Incremental technique for resin composite restorations requires multiple physical contacts between spatulas, tooth cavities and the restorative material recipient. Thus, the decontamination of the tip’s of spatula by chemical agents between each resin composite increment placement is important to reduce chances of potential cross-contamination. Objectives: To evaluate the efficacy of different solutions for decontamination of tips of spatula used in restorative procedures and to establish a decontamination standard protocol. Material and Methods: Spatulas were sterilized in autoclave at127°Cfor 20 minutes and then contaminated with: 1) a suspension of half 1.0 MacFarland scale turbidity of different microorganisms, 2) the pool of equal amounts of these microorganisms; except for the control group. Decontamination techniques consisted of rubbing the tip of the spatulas (1 to 5 consecutive times) using a 2ml 70% ethanol or 2% glutaraldehyde embedded gauze. After decontamination, spatulas were immersed in thioglycolate broth and incubated for 48 hours at37°C. Broth with visible microbial detection was submitted to bacterial identification by Gram stain. Results: Low uniformity of rubbings number was observed to eliminate different microorganisms due to different tested disinfectant agents. Four or five rubbings were needed to decontamination of the tested microorganisms using 70% ethanol. Three rubbings using 2% glutaraldehyde were able to eliminate tested microorganisms. Conclusion: The results demonstrated that 70% ethanol by friction, counting four or five rubbings, was effective to decontaminate spatula’s tip.