Aim: The aim of this study was to evaluate the pH of three different sealers—BioRoot RCS, Sealapex, and AH-plus at different time periods. Materials and methods: The diluted sealer was transferred to a clean dry test tube. The pH of the root canal sealers was assessed using a digital pH meter. The measurement was carried out at 37°C fluid temperature for all the samples to simulate the oral temperature. The pH measurements were carried out at the end of mixing time (0 hours), 4 hours, 8 hours, and at 24 hours before renewal of the test liquids. Statistical analysis was done using t test—comparison of two independent means. The statistical analysis was done using the post hoc tests. Results: The results of the study show that all the three sealers tested were alkaline in nature throughout the test periods. All the sealers showed a significant change during the time periods (p < 0.05). AH-plus showed a gradual reduction in pH within the range of 10–10.5. There was a statistically significant change at all time intervals between AH-plus and BioRoot RCS (p < 0.05), but no difference between Sealapex and BioRoot RCS or AH-plus (p > 0.05). Conclusion: All the three sealers used in this study are alkaline in nature, with BioRoot RCS showing the highest pH and AH-plus displaying the lowest with a significant difference between the two. There was no significant difference between BioRoot RCS and Sealapex at all time periods. Clinical significance: The antimicrobial activity of sealer is directly related to pH of sealer. The pH of three sealers—BioRoot RCS, Sealapex, and AH-plus are in the alkaline range up to 24 hours. Bioceramic sealer and Sealapex show the highest pH with no significant difference between them.