The subfamily Phlebotominae is represented by insects commonly known as sand flies widely distributed in tropical and subtropical regions, with the Neotropical region having the greatest diversity. Only New Zealand or the Pacific Islands, excepting New Caledonia, do not present records. One means of access to the more than 1,060 described species is through the specimens deposited in biological collections, and the availability of associated data on online platforms for consultation on the part of students and researchers. This being the case, this study aimed catalogue the sand flies deposited in the Invertebrate Collection of the MZUSP (University of São Paulo Museum of Zoology). Twenty-nine species designated as type material were found being of nine holotypes, one “allotype” and 90 paratypes described from five countries in the Americas: Panama, Honduras, Jamaica, Mexico, and Brazil. In addition to these, 1,346 non-type specimens, from 12 Brazilian states, are deposited in the collection and have been included here in list form. The MZUSP sand fly collection currently houses specimens from 152 species, which represents 28% (546) of the Phlebotominae fauna of the Americas.