Objectives: This study was carried-out to evaluate and compare the efficacy of various irrigants when used singly or in combination on the microhardness of root canal dentin. Study Design: A total of 50 root-halves were randomly divided into 5 groups immediately after the initial baseline microhardness measurements and treated with: Group-1; 7.5%Ethylenediaminetetraaceticacid (EDTA) + 2.5%sodium hypochlorite (NaOCl), Group-2; 7.5%ethyleneglycol-bis[b-aminoethylether]-N,N,N0,N0-tetraaceticacid (EGTA) + 2.5%NaOCl, Group-3; 7.5%trans1,2diaminocyclohexane NNN',N'tetraaceticacid (CDTA) + 2.5% NaOCl, Group-4; 7.5%EDTA + 2.5% Ethylenediamine (EDA), and Group-5; 1/1 (v/v) EDTA-EDA mixture + 1/1 (v/v) EDTA-EDA mixture. Fifty mL of each solution was used for 1 minute. The reference and post-treatment microhardness values were measured with a Vickers indenter under 80- gram load, 15-second dwell time. Data were analyzed by two-way ANOVA and Bonferroni tests (p=0.05). Results: All solutions decreased microhardness of dentin (p< 0.05). There was statistically significant difference between each group, except Group-1 and 4, after 1st solution application. While Group-2 resulted in a greater re - duction of dentin hardness, Group-5 caused the least change in microhardness values, after 1st solution application (p< 0.05). No statistical difference was observed between Groups 1-4, after 2nd solution application. However, Group-5 showed a significant difference compared with all other groups, after 2nd solution application (p< 0.05). Conclusions: Under the experimental conditions, all tested solutions reduced the microhardness of the root canal dentin. EGTA was the most efficient chelating agent. EDTA-EDA single mixture has led to least change on the microhardness of root dentin