Plant sterols and their derivatives are minor compounds that have been extensively studied in vegetable oils, mainly in olive oil, where they are closely related with its identity. The objective of this work is to determine the content of free and esterified steryl glucosides and their profiles in olive oil in relation to different geographical situation of olive orchards, cultivar, farming modality, and sampling time. The orchards under study were located in the outer ring of the submetropolitan area of Madrid (Spain), where olives from Cornicabra, Manzanilla Cacereña, Manzanilla Castellana, and Picual varieties were grown under traditional and organic modes, and harvested in four different samplings. Conclusions state that cultivar, farming mode, and light exposure do not have outstanding effects, whereas pedoclimate might affect the steryl glucoside presence in a substantial way. Further studies are being carried out presently in order to confirm such statement. Also glucoside derivative profiles are discussed, and reasons for differences with results in previous studies pointed out.