Beneficial dietary and pro-health values, have resulted in the increasing consumption importance of barley. Therefore, genotypes and cultivation methods are sought that guarantee high functional value of grain. The aim of the study was to assess the content of phenolic acids, flavonoids, chlorophylls, anthocyanidins, phytomelanin and antioxidant activity of grain of three barley genotypes depending on agricultural technology. Two of them are primary genotypes with dark grain pigmentationHordeum vulgareL. varnigricansandH.vulgareL. var.rimpaui, the third is a modern cultivar ’Soldo’H.vulgarewith yellow grain, which is the control sample. Evaluated the effect of foliar application of a amino-acids biostimulant on the functional properties of grain under the conditions of organically and conventionally cultivations. The results indicated a higher antioxidant activity and the concentration of phenolic acids, flavonoids and phytomelanin in the black-grain genotypes. Organic cultivation and application of amino acids had increased the content of phenolic compounds in grain. The antioxidant activity was correlated with the content of syringic acid, naringenin, quercetin, luteolin and phytomelanin. Organically cultivation and the foliar application of an amino acid biostimulant improved the functional properties of barley grain, in particular the original, black-grained genotypes.