To evaluate biochemical changes of inner ear fluid following perilymphatic fistula (PLF), free amino acid (FAA) profiles of perilymph in experimental PLF were determined using high performance liquid chromatography (HPLC). Thirty-five guinea pigs were anesthetized and prepared as PLF models by perforating the round window membrane (RWM) of the left ear. Right ears served as controls. Samples (2 microliters) were aspirated from scala tympani through a RWM perforation. Animals were divided into two groups according to time of sampling following PLF induction: 2-week group (n = 17) and 4-week group (n = 18). Compound action potential (CAP) evoked by 1, 2, 4 and 8 kHz tone bursts were measured using a round window electrode from the left ear before PLF induction and from both ears before final sampling. RWM perforations were completely closed at the time of the final sampling in 8 of 17 animals from the 2-week group, and 15 of 18 animals from the 4-week group. In comparison with that in control ears, concentrations of FAA throughout all profiles was dramatically elevated in the PLF ears with a healed RWM perforation. Most PLF ears with persistent RWM perforation showed minimal differences between 2-week and 4-week groups. No remarkable CAP threshold changes were found at any frequencies tested following PLF induction in both the 2-week and 4-week groups. The unchanged 8 kHz threshold suggests that FAA concentration increases only at the basal end of the cochlea. FAAs accumulate within the basal end of scala tympani in ears with a healed RWM.(ABSTRACT TRUNCATED AT 250 WORDS)