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The X-RAD 320 preclinical irradiator was characterized from Monte Carlo simulations using the DOSXYZnrc code from a point source of isotropic radiation that emits energy spectra generated with the SpekCalc program. Those simulations were carried out in a water phantom for different beam qualities and field sizes between 0.5 x 0.5 and 15 x 15 cm2 for which 3D dose matrices were obtained. Relative uncertainties associated with the simulations of up to ± 1.39% were found for depths less than 5 cm. The results obtained for the depth dose curves, dose profiles and output factors for the different beam qualities correspond to the expected behavior.