Depletion kinetics and concentration- and time-dependent toxicity of a tertiary mixture of amitraz and its major hydrolysis products in honeybees

Although amitraz is one of the acaricides most commonly applied within beehives, to date, its time-dependent oral toxicity in honeybees has not been investigated, due to amitraz's instability in aqueous media. In aqueous media such as honey, amitraz rapidly forms a continuously changing tertiary mixture with two of its major hydrolysis products, DMF and DMPF. The contribution of each hydrolysis product to the overall oral toxicity of this acaricide is not known. Therefore, we aimed to characterize the depletion and formation kinetics of amitraz and its hydrolysis products in 50% sucrose solution provided to caged honeybees, including the calculation of the 50% lethal oral concentration (LC₅₀) of amitraz. We sought to determine the contribution of each component of the mixture to the overall observed toxicity. We also investigated the time- and concentration-dependent toxicity of the amitraz mixture and its hydrolysis products. A novel approach based on the analysis of the areas under the depletion and formation curves of amitraz and its hydrolysis products revealed that DMPF, amitraz and DMF accounted for 92%, 7% and 1% (respectively) of the overall toxicity of the mixture. The chronic oral LC₅₀ of amitraz was 3300 μmol/L, of similar magnitude as that of the non-toxic hydrolysis product DMF. The toxicity of DMPF and the mixture decreased over time; whereas the toxicity of DMF increased over time. Amitraz's instability in aqueous media and the highly toxic profile of DMPF, suggest that DMPF is the actual toxic entity responsible for amitraz's toxicity toward honeybees.