Background/objective: The hybrid root canal preparation technique is better in a single visit as compared to multiple visit endodontic therapy in reducing the incidence of post endodontic pain. Relationship of post obturation pain and canal preparation has been studied in the past for the better outcomes of the treatment and for the patient’s belief in the treatment. The objective of this study was to compare the frequency of post obturation pain after single versus multiple visits endodontic therapy by using hybrid root canal preparation technique. Materials and methods: This randomized clinical trial (RCT) was carried out in the Department of Operative Dentistry, College of Dentistry, BAMDC, Multan, Pakistan, between 15th December 2017 and 14th June 2018. After approval from the Institutional Ethical Review Committee, a total of 140 patients with acute pulpitis and non-vital asymptomatic maxillary central incisor to first molar teeth were divided into 2 equal groups. In both groups, canal preparation was done by following hybrid canal preparation technique. Group A patients were treated in a single visit and Group B patients were treated in 2 visits. Patient’s pain response was recorded after 24 hours of treatment, using a Visual Analogue Scale. Data were analyzed using SPSS version 20.0. Results: After 24 hours, when post-operative pain was compared in two groups, it was found that in one visit Group A post obturative pain was significantly less as compared to the 2 visit Group B. In one visit Group A, 18 (25.7%) patients reported pain as compared to the two visits Group B where 38 (54.3%) patients complained of pain. When both the groups were compared by Chi-square test p-value was found 0.001, which was highly significant. Conclusion: Single visit endodontic therapy is better than multiple visits endodontic, in relation to post obturation pain when hybrid root canal preparation technique was used.