The present work focused on the quality and the chemical composition of monovarietal virgin olive oil from the Sigoise variety grown in two different locations in Tunisia, viz., a sub-humid zone (Béjaoua, Tunis) and an arid zone (Boughrara, Sfax). In addition to the quality characteristics (acidity, peroxide value, and the spectrophotometric indices K232 and K270) and the chemical composition (content of fatty acids, antioxidants, and volatile compounds) of the oil, the fruit characteristics of the olives were studied. Except for the content of the majority of the fatty acids, there were significant differences observed in the oil composition of olives that were cultivated in different locations. The content of total phenols and lipoxygenase (LOX) oxidation products was higher for olives grown at the higher altitude, whereas that of alpha-tocopherol, carotenes, and chlorophylls was higher for olives from the Boughrara region (lower altitude). Moreover, olives produced at the higher altitude showed a higher ripeness index and oil content than those cultivated at the lower altitude.