The olive oil industry represents an important productive sector in the Mediterranean basin countries. Olive stone is an essential by-product generated in the olive oil extraction industries and it represents roughly 10% by weight of the olive fruit. The seeds of pickled olives are also a significant waste product. In the present study, we have investigated the possibility of the use of differential scanning calorimetry for the thermal characterization of seeds from green and black pickled olives from Croatia. The differential scanning calorimeter (DSC) with a normal pressure cell equipped with a cooling system was used to determine the thermal properties of seeds from olives. The following analyses were also performed: the determination of calorific values in a pressure bomb calorimeter, the determination of initial water content, the determination of changes of water content during drying at the temperatures of 30 °C, 50 °C and 80 °C, the determination of a percentage content of seeds mass to the mass of the whole olives, and the determination of ash content. Seeds from olives are characterized by very good parameters as a biomass. The analyzed olive seeds were characterized by low water content, low ash content, and a relatively high caloric value.