Since doxorubicin (Adriamycin) is known to bind on membranous negative surface charges, its effect on the electrochemical composition of the cochlear fluids was studied in rats. Doxorubicin was infused into the cerebrospinal fluid via the lateral cerebral ventricle. The endocochlear resting potential was recorded, and endolymph and perilymph of the scala vestibuli were collected from the basal cochlear turn before and 1, 2, and 4 h after the drug application. Na+, K+, and Cl- concentrations and osmolality of the endolymph and perilymph were measured in 1 nl aliquots. In perilymph, Cl- concentration increased 4 h after doxorubicin treatment to reach a concentration 8 mM higher than recorded initially. In endolymph, the endocochlear potential decreased by 5 mV/h while its K+ concentration and osmolality increased by about 3 mM/h and 4 mosmol/kg H2O per hour, respectively. These results suggest that the negative surface charges demonstrated on Reissner's membrane may play a role in the homeostasis of endolymph.