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This work is a comparative study of the effect of two different annealing methods on copper oxide (CuO) nanoparticle properties obtained previously by direct precipitation method. The prepared samples were annealed in air at various temperatures (300, 400, 500°C) for 1 hour. Then they were characterized by employing scanning electron microscopy (SEM), X-ray diffraction (XRD), UV-Visible, and Fourier Transform Infrared (FT-IR) spectroscopy. The main results revealed an increase in the grain size in both methods as the annealing temperature increases. It reaches 30.93 nm in RTA and 26.75 nm in STA at 500°C. XRD spectra showed, in the case of RTA at 500 °C, a significant decrease in the intensity corresponding to the (002) and (111) orientations. This result indicated that beyond 400 °C, one hour of RTA is not suitable for enhancing CuO nanoparticle crystallinity compared to STA. The optical analysis demonstrated that the energy of the optical band gap in STA is higher than that in RTA. It reaches 2,88 eV at 500°C using RTA which is close to the gap value of CuO in the range of 1.8–2.8 eV. FT-IR results showed, for both methods, the presence of characteristic peaks of the Cu-O bonds in the monoclinic CuO structure without any trace of Cu2O structure. Nevertheless, samples exposed to RTA for one hour are more susceptible to absorbing species of C=O bond (C=O bond is due to the atmospheric CO2 absorption and the presence of organic impurities from the synthesis process) than those of STA. Hence, RTA at 500 °C is far from producing CuO nanoparticles with preferred characteristics; it needs further research to examine the effect of higher temperature by controlling the annealing time.