Anurans exhibit a diverse acoustic repertoire, and the release call is emitted when an individual is improperly clasped, often by males mistakenly grasping other males or unreceptive females during amplexus. The release call also serves as a crucial source for taxonomic resolution. However, for certain widespread hylid genera like Trachycephalus, knowledge about their release calls is limited or non-existent. Trachycephalus comprises 18 species, yet only one has its release call described. This study presents the first description of the release call of T. typhonius from a population in the Cerrado of Central-Western Brazil. Its release call is composed of a multipulsed note, distinguishing it from the only other known call in the genus (T. ‘vermiculatus’). Given the differences in the release call parameters between these species, such variations could serve as diagnostic criteria for distinguishing the two lineages. Additional descriptions of release calls, including from more T. typhonius and T. ‘vermiculatus’ populations, are essential for species diagnosis and taxonomic classification.