The aim of this study was to investigate the biochemical composition and antioxidant activity of extra virgin olive oils (EVOOs) produced from different olive cultivars in the two coastal areas of Turkey; Aegean and Mediterranean regions. Samples were divided into two groups according to the olive-growing region and were classified by soft independent modeling of class analogy (SIMCA) algorithm based on chemical composition of EVOOs. Oleic acid was determined as the major fatty acid in both regions and varied from 46.7 to 71.6%. beta-Sitosterol was the main sterol compound, and trioleoylglycerol (OOO) was determined as the main triacylglycerol (TAG) in all samples with >32% ratio. Memecik cultivar (263.4 mg/kg oil) and Gemlik cultivar (279.5 mg/kg oil) had the highest alpha-tocopherol content in Aegean and Mediterranean regions, respectively. Olive oil produced from Gemlik cultivar had the highest total phenolic content and antioxidant capacity in all samples.