Essential oil (EO) composition, phenolic content, and antioxidant activity were investigated in 17 P. abrotanoides populations collected from different geographical regions in Iran. The highest (3.61%) and lowest (1.25%) essential oil yields were measured in populations from Semnan Province (PSESM ) and PISKS from Isfahan Province, respectively. GC/MS analysis identified camphor (4.05 - 35.94%), 1,8-cineole (7.15 - 24.34%), borneol (0 - 21.75%), and α-pinene (2.05 - 10.33%) as the main constituents of Perovskia essential oil. Cluster analysis classified the studied populations into four different groups: (I) high camphene, (II) high camphor/1,8-cineole, (III) high borneol/δ-3-carene, and (IV) high α-cadinol/trans-caryophyllene. The highest flavonoid and phenolic contents were detected in PISAK from Isfahan Province (4.09 ± 0.05 mgQE/gDW, 58.51 ± 1.63 mgGAE/gDW) and PKRGS from Khorasan Province (3.80 ± 0.002 mgQE/gDW, 66.86 ± 0.002 mgGAE/gDW). DPPH and reducing power activity model systems identified PMASA and PKRKL as the populations with the highest antioxidant activity. Finally, the data obtained represented valuable information for introducing elite populations with EO components favorable to pharmaceutical and industrial applications.