Purpose: Sex determination can be done by morphological analysis of different parts of the body. The mastoid region, with its anatomical location at the skull base, is ideal for sex identification. Statistical shape analysis provides a simultaneous comparison of geometric information on different shapes in terms of size and shape features. This study aimed to investigate the geometric morphometry of the inter-mastoid triangle as a tool for sex determination in the Iranian population. Materials and Methods: The coordinates of 5 landmarks on the mastoid process on the 80 cone-beam computed tomographic images (from individuals aged 17-70 years, 52.5% female) were registered and digitalized. The Cartesian x-y coordinates were acquired for all landmarks, and the shape information was extracted from the principal component scores of generalized Procrustes fit. The t-test was used to compare centroid size. Cross-validated discriminant analysis was used for sex determination. The significance level for all tests was set at 0.05. Results: There was a significant difference in the mastoid size and shape between males and females (P<0.05). The first 2 components of the Procrustes shape coordinates explained 91.3% of the shape variation between the sexes. The accuracy of the discriminant model for sex determination was 88.8%. Conclusion The application of morphometric geometric techniques will significantly impact forensic studies by providing a comprehensive analysis of differences in biological forms. The results demonstrated that statistical shape analysis can be used as a powerful tool for sex determination based on a morphometric analysis of the inter-mastoid triangle.