This study presents a rapid and sequential radiochemical separation method for Pu and Am isotopes in radioactive waste samples from the nuclear power plant with anion exchange resin and TRU resin. After radionuclides were leached from the radioactive waste samples with concentrated HCl and HNO3, the sample was allowed to evaporate to dryness after filtering the leaching solution with 0.45 micron filter. The Pu isotopes were separated in HNO3 medium with anion exchange resin. For leaching solution passed through anion exchange column, the Am isotopes were separated with TRU resin. The purified Pu and Am isotopes were measured by alpha spectrometer, respectively, after micro-precipitation of neodymium. The sequential radiochemical separation of Pu and Am isotopes in radioactive waste samples using anion exchange resin and TRU resin was validated with ICP-MS system.