The study evaluated physicochemical properties of bio-waste as a potential biofuel in the form of leaves from ‘Regent’ grapevines grown on six different rootstocks and a control grown on its own roots for three years of cultivation. An elemental analysis was carried out, determining the content of carbon, hydrogen, nitrogen, and sulphur in the leaves tested. A technical analysis of the biofuel was also carried out to determine the content of moisture, volatile matter, and ash. The calorimetric method was used to determine the higher heating value for the material. Fixed carbon and oxygen carbon was calculated based on the elemental and technical analyses. The study showed that the type of rootstock and the year of cultivation influence the amount of leaves obtained from the cultivation area. Leaf entrustment per hectare ranged from 1,140,868.02 in rootstock 161-49 to 1,265,286.7 Mg∙ha–1 in rootstock SO4. Regardless of the year of the study, shrubs grafted on 125AA rootstock and the control had the highest combustion heat of 17.5 MJ∙kg–1 and 17.6 MJ∙kg–1 respectively, while 5BB rootstock had the lowest combustion heat (16.4 MJ∙kg–1). Statistical analysis showed no significant effect of test year on the elemental and technical parameters evaluated. It was observed that regardless of the evaluated parameter and the type of rootstock in most parameters, the values in 2022 were the highest, while in 2021 they were the lowest.