In the present study, the yield, the chemical composition, and the antioxidant activities of the essential oils (EOs) of eight medicinal and aromatic plants (MAPs) cultivated under two environmental conditions characterized by a different altitude (namely mountainous and plain) were evaluated. Cultivation at different environmental conditions resulted in significant differences in the chemical composition and antioxidant activity for most of the studied species. In particular, high altitudes resulted in increased phenolic compounds’ content and antioxidant activity for artemisia plants, while specific parameters increased in the case of spearmint (total phenols) and rosemary (flavonoids). In contrast, in pelargonium, all the tested parameters were positively affected in the plain area, whereas, for laurel and sage, only flavanols remained unaffected. EO yield in mountainous pelargonium and spearmint decreased while, in mountainous laurel, pelargonium and spearmint increased when compared to plain areas. In addition, the major EO constituents’ content for most of the species were affected by environmental conditions. The 2,2-diphenyl-1-picryl-hydrazyl-hydrate (DPPH) and ferric reducing antioxidant power (FRAP) were variably correlated with total phenols, flavonoids, and flavanols, depending on the species and the altitude. Lastly, in limited cases, antioxidant activity (DPPH or FRAP values) was positively correlated with some EO components (e.g., borneol and β-pinene in artemisia and laurel plants grown in the plain, respectively, or 1,8-cineole in mountainous grown verbena plants). In conclusion, environmental conditions (altitude) affected antioxidants’ content and EO yield and composition of the studied MAPs. These findings can be used to introduce cultivation of MAPs in specific ecosystems for the production of high added value products.