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This review is devoted to consideration of physicochemical methods for detection of pharmaceutically active substances, in particular, phosphodiesterase-5 (PDE-5) inhibitors, present in dietary supplements. The necessity of using the most informative approaches for the identification of PDE-5 inhibitors registered as drugs and their structurally modified analogs with unknown safety and efficacy in the composition of dietary supplements is emphasized. © 2022, Springer Science+Business Media, LLC, part of Springer Nature.