Browsing by Author "Rebola, Marlene"
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- Chlorinated Polycyclic Aromatic Hydrocarbons Associated with Drinking Water Disinfection: Synthesis, Formation under Aqueous Chlorination Conditions and Genotoxic EffectsPublication . Pinto, Miguel; Rebola, Marlene; Louro, Henriqueta; Antunes, Alexandra M.M.; José, Silvia S.; Rocha, Maria; Silva, Maria João; Cardoso, Ana SofiaPolycyclic aromatic hydrocarbons (PAHs) are among the most persistent and toxic organic micropollutants present in water and several of them are mutagenic and carcinogenic. Although it has been shown that chlorinated derivatives of PAHs (Cl-PAHs) may be formed during the water chlorination procedure, little is known about their potential genotoxic and carcinogenic effects. The objectives of the present work were to prepare and characterize the major chlorinated derivatives of benzo[a]pyrene (BaP) and fluoranthene (Fluo), to develop an analytical methodology for their quantification in water samples and to analyse their potential genotoxicity. Chlorinated standards were prepared by a newly developed two phase method (water/n-hexane) using sodium hypochlorite. 6-Chloro-benzo[a]pyrene was selectively obtained from BaP, while 1,3- dichloro-fluoranthene and 3-chloro-fluoranthene were obtained from Fluo. All products were isolated and characterized by nuclear magnetic resonance and mass spectrometry. The formation of BaP- and Fluo-chlorinated derivatives under aqueous chlorination conditions was observed using a SPE-HPLC-FLD methodology. In addition, the cytotoxic and genotoxic activities of the three chlorinated derivatives were analyzed in comparison to their parent compounds, in a human-derived hepatoma cell line using the neutral red uptake and comet assays, respectively. The results showed that, at the equimolar doses of 100 and 125 μM, 6-Cl-BaP was able to induce a significantly higher level of DNA damage than BaP, suggesting a more potent genotoxic effect. In contrast, neither Fluo nor its chlorinated derivatives were genotoxic in the same cell line. The identification of new and possibly hazardous water chlorination by-product from PAHs emphasizes the need to minimize total organic carbon content of raw water and the implementation of safer water disinfection methods.
- Contaminantes da água com potencial genotóxico: cianotoxinas e subprodutos de desinfeção da águaPublication . Dias, Elsa; Cardoso, Ana Sofia; Louro, Henriqueta; Santos, Telma; Rebola, Marlene; Pinto, Miguel; Santos, Sílvia; Antunes, Alexandra; Pereira, Paulo; Silva, Maria JoãoA garantia da qualidade da água destinada ao consumo humano é, atualmente, uma preocupação universal, em termos de saúde pública. Assim, a presença de contaminantes de natureza microbiológica ou química, pode constituir um perigo para a saúde humana que importa avaliar, particularmente no que diz respeito a efeitos carcinogénicos. O presente trabalho representa uma abordagem multidisciplinar, abrangendo a caracterização de efeitos citotóxicos, genotóxicos e potencialmente carcinogénicos de toxinas cianobacterianas (microcistinas) e de subprodutos de desinfeção da água (derivados clorados de benzo[a] pireno e fluoranteno). As microcistinas têm sido detetadas em reservatórios de água doce, sendo sobretudo conhecidas pela sua hepatotoxicidade aguda. O nosso trabalho tem vindo a demonstrar que, por um lado, estas toxinas possuem também atividade citotóxica e genotóxica e que, por outro, essa atividade pode ser extensiva a células de outros órgãos, para além do fígado. Estes resultados suportam a hipótese, ainda alvo de alguma controvérsia científica, de que as microcistinas são agentes genotóxicos e potencialmente carcinogénicos. Os derivados clorados de hidrocarbonetos aromáticos policíclicos (HAP) podem formar-se pela reação do cloro, usado na desinfeção da água, com resíduos de HAP presentes na água, sendo que os seus efeitos biológicos são praticamente desconhecidos. O nosso trabalho compreendeu a preparação de três derivados clorados a partir do benzo[a]pireno (BaP) e fluoranteno (Fluo), a saber: 6-Cl-BaP, 3-Cl-Fluo e 1,3-Cl2-Fluo, tendo-se confirmado a sua formação em condições semelhantes às utilizadas em Estações de Tratamento de Águas. Mostrámos ainda que o 6-Cl-BaP apresenta um efeito genotóxico superior ao do BaP, o que veio justificar o alargamento deste estudo a novos subprodutos de desinfeção da água. No seu conjunto, os dados de ambos os estudos contribuíram para gerar evidência científica relativamente ao potencial impacto de contaminantes da água na saúde humana, com importantes implicações para a avaliação de risco e implementação de medidas preventivas, com vista a uma maior segurança na utilização dos recursos hídricos nacionais.
- Formação de subprodutos de desinfeção da água e avaliação do seu potencial genotóxico: o caso dos hidrocarbonetos aromáticos policíclicosPublication . Pinto, Miguel; Rebola, Marlene; Louro, Henriqueta; Antunes, Alexandra M.M.; José, Sílvia S.; Rocha, M. Raquel; Silva, M. João; Cardoso, Ana Sofia
- Formation of emerging disinfection byproducts in water and evaluation of potential genotoxic effects: the case of chlorinated polycyclic aromatic hydrocarbonsPublication . Rebola, Marlene; Pinto, Miguel; Louro, Henriqueta; Antunes, A.M.M.; José, S.S.; Rocha, M.R.; Silva, Maria João; Cardoso, Ana SofiaDisinfection byproducts (DBPs) are formed when disinfectants used in water treatment plants (WTPs) react with natural (or anthropogenic) organic matter present in the source water. Many studies have addressed health risks posed by a life-time exposure to DBPs through chlorinated drinking water or through dermal or inhalation exposure routes. Experimental studies have revealed genotoxic and carcinogenic effects of some DBPs and epidemiological studies evidenced potential associations between chlorinated drinking water and bladder or colorectal cancer. In addition, a possible link between chlorinated drinking water and reproductive/developmental effects has been hypothesized. Many DBPs have been identified in treated water, which justifies the growing concern about the potential health effects of emerging unregulated DBPs, some of which appear to be more genotoxic, in some assays, than the regulated DBPs. Polycyclic aromatic hydrocarbons (PAHs) are among the most persistent contaminants detected in environmental samples such as river sediments and tap water. Water chlorination can lead to the formation of chlorinated derivatives of PAHs (Cl-PAHs) and the few available toxicological studies have shown that Cl-PAHs possess greater mutagenicity than the corresponding parent PAHs. The present study had two main objectives: 1) identification of the major chlorinated derivatives of benzo[a]pyrene (BaP) and fluoranthene (Fluo) formed as chlorination by-products and 2) evaluation of their potential hazard to humans, through the characterization of their potential genotoxic effects in a human cell line. To synthesize chlorinated standards of PAHs, a newly two phase (water/n-hexane) method was developed for BaP and Fluo. 6-Cl-BaP was obtained as the major chlorination product of BaP, and 3-Cl-Fluo and 1,3-Cl2-Fluo of Fluo. The formation of these BaP and Fluo chlorinated derivatives was also observed under WTPs chlorination conditions after at 0.5 until 24 h of exposure. The effects of equimolar concentrations of 6-Cl-BaP vs. BaP and of 3-Cl-Fluo/1,3-Cl2-Fluo vs. Fluo on cell viability and DNA integrity were assessed by the neutral red uptake (NR) and the comet assay, respectively. Exposure of HepG2 cells to a dose-range of 6-Cl-BaP and BaP showed that both compounds are cytotoxic above 50 µM and that, at the equimolar doses of 100 and 125 µM, 6-Cl-BaP is able to induce a significantly higher level of DNA damage than BaP. On the other hand, no changes of cell viability were observed after exposure to several concentrations of Fluo and its derivatives. Likewise, none of the compounds was able to significantly induce DNA damage. In conclusion, the present data confirmed that chlorinated derivatives of BaP and Fluo are formed during WTPs chlorination procedures and allowed the identification of their major chlorinated derivatives that should be further analysed in drinking water. On the other hand, the results from the comet assay evidenced a higher DNA damaging effect of Cl-BaP comparatively to its parent compound, suggestive of a more potent genotoxic effect. In spite of the negative results found for Fluo and its chorinated products, further genotoxicity studies are still needed to allow a definite conclusion. Although health risks of DBPs are small compared to health risks of waterborne diseases, the identification of hazardous Cl-PAHs in water emphasizes the need of development of new and safer water disinfection methods.