Atrazine (ATR), a pesticide extensively applied in the United States, and its main metabolites, desethylatrazine (DEA), desisopropylatrazine (DIA), and diaminochlorotriazine (DACT), have been widely detected in surface water in agricultural regions. While the effects of ATR and its metabolites are not fully understood, several long-term adverse effects have been observed in aquatic species, mice, rats, rabbits, and other organisms after exposure to ATR and its metabolites. We are developing and validating an extraction and quantitation method for ATR, DEA, and DIA involving “quick, easy, cheap, effective, rugged, and safe” (QuEChERS) extraction and liquid chromatography-mass spectrometry (LC-MS) detection. The method will be used to quantitate ATR, DEA, and DIA in muscle and hepatopancreas tissue of the virile crayfish, <i>Faxonius virilis</i>, a keystone species in freshwater environments. The method includes the addition of a stable-isotope internal standard, deuterated atrazine (ATR-d5), to improve the accuracy and precision. Figures of merit including the detection limit, the limit of quantitation, and the dynamic range were determined. Additionally, repeatability was assessed with both intraday and interday precision measurements, and spike recovery tests of ATR and its metabolites were performed. In preliminary studies, atrazine and metabolites have been detected in tissue isolated from the crayfish. The validated method will be used to quantitate levels of ATR and metabolites in tissue following treatment with environmentally-relevant ATR concentrations (80 and 300 ppb), negative control (0 ppb), or positive control (1000 ppb) for 15 days. The levels will be compared with physiological and behavioral changes previously observed in crayfish in order to investigate the relationship between ATR and metabolite levels in tissue and observed physiological and behavioral effects.

Development and validation of a method using QuEChERS and liquid chromatography-mass spectrometry for determination of atrazine and its metabolites in crayfish

2019 Spring National Meeting

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