An algorithm was developed to evaluate equilibrium constants for testosterone (Te) and sex hormone-binding globulin (SHBG) or albumin from serum free testosterone (FTe) measurements performed in a panel of 30 healthy elderly men by means of a near-reference method, i.e., symmetric dialysis (affinity constants: SHBG-Te, 1.13 x 10(9) L/mol; albumin-Te, 4.4 x 10(4) L/mol). Using these estimates, a free testosterone index (FTeI) was calculated from total Te and SHBG concentrations in a further 35 elderly men. This FTeI perfectly matches with actually measured free testosterone concentrations by symmetric dialysis in this second group, with a mean ratio index/measurement of 0.998+/-0.016 (SEM). The efficacy of the algorithm, which represents a simple alternative to previous cumbersome methods for estimation of equilibrium constants, is thereby demonstrated.