Nano or not, what difference does it make? A stable isotope labeling approach to assess fate and bioavailability of metallic engineered nanoparticles in soil

By the time they enter soils, engineered nanomaterials (ENM) have undergone physicochemical transformations and may no longer resemble the pristine materials which have been thoroughly investigated during two decades of nanotoxicology research. Is the behavior of environmentally relevant chemical forms of ENM different from that of other metallic species present in soils? Are they more available to soil organisms than their naturally occurring counterparts? The present study aimed at answering these questions, through the use of isotopically enriched ENMs, which could be traced at low concentrations in soil microcosms, despite high natural metallic background. The relevance of the chemical forms was ensured by introducing isotopically enriched 109Ag, 68ZnO and 46TiO2 ENM to a wastewater treatment plant and using the resulting sewage sludge (final sink for most ENM) as amendment in soil microcosms with earthworms. The sludge application rate to soil was similar to that used in agriculture and the experiment lasted for a month. Protocols using inductively coupled plasma mass spectrometry were developed for determination of isotope ratios in complex matrixes, such as soil and organisms. For 109Ag ENM, the dissolved fraction in soil (i.e. the most easily accumulated in organisms) was extremely low and comparable to that of Ag naturally present in soil, and transfer factors to earthworms were similar to those of natural Ag. For 46TiO2 ENM, the transfer to earthworms was negligible, similarly to what was observed for natural Ti. While no difference in behavior and bioavailability was observed between ENM and their naturally occurring counterparts for Ag and Ti, different results were obtained for Zn. The dissolved fraction for 68ZnO ENM was 3-5 times higher than for Zn forms naturally present in soil, and transfer factors to earthworms twice those of natural Zn. Overcoming long-standing challenges related to environmental relevance of chemical forms and concentrations in nanotechnology studies, the approach provides valuable insight into behavior and impacts of environmentally relevant forms of ENM in soils.