Reversible acetylcholinesterase inhibitors are an important group of drug compounds that are used medicinally to treat Alzheimer's disease. In this study, dissociation constant values (pK(a)) of some reversible acetylcholinesterase inhibitors (donepezil, rivastigmine and galantamine) having different functional groups were determined in different percentages of acetonitrile (MeCN)-water and methanol (MeOH)-water binary mixtures using reversed phase liquid chromatography (RPLC) method. In this way, a complete description of the retention behavior of each solute in the space defined by the pH and organic modifier percentages variables was obtained. The linear relationships established between retention factors of the species and the polarity parameter of the mobile phase (E-T(N)) was proved to predict accurately retention in liquid chromatography (LC) as a function of the acetonitrile and methanol content. In result, the estimated aqueous pK(a) values of studied basic compounds were assessed by comparing them with literature data.