The type series of Leporinus paranensis includes two distinct species, one of which is herein described as new. Leporinus paranensis is redescribed based on its holotype, two paratypes, and additional specimens recently sampled in tributaries of the Grande river, in the north portion of the upper Paraná basin in Brazil. A new species of Leporinus is described based on specimens collected in all major tributaries of the upper Paraná basin in Brazil, and tributaries of the Paraná river in Paraguay. Both species share the presence of three unicuspid teeth on the premaxillary and four on dentary, terminal mouth and three dark midlateral blotches on the body. The new species is distinguished from L. paranensis based on the number of scale series around the caudal peduncle (12 vs. 16). The variation in the body shape of both species were compared through a Principal Component Analysis and showed that they share a similar body shape. Species delimitation analyses using DNA Barcode were applied to compare samples of the new species to congeners and corroborated the uniqueness of the new species. In addition, molecular data revealed that L. bahiensis, L. octofasciatus, and L. taeniatus are possibly closely related to the new species. The conservation status of L. paranensis and the new species are discussed using the IUCN criteria.