Safer by design (SBD)modifications of nanomaterials (NMs)have been pursued, aiming to maintain functionality and yet reduce hazard and support sustainable nanotechnology. The present case study involves copper oxide nanomaterials (CuO NMs)used in paint that have been surface modified by a SBD approach to particles coated with citrate (CIT
−
), ascorbate (ASC
−
), polyethylenimine (PEI
+
), and polyvinylpyrrolidone (PVP). We assessed the effect of the 4 different surface modified (CIT, ASC, PVP and PEI)NMs plus the pristine non-coated (PRI NM)and a Cu salt (CuCl
2
), using the soil multispecies test system (samples at 28–56–84 days). Further, the species were tested individually, and Cu was measured in the test media (soil and soil solution)and organisms. There was a potential relationship with zeta potential, and toxicity of CuO NMs was as follows: -PEI (+28 mV)caused the least impact, -ASC and -CIT (-17 mV, -18 mV)the most, while PVP and PRI (-8 mV, -9 mV)caused an intermediate response. Differences were not explained by the contribution of soluble Cu. Coating interfered with the release of Cu
2+
and/or the activation of copper regulators and detoxification mechanisms in the organisms, i.e. time to reach some kind of stability in organisms' uptake was shorter for -ASC and longer for -PVP during prolonged time. Thus, one of the main findings is that NMs hazard assessment requires long term testing to understand predicted effects across materials. Further, the coverage using a multispecies approach offers increased relevance and a more ecosystem qualified response.
