SOLVENT EFFECT ON TESTOSTERONE-ANTITESTOSTERONE INTERACTION

The inhibition of the binding between testosterone and antitestosterone antiserum caused by organic solvents was studied at pH 7.4, 298 K. Inhibition curves were obtained at variable ranges of molar fractions for the following solvents: methanol (range 0-0.4), ethanol (0-0.317), 1-propanol (0-0.082), 2-propan-ol (0-0.260), t-butanol (0-0.223), ethylenglycol (0-0.189), 2-methoxyethanol (0-0.036), 2-butoxyethanol (0-0.063), 1,4-dioxan (0-0.124), tetrahydrofuran (0-0.238) and acetonitrile (0-0.392). Steroid-antibody binding decreases with increasing molar fraction of solvent in the reaction mixture for all but tetrahydrofuran and acetonitrile, which enhance binding at low molar fraction then cause a sharp inhibition. Molar fraction of solvent that causes a 50% binding inhibition is uncorrelated to some solvent properties (i.e. dielectric constant, polarity index, dipole moment) but is inversely correlated to the molecular mass of the solvent. The correlation becomes better by taking into account the lenght of the solvent molecule, or the Randic molecular connectivity index, suggesting that binding inhibition could be related to the lenght of the solvent molecules that displace water around the steroid molecule. However, the increase of binding observed at low molar fraction with tetrahydrofuran and acetonitrile, together with very different shapes of inhibition curves suggest that a molecular mechanism based on the differential solvation of the steroid by solvent and water molecules must be taken into account to explain adequately the solvent effect on testosterone-antitestosterone interaction.