The Challenge of Studying TiO<sub>2</sub> Nanoparticle Bioaccumulation at Environmental Concentrations: Crucial Use of a Stable Isotope Tracer

Abstract

The ecotoxicity of nanoparticles (NPs) is a growing area of research with many challenges ahead. To be relevant, laboratory experiments must be performed with well-controlled and environmentally realistic (i.e., low) exposure doses. Moreover, when focusing on the intensively manufactured titanium dioxide (TiO<sub>2</sub>) NPs, sample preparations and chemical analysis are critical steps to meaningfully assay NP’s bioaccumulation. To deal with these imperatives, we synthesized for the first time TiO<sub>2</sub> NPs labeled with the stable isotope <sup>47</sup>Ti. Thanks to the <sup>47</sup>Ti labeling, we could detect the bioaccumulation of NPs in zebra mussels (D<i>reissena polymorpha</i>) exposed for 1 h at environmental concentrations via water (7–120 μg/L of <sup>47</sup>TiO<sub>2</sub> NPs) and via their food (4–830 μg/L of <sup>47</sup>TiO<sub>2</sub> NPs mixed with 1 × 10<sup>6</sup> cells/mL of cyanobacteria) despite the high natural Ti background, which varied in individual mussels. The assimilation efficiency (AE) of TiO<sub>2</sub> NPs by mussels from their diet was very low (AE = 3.0 ± 2.7%) suggesting that NPs are mainly captured in mussel gut, with little penetration in their internal organs. Thus, our methodology is particularly relevant in predicting NP’s bioaccumulation and investigating the factors influencing their toxicokinetics in conditions mimicking real environments

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