The aim of this study was to use Cone-beam Computed Tomography (TCHC) to investigate the anatomical configuration, prevalence, and distribution of the Second Mesiobuccal (MB2) canal in upper molars of a Chilean subpopulation, considering variables such as the presence of MB2 canal, Vertucci classification, gender, and age. Cross-sectional observational study. The sample consisted of TCHC scans taken at the dental clinic of Universidad Andres Bello (Vina del Mar). The sample size was calculated using the formula for known population, resulting in 262 scans. The examiners were calibrated using Cohen's Kappa coefficient, then the variables were analyzed through an imaging study using I-CAT Vision software. 439 first and second upper molars were analyzed. The prevalence of MB2 canal in first molars was 63.74 %, while in second molars, it was 20.04 %. The prevalence in first molars was higher in males (73.86 %) than in females (58.62 %), while in second molars, it was 15.81 % in females and 28.41 % in males. Regarding age, in first molars the difference was significant between the age range of 18 to 40 years (66.49 %). Regarding the type of configuration according to Vertucci, 70 % of the first molars had Type II configuration, and 23.65 % had Type IV, with similar results in second molars. The present study demonstrated that MB2 canals are frequent in the analyzed population, especially in first molars, and Type II configuration is the most prevalent. Additionally, a higher prevalence was observed in males and in the age range of 18 to 40 years. These findings provide relevant information about root anatomy in the studied population and can contribute to improving treatment outcomes.