The purpose of this in vitro study was to evaluate the effectiveness of sodium hypochlorite and glutaralclehyde for rapid sterilization of gutta-percha cones contaminated with Bacillus subtilis ATCC 6633. Cones were treated with either 2% glutaralclehyde or 2.5% sodium hypochlorite. The gutta-percha cones were sterilized with a 2.5% sodium hypochlorite solution throughout the test. Glutaralclehyde did not decontaminate the gutta-percha cones even after 15 minutes of contact. All positive controls showed positive results during the first 24 hours. Negative controls were followed up for 24 and 48 hours and demonstrated the efficiency of previous sterilization. No microbial growth was observed in the groups tested. Even though gutta-percha cones are usually sterile during storage, they can be easily contaminated if incorrectly manipulated. NaOCl at 2.5% concentration is an effective agent for decontamination of gutta-percha cones at no additional cost.