Aim: To evaluate and compare the dentinal tubule penetration of glycerine, propylene glycol and ringer's lactate solution. Materials and methods: Forty-five single-rooted human mandibular premolars with mature apices were decoronated keeping a length of 12 mm of a root. Biomechanical preparation till rotary protaper F2 and irrigation according to the protocol of all the roots was performed. The specimens were randomly assigned into three random groups (n = 15) of glycerine, propylene glycol and ringer lactate mixed with safranin o dye. Roots were split into two halves using disks. The evaluation of the area of penetration of glycerine, propylene glycol, and ringer lactate solution was performed using stereomicroscope and Image J software. For analysis single way ANOVA test followed by post hoc test was used. The level of significance was set at p <0.05. Results: The maximum depth of dye penetration with propylene glycol was significantly greater than glycerine and ringer lactate solution when the level of significance was set at p <0.05 Conclusion: Propylene glycol penetration in the dentinal tubules was more effective than glycerine and ringer lactate solution indicating its potential use in delivering intracanal medicaments Clinical significance: The low surface tension vehicles help in propagating the intracanal medicaments into the canal irregularities to aid in thorough disinfection of the canals.