DESENVOLVIMENTO DE PROTOCOLO PARA ANÁLISE DE ATRAZINA E METABÓLITOS EM EMBRIÕES DE AVE

Abstract

Animal contamination with atrazine (ATZ) is associated with toxic effects, since atrazine is reported as an endocrine disruptor in adults, both males and females, as well as in embryos of various animal classes, affecting hormonal regulation and, consequently, the developmental patterns of these individuals. Therefore, since chicken embryos (Gallus gallus) are model animals for studies involving embryonic development, the aim of this work was to develop a protocol for the study of atrazine bioaccumulation in bird embryos. The methodology used was based on in ovo treatment of three groups of embryos: a control group with injections of distilled water, a group treated with atrazine solution injections at 500 µg/L, and a group treated with atrazine injections at 1.000 µg/L. Embryo collection occurred 5 or 7 days after incubation. Embryos were prepared for analysis using the QuEChERS methodology (an acronym for Quick, Easy, Cheap, Effective, Robust, and Safe), followed by analysis through high pressure liquid chromatography with a diode array detector (HPLC-DAD). In house analytical validation of the methodology was performed. The limit of detection (LOD) for ATZ, desisopropylatrazine (DIA) and desethylatrazine (DEA) were 5 µg/L, 11 µg/L and 7 µg/L, and limit of quantification (LOQ) were 16 µg/L, 35 µg/L and 22 µg/L respectively. The proposed methodology presented satisfactory results with good repeatability less than 18%. Regarding the results obtained, atrazine and its metabolites, DIA and DEA, were not detected in embryos at any of the administered concentrations. Mortality and malformation rates were also evaluated, but no significant discrepancies were able to associate the effects of atrazine with deviations from the embryonic developmental pattern. Future studies should be conducted to confirm if bird embryos are incapable of bioaccumulating atrazine and to ensure that the use of this pesticide does not affect the health of animals at the developmental level.