A high performance liquid chromatography procedure, using anion exchange chromatography of the gliadin proteins at pH 10·4, has been devised as an alternative to gel electrophoresis for identification of wheat varieties. Varietal identification was, thus, possible in less than one hour. Sample preparation and extraction took about 30 min. The sample for chromatography could be prepared directly by extraction of flour with the starting buffer, indicating that a lengthy purification of the gliadin fraction is not necessary. Chromatography required approximately 20 min, and regeneration of the column was achieved in 5–10 min. Chromatographic separation of the gliadin proteins was less effective at pH values below 10·4. In initial experiments, 13 genotypes, including two varieties that are difficult to differentiate by electrophoresis, were examined and each was readily differentiated by the technique. Analysis of both purified gliadins and total flour extracts indicated that the results were independent of environmental factors.