Nowadays, soil contamination by risk elements from industry is a serious environmental problem. As one of the factors, it is necessary to monitor the biological availability of potentially toxic elements from soil to plants. The aim of the study is to evaluate the biological availability of the aboveground parts of barley biomass in the soil contaminated by PTEs. Biological availability was monitored for the risk elements Mo, Sr, As, and Cd. Moreover, the mutual interaction between individual soil treatments with the addition of PTEs and the content of selected risk elements (Mo, Mn, Sr, Cr, As, Cd, Pb, Cu, Zn) in barley straw was studied. Non-parametric Mann-Whitney U test was used for the assessment of statistically significant change in the biological availability of PTEs for barley straw. In many cases, a positive or negative correlation was determined between the bioavailability of metals in different variants of contamination. Mo, Sr, As, or Cd positively affected the intake of Zn into barley straw at increased concentrations in the soil. Of all the variants of contamination, Cd brought about the greatest support for the intake of the elements, which led to an increase in the intake of Mo, Sr, Cd, and Zn. On the other hand, the same element acted as a blocker of the largest number of PTEs compared to other variants of contamination, namely Cr, As, Pb, and Cu. For PTEs, Mo, Sr, and Cd confirmed As a blocker of intake.