SummaryThis study evaluates the feasibility of using near-infrared (NIR) spectroscopy as a rapid and environmentally friendly technique for validation and prediction of the total phenolic content (TPC) and antioxidant activity (AOA) indices (as 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging, inhibition time (IT) of the Briggs-Rauscher oscillating reaction, and relative antioxidant capacity (RAC)) of berry fruit extracts. The analysed berry samples originated from Croatia (blackberries, wild blueberries, raspberries, red currants and strawberries) and Bulgaria (wild blueberries, raspberries and strawberries). Principal component analysis and partial least squares (PLS) regression were used from the set of chemometric tools in distinguishing and validating the measured berry fruit extract. ANOVA and PCA showed no signifi cant impact of the origin and freshness of the samples. PLS models were developed to validate the relationship of NIR spectra with TPC and AOA of berry fruits. Representativeness of the models was expressed with the R 2 and the ratio of performance to deviation. Calculated R 2 values were above 0.84 and the ratio of performance to deviation was between 1.8 and 3.1, indicating adequacy of the PLS models.