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The purpose of this study was to determine the position of the apical foramen (AF) in relation to root surfaces of human permanent teeth using cone-beam computed tomographic (CBCT) imaging and novel advanced imaging analysis software (e-Vol DX; CDT Software, Bauru, SP, Brazil).The AF position was determined on CBCT scans viewed and analyzed using e-Vol DX of 1400 teeth (422 patients) according to the root surface as follows: buccal, mesiobuccal, mesial, mesiolingual/palatal, lingual/palatal, distolingual/palatal, distal, distobuccal, and central. Categoric variables were described as frequencies and percentages. Frequencies were reported with their confidence intervals (95%). Categoric variables were analyzed using the chi-square test with Yates correction. The level of significance was set at α = 0.05.The most frequent AF position in maxillary anterior teeth was central (46%–60%). The AF in mandibular central incisors was buccal in 44% of the cases. In maxillary first and second premolars, 39.98% and 42.56% of all AFs were central. In maxillary first and second molars, 46.12% and 57.49% of all AFs were central. The most frequent AF position in mandibular first and second premolars was central (42.85% and 50.98%). In mandibular first molars, 48.72% of all AFs were central.The AF position in human permanent teeth was central in 48.95% and 42.08% of the maxillary and mandibular teeth. CBCT images analyzed by e-Vol DX can be used to determine the true anatomic position of the AF and can be a useful tool for the treatment planning of nonsurgical and surgical endodontic treatments.