This study provides new records, geographical distribution extensions and a checklist of the current ctenids species in Colombia based on the review of four arachnological collections and published literature. A total of 15 new records for Ctenidae in Colombia are reported; nine of these species are new records for the country and the distribution of the remaining six is expanded. The genus Centroctenus Mello-Leitão, 1929 (C. ocelliventer Strand, 1909) is recorded for first time in Colombia and Cupiennius coccineus (F.O. Pickard-Cambridge, 1901) for South America. Due to the strategic geographic position of Colombia, which is a transition zone between Southern and Central American biotas, species inventories in different localities are important to fill distributional gaps. The number of known species of ctenids in Colombia is increased from 16 to 25 and these data will be useful for future studies in taxonomy, systematics and biogeography of this family.