The protonation constant values of mercaptosuccinic acid, L-dopa and 1, 10 -phenanthroline were determined in dimethylformamide (0-60%v/v) at 303.0 K at ionic strength of 0.16 mol L -1 using pH-metric technique. The protonation constants were calculated with the computer program MINIQUAD75 and selection of the best fit models was based on the statistical parameters. The log K values were found to increase of the organic solvent content. The linear variations of the protonation constants with the reciprocal of the dielectric constant of the medium have been attributed to the dominance of electrostatic forces. Distribution of species, protonation equilibria and effect of influential parameters on the protonation constants have also been presented.