We measured free thyroxin (FT4) and free triiodothyronine (FT3) in serum of patients taking the anti-epileptic drugs phenytoin and carbamazepine, both by equilibrium dialysis procedures and analog-type radioimmunoassays. By either assay, the mean concentration of FT4 was significantly decreased in patients receiving either drug, whereas their FT3 concentrations were normal or only slightly decreased. Adding therapeutic concentrations of these drugs in vitro to control sera had a small or no incremental effect on FT4 and FT3 as measured by either method, but adding greater concentrations of the drugs in vitro markedly increased the concentrations of the free hormones. These results indicate that the main mechanism of the decrease in concentrations of free thyroid hormones in serum during therapy with anticonvulsant drugs is not the displacement of hormones from their binding to plasma proteins. We also determined, using a new and sensitive immunoradiometric assay, that patients taking carbamazepine, but not those taking phenytoin, had significantly less thyrotropin in the serum.