Selenium (Se) biofortification of barley is a suitable strategy to increase the Se concentration in grain. In the present paper, the suitability of this Se-biofortified grain for making Se-enriched beer is analyzed. The aim of the present study was to evaluate the effect of different Se fertilizer doses (0, 10, and 20 g of Se ha(-1)) and forms (sodium selenate or sodium selenite) on the Se loss during the malting and brewing processes and Se speciation in grain, malt, wort, and beer. Samples were analyzed using inductively coupled plasma-mass spectrometry (ICP-MS) and high-performance liquid chromatography (HPLC)-ICP-MS for total Se and speciation. Mashing-lautering was the process with the greatest Se loss (83.8%). After malting and brewing, only 7.3% of the initial Se was retained in beer, mainly in selenite form. Even so, the fertilizer application of sodium selenate at 20 g ha(-1) increased the total Se concentration almost 6-fold in the final beer in comparison to the use of grain derived from unfertilized barley. The present paper provides evidence that the use of Se-biofortified barley grain as a raw material to produce Se-enriched beer is possible, and the results are comparable to other methods in terms of efficiency.