The exposure to nanoparticles (NPs) represents a severe problem to human health, because they are becoming more widely used and their number of applications continually increases. Particles containing copper are emitted from smelters, iron foundries, power stations and municipal incinerators (WHO, 1998), as well as from brake linings during braking, Kukutschova et al. (2011). Copper and copper oxide NPs are frequently used as catalysts, heat transfer fluids in machine tools (Kim et al., 2011), inks, anode material in lithium-ion batteries (Guo et al., 2002) and many others. Even though CuO NPs were found highly toxic, Karlsson et al. (2008) and it is likely that NPs enter human body via respiratory tract, the inhalation exposure experiments of CuO NPs with laboratory animals are still rather rare (Pettibone et al., 2008 and Lebedova et al., 2016). The exposure chamber for long lasting inhalation experiments was constructed at the Institute of Analytical Chemistry of the CAS (Večeřa et al., 2011) and some methods of NPs generation for these experiments were already tested in our laboratory (Moravec et al., 2015 and Moravec et al., 2016a). The generation of Cu/Cu2O NPs by thermal decomposition of copper acetylacetonate (CuAA) was reported by Moravec et al. (2016b) and here we present the results of long lasting generation of NPs by oxidation of CuAA.