Background: Microleakage is one of the major causes of endodontic treatment failure. Many microleakage testing techniques have been introduced through the years, but there has been no agreement as to which technique gives the most accurate results. The objective of this research was to assess the accuracy of fluid filtration when analyzing the apical sealing capability of the aggregate of mineral trioxide, Biodentine, and total putty. Materials and methods: A sample of 40 human central incisors was collected. The samples were decoronated at cementoenamel junction followed by preparing the canal up to size 80. Using the lateral compaction method, the obturation was done with AH plus sealer. The apical 3 mm root was resected after which ultrasonic retro tips have been utilized to make the retrograde preparation. Apical microleakage evaluation for each of the specimens was performed utilizing the fluid filtration method at specific time intervals, i.e., immediate, 1 week, 1 month, 3 months, and 6 months. Results: The outcomes of the current study inferred that MTA revealed better apical sealing properties than that of Biodentine and total putty. Overall, comparative analysis of microleakage immediately and post restoration at different time intervals was done using one-way ANOVA ‘F’ test that was highly significant, p <0.001. Pairwise comparison was done using Tukey\'s post-hoc test p <0.001. Conclusion: Thus, the results of the present study conclude that MTA with its superior sealing ability might provide long-term sealing benefits to the root canal system than that of Biodentine and total putty. Key message: In this study, MTA has shown a better long-term sealing ability as compared to Biodentine and total putty. In a surgically attempt to repair the tooth, it is necessary to select a material with promising long-term sealing ability, MTA as a root-end filling material can be suggested as a better alternative to the other bioactive materials.