Due to the advantage of RP-HPLC with a variety of compositions of mobile phases, experiments on water-soluble charged species were examined. The samples were mononucleotides (5-CMP, 5-UMP, 5-GMP, 5-IMP, 5-AMP), and the buffers used were sodium phosphate monobasic and acetic acid. The concentrations of buffers ranged from 0.01 to 10 mM, while that of the methanol, an additive to the mobile phase was 5 to 20 vol.%. To predict the retention factor of a sample in terms of its methanol composition (M, vol.%) and buffer(C(sub)B, mM), the following nonlinear equation is suggested, k= $frac{a+b C_B}{(1+c C_B) M^d}($ where a, b, c, and d were experimentally determined constants. The regression coefficients were above 0.96, and the agreement between experimental and calculated retention factors were relatively good.