The Cerrado is recognized as an important global conservation “hotspot” due to its remarkable biodiversity and the high rate of threats to unique ecosystems present in this region. It is the second largest biome in Brazil and is characterized by a great diversity of plant physiognomies, including fields, savannas, riparian forests, and seasonal humid areas. The objective of this study was to evaluate the vegetation cover of a Conservation Unit belonging to the Cerrado biome, before and after the occurrence of a fire event, applying the Normalized Difference Vegetation Index (NDVI) in the burned area polygon (scar of fire). The burned area was obtained through segmentation using ArcGIS software version 10.8, with the aim of improving the obtainment of the effectively burned area, through the use of images with better spatial resolution, while the NDVI maps of the burned area were obtained by crossing the Sentinel-2 satellite bands, in scenes captured in September 2019 in pre- and post-fire events. To measure biomass loss resulting from the fire, the pre-fire NDVI image was subtracted from the post-fire NDVI image, resulting in a difference image. The latter was categorized into five classes: low, moderate, medium, high, and very high. The results indicate a more severely burned area of approximately 202 hectares, also indicating differences in NDVI values before and after the fire, with a decrease of 32% in maximum values and 13% in minimum values; and as expected, NDVI values were higher before the fire than after the fire. With regard to biomass loss in the most severely burned area of the park, the moderate class stood out as the most representative, covering 35.8% of the total, followed by the medium class with 25.6% and the very high class, the most extreme, with 3.9%. It is concluded that even in the most severely burned area, the fire had moderate proportions with an impact distributed throughout the physiognomies present in the Park.