Essential oils (EOs) are compounds that are biosynthesized autonomously by aromatic plants as their secondary metabolites. Vitex agnus-castus plant leaves, flowers and fruits are of pharmacological importance. In this study, the chemical composition determination of the essential oils of the Vitex agnus-castus flowers with different colors (white, pink, purple) growing in Aydin ecological conditions was carried out with the extraction using Clevenger hydro-distillation apparatus and then the essential oil composition was determined by Gas Chromatography-Mass Spectrometer (GC-MS). As a result of the study, ‘Çakmar’ pink-flowered population had the main component (-)-allospathulenol (14.59%), ‘Çakmar’ purple-flowered and ‘Koçarli’ purple-flowered populations had the main component pyrrolo (3,2,1-jk) carbazole (14.42% and 14.68%, respectively), while ‘Çakmar’ white flowered, ‘Koçarli’ white flowered and ‘Koçarli’ pink flowered populations had the main component caryophyllene (15.54%, 12.64%, and 19.05 respectively).