For the first time the effects of different sampling periods and their interaction with five major autochthonous Croatian Istrian olive cultivars and the Italian cultivar ‘Leccino’ on the quantity and composition of olive leaf phenolic compounds and mineral nutrients were investigated. For that purpose, olive leaves were sampled in two collecting periods, in October and March, coinciding with the harvesting and pruning periods, respectively. All selected cultivars had a higher oleuropein leaf content in the pruning collecting period, with the highest levels noted for the ‘Leccino’ and ‘Buža’ cultivars. Cultivar significantly affected almost all the investigated phenols, with higher concentrations of these valuable compounds in the pruning than in the harvesting period. Differences observed in leaf mineral composition were closely related to the differences in phenolic profiles and were significantly affected by genotype. Some of the studied mineral nutrients, such as P, Cu and B, were found to be significantly correlated with the most abundant olive leaf phenolic compounds, oleuropein and verbascoside.