The aim of this study is to investigate at what extent the results of standard nanoparticle (NP) toxicity testing methodologies are affected by the different exposure procedures on soil organisms. In this view, differences in physicochemical properties of ZnO NPs (<100 nm), ZnO bulk (<200 nm) and ionic Zinc (ZnCl2) and their ecotoxicological potential toward Lepidium sativum were investigated with respect to three different spiking methods. Results show that the spiking procedures give homogeneous distribution of the testing nanomaterial in soil but the physicochemical and ecotoxicological properties of the testing species differ according to the spiking procedure. Dry spiking produced the highest ZnO solubility whereas spiking through dispersions of ZnO in water and in aqueous soil extracts produced the lowest. At the same time, the ecotoxic effects showed different trends with regard to the spiking route. The need for a definition of agreed methods concerning the NP spiking procedures is, therefore, urgent. © 2015, Springer Science+Business Media Dordrecht.

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