O destino de grande parte da matéria orgânica presente nos oceanos é o sedimento marinho. Essa matéria orgânica, que vai sendo armazenada ao longo dos anos, produz uma reserva única de informações sobre os processos biogeoquímicos ocorridos no passado e como esses processos responderam às mudanças ambientais. Certas moléculas orgânicas são sintetizadas apenas por organismos marinhos ou somente por plantas superiores terrestres. A abundância relativa desses compostos em sedimentos marinhos pode ser usada para determinar a contribuição de matéria orgânica terrestre ou marinha que foi depositada. Outros compostos, cujas fontes naturais são raras ou inexistem, encontrados em sedimentos, refletem a influência antrópica no meio. Há ainda compostos, com origem biológica especifica, cujas estruturas são influenciadas pelo controle enzimático dos organismos que os produzem, que são influenciados pelo meio. Esses compostos, cuja presença pode fornecer evidências inequívocas da sua fonte ou informações sobre condições oceanográficas recebem o nome de marcadores biológicos e são comumentemente abreviados como biomarcadores. Esse trabalho produziu uma metodologia analítica para identificação e quantificação de biomarcadores como os hidrocarbonetos alifáticos, hidrocarbonetos policíclicos aromáticos, esteróis e alquenonas através de uma única extração lipídica. Essa metodologia foi avaliada por diversos critérios analíticos e validada através de materiais de referência e de exercícios de comparação entre laboratórios. A metodologia foi aplicada em um testemunho de sedimento marinho obtido em Cabo Frio. A quantificação das alquenonas nas amostras do testemunho permitiu estimar a variação da temperatura da superfície oceânica ao longo dos anos e correlacioná-la com variações na intensidade da ressurgência, além de permitir verificar como os demais biomarcadores variaram em função da variação da temperatura do meio onde foram sintetizados. / The fate of most organic matter in the oceans is the marine sediment. This preserved organic matter became an unique reservoir of information about the operation of biogeochemical processes in the geological past, and how these processes responded to environmental changes. Some molecules are only synthesized by sea organisms, others only by higher plants. The relative abundance among these compounds in sea sediments can be used to determine the source of the organic matter. Other organic compounds, without natural sources, are an indicative of anthropogenic influence. The marine sediment also can accumulate compounds with the specific biological sources, which structures are influenced by the enzymatic control of the precursor organisms. All of these compounds, which can provide clear evidences of its sources or information about oceanographic conditions, are called biological markers or biomarkers. The present work put out an analytic method to identify and quantify biomarkers like aliphatic hydrocarbons, PAHs, sterols and alkenones with only one lipidic extraction. This method was evaluated by several analytic criteria and validated by reference material and by interlaboratory comparison study. The method was used to analyze a marine sediment core from Cabo Frio. The alkenones quantification in this core allowed not only appreciate changes in the sea surface temperature (SST) along the years and correlate them with changes in the upwelling, but also verify how the other biomarkers responded to SST changes.

O destino de grande parte da matéria orgânica presente nos oceanos é o sedimento marinho. Essa matéria orgânica, que vai sendo armazenada ao longo dos anos, produz uma reserva única de informações sobre os processos biogeoquímicos ocorridos no passado e como esses processos responderam às mudanças ambientais. Certas moléculas orgânicas são sintetizadas apenas por organismos marinhos ou somente por plantas superiores terrestres. A abundância relativa desses compostos em sedimentos marinhos pode ser usada para determinar a contribuição de matéria orgânica terrestre ou marinha que foi depositada. Outros compostos, cujas fontes naturais são raras ou inexistem, encontrados em sedimentos, refletem a influência antrópica no meio. Há ainda compostos, com origem biológica especifica, cujas estruturas são influenciadas pelo controle enzimático dos organismos que os produzem, que são influenciados pelo meio. Esses compostos, cuja presença pode fornecer evidências inequívocas da sua fonte ou informações sobre condições oceanográficas recebem o nome de marcadores biológicos e são comumentemente abreviados como biomarcadores. Esse trabalho produziu uma metodologia analítica para identificação e quantificação de biomarcadores como os hidrocarbonetos alifáticos, hidrocarbonetos policíclicos aromáticos, esteróis e alquenonas através de uma única extração lipídica. Essa metodologia foi avaliada por diversos critérios analíticos e validada através de materiais de referência e de exercícios de comparação entre laboratórios. A metodologia foi aplicada em um testemunho de sedimento marinho obtido em Cabo Frio. A quantificação das alquenonas nas amostras do testemunho permitiu estimar a variação da temperatura da superfície oceânica ao longo dos anos e correlacioná-la com variações na intensidade da ressurgência, além de permitir verificar como os demais biomarcadores variaram em função da variação da temperatura do meio onde foram sintetizados. / The fate of most organic matter in the oceans is the marine sediment. This preserved organic matter became an unique reservoir of information about the operation of biogeochemical processes in the geological past, and how these processes responded to environmental changes. Some molecules are only synthesized by sea organisms, others only by higher plants. The relative abundance among these compounds in sea sediments can be used to determine the source of the organic matter. Other organic compounds, without natural sources, are an indicative of anthropogenic influence. The marine sediment also can accumulate compounds with the specific biological sources, which structures are influenced by the enzymatic control of the precursor organisms. All of these compounds, which can provide clear evidences of its sources or information about oceanographic conditions, are called biological markers or biomarkers. The present work put out an analytic method to identify and quantify biomarkers like aliphatic hydrocarbons, PAHs, sterols and alkenones with only one lipidic extraction. This method was evaluated by several analytic criteria and validated by reference material and by interlaboratory comparison study. The method was used to analyze a marine sediment core from Cabo Frio. The alkenones quantification in this core allowed not only appreciate changes in the sea surface temperature (SST) along the years and correlate them with changes in the upwelling, but also verify how the other biomarkers responded to SST changes.

A method to detect nitrated polycyclic aromatic hydrocarbons (nitro-PAHs) in the air has been developed using gas chromatography with an atmospheric pressure chemical ionisation source, coupled to a triple quadrupole mass spectrometer (GC-APCI-MS/MS). An attempt to create a GC method using negative chemical ionisation (NCI) and a high-resolution mass spectrometer, Orbitrap, (GC-NCI-MS) was made as well without success. Nitro-PAHs are a group of widespread contaminants considered as possible mutagens and carcinogens. They are formed from incomplete combustion of organic matter as well as secondary reactions between PAHs and nitrogen dioxide (NO2) in the air. Three air samples (outdoor, in an air vent and indoor), both particulate and gaseous phase, collected in central Örebro during the spring were extracted and analysed for nitro-PAHs on GC-APCI-MS/MS and for PAHs on GC-MS in EI (electron ionisation) mode. Several studies have reported lower amounts of nitro-PAHs than PAHs in the air, which can be seen in this report as well. The levels of detected nitro-PAHs (1-nitronaphalene and 9-nitroanthracene) and PAHs (naphthalene, anthracene, fluoranthene and pyrene) in the different samples were 10.2-119 pg/m3 and 291-77700 pg/m3, respectively. Neither group of analytes were found in the particulate phase. The highest concentrations of both nitro-PAHs and PAHs were found indoors, suggesting that other sources than emissions from traffic are responsible.

Large polycyclic aromatic hydrocarbons with molecular mass exceeding 278 Da are rarely object of study, in spite of the fact that some compounds from this group are proved carcinogens. These compounds belong to the group of persistent organic compounds and they are considered as dangerous environmental contaminants. Diploma thesis is focused on the evaluation of the occurence of large PAHs in technological and waste waters produced in black pitch processing in the DEZA Valašské Meziříčí factory. The method of their analysis based on reversed-phase liquid chromatography with non-aqueous gradient; UV-VIS and fluorescence detection will be used. This method will enable the study of changes of large PAHs distribution at various stages of waste water treatment process.

The environmental fate of polycyclic aromatic hydrocarbons (PAH) is a significant issue, raising interest in bioremediation. However, the physio-chemical characteristics of PAHs and the physical, chemical, and biological properties of soils can drastically influence in the degradation. Moreover, PAHs are toxic and carcinogenic for humans and their rapid degradation is of great importance. The process of degradation of pollutants can be enhanced by manipulating abiotic factors. The effect of soil pH on degradation of PAHs with a view to manipulating soil pH to enhance the bioremediation of PAH’s was studied. The degradation rate of key model PAHs (Phenanthrene, Anthracene, Fluoranthene, and Pyrene) was monitored in J Arthur Brower’s topsoil modified to a range of pH between pH 4.0 and pH 9.0 at half pH intervals. Photo-catalytic oxidation of PAHs in the presence of a catalyst (TiO2) under UV light at two different wavelengths was studied. The degradation of PAHs during photo-catalytic oxidation was carried out at varying soil pH, whilst the degradation rate of each individual PAH was monitored using HPLC. It was observed that pH 6.5 was most suitable for the photo-degradation of all the PAHs, whilst in general acidic soil had greater photo-degradation rates than alkaline soil pH. Photo-degradation of PAHs at 375 nm exhibited greater degradation rates compared to 254 nm. Phenanthrene at both the wavelengths had greater degradation rate and pyrene has lower degradation rate of the four PAHs. Pure microbial cultures were isolated from road-side soil by shaken enrichment culture and characterized for their ability to grow on PAHs. Bacterial PAH degraders, isolated via enrichment were identified biochemically and by molecular techniques using PCR amplification and sequencing of 16S rDNA. Sequences were analyzed using BLAST (NCBI) and their percentage identity to known bacterial rDNA sequences in the GeneBank database (NCBI) was compared. The 6 bacterial strains were identified as Pseudomonas putida, Achromobacter xylosoxidans, Microbacterium sp., Alpha proteobacterium, Brevundimonas sp., Bradyrhizobium sp. Similarly, fungal PAH degraders were identified microscopically and with molecular techniques using PCR amplification and sequencing of 18S rDNA and identified as Aspergillus niger and Penicillium freii. Biodegradation of four PAHs with two and four aromatic rings were studied in soil with inoculation of the six identified bacteria and two identified fungi over a range of pH. It was observed that pH 7.5 was most suitable for the degradation of all the PAHs maintained in the dark. A degradation of 50% was observed in soil pH 7.5 within first three days which was a seventh of the time taken at pH 5.0 and pH 6.5 (21 days). Greater fungal populations were found at acidic soil pH and alkaline soil pH, in comparison with neutral pH 7.0. Pencillium sp. was found to be more prevalent at acidic pH whilst Aspergillus sp. was found to be more prevalent at pH 7.5-8.0. Bacterial populations were greater at pH 7.5 which was highly correlated with soil ATP levels. It was therefore evident that the greatest rates of degradation were associated with the greatest bacterial population. Soil enzyme activities in general were also greatest at pH 7.5. The converse effect of pH was found with fastest rate of photo-catalytic degradation at the optimal conditions were observed at acidic condition in soil pH 6.5 whilst, the results obtained during biodegradation at the optimal conditions exhibits fastest rate of degradation at alkaline conditions particularly at pH 7.5. Thus, manipulation of soil pH to 7.5 has significant potential to dramatically increase the degradation rate of PAHs.

Petroleum hydrocarbons are the most widespread contaminants in the environment. Interest in the biodegradation of polycyclic aromatic hydrocarbons and compounds (PAHs/PACs) is motivated by their ubiquitous distribution, their low bioavailability, high persistence in soils and their potentially deleterious effects to human health. Identifying the diversity of microorganisms that degrade PAHs/PACs can be utilised in the development of bioremediation techniques. Understanding the mechanisms of bacterial populations to adapt to the presence of pollutants and the extent that lateral transfer of key functional genes occurs, will allow the exploitation of microbial PAC/PAH-degradative capabilities and therefore enhance the successful application of bioremediation strategies. A key aim of this study was to isolate and identify PAC-degrading bacteria for potential use in future bioremediation programmes. A series of PAC enrichments were established under the same experimental conditions from a single sediment sample taken from a highly polluted estuarine site. Distinct microbial community shifts were directly attributable to enrichment with different PAC substrates. The findings of this study demonstrate that five divisions of the Proteobacteria and Actinobacteria can degrade PACs. By determining the precise identity of the PAC-degrading bacteria isolated, and by comparing these with previously published research, this study showed how bacteria with similar PAC degrading capabilities and 16S rRNA signatures are found in similarly polluted environments in geographically very distant locations e.g. China, Italy, Japan and Hawaii. Such a finding suggests that geographical barriers do not limit the distribution of key PAC-degrading bacteria. This is significant when considering the diversity and global distribution of microbes with PAC-degradative capabilities and the potential for utilising these microbial populations in future bioremediation strategies. In the laboratory, enrichment of bacteria able to utilise PAHs has commonly been performed in liquid media, with the PAH dissolved in a carrier solvent. This study found the presence of a carrier solvent significantly affects the resultant microbial population. Although the same sediment sample was used as the bacterial source in all enrichments, different bacterial strains were obtained depending upon the presence of the carrier solvent and the PAH. This is important when considering appropriate methodology for the isolation of PAH-degrading bacteria for future bioremediation programmes. Additionally, the species comprising the resultant population of the enrichment when a carrier solvent was present were similar to previously reported PAH-degrading species. Such a finding necessitates review of previously reported PAH-degrading bacterial species that have been isolated and identified from enrichments using a carrier solvent. Understanding how bacteria acclimatise to environmental pollutants is vital for exploiting these mechanisms within clear up strategies of contaminated sites. Two major lineages of the α subunit of PAH dioxygenases were identified: Actinobacteria and Proteobacteria. Comparison of the α subunit phylogeny with the 16S rRNA phylogeny implies that the PAH-dioxygenases evolved prior to the separation of these phyla or that lateral transfer occurred in the very distant past. No evidence for lateral transfer of the α subunit between the Actinobacteria and Proteobacteria was found in the phylogenetic analyses of this research. Multiple lateral transfer events were inferred between the species of the Actinobacteria and between the classes of the Proteobacteria. The clustering of the taxa within the α subunit phylogeny indicates that lateral transfer of the α subunit gene occurred after the separation of the classes of Proteobacteria and also after the speciation of the γ-Proteobacteria. These findings reveal how bacteria have acclimatised to PAH pollutants through multiple lateral transfer events of a key PAH-degradative gene. This knowledge of the transfer of genetic material will broaden our prospects of exploiting microbial populations.

In 1990, creosote contamination was discovered at the location of a railroad tie treatment facility active in the 1950s until 1973. In 1997, a phytoremediation field study was implemented with the planting of 1,026 hybrid poplar trees and 36 cells of vegetated and unvegetated grass and legume treatments. The hybrid poplar tree phytoremediation system was designed to control infiltration and groundwater flow and enhance subsurface remediation. The grass phytoremediation system was designed to control erosion and enhance surface soil remediation. The overall objectives of this study were to: (1) assess the extent of subsurface remediation, (2) determine the mechanisms of remediation attributable to the hybrid poplar tree phytoremediation system and microbial degradation, (3) assess the effects of the grass phytoremediation system on surface soil remediation, and (4) determine the mechanisms of surface soil remediation resulting from the grass phytoremediation system. / Master of Science

Cachaça é a terceira das bebidas alcoólicas fermento-destilado mais consumida no mundo, com uma produção de aproximadamente 2,5 litros de bilhões por ano. Tradicionalmente, a cachaça é produzida a partir da destilação do mosto fermentado da cana-de-açúcar. Porém, ela pode sofrer a contaminação por HPAs (Hidrocarbonetos Policíclicos Aromáticos) quando a cana-de-açúcar utilizada na sua produção é queimada antes da sua colheita. Objetivando a distinção entre as amostras de cachaça produzidas a partir da cana-de-açúcar colhida após a queima ou não dos canaviais, foram analisados 15 HPAs em 26 amostras de cachaça produzidas com cana-de-açúcar queimada e 105 amostras de cachaça produzidas com cana-de-açúcar não queimada. As amostras de cachaça foram previamente concentradas por extração em fase sólida (SPE) e analisadas por cromatografia liquida de alta eficiência (HPLC), acoplada a um detector de fluorescência. Este método apresentou uma boa separação cromatográfica para a análise dos seguintes HPAs estudados: naftaleno, acenafteno, fluoreno, fenantreno, antraceno, fluoranteno, pireno, benzo(a)antraceno, criseno, benzo(b)fluoranteno, benzo(k)fluoranteno, benzo(a)pireno, dibenzo(a,h)antraceno, benzo(g,h,i)perileno e indeno(1,2,3-c,d)pireno. O método cromatográfico apresentou bons limites de detecção (5,68E-04 a 45,4 µg L-1), boa porcentagem de extração e recuperação (81,5 ± 9 % a 113 ± 5 %) e uma boa reprodutibilidade (> 90,0 %). As amostras produzidas a partir de cana-de-açúcar queimada apresentaram teores médios totais dos HPAs de 21,1 µg L-1, enquanto que as amostras produzidas com cana-de-açúcar não queimada apresentaram teores médios totais dos HPAs de 1,91 µg L-1. Os dados analíticos foram tratados por meio de análise multivariada (PCA, FA, PLS, DA, LDA, QDA e CDA), possibilitando uma boa distinção entre as amostras produzidas com cana queimada e não queimada. Os resultados dos modelos estatísticos revelaram uma elevada probabilidade (85,0 %) na diferenciação entre estes dois grupos de cachaça / Cachaça is the third most consumed fermented alcoholic beverage in the world, with a production of nearly 2.5 billion liters a year. Traditionally, cachaça is made from the distillation of the fermented sugar cane juice. However, it may be contaminated by PAHs (Polycyclic Aromatic Hydrocarbons) when the sugar cane used for its production is burned before harvesting. Aiming at distinguishing spirits made from burned and non-burned sugar cane, 15 PAHs in 26 cachaças samples obtained from non-burned and 105 cachaça samples obtained from burned sugar cane have been analyzed. The cachaça samples were previously concentrated by solid phase extraction (SPE) and analyzed by high performance liquid chromatography (HPLC), coupled with a fluorescence detector. This method presented good chromatographic separation for the analysis of the following PAHs: naphthalene, acenaphthene, fluorene, phenanthrene, anthracene, fluoranthene, pyrene, benz(a)anthracene, chrysene, benz(b)fluoranthene, benz(k)fluoranthene, benz(a)pyrene, dibenz(a,h)anthracene, benz(g,h,i)perylene, and indeno(1,2,3- c,d)pyrene. The chromatographic method showed good detection limits (5.68E-04 to 45.4 ? g L-1), good index for the clean up percentage and recovery (from 81.5 ± 9% to 113 ± 5%) and a good reproducibility (> 90.0%). The samples obtained from burned sugar cane presented 21.1 ? g L-1 total PAHs average contents while the ones obtained from non-burned sugar cane showed PAHs average contents of 1.91 ?g L-1. Such analytical data were treated by means of the multivariate statistical methods (PCA, FA, PLS, LDA, QDA and CDA), thus allowing good distinction among the samples obtained from burned and non-burned sugar cane. Results of the Abstract IQSC-USP statistical models presented high probability (85.0%) for the distinction between these two groups of spirits.

Os hidrocarbonetos policíclicos aromáticos (HPAs) por serem considerados contaminantes persistentes, pela sua ubiqüidade no meio ambiente e pelo reconhecimento de sua genotoxicidade, têm estimulado atividades de pesquisa com o objetivo de determinar e avaliar as origens, transporte, transformação, efeitos biológicos e a acumulação em compartimentos de ecossistemas aquático e terrestre. Nesta dissertação, a matriz estudada foi o sedimento coletado na represa do Parque Pedroso em Santo André, SP. A técnica analítica empregada foi a cromatografia líquida em fase reversa com detector UV/Vis. O tratamento dos dados foi estabelecido durante o processo de desenvolvimento da metodologia para que houvesse confiabilidade nos resultados. As etapas envolvidas foram avaliadas usando o conceito da Validação de Ensaios Químicos. Os parâmetros selecionados para a validação analítica foram Seletividade, Linearidade, Intervalo de Trabalho, Sensibilidade, Exatidão, Precisão, Limite de Detecção, Limite de Quantificação e Robustez. Esses parâmetros apresentaram resultados satisfatórios, o que permite a aplicação da metodologia desenvolvida, sendo ainda um método simples que permite a minimização da contaminação e da perda de compostos por excesso de manipulação. Para os HPAs estudados não foram encontrados resultados positivos, acima do limite de detecção, em nenhuma das amostras coletadas na primeira fase. Já na segunda coleta foram encontradas pequenas alterações principalmente em acenaftileno, fluoreno e benzo[a]antraceno. Embora a área se encontre preservada, já se notam pequenos sinais de contaminação. / The polycyclic aromatic hydrocarbons (PAHs), by being considered persistent contaminants, by their ubiquity in the environment and by the recognition of their genotoxicity, have stimulated research activities in order to determine and evaluate their sources, transport, processing, biological effects and accumulation in compartments of aquatic and terrestrial ecosystems. In this work, the matrix studied was sediment collected at Pedroso Park\'s dam at Santo André, SP. The analytical technique employed was liquid chromatography in reverse phase with a UV/Vis detector. Statistics treatment of the data was established during the process of developing the methodology for which there was reliable results. The steps involved were evaluated using the concept of Validation of Chemical Testing. The parameters selected for the analytical validation were selectivity, linearity, Working Range, Sensitivity, Accuracy, Precision, Limit of Detection, Limit of quantification and robustness. These parameters showed satisfactory results, allowing the application of the methodology, and is a simple method that allows the minimization of contamination and loss of compounds by over-handling. For the PAHs tested were no found positive results, above the limit of detection, in any of the samples collected in the first phase. But, at the second collection, were found small changes mainly acenaphthylene, fluorene and benzo[a]anthracene. Although the area is preserved, it is possible to realize little signs of contamination.

Polycyclic aromatic hydrocarbon (PAH) compounds, which are common to coal tar and coal-tar-based products, are ubiquitous environmental contaminates with carcinogenic effects to human health and aquatic life. Transport of PAHs via solid phase particulate motion, gaseous phase volatilization, and aqueous phase dissolution into urban waterbodies of PAH compounds from coal-tar-based pavement sealant products has been studied. Pavement rejuvenators are products applied to increase the usable life of pavement. Coal-tar-based rejuvenators contain a significantly larger mass fraction of coal-tar with respect to coal-tar-based sealants, but pavement rejuvenators have not been as extensively studied as pavement sealants. Chemical analysis of detached pavement material treated with coal-tar-bases, asphalt-based, and bio-based rejuvenators was conducted with gas chromatography – mass spectrometry (GC/MS) analysis for 16 PAH compounds and two aromatic heterocyclic organic compounds following extraction with methylene chloride. Detached pavement material was collected from 19 simulated asphalt surface abrasion experiments that used a model mobile load simulator (MMLS) test apparatus that replicated surface challenges from vehicular traffic. The MMLS test apparatus configuration allowed asphalt disc samples treated with different rejuvenation products, to be tested and for detached material to be collected and quantified prior to GC/MS analysis. Test cases evaluated the influence of rejuvenation product type and cure time, as well as the effect of sand application (simulating sand application during slippery winter storm conditions) had on detached particulate and ultimate PAH compound loading. The average mass of particulate detachment from samples following a 48 hour cure time, for the asphalt-based and coal-tar-based rejuvenator products were 0.347 g and 0.480 g, respectively. This mass of detached material was lower than that from pavement treated with bio-based rejuvenator and the control (not treated), which had 4.858 g and 2.567 g of detached particulate material, respectively. When the product cure time was increased to three weeks, which was significantly long enough to capture effects of compound volatilization, average particulate detachment increased to 0.882 g for the coal-tar-based rejuvenator and decreased for the bio-based rejuvenator to 2.600 g. Six tests performed with a single application of winter storm sand after a 48 hour product cure time showed an increase in average particulate detachment to 1.450 g and 0.617 g for pavement treated with the asphalt-based and coal-tar-based rejuvenators, respectively. Conversely, under the same conditions, a reduction in average detached particulate to 3.749 g was observed for the bio-based product. Detached particulate material quantities for each test case were used with the respective cumulative concentration of 16 PAH compounds quantified to make an assessment on the potential PAH compound contamination via solid phase particle transport. The average PAH compound concentration in particulate detached from pavement treated with a coal-tar-based rejuvenator was 3062.8 mg PAH per kg of particulate. This was an order of magnitude higher than the average PAH concentration measured in particulate detached from the two control pavement samples and the two asphalt-based samples after a 48 hour cure time, which were 322.1 and 508.1 mg PAH per kg detached particulate, respectively. PAH compound concentrations were also normalized by the surface area of pavement treated with a rejuvenator to determine the potential PAH compound contamination per unit area. Normalized results for each rejuvenator type were averaged to make an overall evaluation of the potential rejuvenator specific PAH compound loading. The coal-tar-based, bio-based, and asphalt-based rejuvenators had a normalized cumulative solid-phase PAH compound release of 2.35, 0.88, and 0.17 mg PAH per square foot of pavement rejuvenated, respectively. In addition, carbazole was quantified in all pavement samples treated with the coal-tar-based rejuvenator at an average concentration of 125.6 mg carbazole per kg detached particulate. Acridine was quantified in detached particulate from five of seven coal-tar-based test performed at an average concentration (excluding non-detection samples) of 42.1 mg acridine per kg detached particulate.