Simultaneous determination of free thyroxine and capacity of thyroxine-binding globulin

A simple method is described for the simultaneous determination of capacity thyroxine-binding of globulin (TBG) and free thyroxine concentration (FT4). The ratio of bound to free T4 (B/F) is first determined for two total-T4 concentrations using a Sephadex G-25 competitive-binding technique. TBG capacity and FT4 can both the calculated assuming a known value of affinity constant of TBG. The method is linear over a twenty-fold serum dilution. FT4 calculated is identical to that calculated using the method of Irvine. TBG capacity is shown to be linearly correlated to TBG concentration as determined by radioimmunoassay and is consistent with a molecular weight of 69,000 Daltons and one T4 binding site per molecule. FT4 is found to correlate with the free thyroxine index in a complicated way, depending on the degree of TBG saturation.