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Aggregation process of riboflavin molecules in binary mixtures water - dioxane, water - DMSO, and ethanol - isobutanol, were investigated using spectroscopic methods and quantum-chemical calculation. It was shown that at a constant concentration of riboflavin and different ratios of binary mixtures, a deformation of the electronic absorption spectra with a hypochromic effect is observed. The observed changes are caused by the formation of a hydrogen bond and dipole-dipole interaction between riboflavin molecules, which is accompanied by