Crop seeds are the main staples in human diet, especially in undeveloped countries. In any case, the diet needs to be rich not only in macro-nutrients like carbohydrates and protein, but also in micro-nutrients. Nevertheless, both the macro- and micro-nutrients presented in seeds largely vary in consequence of field and environment conditions. In this research, 60 lines of a barley RILs population segregating for the SSR marker Hvm74, which is genetically linked to the GPC (grain protein content) locus (HvNAM-1), were studied in 4 environments (two growing years and two field managements) by carrying out a comprehensive profile of seed macro- (starch, total nitrogen and total soluble protein) and micro-nutrients (phytate, phenolics, flavonoids, Pi, Zn and Fe). Under field conditions, all the components were largely affected by the environment, but TN (total nitrogen) exhibited high genotype contribution, while micro-nutrients displayed higher genotype × environments interactions (GEI) than macro-nutrients. In order to approach the effects of carbon-nitrogen (C–N) balance on other seed components, two C/N ratios were calculated: C/TN (CNR1) and C/TSP (CNR2). CNR2 exhibited stronger negative correlations with all micro-nutrients. Hence, the significant GEI and its negative relationships with CNR2 highlighted the different characters of micro-nutrients in barley seeds.