This dissertation focuses on the development of analytical methodology for the analysis of polycyclic aromatic hydrocarbons (PAHs) in water samples. Chemical analysis of PAHs is of great environmental and toxicological importance. Many of them are highly suspect as etiological agents in human cancer. Among the hundreds of PAHs present in the environment, the U.S. Environmental Protection Agency (EPA) lists sixteen as "Consent Decree" priority pollutants. Their routine monitoring in environmental samples is recommended to prevent human contamination risks. A primary route of human exposure to PAHs is the ingestion of contaminated water. The rather low PAH concentrations in water samples make the analysis of the sixteen priority pollutants particularly challenging. Current EPA methodology follows the classical pattern of sample extraction and chromatographic analysis. The method of choice for PAHs extraction and pre-concentration is solid-phase extraction (SPE). PAHs determination is carried out via high-performance liquid chromatography (HPLC) or gas chromatography/mass spectrometry (GC/MS). When HPLC is applied to highly complex samples, EPA recommends the use of GC/MS to verify compound identification and to check peak-purity of HPLC fractions. Although EPA methodology provides reliable data, the routine monitoring of numerous samples via fast, cost effective and environmentally friendly methods remains an analytical challenge. Typically, 1 L of water is processed through the SPE device in approximately 1 h. The rather large water volume and long sample processing time are recommended to reach detectable concentrations and quantitative removal of PAHs from water samples. Chromatographic elution times of 30 - 60 min are typical and standards must be run periodically to verify retention times. If concentrations of targeted PAHs are found to lie outside the detector's response range, the sample must be diluted (or concentrated), and the process repeated. In order to prevent environmental risks and human contamination, the routine monitoring of the sixteen EPA-PAHs is not sufficient anymore. Recent toxicological studies attribute a significant portion of the biological activity of PAH contaminated samples to the presence of high molecular weight (HMW) PAHs, i.e. PAHs with MW ≥ 300. Because the carcinogenic properties of HMW-PAHs differ significantly from isomer to isomer, it is of paramount importance to determine the most toxic isomers even if they are present at much lower concentrations than their less toxic isomers. Unfortunately, established methodology cannot always meet the challenge of specifically analyzing HMW-PAHs at the low concentration levels of environmental samples. The main problems that confront classic methodology arise from the relatively low concentration levels and the large number of structural isomers with very similar elution times and similar, possibly even virtually identical, fragmentation patterns. This dissertation summarizes significant improvements on various fronts. Its first original component deals with the unambiguous determination of four HMW-PAHs via laser-excited time-resolved Shpol'skii spectroscopy (LETRSS) without previous chromatographic separation. The second original component is the improvement of a relatively new PAH extraction method - solid-phase nanoextraction (SPNE) - which uses gold nanoparticles as extracting material for PAHs. The advantages of the improved SPNE procedure are demonstrated for the analysis of EPA-PAHs and HMW-PAHs in water samples via GC/MS and LETRSS, respectively.

Fort McMurray has experienced significant environmental disruptions, raising concerns about chemical releases that affect the environment, particularly berries. This thesis explores the relationship between environmental factors and the nutritional quality of pin cherry (Prunus pensylvanica L.F.) and common blueberry (Vaccinium myrtilloides Michx) fruits and soil in Fort McMurray (reclaimed and natural lands). Samples were collected in August 2022 and analyzed for chemicals and quality variables. The results revealed concentration variations among samples, with hydrocarbons higher in reclaimed areas and blueberries than others and soil surpassing those in fruits. Copper and iron in fruits exceeded regulatory limits. A strong association between soil chemicals and alkylated hydrocarbons in fruits explained most differences. Soil trace elements and properties were primary environmental drivers, while hydrocarbons were secondary influencers, evident mainly in reclaimed berry environments. Antioxidant-focused nutritional quality in reclaimed berries was predominantly influenced by these key drivers in soil, requiring consistent monitoring.

[pt] Um grande vazamento de óleo bruto ocorreu na costa brasileira em 2019 afetando uma extensão de 4.334 km de faixa litorânea, 11 estados e várias localidades. Com isso, moradores locais entraram em contato direto com óleo cru ficando expostos à contaminação por hidrocarbonetos policíclicos aromáticos (HPA). Os HPA são compostos orgânicos carcinogênicos de origem petrogênica e pirogênica, que são ubíquos e persistentes. Quando seres humanos são expostos a esses contaminantes, o organismo pode metabolizar e gerar metabólitos desses HPA com prováveis efeitos negativos à saúde humana. Com isso, este trabalho visou analisar a exposição de uma comunidade de pescadores (localizada em Canavierias, BA) aos HPA provindos de derrames de petróleo através da análise de metabólitos de HPA em urina. Para isso, as 44 amostras foram extraídas por extração em fase sólida (SPE) e analisadas por cromatografia líquida acoplada a espectrômetro de massas triplo quadrupolo (HPLC-MS/MS). Este trabalho também contempla a comparação de diferentes métodos de extrações em 18 amostras de urina selecionadas. No Centro de Pesquisas, Desenvolvimento e Inovação Leopoldo Américo Miguez de Mello (CENPES) realizou-se extração em fase sólida e a extração líquido-líquido, enquanto na PUC-Rio foi realizada a extração por SPE. Os resultados referentes às 44 amostras analisada na PUC-Rio, demonstraram que 4 dos 11 metabólitos de interesse foram identificados e quantificados. Para o 1-OH-Nap a faixa de concentração foi (menor que LQ – 44 n mL(-1) ), (menor que LQ-21 ng mL(-1) ) para o 2-OHNap, (menor que LQ -5,4 ng mL(-1) ) para o 3,9-OH-Phe e (menor que LQ- 0,76 ng mL(-1) ) para o 6-OH-Chr. Ao comparar os resultados obtidos com trabalhos da literatura, observou-se que os moradores da comunidade de Canavieiras apresentaram valores baixos de concentração dos metabólitos que foram quantificados. Mesmo assim, é importante a existência de trabalhos voltados para o monitoramento destes compostos já que não somente o ambiente como também apresentam-se como um risco à saúde humana. / [en] A major crude oil spill occurred off the coast of Brazil in 2019 affectinga 4,334 km stretch of coastline, 11 states and several localities. As a result,residents came into direct contact with crude oil and were exposed tocontamination by polycyclic aromatic hydrocarbons (PAHs). PAHs arecarcinogenic organic compounds of petrogenic and pyrogenic origin, whichare ubiquitous and persistent. When humans are exposed to thesecontaminants, the organism can metabolize and generate metabolites ofthese PAHs with probable negative effects on human health. Therefore,this work aimed to analyze the exposure of a fishing community (located inCa-navierias, BA) to PAHs from oil spills through the analysis of PAHmetabolites in urine. For this, the 44 samples were extracted by solid phaseextraction (SPE) and analyzed by liquid chromatography coupled to triplequadrupole mass spectrometer (HPLC-MS/MS). This work also includes thecomparison of different extraction methods in 18 selected urine samples.Solid phase extraction and liquid-liquid extraction were performed at theLeopoldo Américo Miguez de Mello Research, Development and InnovationCenter (CENPES), while SPE extraction was performed at PUC-Rio. Theresults for the 44 samples analyzed at PUC-Rio showed that 4 of the 11metabolites of interest were identified and quantified. For 1-OH-Nap theconcentration range was (less than LQ - 44 n mL(-1)), (less than LQ-21 ng mL(-1)) for 2-OH-Nap,(less than LQ -5.4 ng mL(-1)) for 3,9-OH-Phe and (less than LQ- 0.76 ng mL(-1)) for 6-OH-Chr.When comparing the results obtained with literature works, it was observedthat the residents of the community of Canavieiras had low concentrationvalues of the metabolites that were quantified. Even so, it is important theexistence of works focused on the monitoring of these compounds since notonly the environment but also present themselves as a risk to human health.

This study aims to determine the concentration of 16 Polycyclic Aromatic Hydrocarbons (PAHs) in urban soils from West Yorkshire in order to determine what the factors are controlling their distribution and abundances. Although PAHs have been reported before from soils and sediments, the majority of these studies have come from China, sometimes with contrasting results, which emphasises the need to obtain equivalent data from other areas. Therefore this work provides the first measurements of their type from the area studied. Soil samples were collected from one hundred sites across an area from Bradford to Leeds on two occasions, one in autumn and one in the following summer. The soil samples were analyzed by X-ray diffraction and all found to have similar mineralogical composition, which was mainly silica and calcite. Trials using iodine as a marker for PAHs showed there were notable interactions between the minerals and PAHs, with calcium carbonate absorbing PAHs much more than silica. There is a negative correlation between the soil organic content (determined by loss on ignition) and PAHs, which confirms the PAH-mineral interaction. Gas chromatography with mass spectrometry (GC-MS) using targeted selected ion monitoring was used to determine and quantify PAHs in the one hundred soil samples with the aid of PAH external standards. The results showed highest concentrations of total PAHs in the same sample from Leeds from the autumn (1,525 ng/g) and in the summer (1,768 ng/g). In Bradford there was only moderate pollution of PAHs, the maximum being 122 ng/g. However the majority of data from Bradford showed lower levels of pollution in both summer and autumn. On the basis of prior published information, the ratio of these compounds has been used to help in identifying sources. In the samples collected from Bradford in both seasons and Leeds in autumn the PAH pollution originated from pyrogenic, biomass and petroleum combustion, however in the summer the source appeared more to be from a petrogenic source. These ratios in the samples which were collected from the area between Bradford and Leeds implied pyrogenic, biomass source of pollution in the autumn, but in the summer another source of organic compounds was indicated namely petroleum combustion. When the locations were resampled nine months later, after taking into account within-site variability, there was a strong indication that the PAH concentrations were higher. This might have been due to a seasonal effect, but when a further (third) subsample was taken at a later date it showed a further increase in PAH level which suggests the effect is accumulative rather than seasonal. The results were analysed using Principal Component Analysis (PCA) to determine whether the type of road had an effect on the concentration of the 16 PAHs compounds, but it was concluded that there was no such effect. However, the distance from the soil sample to the nearest road did have an effect on the concentration of the 16 PAH compounds, especially in soil samples having the shortest distance to the road. Traffic volume was also tested and found to influence the PAH concentrations. It is notable that, comparing the groupings from autumn with those from summer by cluster analyses, they largely had the same compounds grouping together in both cases; only two compounds differed at all in where they occurred in the clusters, with consistent patterns of grouping found for the other compounds. These analyses indicate that PAH compounds behave in a consistent way amongst groups of PAH compounds. The grouping of PAHs appears linked to their sources rather than number of rings or molecular weight. / Libyan Government and Embassy

No description available.

No description available.

Soil that is heavily contaminated with polycyclic aromatic hydrocarbons (PAHs) is often found at the sites of former gasworks and wood-impregnation plants. Since PAHs are toxic these sites represent a hazard to human health and the environment, and therefore they need to be treated, preferably by a method that destroys the contaminants, and thus eliminates the problem permanently. However, during biological and chemical degradation of PAHs other toxic compounds may be formed. If these transformation products are sufficiently persistent they could potentially accumulate during remedial processes. In the work underlying this thesis the degradation and transformation of PAHs were studied in three remedial processes: viz. a pilot-scale bioslurry reactor, microcosms with wood-rotting fungi and lab-scale treatments with Fenton's reagent. A group of transformation products referred to as oxygenated-PAHs (oxy-PAHs) was found to be particularly important, as these compounds are toxic and were shown to be relatively persistent in the environment. The oxy- PAHs were, for instance, found at significant concentrations in the gasworks soil used in most of the studies. This soil was highly weathered and had therefore been depleted of the more readily degradable compounds. In addition, experiments in which earthworms were exposed to the gasworks soil showed that the oxy-PAHs were more easily taken up in living organisms than PAHs. To facilitate the studies, new extraction and fractionation methods were developed. For instance, pressurized liquid extraction (PLE) was investigated for its reliability and efficiency to extract PAHs and oxy-PAHs from soil. Furthermore, a selective PLE-method was developed that can simultaneously extract and separate the PAHs and oxy-PAHs into two different fractions. This was accomplished by adding a chromatographic material (silica or Florisil) to the extraction cell. Under certain conditions all three remedial processes resulted in increasing amounts of oxy- PAHs in the soil. For example, 1-acenaphthenone and 4-oxapyrene-5-one accumulated in the bioslurry reactor. Similarly, in the soil inoculated with a white-rot fungus 9-fluorenone, benzo[a]anthracene-7,12-dione, 4-hydroxy-9-fluorenone and 4-oxapyrene-5-one accumulated. Finally, in an ethanol-Fenton treatment the concentration of some PAH-quinones increased in the soil. The results show that it might be necessary to monitor oxy-PAHs as well as PAHs during the remediation of PAH-contaminated sites. Otherwise, the soil may be considered detoxified too early in the process. In the long term it would be desirable to include analyses with sufficient marker compounds to follow the possible production and elimination of the oxy-PAHs. However, until such compounds can be identified it is suggested that contaminated soil should be screened for oxy-PAHs in general. The selective PLE-method presented in this thesis could be a useful tool for this.

Les adduits exocycliques à l’ADN paraissent être la conséquence indirecte, sous la médiation de la peroxydation lipidique, du stress oxydant cellulaire induit par les composés aromatiques nitrés (CANs) de l’environnement. Ces derniers, formés le plus souvent in situ dans les environnements complexes sont un sujet de préoccupation croissante en santé environnementale. Le but étant de comprendre les mécanismes de formation et de dégradation de ces adduits afin d’en apprécier leur place dans la toxicité des CANs et leur intérêt en tant que biomarqueurs du stress oxydant induit par l’environnement.Nous avons réalisé la synthèse de deux adduits exocycliques : le 1,N²-éthéno-2’-déoxyguanosine (εdG) et le 1,N²-propano-2’-déoxyguanosine (pdG-HNE) et étudié leurstabilité en présence d’une oxydation radicalaire. Il est apparu que le pdG-HNE semble être lemeilleur candidat en tant que biomarqueur du stress oxydant.Pour approcher au mieux la chimie du vivant, nous nous sommes posés la question dela stabilité de ces adduits en milieu cellulaire. Une étude préliminaire de génotoxicité a étéréalisée et montre que seuls les hydrocarbures aromatiques polycycliques nitrés présententune potentialité mutagène significative. Nous avons donc étudié le pyrène et ses dérivés nitrés(1-Nitropyrène/1,3-Dinitropyrène/1,6-Dinitropyrène/1,8-Dinitropyrène) afin d’étudier leurcapacité à générer des adduits dans les lymphocytes humains.Nos résultats montrent que le 1-Nitropyrène génère in vitro des adduits stables dans letemps ; ce qui pose la question de leur réparabilité par les systèmes cellulaires et de leurspotentialités cancérogènes pour l’homme. / Exocyclic DNA adducts seem to be the indirect consequence, mediated by lipidperoxidation, of oxidative stress induced by nitro aromatic compounds (NACs). The latterusually formed in situ in environmental complex mixtures are a matter of concern inenvironmental health. The aim is to understand the mechanisms of formation and degradationof these adducts to assess their place in toxicity of NACs and their importance as oxidativestress biomarkers induced by the environment.We synthesized two exocylic adducts: 1, N²-etheno-2’-deoxyguanosine (εdG) and 1,N²-propano-2'-deoxyguanosine (pdG-HNE) and studied their stability when a radicaloxidation is present. It appeared that pdG-HNE seems a suitable biomarker of oxidative stress.To come close to life chemistry, we were wondering whether these adducts are stablein cellular environment. A preliminary study of genotoxicity was carried out and showed thatonly nitro polycyclic aromatic hydrocarbons have a significant mutagenic potency. Thereforewe studied pyrene and nitropyrenes (1-Nitropyrene/1,3-Dinitropyrene/1,6-Dinitropyrene/1,8-Dinitropyrene) to examine their ability to produce adducts in human lymphocytes.Our results show that 1-Nitropyrene give rise to stable adducts in vitro, which raisesthe question of their repairability by cellular systems and their potential carcinogenic tohumans.

Les hydrocarbures aromatiques polycycliques (HAP) sont des polluants organiques, ubiquistes, potentiellement toxiques et cancérigènes. Dans les sols, la dégradation des HAP est principalement due à l'activité microbienne. Certaines études ont montré que la biodégradation des HAP pouvait être augmentée dans la rhizosphère des plantes où le nombre et l'activité microbienne sont stimulés, grâce aux exsudats racinaires. Cependant les bénéfices des plantes ne sont pas toujours observés, et les exsudats pourraient aussi modifier la biodisponibilité des HAP. Les objectifs de ce travail ont été de mieux comprendre ces interactions sol-plante-microorganismes qui conditionnent le devenir des HAP dans la rhizosphère en suivant notamment (i) les bactéries possédant les gènes codant une HAP-dioxygènase, (ii) les espèces bactériennes impliquées dans la dégradation du phénanthrène, et (iii) la disponibilité et la biodégradation des HAP dans des terres industrielles historiquement contaminées.Les expériences ont été conduites dans des dispositifs à compartiments, lesquels permettent une diffusion des exsudats racinaires dans le sol tout en retenant physiquement les racines, puis en microcosmes avec un ajout d'exsudats racinaires naturels produits à partir d'une culture hydroponique de ray-grass (Lolium perenne, L). Les expériences ont été réalisées dans un premier temps avec du sable en ajoutant du phénanthrène (PHE) et un inoculum bactérien issu d'un sol d'une ancienne cokerie puis directement avec des sols historiquement contaminés en HAP. Les nombres de copies de gènes codant pour l'ADNr 16S et pour des HAP-dioxygènases ont été quantifiés par PCR en temps réel pour estimer la proportion de bactéries dégradantes. Les structures des communautés ont été comparées par électrophorèses (TTGE). En plus de l'analyse des 16 HAP totaux, une extraction non exhaustive des HAP a été réalisée à la cyclodextrine pour en estimer la disponibilité. L'utilisation de la méthode SIP (stable isotope probing) avec du 13C-phénanthrène a permis d'identifier les bactéries directement impliqués sa dégradation dans un sol historiquement contaminé. Les expériences en dispositifs à compartiments ont confirmé que la dissipation du phénanthrène est plus importante lorsque la distance aux racines est plus faible, et montrent que le nombre de copies de gène 16S et de gène de HAP-dioxygénase varie avec l'âge des plantes et du temps de contact des compartiments latéraux avec le tapis racinaire. Mais elles montrent aussi que la dissipation du phénanthrène n'est pas plus importante dans les pots plantés, tandis que dans les expériences en microcosmes une inhibition de la dissipation du PHE a même été observée en présence d'exsudats. La présence d'exsudats racinaires a profondément modifié la structure des communautés dégradant les HAP, et l'expérience SIP a permis d'identifier les bactéries directement impliquées dans la dégradation du 13C-phénanthrène et de montrer qu'elles étaient différentes en présence ou non d'exsudats. En présence d'exsudats, la proportion des bactéries dégradantes dans la population totale est passée de 1 % dans la terre d'origine et dans les traitements sans exsudats à plus de 10 %. Même si les exsudats racinaires ralentissent la dissipation du phénanthrène, en fournissant une source de carbone plus facilement métabolisable, ils ont augmenté la quantité de HAP extractibles à la cyclodextrine dans deux des trois sols historiquement contaminés, suggérant un effet de ceux-ci sur la biodisponibilité des HAP / Polycyclic Aromatic Hydrocarbons (PAH) are organics pollutants, ubiquitous, toxics and potentially carcinogenic. In soil, PAH degradation is mainly attributed to microbial organism. Several studies have thus reported enhanced PAH degradation in soil in the presence of plants. Rhizospheric soil increase the number et the activity of microorganisms in soil by the release of roots exudates. However, bene?cial effects of plants in the remediation are not always observed and roots exudates could be limited PAH biodegradation. The object of this study was to investigate the fate of PAHs in rhizosphere, following (i) the PAH-dioxygenase genes DNA to quantify the PAH-degrading bacteria, (ii) species implicated in phenanthrene biodegradation, and (iii) PAH availability and biodegradation from industrial soils.Different experimental devices have been designed to study detailed processes in the rhizosphere. First is a compartments devices were a nylon mesh permits diffusion of plant soluble substances towards the adjacent root free compartment as a rhizosphere. Secondly microcosms were enriched with natural roots exudates from hydroponic culture of ray-grass (Lolium perenne L.). In first time, experiments were conducted using sand and bacterial inoculum from an industrially PAH-contaminated soil and then directly with a soil historically contaminated by PAH. The Real-Time PCR quantification of 16S rRNA gene copy and of functional PAH-RHD? genes permitted to assess the proportion of a degrading bacteria. Bacterial community structure was approached from Temporal Thermal Gradient gel Electrophoresis (TTGE) fingerprinting, and bands sequencing. Nonexhaustive cyclodextrin-based extraction technique provided a estimate of the ?labile? or available pool of PAH in soil. Use of stable isotope probing (SIP) technique with [13C]phenanthrene allowed a bacterial identification of directly implicated in industrial soil.The presence of exudates modified microbial community of PAH-degrading bacteria. SIP experiment showed that 13C-labelled PHE-degrading bacteria was different depending on the exudates input. Many species having to degrade phenanthrene were able to use exudates. Presence of root exudates increased the proportion of PAH-RHD? genes compared to the bulk soil at the beginning and in microcosms without exudates (respectively 10% and 1 %). However, phenanthene dissipation in sand or soil were weaker with root exudates and aged PAH concentrations has not shifted during incubation time. Nevertheless, the root exudates increased the PAH labile fraction extract with cyclodextrin solution into two in three soils historically contaminated

2-Nitrofluorene is a model representative of nitrated polycyclic aromatic hydrocarbons (NPAH) which belongs to a group of mutagens and carcinogens. Interaction of DNA with genotoxic 2-nitrofluorene was monitored by an electrochemical DNA biosensor made of a glassy carbon electrode (GCE) and low molecular weight DNA from salmon sperm. Techniques used are electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV) and square-wave voltammetry (SWV). Using the EIS technique, no damage to DNA, which would cause strand breaks in DNA, was observed, whereas using the CV technique, the intercalation of NF to the structure of DNA was observed, leading to the formation of a NF-DNA complex. The intercalation results in a reduction of electroactive sites which can be oxidized. It was verified using the SWV technique, by which a decrease of the peak heights of adenosine and guanosine was observed. Because of the dangerous effect of NF on the structure of DNA, an electroanalytical method for its determination was developed. An applicability of the method was successfully tested on a model sample of sand. For the development of the technique, differential pulse voltammetry (DPV) was used in a mixture of the Britton-Robinson buffer of pH 7.0 and ethanol in a ratio of 7:3 (v/v) and with a periodic...