This work describes a method for the separation and purification of Ac-225 from a Th-229 source. The procedure is based on the combination of ion exchange and extraction chromatographic methods in nitric acid media and allows the preparation of carrier-free, clinical grade Ac-225 with an overall yield exceeding 95%. Quality control of the product is performed using radiometric (alpha, gamma spectrometry) and mass spectrometric methods. The Ac-225 product can be loaded on a radionuclide generator for the preparation of Bi-213 for preclinical and clinical studies of targeted alpha therapy of cancer and infectious diseases.