Ecological regions aggregate habitats with similar biophysical characteristics within well-defined boundaries, providing spatially consistent platforms for monitoring, managing and forecasting the health of interrelated ecosystems. A major obstacle to the implementation of this approach is imprecise and inconsistent boundary placement. For globally important mountain regions such as the Eastern Arc (Tanzania and Kenya), where qualitative definitions of biophysical affinity are well established, rule-based methods for landform classification provide a straightforward solution to ambiguities in region extent. The method presented in this paper encompasses the majority of both contemporary and estimated preclearance forest cover within strict topographical limits. Many of the species here tentatively considered ‘near-endemic’ could be reclassified as strictly endemic according to the derived boundaries. LandScan and census data show population density inside the ecoregion to be higher than in rural lowlands, and lowland settlement to be most probable within 30 km. This definition should help to align landscape scale conservation strategies in the Eastern Arc and promote new research in areas of predicted, but as yet undocumented, biological importance. Similar methods could work well in other regions where mountain extent is poorly resolved. Spatial data accompany the online version of this article.