In order to evaluate changes in extra- and intracellular pH in the brain during hypoglycemia rats were injected with insulin and pH changes evaluated when the EEG showed a slow-wave-polyspike pattern ('precoma'), or when EEG activity had ceased for 5, 15 or 30 min ('coma'). Extra- and intracellular acid-base changes were evaluated from pCO2 and HCO3-concentrations. In order to allow calculation of intracellular pH ( and HCO3- concentrations) changes in extracellular fluid volume were estimated by measurements of cortical tissue impedance. The main results were as follows. (1) At constant arterial pCO2 and CSF HCO3- concentration either rose (15 min of coma) or remained unchanged (all other groups). However, since the cerebrovenous (and tissue) pCO2 fell, all groups except one (30 min of coma) showed a significant increase in extracellular fluid pH. (2) During severe hypoglycemia, and especially when EEG activity ceased, cortical impedance increased markedly. Calculations with the help of the Rayleigh and Maxwell equations showed that the extracellular fluid volume was reduced to about 50% of control. (3) Intracellular pH increased significantly in precoma and in coma of 15 and 30 min duration. However, pH in the 5 min coma group was significantly lower (but no different from control). (4) In general, the increase in intracellular pH is consistent with previous findings that hypoglycemia is associated with oxidation of endogenous acid metabolites. However, the data suggest that in the initial period of coma acids accumulate by some unidentified mechanism.