Cereal Chem 65:28-40   |  VIEW ARTICLE

Identification of Wheat Cultivars by Reversed-Phase High-Performance Liquid Chromatography of Storage Proteins.

B. A. Marchylo, D. W. Hatcher, and J. E. Kruger. Copyright 1988 by the American Association of Cereal Chemists, Inc. 

Wheat storage proteins comprised of the gliadins and high and low molecular weight glutenins were resolved by reversed-phase high-performance liquid chromatography (RP-HPLC). Resultant chromatograms or protein "fingerprints" were used to identify 18 registered and one nonregistered Canadian and eight nonregistered American spring wheat cultivars. Six commercially available HPLC columns differing in column dimensions, hydrocarbon chain length, and particle and pore size were evaluated, and the effect of column-to-column variability and column stability on resolution and selectivity are discussed . The effect of column temperature on the separation of storage proteins was studied, and this was assessed in relation to quantitation of chromatograms and the cultivar identification process. Reproducibility of peak areas is reported. On a qualitative basis, all registered cultivars were distinguishable from nonregistered cultivars. Quantitation of chromatograms derived from RP-HPLC analysis of ground grain extracts of admixtures of a registered and nonregistered cultivar was carried out. Highly significant linear correlations were obtained when peak areas or relative peak areas were plotted against cultivar concentration. This provided a basis for quantitative computer analysis of admixtures.