The number of described species of Pseudopaludicola has increased at unprecedented rates over the past two decades. This increase was mainly driven by the sampling effort in regions historically neglected and the use of evidence from bioacoustic, genetic, and morphological datasets combined. Here, we describe a new species of Pseudopaludicola from a transitional zone between Amazonia and Cerrado in western Tocantins State as revealed through morphological, molecular, and bioacoustic analysis. Pseudopaludicola javae sp. nov. is distinguished from its congeners by its small size; knobbed terminal phalanges; smooth upper eyelids and heel; relatively short hind limbs; smooth, whitish, and subgular inflated vocal sac with few melanophores around the jaw; and advertisement call composed of an irregular series of multipulsed notes with 10-17 nonconcatenated pulses separated by intervals of 3-55 ms, emitted at a rate of 27-51 pulses/sec. The new species occurs in four Tocantins municipalities, all in the Araguaia-Tocantins interfluve. We also provide the first records of Pseudopaludicola jazmynmcdonaldae outside the type locality (Caseara Municipality, Tocantins State), extending its distribution nearly 245 km southward from Caseara. Pseudopaludicola javae sp. nov. is the second species of Pseudopaludicola described recently from western Tocantins.