Objective: This study aimed to evaluate the in vitro effects of BioPure™ MTAD® and 17% EDTA on the removal of the smear layer on instrumented teeth with the aid of 1% NaOCl. Methodology: Human single-rooted teeth (maxillary incisors and canines) (n=25) were randomly divided into two Experimental Groups (n=10/group) and a Positive Control Group (n=5/group). After the instrumentation, the final irrigation was performed with 10 mL of 17% EDTA/1% NaOCl solution in Experimental Group 1 for 1 minute and with BioPure™ MTAD® in Experimental Group 2. Next, the teeth were cleaved, and the degree of cleanliness of the apical dentin surface was evaluated using scanning electron microscopy (SEM). Results: This study revealed that the 17% EDTA/1% NaOCl solution was more effective in removing the smear layer in the apical third of the root canal compared to MTAD (p<0.05). However, statistical analysis of the results revealed that 17% EDTA was more effective in removing the smear layer 6 mm from the apex in EXPG 1-6, which indicates that the more distant the smear layer is from the apex, the more effective is the chelating solution in removing the smear layer. Conclusion: The two tested substances were not fully effective in the complete removal of the smear layer in the apical third of the root canal, although 17% EDTA/1% NaOCl solution showed greater efficacy when compared to BioPure™ MTAD®, particularly at 6 mm from the apex.