Despite the potential of sesquiterpene hydrocarbons in olive oil authentication, their metabolism in Olea europaea is poorly understood, and little is known about their biochemical regulation in olives as a function of ripening. To ascertain some metabolic relationships between sesquiterpene hydrocarbons and olive ripening, the content of sesquiterpene hydrocarbons was assessed in virgin olive oils from two olive varieties grown in the same geographical area and produced at different harvesting periods. During the ripening, the accumulation of sesquiterpenes in the olive itself, and thus in the oil, differed according to their molecular structure: bicyclic sesquiterpenes, showed decreasing concentrations the later the harvest, while acyclic farnesene-like compounds progressively increased through the olive ripening process. This is first evidence that the accumulation of sesquiterpene hydrocarbons in olive, and hence in olive oil, is modulated during ripening. Therefore, the degree of ripening of olives should be taken into consideration when considering the sesquiterpenic profile of virgin olive oil for their authentication.