To investigate the mean tooth length and root canal configuration of human maxillary and mandibular premolars in the population of Hyderabad, Pakistan, using Cone Beam Computed Tomography (CBCT). Observational study. Place and Duration of the Study: Department of Operative Dentistry and Endodontics, Liaquat University of Medical and Health Sciences, Jamshoro, Sindh, Pakistan, from December 2022 to March 2023. Using retrospective analysis of CBCT scans, tooth length, the number of roots, and root canal configuration of 536 mandibular and maxillary premolars were assessed in the subpopulation of Pakistan. The data were analysed employing descriptive statistics, Chi-square tests, and One-Way ANOVA to determine the association between various factors and root canal morphology. Vertucci Type 1 (one canal) was the most common configuration in maxillary premolars, with frequencies ranging from 46.3 to 52.2%. The mean length of all 8 premolars ranged from 20.56 mm to 21.58 mm. C-shaped canal configurations were observed in a small percentage of maxillary and mandibular premolars, with C1 being the most common. No significant association was found between gender and canal configuration or tooth length, and in tooth lengths among different canal configurations. Vertucci Type 1 was the most common configuration in maxillary premolars, followed by Vertucci Type 4. Single-rooted premolars were more prevalent, while two-root configurations were less common. C-shaped canal configurations were observed in a small percentage of premolars, with higher frequency in mandibular premolars. Cone beam computed tomography, Dental anatomy, Endodontics, Premolars, Root canal morphology.