São Paulo state, Brazil, is one of the main areas of sugar cane agriculture in the world. Herbicides, in particular, ametryn, are extensively used in this extensive area, which implies that this herbicide is present in the environment and can contaminate the surface water by running off. Thereby, residues of ametryn were analyzed in samples of river water an river sediment and in freshwater bivalves obtained from the rivers Sapucaí, Pardo and Mogi-Guaçu in São Paulo State, Brazil. Samples were taken in the winter of 2003 and 2004 in two locations in each river. The specimens of freshwater bivalves collected and analyzed were Corbicula fluminea, an exotic species, and Diplodon fontaineanus, a native species. Additionally, the evaluation of the ability of bioconcentration and depuration of ametryn by the freshwater bivalve Corbicula fluminea was also performed. Ametryn concentrations in the samples were measured by liquid chromatography coupled to mass spectrometry. Residues of ametryn in water (50 ng/L) and in freshwater bivalves (2-7 ng/g) were found in the Mogi-Guaçu River in 2004, and residues in river sediments were found in all rivers in 2003 and 2004 (0.5-2 ng/g). The observation of the aquatic environment through the analysis of these matrixes, water, sediment, and bivalves, revealed the importance of the river sediment in the accumulation of the herbicide ametryn, which can contaminate the biota.
The bioaccumulation of atrazine in bivalves is expected due to its wide use in agriculture and its moderate water solubility (33 mg/l). To assess the capacity of bioaccumulation, we performed a laboratory study in which two bivalves species (Anodontites trapesialis and Corbicula fluminea) were exposed to atrazine (concentrations of 0.06 to 0.34 microg/ml) during 48 h. The results showed that both bivalve species were able to bioaccumulate atrazine in their tissues. In another experiment, groups of bivalves from the specie A. trapesialis were exposed to atrazine at the concentration of 1 mug/ml and different organs of these animals were analyzed. The visceral mass, foot and mantle plus siphon showed the highest values of atrazine bioaccumulation.
Foram descritos métodos analíticos empregando cromatografia líquida acoplada a espectrometria de massas (LC-MS-MS) para a determinação de ametrina em água superficial, sedimento de rio e no molusco bivalve Corbicula fluminea. A técnica de extração líquido-líquido foi utilizada para a preparação das amostras. As análises das amostras foram feitas em coluna RP-18 de fase reversa com detecção no modo electrospray positivo e Multi Reaction Monitoring (MRM). O limite de quantificação de ametrina foi 20 ng L -1 , 0,1 ng g -1 e 0,5 ng g -1 para amostras de água, sedimento e bivalves, respectivamente. Na etapa de validação dos métodos foram avaliadas também a linearidade, a precisão, a exatidão e a recuperação. Os resultados obtidos na validação do método estão de acordo com os limites internacionais e com a literatura, sugerindo que o método desenvolvido é adequado para a quantificação de ametrina em amostras de água superficial, sedimento de rio e nos bivalves Corbicula fluminea. Estes métodos foram aplicados na análise de amostras coletadas no Rio Mogi-Guaçu e Rio Pardo, São Paulo, Brasil. O nível mais elevado de ametrina foi 1,44 ng g -1 , encontrado nas amostras de bivalves do Rio Mogi-Guaçu.Analytical methods employing liquid chromatography-mass spectrometry (LC-MS-MS) for ametryn determination in river water, river sediment and in freshwater bivalve mussel Corbicula fluminea are described. Liquid-liquid extraction was used for sample preparation. The analyses of ametryn in samples were performed on a reversed-phase RP-18 column with MS detection in positive electrospray and multi reaction monitoring modes. The quantitation limit of ametryn was 20 ng L -1 , 0.1 ng g -1 and 0.5 ng g -1 , for water, sediment and bivalve samples, respectively. Linearity, precision, accuracy and recovery were also reported. The results obtained for method validation are within the international limits and in accordance with literature, suggesting that the developed methods are suitable for the quantitation of ametryn in river water, river sediment and in bivalve Corbicula fluminea. These methods were applied for the analysis of samples from Mogi-Guaçu River and Pardo River, São Paulo State. Results indicated that the highest level of ametryn was 1.44 ng g -1 in bivalve samples from Mogi-Guaçu River.
A method for the analysis of atrazine in freshwater bivalves has been developed. The method involves a simple liquid-liquid extraction procedure followed by high-performance liquid chromatography analysis using a C 18 column, a methanol-water (60 : 40, v=v) mobile phase, a flow-rate of 1 mL=min, and UV detection at 230 nm. After optimization of the conditions, the method was validated, showing 81.7% atrazine recovery. Performance data for the method such as linearity, precision, and quantitation limit are also reported.
Sonhar mais um sonho impossível, Lutar quando é fácil ceder, Vencer o inimigo invencível, Negar quando a regra é vender. UNIVERSIDADE DE SÃO PAULO FFCLRP-DEPARTAMENTO DE BIOLOGIA PROGRAMA DE PÓS-GRADUAÇÃO EM BIOLOGIA " "B Bi io oa ac cu um mu ul la aç çã ão o d do o h he er rb bi ic ci id da a a at tr ra az zi in na a p pe el la as s e es sp pé éc ci ie es s d de e b bi iv va al lv ve es s l li im mn ni ic co os s A An no od do on nt ti it te es s t tr ra ap pe es si ia al li is s ((L La am ma ar rc ck k, , 1 18 81 19 9)) e e C Co or rb bi ic cu ul la a f fl lu um mi in ne ea a ((M Mü ül ll le er r, , 1 17 77 74 4)). ." " Analu Egydio Jacomini Dissertação apresentada à Faculdade de Filosofia, Ciências e Letras de Ribeirão Preto-USP, como parte das exigências para obtenção do título de MESTRE EM CIÊNCIAS-Área: BIOLOGIA COMPARADA.
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