The aim of this study was to elucidate underlying biochemical mechanisms of drought induced in spruce species on Kopaonik mountain by application of antioxidant assays for estimation of oxidative stress in spruce needles that were characterized as healthy as well as those that exhibit symptoms of severe drought. Results of radical scavenger capacities against DPPH and ABTS radicals, showed increased values in drought affected needles thus exhibiting higher antioxidant properties. With further analysis we proved that drought conditions induced significantly increased biosynthesis of total phenolic compounds (TPC) as well as flavonoids in drought samples comparing to the controls. Drastically elevated level of lipid peroxidation indicated that drought induced oxidative stress. Furthermore, under drought condition free proline accumulation was significantly higher comparing to the healthy controls. All obtained data indicate that application of biochemical assays could serve as a diagnostic tool for recognition of early symptoms of drought and could serve as a criterion for selection of individuals that are more tolerant to drought stress.