The Papaveraceae family is one of the most important alkaloid containing families. It contains isoquinoline alkaloids including aporphines, protopines, protoberberines and proaporphines. The genus Glaucium (Papaveraceae) comprises about 25 species of herbaceous plants in the world and Aporphinoids are the greatest number of isolated alkaloids from this genus. S-(+)-Glaucine (C21H25NO4) is the main alkaloid component of the Papaveraceae family, which are collected during the blooming period. This compound belongs to the group of aporphine alkaloids and is used in medicine as an antitussive agent. In the current research, we studied leaf Glaucine composition of nine Glaucium species from Iran. These species were harvested from the northern and western provinces of the country and their leaf ethanolic extracts were subjected to HPLC, for detection the types and amounts of their Glaucine compounds. The concentration of these Glaucine compounds differed between the species. The highest amount of Glaucine compounds were detected in G. corniculatum var corniculatum and G. corniculatum var flaviflorum, while G. flavum var flavum had the lowest one. The evaluated species divided into two distinct group in WARD tree. All of evaluated individual belonged to the same species of the genus, therefore our findings revealed that the Glaucine data were useful at species level for identification of the species.