Abstract. During the mid-19th century, the German naturalist Hermann Karsten conducted a 12-year exploration (1844–1856) in the territories of Ecuador, New Granada (now Colombia) and Venezuela, allowing him to produce important botanic, geographic and geologic descriptions with valuable information that permits us to refer to him as a pioneer in many of these topics. With his return to Europe, abundant geological, paleontological and living plant specimens were brought and housed in European museums and botanical gardens. The Karsten collection included an important invertebrate collection from the Cretaceous of the Andes of Colombia and Venezuela, which was studied and published by himself and the renowned German paleontologist Leopold von Buch, filling a large void in the knowledge about ancients faunas. H. Karsten's vertebrate collection was never illustrated or subjected to a detailed taxonomic study, being mentioned in scientific publications in a repetitive manner and with incorrect taxonomic and provenance information. More than 160 years after they were collected, we carried out a taxonomic revision of all H. Karsten's vertebrate specimens from Colombia and Venezuela, which are housed in the Museum of Natural History in Berlin. These specimens are represented by cranial and postcranial elements of megafauna, which include Megatheriidae, Mylodontidae and Glyptodontidae (Xenarthra), Toxodontidae (Notoungulata), Gomphotheriidae (Proboscidea), and many other indeterminate mammal remains. This revision is intended to clarify the taxonomy and provenance of the specimens, emphasizing the historical importance of this fossil collection and its significance for the paleontology of the region.