Little attention has been paid to the effect of long-term fertilizer application on concentrations of elements in grain of barley produced on the metal non-contaminated agricultural soil. In 2010, we analyzed yield and concentrations of elements in grain of spring barley in unfertilized control, mineral fertilizer application (N₄P₂K₂ – 70, 60 and
100 kg N, P and K per ha) and combinations of farmyard manure or poultry litter with mineral fertilizer (FMN₄P₂K₂ and PLN₄P₂K₂) treatments in the Ruzyně Fertilizer Experiment established on Luvisol in 1955 in Prague (Czech Republic). The yield of grain ranged from 4.03 to 9.74 t/ha in the control and FMN₄P₂K₂ treatment. There was a positive effect of fertilizer application on concentrations of nitrogen, phosphorusand potassium, but no effect on concentrations of calcium and magnesium. With the exception of iron, concentrations of micro (copper and zinc) and risk elements (arsenic, cadmium, chromium, lead, manganese and nickel) were not significantly affected by the fertilizer treatments. Long-term use of organic and mineral fertilizers with appropriate application rates does not represent any risk for contamination of barley grain by risk elements on mineral rich and metal non-contaminated agricultural soils.