Korea Atomic Energy Research Institute (KAERI) is planning to disposal of the radioactive contaminated cement waste form to the final disposal facility. The final disposal facility require evaluation of immersion, compressive strength, and radionuclide inventory of radioactive wastes to meet the acceptance criteria for safe disposal. According to the LILW acceptance criteria of the Nuclear Safety and Security Commission ok Korea (NSSC), the disposal limit radioactivity of 129I (3.70×101 Bq/g) is lower than other radionuclides. 129I emits low energy as its disposal limit is low, so it is difficult to analyze in the presence of 137Cs and 60Co which emit high energy. Therefore, it is essential to an accurately separate and analyze iodine in radioactive waste. In this study, we focused on the determination of 129I in cement waste form containing 137Cs, 60Co. We added 1 g of 129I(11.084 Bg), 137Cs(1,300 Bq) and 60Co(402 Bq) to cement waste form, respectively. The separation of 129I in cement waste form was carried out using an acid leaching method. And, we confirmed the specific activity of 137Cs and 60Co at each separation step. It was observed that an acid leaching method showed the remove efficiency 137Cs(99.97%) and 60Co(99.94%), respectively. In addition, 129I was also analyzed at approximately 96.44% in simulated contaminated cement waste form. In conclusion, through this experiment, it was confirmed that 129I could be successfully separated and analyzed by using the acid leaching method in cement waste form containing excessive 137Cs and 60Co.