Selenium (Se) is an important element associated with the enhancement of antioxidant activity in organisms. Potato is very suitable for fertilisation with Se (biofortification). The experiment was performed to examine the effect of foliar application of Se as sodium selenite (200 or 400 g Se/ha) at the tuberisation stage on a spectrum of amino acids in tubers of varieties. The trends of amino acids were consistent in both years of the study. Application of Se increased the relative content of total essential (EAA) and non-essential (NEAA) amino acids relative to the controls (Karin: EAA 16.81-21.73% and NEAA 14.18-18.63%; Ditta: EAA 4.71-13.00% and NEAA 5.78-6.49%). The increase in the content of phenylalanine (Phe) was particularly significant (up to 48.9%) when also the contents of aspartic acid (Asp), glutamic acid (Glu), threonine (Thr), and tyrosine (Tyr) increased significantly compared with the controls. The results of changes in the content of isoleucine (Ile), leucine (Leu), lysine (Lys), methionine (Met), valine (Val), alanine (Ala), arginine (Arg), proline (Pro), cysteine (Cys), glycine (Gly), histidine (His), and serine (Ser) were also discussed. The highest dose of selenium is shown as a stress factor. Its toxic effects resulted in a change of amino acid contents.