addicional. L’ús dels mapes i espectres resolts proporciona informació molt diversa, com és ara la identificació, la quantificació i la caracterització de l’heterogeneïtat dels constituents de la imatge o la interpretació global i local d’un procés. Els mapes resolts han estat també una informació de partida excel·lent en altres tipus d’anàlisi, com la segmentació de la imatge o en procediments de superresolució, orientats a millorar la resolució espacial de les imatges instrumentals. La combinació de l’anàlisi multiconjunt de resolució i segmentació s’ha revelat de gran utilitat per a distingir poblacions de mostres de teixits biològics amb diferents estats patològics. Per últim, s’ha proposat un procediment per a la fusió i anàlisi d’imatges adquirides amb diferents tècniques espectroscòpiques i de diferent resolució espacial mitjançant una nova variant del mètode MCR-ALS per a estructures multiconjunt incompletes, que permet aprofitar la informació complementària de les tècniques acoblades i preservar la màxima resolució espacial. / Hyperspectral images are unique instrumental measurements that contain chemical (spectral) information and detailed knowledge of the distribution of the sample constituents on the sample surface scanned. This thesis is mainly oriented to know in depth the nature of this instrumental measurement in order to design and adapt specific chemometric tools that help in the proposal of general protocols for the interpretation of hyperspectral images according to the spectroscopic technique used and the chemical problem of interest. Particularly, much work has been focused on the study of the potential of multivariate curve resolution-alternating least squares, MCR-ALS, for the analysis of hyperspectral images. This algorithm provides distribution maps and pure spectra for the image constituents from the sole information contained in the raw measurement. Within this framework, individual analysis of images and image multiset analysis on data structures formed by images collected with the same technique or by images coming from different spectroscopic platforms have been explored. From the study of Raman and IR hyperspectral images linked to different chemical problem typologies, data analysis protocols have been proposed that include preprocessing of original data, recovery of distribution maps and pure spectra of image constituents and postprocessing of resolved maps and pure spectra to obtain further information. Resolved distribution maps and pure spectra provide diverse information, such as identification, quantification and heterogeneity characterization of the image constituents or the global and local description of a process. The use of resolved distribution maps has proven to be an excellent starting point for other kinds of analysis, such as image segmentation or super-resolution algorithms, oriented to improve the spatial resolution of experimental hyperspectral images. Combined multiset resolution and segmentation analysis has been shown to be very useful for the differentiation of populations of biological tissues with different pathological conditions. Finally, a strategy for data fusion of hyperspectral images from different spectroscopic platforms and different spatial resolution has been proposed. This approach uses a new variant of MCR-ALS for incomplete multiset structures that takes advantage of the complementary information provided by the different spectroscopic techniques without losing spatial resolution.

Fullerenes or Buckyballs are a group of carbon nanoparticles, classified as engineered nanoparticles. The advent of fullerene nanoparticles in commercial, industrial and biomedical applications raises concern about their potential ecological and human health risks. Fullerenes have not been regulated, although the European Commission (European Commission, 2002), the European Parliament (European Parliament, 2008) and the US Environmental Protection Agency (EPA, 2015) have prioritised legislation on nanomaterials handling and disposal. Given the current lack of studies regarding their presence, fate and behaviour in consumer products and environmental matrices and their associated human and environmental risks it is becoming increasingly important to be able to characterise and quantitate fullerene nanoparticles, especially derivatives, in a wide range of matrix types. To fill these knowledge gaps, one of the objectives of this thesis was the development of analytical methodologies for the determination of pristine and surface modified fullerenes by ultra-high performance liquid chromatography coupled to MS (UHPLC-MS) and by nonaqueous capillary electrophoresis (NACE) and micellar electrokinetic capillary chromatography (MECC) with UV-Vis detection in different environmental matrices (water and sediment samples) and cosmetic products, respectively. In addition, in this thesis the size and shape characterisation of surface modified fullerenes in aqueous solutions of different pH and ionic strength values was studied by combining several techniques (CE, AF4-MALS and TEM). Regarding the analysis of fullerenes, the use of a sub-2 µm C18 column, toluene-methanol as a mobile phase and of APPI ionisation source allowed to develop a UHPLC method coupled to MS/(MS) for the analysis of five pristine (C60-C84) and three surface modified fullerenes (PCBM, PCBB and C60-pyrr) in less than 4.5 min showing high sensitivity and selectivity. Furthermore, the employment of H-SIM mode for pristine fullerenes (mass resolving power >12,500 FWHM), and SRM mode for fullerene derivatives, allowed achieving MLODs lower than most of those previously reported in the literature. For the extraction of the water samples, liquid-liquid extraction (LLE) with toluene and the addition of salt is proposed obtaining recoveries higher than 83 %. For sediments we propose the use of pressurised liquid extraction (PLE) performed at a high extraction temperature (150 °C) using one extraction cycle of 10 min achieving good recoveries (70-92 %). The developed methodology allowed us to report for the first time the presence of C60-pyrr, PCBM and PCBB in sediments (2.0 - 8.5 ng Kg-1 levels) and of PCBM and PCBB in pond water samples (0.1- 5.1 pg L-1 levels). Two CE methods, a non-aqueous (NACE) and a micellar (MECC) method have been developed. LOQs at mg L-1 levels were obtained with both methods making possible their application for the analysis of cosmetic products. Both methods (NACE and MECC) were applied for the quantitation of C60 in cosmetic products and the results are comparable to those obtained by LC-MS (in an anti-aging serum). With respect to the characterisation of fullerene aggregates, the use of AF4-MALS demonstrated that fullerene aqueous solutions contain particles of different aggregation degree in agreement with TEM micrographs. The size determination of the studied compounds by AF4 at different salt concentrations demonstrated that the enhanced aggregation of fullerenes with the increase in the ionic strength could explain the broad, multiple and distorted peaks obtained in MECC which were more obvious at high buffer concentration. The hydrodynamic radii of polyhydroxy-fullerenes increased more than 5 times and those of the carboxy-fullerene derivatives up to 180 nm (3rd peak) (for C60-pyrr tris acid) for 0.1M NaCl. The propensity of fullerenes to aggregate in both aqueous solutions and organic solvent mixtures justify the electrophoretic peak profiles observed in MECC (i.e., broad peaks at high electrolyte concentration) and their higher retention in C18 columns using toluene-acetonitrile compared to toluene-methanol mobile phases (due to the formation of bigger aggregates), respectively. / Los fulerenos son cada vez más utilizados en medicina (administración de fármacos), en procesos ambientales (remediación) y en la industria (células solares) debido a sus propiedades estructurales y electrónicas únicas. En este contexto, es importante aumentar el conocimiento actual con respecto a las características, el comportamiento, el destino y la toxicidad de los fulerenos, una nueva clase de contaminantes emergentes orgánicos. Por lo tanto, es cada vez es más importante ser capaz de caracterizar y cuantificar las nanopartículas de fulerenos en una amplia gama de tipos de matriz y para este fin, se necesita el desarrollo de métodos analíticos para su análisis cuantitativo y cualitativo. Además de la determinación de las concentraciones de fulerenos en matrices complejas, se requiere su caracterización en términos de grado de agregación, distribución del tamaño y morfología de la superficie en soluciones acuosas de diferentes características (fuerza iónica y pH) con el fin de proporcionar herramientas para establecer su riesgo. En esta tesis se han establecido metodologías para el análisis de ocho fulerenos en muestras ambientales (agua y sedimentos) por cromatografía líquida acoplada a espectrometría de masas y se ha detectado por la primera vez la presencia de tres derivados de fulerenos. Además, se ha desarollado metodología para el análisis de fulerenos hidrofobicos y solubles en agua mediante la electroforesis capilar en medio no-acuoso (NACE) y la cromatografía capilar electrocinética micelar (MECC) y los metodos se han aplicado para la determinación de C60 en productos cosmeticos. Por último, se ha estudiado el comportamiento de agregación de los fulerenos en soluciones acuosas de diferentes características (pHs, fuerzas iónicas) mediante diferentes técnicas (electroforesis capilar (CE), asymetrical flow-field flow fractionation (AF4), microscopia electrónica de transmisión (TEM)). En este contexto se han determinado los tamaños de los agregados formados en las condiciones evaluadas, y la morfología de las partículas.

Dissertação (mestrado) - Universidade Federal de Santa Catarina, Centro Tecnológico. / Made available in DSpace on 2012-10-18T02:43:26Z (GMT). No. of bitstreams: 0Bitstream added on 2014-09-25T19:22:19Z : No. of bitstreams: 1 170491.pdf: 7716714 bytes, checksum: b035fa49eb5e084aaa14271437cc0570 (MD5) / O ensino da matemática nas escolas brasileiras dissocia os conteúdos dos campos numéricos e algébricos, quando deveria ocorrer exatamente o contrário. Desde os primeiros anos de vida, a criança já distingue formas, revelando uma geometria espontânea. A proposta deste trabalho é apresentar a concepção de um Ambiente Hipermídia para aprendizagem da Geometria Analítica procurando resgatar o fascínio das conquistas obtidas pela criança, antes de ingressar na escola e como manter o interesse por novas conquistas geométricas no campo estrutural e dimensional. Espera-se, com este trabalho, dar um novo alento à geometria e fornecer aos professores uma nova ferramenta de auxílio no processo ensino-aprendizagem.

Orientador: Denise Bevilaqua / Banca: Miguel Ruiz / Banca: Monica Cristina Teixeira / Resumo: A diminuição gradativa, porem constante das reservas mundiais de metais no mundo, tem forçado o desenvolvimento de novas tecnologias para extração e aproveitamento destes bens minerais. A redução de custos e as novas diretrizes ambientais são o maior impulso para a busca destas novas tecnologias.A biohidrometalurgia, mais especificamente, a biolixiviação dentro deste contexto assume uma importância significativa. Os custos de operação deste processo são muito inferiores aos processos convencionais, além de ser ambientalmente mais aceito, uma vez que não gera gases tóxicos. As próprias bactérias, presentes nestas ocorrências minerais, produzem os insumos necessários para a lixiviação, ácido e íons férricos. A principal bactéria envolvida neste processo e também a mais estudada é o Acidithiobacillus ferrooxidans. Esta é a única bactéria descrita capaz de utilizar íons ferrosos e compostos reduzidos de enxofre como fonte energética. A calcopirita, mineral de cobre mais abundante na face da Terra, apresenta grande refratariedade ao ataque químico e bacteriano, e ainda não há nenhum processo industrial operando em escala comercial.Neste trabalho foram investigados o efeito da adição de diferentes quantidade de pirita, sulfeto de ferro (FeS2) no aumento da solubilização de cobre deste sulfeto. A metodologia foi estabelecida baseandose no mecanismo galvânico de dissolução dos minerais. Os estudos foram realizados através de ensaios respirométricos e de lixiviação em frascos agitados. A presença de bactéria e a adição de ionscloretos também foram avaliadas. Os ensaios respirométricos apresentaram curvas iniciais de consumo de oxigênio sem diferenças significativas para todas as condições analisadas. Os valores finais de consumo de oxigênio variaram. As condições que continham calcopirita/pirita nas ... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: The gradually depletion of world reserves, however, constant has forced the development of new technologies for extraction and exploitation of these minerals. Cost reduction and new environmental guidelines are the biggest impulse to the search for these new technologies. The biohydrometallurgy, specifically, the bioleaching in this context is of significant importance. The operating cost of this process is much lower than conventional process, in addition to being more environmentally friendly, since it does not generate toxic gases. The bacteria themselves, present in these mineral occurrences produce the inputs needed for leaching acid and ferric ions.The main bacterium involved on this process and also the most studied is the Acidithiobacillus ferrooxidans. This is the only bacterium described able to use ferrous ions and reduced sulfur compounds an energy source. The chalcopyrite, the most abundant copper mineral in the earth, is refractory to the chemical and bacterial attack, and there is no industrial process operating on a commercial scale yet.In this work were investigated the effect of adding different amounts of pyrite, ion sulfide (FeS2) to increase the solubility of copper sulfide. The methodology was established based on galvanic mechanism of dissolution of minerals. The studies were conducted through respirometric tests and bioleaching experiments in shake flasks. The presence of bacteria and the addition of chloride ions were also evaluated.The respirometric tests showed initial curves of oxygen consumption with no significant differences for all conditions examined. The final values of oxygen consumption diverse. The conditions contained chalcopyrite/pyrite in rations 1:3 and 1:1 had values greater than 400 μL of O2 consumed, while other conditions inoculated, these values... (Complete abstract click electronic access below) / Mestre

Amostras de óleo vegetal foram preparadas na forma de microemulsão de água em óleo (w/o) utilizando dodecil sulfato de sódio (SDS) como surfactante e um álcool como co-surfactante. As microemulsões foram caracterizadas através de medidas de viscosidade, índice de refração, condutividade elétrica, espalhamento de luz dinâmico e voltametria. Ensaios preliminares para a quantificação de analitos por análise direta foram realizados por voltametria linear e eletroforese capilar. As microemulsões de água em óleo de soja apresentaram gotículas de dimensões nanométricas e estabilidade termodinâmica dependente da temperatura, da concentração dos eletrólitos dissolvidos e da natureza do co-surfactante empregado. Por outro lado, o raio hidrodinâmico das gotículas (Rh) diminui com o aumento da temperatura. Quanto aos valores de condutividade, maiores do que os obtidos para o óleo de soja e para a água deionizada, aumentam com a temperatura, na faixa de 20 a 65 oC, e com o teor de água na microemulsão, entre 5 e 7,5 % de água. A maior estabilidade termodinâmica foi alcançada para uma microemulsão contendo 40,0 % de óleo, 43,2 % de pentanol, 10,8 % de SDS e 6,0 % de água, em massa, na razão 1:4 [SDS]:[álcool] Medidas voltamétricas com um ultramicroeletrodo de Pt em microemulsões contendo ferroceno ou ácido oléico dissolvidos evidenciaram a dependência linear das correntes limite anódica e catódica com a concentração da espécie eletroativa. Já a oxidação do ferroceno por voltametria cíclica, usando o mesmo ultramicroeletrodo, mostrou que a diminuição dos coeficientes de difusão nestes meios permite realizar medidas em estado transiente, empregando velocidades convencionais de varredura em potencial. Experimentos por cromatografia eletrocinética em microemulsão reversa (RMEEKC) com n-pentanol como fase contínua permitiram a separação de solutos neutros e aniônicos em amostras de óleo vegetal e gordura animal. Finalmente, foi desenvolvido um procedimento rápido para a separação e identificação dos biofenóis presentes em óleos, utilizando como eletrólito de corrida uma mistura de metanol e 1-propanol contendo KOH. Os resultados evidenciam a possibilidade de análise direta de óleos vegetais empregando métodos eletroanalíticos em microemulsões w/o, e métodos por eletroforese capilar, quer em microemulsões, quer em meio não aquoso.