Diosmin is a naturally occurring flavone glycoside used in the treatment of venous diseases. In this review, we present the clinical aspects of the use of diosmin preparations in venous stasis, hemorrheologic disorders and vein wall remodeling. Because of its multiple applications in biology and its many therapeutic activities, research on isolation and identification of diosmin is of high relevance. The aim of this review is to present an overview of techniques of isolation and separation of diosmin in plant material, pharmaceutical formulations such as Daflon, Diosed and Dioven tablets, and biological fluids.