Iran, as a multi-ethnic country, possesses significant linguistic and religious diversity, particularly in its border regions. This diversity, coupled with the country’s sensitive geopolitical position, provides a fertile ground for the emergence and persistence of ethno-nationalist movements. Despite governmental efforts to promote national integration, certain ethnic groups in border areas continue to demand a redefinition of their identity and status within the socio-political structure of the state. The present study aims to examine the role of border situation in reinforcing separatist tendencies in Ardabil Province—a region characterized by its unique ethnic composition and adjacency to foreign states. This research is applied in nature and has been conducted using a descriptive–analytical method. Data were collected through a researcher-designed questionnaire and analyzed using SPSS and ArcGIS software. Information was gathered through a combination of library-based and fieldwork methods. The findings reveal that the sense of separatism among residents of the northern and border counties of Ardabil Province is significantly higher compared to those living in the central and southern areas. These results highlight the substantial role of border situation in strengthening identity-based and separatist tendencies in the border regions of Iran.