The advertisement call of Peltophryne florentinoi, an endemic toad from Zapata Swamp, Matanzas, Cuba, is described in terms of temporal properties and dominant frequency. A comparison with the advertisement calls produced by the other three species of large toads in Cuba (P. peltocephala, P. fustiger and P. taladai) is also provided. Like other large toads from Cuba, P. florentinoi produces advertisement calls with long duration (> 10 s), low dominant frequency (< 1 kHz), and a series of complex pulses that are composed of two to five sub-pulses. All temporal features of the P. florentinoi call are significantly different from P. fustiger calls: number of pulses, pulse duration, and pulse rate separates it from P. peltocephala. P. florentinoi differs from P. taladai in call duration, number of pulses, and pulse rate. The occurrence of different types of complex pulses differed in all species studied. Typically, P. florentinoi pulses contain three sub-pulses, P. taladai four, and P. fustiger and P. peltocephala contain two sub-pulses. The dominant frequency of the advertisement call of P. florentinoi did not differ from the other species' calls. Call duration and pulse rate are the most useful acoustic properties to distinguish P. florentinoi advertisement calls from those of the other three large Cuban toads.