Abstract: Using Xi’an pulsed reactor as neutron source, the production method of ¹⁶¹Tb was established. The influence of composition and mass of the target material on the productivity of ¹⁶¹Tb was investigated. In order to acquire purity ¹⁶¹Tb, an ion exchange chromatography method for Tb separation from the target material was studied. The influence of some factors such as length of resin bed, pH of eluent, amount of matrix Gd³⁺ and temperature on the separation was investigated using Dowex50 cation exchange resin. The results were showed that the composition and mass of the Gd₂O₃ target material obviously influenced the productivity of ¹⁶¹Tb, isotope enriched target material was optical choose for ¹⁶¹Tb production. Conditions such as matrix amount and length of resin bed had few effects on the separation of Tb within a small range, however, pH of eluent and temperature markedly affected the separation. The optimized separation condition for mixture solution of Gd³⁺, Tb³⁺ and Dy³⁺ was that 17.50 cm as length of resin bed, matrix amount of 50 mg, 0.20 mol/L 2-HIBA as elution, pH=3.80 and pH=4.00 for pH of Tb and Dy eluent, repectively, temperature higher than 25 ℃. Under the optimized condition, the recovery of Tb was higher than 95% and decontamination factor of Tb to Gd is higher than 10 000.