Essential oil (EO) is a mixture of low molecular weight constituents that are responsible for its characteristic aroma. These constituents include terpenoid and non-terpenoid hydrocarbons and their oxygenated derivatives. This chapter focuses on the heterogeneous composition of the essential oils. It discusses the usage of essential oil constituents as a key marker of the oil quality, freshness and unique characteristics. It describes the biological activity and synergistic effect of the essential oil constituents as antioxidant, antibacterial, antifungal and anticancer agents.