In the present work is concerned with preparation, separation and structure elucidation of solid ion-pairs of three antifungal drugs Fluconazole (FLZ), Voriconazole (VRZ) and Butoconazole nitrate (BTZ) with Rose Bengal (Rbeng). The solid ion-pairs were prepared and investigated using FT-IR, mass spectral (MS) and thermal analyses (TA), in addition to elemental analysis (EA). The general formulae of the prepared ion-pairs were determined. Moreover, the formulae of prepared ion-pairs were proposed and structurally identified. The biological activities of the separated solid ion-pairs towards five types of bacteria and one type fungi in comparison with their drugs were studied, and they were found to be biologically active more than their parent drugs.