ABSTRACT The maintenance of biodiversity and the capacity of natural systems to provide goods and services for people is affected on different levels by the intensity of human activities on ecosystems. In this study, we apply a Legacy-adjusted Human Footprint Index (LHFI) to evaluate the spatiotemporal variation of anthropic impact in Colombia in 1970, 1990, 2000 and 2015. We identified hotspots of change in LHFI and we evaluated the intensity of anthropic pressures in natural regions and ecosystems. We found that LHFI in Colombia increased between 1970 and 2015. The Andean and Caribbean regions presented the highest levels of anthropic impact, remaining stable over time. Hotspots of change were mainly located in the following regions: Andean (Antioquia, Cauca and Valle del Cauca states), Amazon (Amazonas, parts of Meta, Guaviare and Putumayo states) and Orinoco (Casanare and parts of Meta and Vichada states). In addition, ecosystems that under the IUCN ecosystem risk categories are Critically Endangered (CR) and Vulnerable (VU) were the most affected by a high level of human impact. Spatiotemporal evaluation of the human footprint in Colombia provides new insights about trends in human pressures on ecosystems and constitutes an analytical tool with high potential for harmonizing land use planning and biodiversity conservation.