New ligands obtained in the reaction of o-phenylenediamine and series of aldehydes (L1-L5) were synthesized. Molecular architecture of the synthesized ligands L1-L5 has been determined by the X-ray crystal structure analysis. ¹ H NMR NOE diff. spectra (differential Nuclear Overhauser Effect) of ligands indicate that they have different conformation in the solution in comparison to the solid state (crystal structure). The substrates molar ratio and the reaction time allowed the controlled synthesis of mono substituted o-phenylenediamine or benzimidazole derivatives with no catalyst used. For all compounds the relatively high fluorescence efficiency was observed in the range 0.41–0.83 in MeCN and 0.48–0.96 in CH ₂ Cl ₂ solution. Based on DFT calculations the fluorescence mechanism was associated with HOMO→LUMO transition between aromatic rings and benzimidazole moieties.