The study aimed to conduct a complex assessment of winter barley base lines to select promising parental lines for inclusion in a hybridization program. This article presents data from field and laboratory trials conducted between 2022 and 2024, identifying the most promising winter barley varieties and lines from the collection at the Agricultural Research Center “Donskoy”. In autumn, 119 collection samples were sown at optimal agronomic times using a Wintersteiger Plotseed seeder to select the most promising (valuable) genotypes for use in breeding programs. Field trials and observations were conducted according to the Methodology of the State Variety Testing of Agricultural Crops (2019). The analysis of the studied varieties and breeding lines was performed using an improved methodology for complex assessment of winter barley breeding, developed at the ARC “Donskoy”, based on 14 key characteristics (identifiers). Structuring the data array using a complex scoring index for each sample, following a proposed algorithm to identify typical zones based on yield potential during analysis of the integrated frequency function of a variety or line, resulted in the establishment of four zones: 23 most promising samples (19.33% of the total), 31 promising samples (26.05% of the total), 42 less promising samples (35.29% of the total), and 23 unpromising samples (19.33% of the total). The most promising group included samples such as Paradiz, Banio, Cotanici, and Fakir, which exhibited high scores for a complex of key economically valuable traits and properties. These most promising samples will be included in the hybridization program in 2025 to develop new winter barley varieties with improved economically valuable traits and properties.