Abstract: In order to obtain high quality ⁸⁹Zr nuclide for nuclear medicine imaging, ⁸⁹Zr was produced by irradiating ⁸⁹Y with HM-20S cyclotron. The nuclear reaction excitation function was calculated. The target system was designed and the target was prepared. ⁸⁹Zr was prepared by irradiating target with cyclotron. The radiochemical separation method of the target and the ⁸⁹Zr labeling process were studied. The results show that the optimum energy range of ⁸⁹Zr beam produced by cyclotron was 9-13.5 MeV, while, the maximum reaction cross section was 13.5 MeV energy. The extracted beam current of the cyclotron is 14 MeV, and the residual energy is about 12.86 MeV after attenuation by the vacuum film and the target film. The beam has a Gaussian distribution in the ⁸⁹Y target, and the range is approximately 0.821 mm. The activity of ⁸⁹Zr at the end of bombardment was 22.20-85.45 mCi and the yield was about 36.51-63.30 MBq/(μA·h) when ⁸⁹Y target was irradiated with 15 μA beam for 1.50-3.33 h. UTEVA resin was used to separate ⁸⁹Zr from the irradiated target. The recovery rate of ⁸⁹Zr was 86.27%, and the removal rate of Y(Ⅲ) was 99.80%. Successful ⁸⁹Zr-DFO-NCS labeling was performed with a radiochemical purity of more than 98%.