The polymorphism of prolamin storage proteins was studied in seed samples of 20 historical cultivars of   spring barley (Hordeum vulgare L.) of Czech and Slovak origin, using polyacrylamide gel electrophoresis (PAGE). Only two samples were uniform. Most heterogeneity of prolamin patterns was observed in the oldest accessions. By means of a prolamin identity index it was possible to distinguish sister lines from admixtures within the seed samples. The obtained spectra will be used as additional descriptors for the spring barley core collection of the Collection of Genetic Resources of the Agricultural Research Institute Kroměříž, Ltd.