Nowadays, there is a general concern regarding the increasing global talk about functional foods that respond to our demands and needs as consumers in order to maintain health and body weight through a correctly balanced diet. Cereals are key elements of nutrition and a healthy diet, and they also play a significant role in health promotion due to the useful nutrient content. Therefore, this work aims to identify barley and oat genotypes suitable for human nutrition and to achieve practical results for their widespread use in preventing or treating certain chronic diseases by analyzing the nutritional and physical properties of 52 genotypes of oat and barley conserved in Suceava Gene Bank, Romania. The first part of this manuscript is the presentation of these accessions and the evaluation of their most important properties. For oat and barley cultivars, detailed processing was carried out, involving the computation of variation amplitude, coefficients of correlation and cluster analyses, both for biochemical (protein, lysine and tryptophan contents) and physical (test weight and seed weight) properties. The results indicated high variability between oat and barley varieties. Thus, according to the results, the 26 varieties of oat exhibited almost double the content of lysine compared to barley seeds, while tryptophan had higher values in barley than in oat seeds. Overall, both species play an essential role in human nutrition, barley being important because of its high protein content and higher productivity compared to oats, which, although not as productive, have better quality seeds due to their higher lysine content. The results presented are not only of scientific interest but also have practical implications for agriculture, food safety, nutrition and human health. The documented information will facilitate new studies needed to contribute to improving human nutrition and health.