This study aimed to evaluate root number and morphological variations of the root canal system in maxillary and mandibular premolars among the Saudi subpopulation.A total of 500 cone-beam computed tomography (CBCT) images were assessed, including 2442 maxillary and mandibular premolars. The evaluation encompassed the number of roots and canals and their configuration based on the Vertucci classification. Gender differences were considered, along with the position of premolars and unilateral or bilateral symmetry.Maxillary first premolars predominantly exhibited two roots (82.6%), followed by one root (16.3%) and three roots (0.97%), with a majority having two root canals (83.6%). Regarding maxillary second premolars, the most common morphology was one root (66.6%), with two root canals (40.5%) observed more frequently than one root canal (38.1%). Mandibular first and second premolars mainly featured one root (84.8% and 96.1%, respectively) with one root canal (70.8% and 90.2%, respectively), whereas two root canals were less prevalent (2.3% and 1.8%, respectively). Three canals and three roots were rarely found in mandibular premolars (0.3%). Regarding maxillary premolars, males exhibited significantly higher type IV and mandibular type I root configurations compared with females. Conversely, regarding mandibular premolars, females showed significant mandibular type V and higher type I root configurations compared with males.CBCT imaging facilitated precise assessment of root morphology and root canal configurations in maxillary and mandibular premolars. The present findings can aid dentists in diagnosing, evaluating case difficulty, and devising effective root canal treatments, particularly in patients from Saudi Arabia.