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Tratamento anaeróbio de efluente contendo pentaclorofenol em reator de leito fluidificado / Anaerobic treatment of effluent cointaining pentachlorophenol in a fluidized bed reactorFreire, Flavio Bentes 21 March 2005 (has links)
Remoções quase totais de clorofenóis, por processos anaeróbios, já foram atingidas mediante elevada concentração de matéria orgânica e um número considerável de fontes de carbono. Porém, é necessário que se investigue o desempenho dos reatores submetidos a condições menos idealizadas. Este trabalho apresenta o desenvolvimento de uma metodologia para avaliar um reator anaeróbio de leito fluidificado (RALF) como etapa de pré-tratamento de substrato sintético contendo pentaclorofenol (PCP). Para isso, foram adotadas as seguintes condições experimentais: fonte única de carbono; concentração reduzida de matéria orgânica; biomassa não previamente adaptada, e menores tempos de detenção hidráulica. Para cumprir o objetivo citado, foram desenvolvidos dois reatores, com 2,2 e 16 litros, respectivamente, e foram realizadas etapas de avaliação de materiais suporte, estudo de diversas interações (partícula/fluido, partículas fluido, biopartícula/fluido e biopartículas/fluido), e análise do desempenho do RALF sem PCP e com PCP. Os materiais suporte escolhidos foram a alumina, o basalto e o carvão ativado granular, todos com diâmetros próximos a 3 mm. Eles foram avaliados em ensaios de caracterização física, de microscopia eletrônica de varredura, de fluidodinâmica e de análise de imagens. O carvão apresentou os melhores resultados e foi escolhido para a continuidade do trabalho. As interações foram analisadas através de ensaios fluidodinâmicos, isotermas de adsorção, dentre outros. Nas condições experimentais utilizadas, a biomassa aumentou a densidade da partícula, fato que foi comprovado pelos ensaios para determinação da velocidade mínima de fluidificação, de expansão e porosidade do leito, e também de velocidade terminal da partícula isolada. As isotermas de adsorção demonstraram que a presença de matéria orgânica diminuiu em 9% a adsorção do PCP. O RALF apresentou um comportamento satisfatório no período de operação sem PCP. Nos 105 dias de operação, a eficiência média de remoção de DQO ficou quase sempre superior a 90%, e os outros parâmetros de monitoramento de desempenho apresentaram valores típicos, indicando estabilidade no processo. A presença de PCP no sistema, nas concentrações utilizadas, não alterou a qualidade da biomassa presente, e nem os parâmetros de monitoramento de desempenho, como DQO, alcalinidade, pH e ácidos voláteis. Mesmo sob condições menos idealizadas, foi observado um desempenho satisfatório do reator na remoção do PCP. Em concentrações variando de 1 a 6 mg/L, foram observadas eficiências médias de remoção de 93% e 70%, respectivamente, para os 80 dias de operação nessas condições. Diante dos resultados obtidos, foi possível concluir que a remoção de PCP em um RALF pode ser atingida mediante condições operacionais mais simples. / The almost total removal of chlorophenols by anaerobic processes has been achieved under high concentrations of organic matter and a considerably large number of carbon sources. However, it is necessary to investigate the performance of such reactors under less idealized conditions. The present work shows the development of a new methodology for evaluating the use of an anaerobic fluidized bed reactor (AFBR) in the pretreatment step of a synthetic substrate containing pentachlorophenol (PCP). In order to do that, the following experimental conditions were used: a single source of carbon; reduced concentrations of organic matter; non-previously adapted biomass, and smaller hydraulic retention times. To achieve the main objective, two reactors, one having a volume of 2.2 L and the other one, 16 L, were especially designed; evaluation steps of support material and the study of several interactions (particle/fluid, particles/fluid, bioparticle/fluid, bioparticles/fluid) were carried out, and the performance of the AFBR with and without PCP was analyzed. The three support materials employed were alumina, basalt and granular activated carbon, all having average diameters of 3 mm. These materials were evaluated through physical characterization methods, microscopy, hydrodynamic and image analyses. The activated coal showed the best results and was exclusively employed in the remainder of the work. The interactions were analyzed by hydrodynamics and adsorption isotherms, to name but a few. Under the experimental conditions employed, the biomass made the particle density increase, a fact that was verified in the determination of the minimum fluidization velocity, the bed porosity and expansion as well as the terminal velocity of isolated particles. The adsorption isotherms showed a decrease of 9% in PCP absorption due to the existence of organic matter. The AFBR showed satisfactory results during the operation without PCP. In the 105 days of operation, the average efficiency of COD removal remained almost always over 90%, while the other performance monitoring parameters remained at typical values, indicating the process stability. The presence of PCP under the concentrations used in the system did not seem to affect neither the quality of the biomass nor the performance monitoring parameters, like COD, alkalinity, pH and volatile acids. Even under less idealized conditions, the reactor showed good performance in removing PCP. The average efficiencies of removal attained around 93% and 70% for concentrations of 1 to 6 mg/L respectively. These concentration levels were kept during 80 days. In view of the results obtained, it is possible to conclude that reasonably good efficiency of PCP removal in AFBR can be attained under simpler operational conditions.
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Avaliação da performance de um reator anaeróbio híbrido (RAH) e da atividade das populações de microrganismos anaeróbios na ausência e na presença de Pentaclorofenol (PCP) / not availableMontenegro, Martha de Almeida Prado 01 June 2001 (has links)
O presente trabalho foi desenvolvido com o objetivo de verificar a eficiência de um reator anaeróbio híbrido (RAH) alimentado com uma mistura de ácidos orgânicos acético, propiônico, butírico, e láctico, bem como do álcool metanol, perfazendo uma DQO de 6,88 g/L, e avaliar nessas condições, a degradação do pentaclorofenol (PCP) na faixa de 2,0 a 21,0 mg/L. O RAH apresentou adequada performance na ausência de PCP, tendo sido inoculado com um lodo com AME cerca de 0,57 g DQO-CH4/g SV.d. Durante os 21 meses de operação do RAH na ausência de PCP verificou-se uma remoção média de DQO de 93% produção média de metano de 84%. Através de testes de toxicidade realizadas em batelada com o lodo granulado do RAH, antes da adição de PCP no reator, calculou-se o IC50, cujos valores foram 10, 12 e 13,69 mg/L. Na presença de PCP na faixa de 2,0 a 2,1 mg/L, o RAH apresentou remoção média de DQO de 96,7%, produção média de metano de 85,5% e remoção dos ácidos voláteis próxima a 74% do acético, 93% do butírico e 64% do propiônico. Individualmente, na presença da maior concentração de PCP adicionada, ocorreu decréscimo na remoção dos ácidos voláteis, principalmente do ácido propiônico e na taxa de conversão DQO/biogás. O PCP foi removido do sistema na ordem de 99% pela ação do lodo granulado com predominância do grupo das Archaea metanogênicas, verificada por exames microscópicos e hibridação \"in situ\". Valores da ordem de grandeza microbiana para os microrganismos metanogênicos nos períodos anteriores e posteriores a adição de PCP permaneceram na faixa de 105 e 106 céls./mL, quando cultivados em metanol e lactato mais sulfato, respectivamente. Os resultados sugerem que as Archaea metanogênicas podem estar envolvidas na degradação do PCP. A velocidade de remoção do organoclorado foi igual a 1,07 mg PCP/g SV.d quando a maior concentração de PCP foi estudada (21,0 mg/L). / The present research aimed to verify the efficiency of an Anaerobic Hybrid Reactor (AHR) supplied with a mixture of fatty acids, acetic, propionic, butyric and lactic and methanol as well. The total amount of COD was 6.88 g/L. The performance of the reactor was remarkably stable and efficient during PCP additions at range from 2.0 to 21.0 mg/L. The AHR showed a great performance in the PCP absence, inoculated with sludge with an specific methanogenic (SMA) activity of 0,057 g.COD-CH4/g. VS.d. During the 21 months of operation without PCP, the reduction of COD was around 93% and methane was up to 84% in the biogas. Before PCP addiction, two toxicity batch tests conducted with the granular sludge presented IC50 values around 10.12 mg/L and 13.69 mg/L. In the presence of PCP, at the range of 2.0 to 21.0 mg/L, the efficiency of volatile fatty acids breakdown was 93%, 64% and 74% respectively for butyric, propionic and acetic acids. Individually, at the presence of the higher PCP concentration studied, a decrease in the conversion of COD to biogas and organic acids removal occurred, mainly with propionic acid. PCP total removal of more than 99% was reached by granular sludge activities formed by the total time of reactor operation with a prevalence of methanogenic Archaea, verified under direct microscopy exams and in situ hybridization. Methanogenic cells predominance was noticed with 105 to 106 cells/mL during enumeration on methanol and lactate plus sulfate culture media, respectively. The results suggest that methanogenic Archaea can be involved in PCP degradation. The organochlorine removal rate was 1.07 mg PCP.g-1 VS.d-1 during the highest PCP (21.0 mg/L) concentration addition.
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Avaliação da comunidade e atividade microbiana em reator anaeróbio de leito fixo (RAHLF) operado com pentaclorofenol (PCP), através de métodos cromatográficos, exames microscópicos e técnicas moleculares como PCR, ARDRA e slot-blot / Evaluation of microbial communities and their activities in a horizontal anaerobic immobilized system (HAIS) fed with pentachlorophenol (PCP) by using chromatography, microscopy and molecular techniques of the PCR, ARDRA and slot-blotElizabeth Aparecida Baraldi 06 August 2001 (has links)
Foi estudada a degradação do pentaclorofenol (PCP) em reator aneróbio horizontal de leito fixo (RAHLF) de volume de 2000 mL. O reator foi inoculado com microrganismos oriundos de reatores aneróbios não previamente adaptados a PCP. Atividade microbiana foi monitorada através de técnicas clássicas na presença do organoclorado na faixa de 2,0 a 13 mg/L de PCP. O reator apresentou eficiência de 97% na remoção de DQO e completo desaparecimento do composto de PCP em todas as concentrações testadas. A fração orgânica foi consumida totalmente na primeira terça parte do reator de acordo com os valores determinados de ácidos voláteis, DQD e PCP. Não foi verificada inibição da atividade de culturas microbianas. Os exames microscópicos, fluorescência e varredura, permitiram verificar o predomínio de microrganismos pertencentes ao Domínio Archea. As técnicas moleculares PCR, ARDRA e hibridação slot-blot confirmaram o predomínio do Domínio Archaea e possibilitaram a verificação de alterações na diversidade das populações após adição de 2 mg PCP/L. Conclui-se que o reator sem prévia adaptação do inóculo foi eficiente para o tratamento do PCP, e os microrganismos relacionados às Archaea metanogênicas acetocláticas podem estar envolvidas na degradação deste composto. / The degradation of pentachlorophenol (PCP) was studied in a 2000 mL. Horizontal Anaerobic Immobilized System (HAIS). The reactor was inoculated with microorganisms obtained from an anaerobic reactor without previous adaptation to the PCP. The microbial activity was evaluated by using classic techniques in order to . monitor its behavior during the HAIS fed with a range of PCP between 2.0 to 13 mg/L. The reactor presented 97% of efficiency in the removal of COD and complete decrease of PCP in alI concentrations tested. The total consumption of organic fraction took place mainly in the first third part of the reactor according the values of volatile fatty acids, COD and PCP obtained. Microbial inhibition was not verified in during HAIS operation. Microscopic examinations allowed certifying the Archaea Domain predominance according the morphologies observed. The molecular techniques polimerase chain reaction (PCR), ARDRA and slot-blot hibridation confirmed the predominance of Archaea Domain and alIowed verifying some changes in the population\'s diversity under additions of 2mg PCP/L. The efficiency of PCP decreased in the anaerobic reactor was related to the presence of Archaea Domain, especially the acetoclastic methanogens, whose where probably involved with the organochlorine compound degradation.
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Synthesis and characterization of nitrogen-doped titanium oxide nanoparticles for visible-light photocatalytic wastewater treatmentPelaschi, Mohammad Ali 05 October 2018 (has links)
TiO2 nanoparticles are one of the most suitable materials for photocatalysis, specifically for water and air treatment and removal of a wide variety of organic pollutants such as dyes, aromatic compounds, and chlorinated aromatic compounds. Methods of synthesis of TiO2 are generally categorized in two main classes of wet chemical, and dry methods. Wet chemical methods generally provide a better control over size, size distribution, and shape; all of which significantly affect photocatalytic performance of the produced nanoparticles. Despite its advantages over other semiconductor photocatalysts, wide band-gap of titania restrains its photocatalytic activity to only UV light, which only makes up to 5% of the light reaching surface of the earth. To induce visible-light activity, titania has been doped by different dopants, including transition metal-dopants such as Fe, and Co and non-metal dopants such as N, and C. Nitrogen has been shown to be a better dopant, providing a suitably placed energy state within the band-gap of TiO2, and not suffering from issues related to transition-metal dopants such as low thermal and physical stability and high electron-hole recombination rates. To dope titania with nitrogen, one could add the nitrogen source together with other precursors during synthesis, referred to as wet chemical doping methods, or anneal the synthesized titania nanoparticles under a flow of ammonia at high temperatures, referred to as dry doping methods. While different doping methods have been studied individually, the author maintains that there has been an absence of research comparing the effectiveness of these methods, on photocatalytic performance of N-doped TiO2 within a consistent experiment. In this research TiO2 nanoparticles were synthesized by a facile, inexpensive sol-gel method, and doping was done by wet chemical methods, dry methods, and a combination of both these methods. Visible-light photocatalytic activity of these nanoparticles was evaluated by their efficiency in degradation of methyl orange. The results show wet doping methods increase the efficiency of titania nanoparticles more than dry doping, or combination of both. Further investigation showed that the main reason for higher activity of wet chemically doped nanoparticles is due to their higher available surface area of 131.7 m2.g-1. After normalizing the available surface area, measured by the BET method, it was shown that a combination of wet chemical doping, and dry doping at 600 °C result in the most active nanoparticles, but high temperature dry doping severely decreases the surface area, lowering the overall efficiency of the product. Additionally, N-doped TiO2 nanoparticles were synthesized using a simple hydrothermal method, in which the nitrogen source was used not only to dope, but also to control shape, size, size distribution, and morphology of the titania nanoparticles, and to induce aqueous colloidal stability. It was shown that addition of triethylamine during the synthesis, results in ultra-small, colloidally stable, cubic TiO2 nanoparticles, while using triethanolamine results in formation of TiO2 pallets, assembled into spherical, rose-like structures. The synthesized nanoparticles show impressive efficiency in visible-light removal of phenol, 4-chlorophenol, and pentachlorophenol, achieving 100% degradation of a 100-ppm phenol solution in 90 min, more than 98% degradation of a 20-ppm 4-chlorophenol solution in 90 min, and 97% degradation of a 10-ppm pentachlorophenol in 180 min with 500 ppm loading of the catalyst in all cases. Moreover, synthesized nanoparticles showed no sign of deactivation after 5 consecutive runs, removing 4-chlorophenol, showing their reusability. / Graduate
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Tratamento anaeróbio de efluente contendo pentaclorofenol em reator de leito fluidificado / Anaerobic treatment of effluent cointaining pentachlorophenol in a fluidized bed reactorFlavio Bentes Freire 21 March 2005 (has links)
Remoções quase totais de clorofenóis, por processos anaeróbios, já foram atingidas mediante elevada concentração de matéria orgânica e um número considerável de fontes de carbono. Porém, é necessário que se investigue o desempenho dos reatores submetidos a condições menos idealizadas. Este trabalho apresenta o desenvolvimento de uma metodologia para avaliar um reator anaeróbio de leito fluidificado (RALF) como etapa de pré-tratamento de substrato sintético contendo pentaclorofenol (PCP). Para isso, foram adotadas as seguintes condições experimentais: fonte única de carbono; concentração reduzida de matéria orgânica; biomassa não previamente adaptada, e menores tempos de detenção hidráulica. Para cumprir o objetivo citado, foram desenvolvidos dois reatores, com 2,2 e 16 litros, respectivamente, e foram realizadas etapas de avaliação de materiais suporte, estudo de diversas interações (partícula/fluido, partículas fluido, biopartícula/fluido e biopartículas/fluido), e análise do desempenho do RALF sem PCP e com PCP. Os materiais suporte escolhidos foram a alumina, o basalto e o carvão ativado granular, todos com diâmetros próximos a 3 mm. Eles foram avaliados em ensaios de caracterização física, de microscopia eletrônica de varredura, de fluidodinâmica e de análise de imagens. O carvão apresentou os melhores resultados e foi escolhido para a continuidade do trabalho. As interações foram analisadas através de ensaios fluidodinâmicos, isotermas de adsorção, dentre outros. Nas condições experimentais utilizadas, a biomassa aumentou a densidade da partícula, fato que foi comprovado pelos ensaios para determinação da velocidade mínima de fluidificação, de expansão e porosidade do leito, e também de velocidade terminal da partícula isolada. As isotermas de adsorção demonstraram que a presença de matéria orgânica diminuiu em 9% a adsorção do PCP. O RALF apresentou um comportamento satisfatório no período de operação sem PCP. Nos 105 dias de operação, a eficiência média de remoção de DQO ficou quase sempre superior a 90%, e os outros parâmetros de monitoramento de desempenho apresentaram valores típicos, indicando estabilidade no processo. A presença de PCP no sistema, nas concentrações utilizadas, não alterou a qualidade da biomassa presente, e nem os parâmetros de monitoramento de desempenho, como DQO, alcalinidade, pH e ácidos voláteis. Mesmo sob condições menos idealizadas, foi observado um desempenho satisfatório do reator na remoção do PCP. Em concentrações variando de 1 a 6 mg/L, foram observadas eficiências médias de remoção de 93% e 70%, respectivamente, para os 80 dias de operação nessas condições. Diante dos resultados obtidos, foi possível concluir que a remoção de PCP em um RALF pode ser atingida mediante condições operacionais mais simples. / The almost total removal of chlorophenols by anaerobic processes has been achieved under high concentrations of organic matter and a considerably large number of carbon sources. However, it is necessary to investigate the performance of such reactors under less idealized conditions. The present work shows the development of a new methodology for evaluating the use of an anaerobic fluidized bed reactor (AFBR) in the pretreatment step of a synthetic substrate containing pentachlorophenol (PCP). In order to do that, the following experimental conditions were used: a single source of carbon; reduced concentrations of organic matter; non-previously adapted biomass, and smaller hydraulic retention times. To achieve the main objective, two reactors, one having a volume of 2.2 L and the other one, 16 L, were especially designed; evaluation steps of support material and the study of several interactions (particle/fluid, particles/fluid, bioparticle/fluid, bioparticles/fluid) were carried out, and the performance of the AFBR with and without PCP was analyzed. The three support materials employed were alumina, basalt and granular activated carbon, all having average diameters of 3 mm. These materials were evaluated through physical characterization methods, microscopy, hydrodynamic and image analyses. The activated coal showed the best results and was exclusively employed in the remainder of the work. The interactions were analyzed by hydrodynamics and adsorption isotherms, to name but a few. Under the experimental conditions employed, the biomass made the particle density increase, a fact that was verified in the determination of the minimum fluidization velocity, the bed porosity and expansion as well as the terminal velocity of isolated particles. The adsorption isotherms showed a decrease of 9% in PCP absorption due to the existence of organic matter. The AFBR showed satisfactory results during the operation without PCP. In the 105 days of operation, the average efficiency of COD removal remained almost always over 90%, while the other performance monitoring parameters remained at typical values, indicating the process stability. The presence of PCP under the concentrations used in the system did not seem to affect neither the quality of the biomass nor the performance monitoring parameters, like COD, alkalinity, pH and volatile acids. Even under less idealized conditions, the reactor showed good performance in removing PCP. The average efficiencies of removal attained around 93% and 70% for concentrations of 1 to 6 mg/L respectively. These concentration levels were kept during 80 days. In view of the results obtained, it is possible to conclude that reasonably good efficiency of PCP removal in AFBR can be attained under simpler operational conditions.
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Avaliação da performance de um reator anaeróbio híbrido (RAH) e da atividade das populações de microrganismos anaeróbios na ausência e na presença de Pentaclorofenol (PCP) / not availableMartha de Almeida Prado Montenegro 01 June 2001 (has links)
O presente trabalho foi desenvolvido com o objetivo de verificar a eficiência de um reator anaeróbio híbrido (RAH) alimentado com uma mistura de ácidos orgânicos acético, propiônico, butírico, e láctico, bem como do álcool metanol, perfazendo uma DQO de 6,88 g/L, e avaliar nessas condições, a degradação do pentaclorofenol (PCP) na faixa de 2,0 a 21,0 mg/L. O RAH apresentou adequada performance na ausência de PCP, tendo sido inoculado com um lodo com AME cerca de 0,57 g DQO-CH4/g SV.d. Durante os 21 meses de operação do RAH na ausência de PCP verificou-se uma remoção média de DQO de 93% produção média de metano de 84%. Através de testes de toxicidade realizadas em batelada com o lodo granulado do RAH, antes da adição de PCP no reator, calculou-se o IC50, cujos valores foram 10, 12 e 13,69 mg/L. Na presença de PCP na faixa de 2,0 a 2,1 mg/L, o RAH apresentou remoção média de DQO de 96,7%, produção média de metano de 85,5% e remoção dos ácidos voláteis próxima a 74% do acético, 93% do butírico e 64% do propiônico. Individualmente, na presença da maior concentração de PCP adicionada, ocorreu decréscimo na remoção dos ácidos voláteis, principalmente do ácido propiônico e na taxa de conversão DQO/biogás. O PCP foi removido do sistema na ordem de 99% pela ação do lodo granulado com predominância do grupo das Archaea metanogênicas, verificada por exames microscópicos e hibridação \"in situ\". Valores da ordem de grandeza microbiana para os microrganismos metanogênicos nos períodos anteriores e posteriores a adição de PCP permaneceram na faixa de 105 e 106 céls./mL, quando cultivados em metanol e lactato mais sulfato, respectivamente. Os resultados sugerem que as Archaea metanogênicas podem estar envolvidas na degradação do PCP. A velocidade de remoção do organoclorado foi igual a 1,07 mg PCP/g SV.d quando a maior concentração de PCP foi estudada (21,0 mg/L). / The present research aimed to verify the efficiency of an Anaerobic Hybrid Reactor (AHR) supplied with a mixture of fatty acids, acetic, propionic, butyric and lactic and methanol as well. The total amount of COD was 6.88 g/L. The performance of the reactor was remarkably stable and efficient during PCP additions at range from 2.0 to 21.0 mg/L. The AHR showed a great performance in the PCP absence, inoculated with sludge with an specific methanogenic (SMA) activity of 0,057 g.COD-CH4/g. VS.d. During the 21 months of operation without PCP, the reduction of COD was around 93% and methane was up to 84% in the biogas. Before PCP addiction, two toxicity batch tests conducted with the granular sludge presented IC50 values around 10.12 mg/L and 13.69 mg/L. In the presence of PCP, at the range of 2.0 to 21.0 mg/L, the efficiency of volatile fatty acids breakdown was 93%, 64% and 74% respectively for butyric, propionic and acetic acids. Individually, at the presence of the higher PCP concentration studied, a decrease in the conversion of COD to biogas and organic acids removal occurred, mainly with propionic acid. PCP total removal of more than 99% was reached by granular sludge activities formed by the total time of reactor operation with a prevalence of methanogenic Archaea, verified under direct microscopy exams and in situ hybridization. Methanogenic cells predominance was noticed with 105 to 106 cells/mL during enumeration on methanol and lactate plus sulfate culture media, respectively. The results suggest that methanogenic Archaea can be involved in PCP degradation. The organochlorine removal rate was 1.07 mg PCP.g-1 VS.d-1 during the highest PCP (21.0 mg/L) concentration addition.
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A mechanistic study of organochlorine hepatotoxicitySchroeder, Ilka Elizma 22 May 2012 (has links)
Pentachlorophenol, (PCP) is an organochlorine compound which was first developed in the 1930’s. PCP is said to be the most toxic of the chlorophenols and is classified as a hazardous substance and a probable human carcinogen. PCP has proven to be cytotoxic to a number of cell lines translating to its effect on various organs. The aim of the study was to assess organochlorine-induced hepatotoxicity in a mechanistic manner using an in-house developed procedure. Also, the possible hepatoprotective effect of methanolic extracts of the bark of two medicinal plants, Burkea africana (BA) and Syzygium cordatum (SC), as well as the known hepatoprotective agent, N-acetyl cysteine (NAC), were investigated. In addition to PCP, two of its major metabolites, tetrachloro-1,2-hydroquinone (TCHQ) and tetrachloro-1,4-benzoquinone (TCBQ) were also evaluated. A hepatocarcinoma cell line (HepG2) was used to investigate the effect of these compounds on different parameters of cellular function. Cytotoxicity was assessed using the neutral red uptake assay. Cytochrome P4501A1 (CYP1A1) activity was determined using ethoxy-resorufin-O-deethylation as surrogate. Generation of reactive oxygen species (ROS) was investigated by measuring dichlorofluorescein diacetate cleavage. Effects on mitochondrial membrane potential were determined using JC-1 staining, whilst necrosis was investigated by assessing plasma membrane integrity using propidium iodide (PI)staining. The degree of apoptotic death was determined by quantifying caspase-3 activity. Assays were repeated with an additional 1 h pre-treatment of the cells with either NAC, SC or BA in order to investigate whether these compounds were able to protect against the toxicity induced by PCP and its metabolites. The IC50 values of PCP, TCHQ and TCBQ were 68.0, 144.0 and 129.4 μM, respectively. All three test compounds induced CYP1A1 activity with PCP being the most potent. TCBQ produced extensive ROS generation. TCHQ also induced ROS generation, whilst PCP appeared to have no significant effect on ROS generation. All test compounds caused mitochondrial depolarization. None of the test compounds caused an increase in necrotic cell death. PCP, TCHQ and TCBQ had negligible effects on apoptosis. Both SC and BA alleviated the toxic effects observed in cells treated with PCP. Minor increases in viability occurred in cells pre-treated with plant extracts prior to exposure to both metabolites. NAC, as well as both plant extracts, greatly reduced CYP 1A1 activity induced by PCP. NAC, SC and BA exacerbated CYP1A1 induction in cells exposed to concentrations of TCBQ and TCHQ that initially produced little or no effect on CYP1A1 activity. Contrarily, decreased CYP1A1 activity was observed in cells exposed to concentrations of TCBQ and TCHQ where extensive induction of CYP1A1 occurred. NAC, as well as both plant extracts, suppressed ROS generation in cells exposed to all test compounds. In cells exposed to PCP and TCBQ more extensive mitochondrial depolarization was seen when pre-treated with NAC and plant extracts than when exposed to the compounds alone. Negligible effects were seen in pre-treated cells exposed to TCHQ. BA and SC caused increases in necrotic death in cells exposed to the test compounds. NAC, BA and SC had negligible effects on the changes in caspase-3 activity induced by the test compounds. From the results it is proposed that PCP induces its own metabolism by increasing CYP1A1 activity. It also causes mitochondrial insult which could lead to the opening of the mitochondrial permeability transition pore and subsequent release of cytochrome C, activation of caspases and eventually apoptotic cell death. With regard to TCHQ and TCBQ, results suggest that extensive ROS generation caused damage to various cellular macromolecules and that this could be the main cause of their toxicity. NAC, SC and BA appeared to alleviate toxicity in certain instances. Further investigation is required in order to assess them as possible hepatoprotective agents. Copyright / Dissertation (MSc)--University of Pretoria, 2011. / Pharmacology / unrestricted
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