In this research, a homemade autoclave reactor was used for preparing iron oxide nanoparticles by hydrothermal technique at different reaction times by using ferric chloride, ferrous sulfate, and ammonium hydroxide as raw materials. The XRD characterization showed that the nanoparticles of the samples have high crystallinity with the crystal phase of magnetite, furthermore, the crystal phase of hematite appears clearly as the reaction time increased. The SEM results showed when the time of reaction increased the average particle size increased too from 28.1 to 49.2 nm. That means the reaction time is an effective parameter for the nanoparticle's growth, The EDX spectrum verified the confirmation of iron oxide nanoparticles by the appearance of Iron and Oxygen peaks. The FT-IR results showed that all samples have an absorption peak at about 578 cm-1 corresponding to the Fe-O bond stretching modes of the in magnetite and the peak of hematite appeared as the reaction time increased above 2 hours which was confirmed with XRD results. Finally, the reaction time is a powerful tool for controlling in size and phase of nanoparticle preparation. Keywords: Hydrothermal, Iron Oxide, Reaction time, magnetite, and hematite.