Endodontic therapy is essentially a debridement procedure that requires the removal of the irritants of the canal and periapical tissue if success is to be gained. The debridement may include instrumentation of the canal, placement of medicament and irrigants. Complete cleaning of the root-canal system requires the use of irrigants that dissolve organic and inorganic material. The study aimed to evaluate changes in dentin microhardness after canal irrigation with different solutions. Twenty four freshly extracted human mandibular molars (distal roots with single canals) were used. 10mm root length was taken as standard length. The roots were embedded into auto polymerizing acrylic resin using plastic molds before the canals preparation and micro hardness test. The distal roots were prepared with one shape rotary file. Before the preparation each root was irrigated with 1ml distilled water. Then the roots were divided into four groups according to the final irrigation protocol: Group A: NaOCl 2.5%, Group B: EDTA 17%, Group C: Citric Acid 40%, Group D: Distilled Water. For (Vicker microhardness test) the same load and time 500 g test load for 20 seconds, will be conducted three times at distance 0.5mm from canal lumen ; thus there will be 9 indentations on each specimen surface. An average of the three readings for each test condition will be recorded as the VHN value of a specimen. Comparing all four groups statistically there was no significant difference among them. The mean values were found more reduced in EDTA group followed by NaOCL group, and then Control and Citric Acid groups. All the groups showed reduction in dentin microhardness. EDTA group showed the maximum reduction followed by NaOCL group, and least with Citric Acid group.