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(M/EH025) Frozen mitochondria bioassay to detect toxicity of atrazine and its metabolites. Perin, Guido 1, Iero, Alessandra1, Manente, Sabrina1, Bragadin, Marcantonio1, 1 ABSTRACT- The herbicides atrazine was widely used all over, since some ten years its toxicity has been investigated. Our aim was to monitor whether atrazine and its metabolites in aqueous solutions are toxic for mitochondria. Thus, the frozen mitochondria bioassay was carried out. To perform this toxicity test beef heart mitochondria once prepared are stored at -22 centigrade. During the test (20 min), the oxygen consumption is monitored by a Clark electrode. For atrazine and the metabolites, two different set of measures were carried out in order to determine acute and sub-chronic toxicity. The frozen mitochondria standard procedure was applied during acute toxicity measurements, such to highlight immediate effects of the compounds on the oxygen consumption. However, both atrazine and the metabolites didn't show any acute toxicity effects. The sub-chronic measurements were performed developing a new procedure. In fact, mitochondria were incubated with the toxic compound and successive aliquots of this solution were monitored by the Clark electrode in order to obtain the oxygen trend during long period of time (4 hours). A decrease in the oxygen consumption as sub-chronic toxicity effect is shown when concentration in the range of ppb are tested; environmental concentrations are much lower then the effective concentration, though. Key words: frozen mitochondria, bioassay, acute and subchronic toxicity, atrazine |
