The purpose of this investigation was to use distortion-product otoacoustic emission (DPOAE) testing to address the issue of possible ototoxicity from the use of neomycin/polymyxin B/hydrocortisone otic suspension following tympanostomy tube placement. We retrospectively reviewed our clinical records and identified 36 children (52 ears) who had met our three study criteria: (1) unilateral or bilateral placement of transtympanic ventilation tubes, (2) treatment for 3 to 5 days with neomycin/polymyxin B/hydrocortisone drops, and (3) postoperative evaluation by DPOAE testing. We identified another set of 36 children (52 ears) who had not received these drops and who had not undergone tube placement but who had been evaluated by DPOAE testing to serve as a control group. We found no significant differences in DPOAE amplitudes between the treatment and control groups. These findings are consistent with decades of clinical experience indicating that neomycin/polymyxin B/hydrocortisone otic suspension is safe when used responsibly.