The aim of this study is to determine the location of MB2 in maxillary first molars in relation to the MB1 using the cone‐beam computed tomography (CBCT) imaging. In this retrospective study, electronic health records’ system was utilised to identify patients who acquired CBCT for the maxillary first molar as an imaging adjunct to their endodontic treatment. Data were evaluated, and statistical analysis was performed. For the initial treatment cases, the distance from MB1 to MB2 was 2.06 ± 0.52 mm. This did not change significantly for cases that were being retreated. Statistically significant differences were observed between the distances from MB1 to MB2 in males vs. female teeth with males demonstrating longer distances as compared to females. Variations in anatomy exist between patients but this study established some guidelines for clinicians to locate the MB2 canal orifices. This could potentially help with treating these teeth with minimal failure rates.