DisplayTitle

Separation of Wheat Proteins by Two-Dimensional Reversed-Phase High-Performance Liquid Chromatography Plus Free Zone Capillary Electrophoresis¹

¹Cooperative investigations, U.S. Department of Agriculture, Agricultural Research Service and the Department of Grain Science and Industry, Kansas State University. Contribution 97-145-J, Department of Grain Science and Industry, Kansas State Agricultural Experiment Station, Manhattan, KS 66506. Mention of trademark or proprietary products does not constitute a guarantee or warranty by the U.S. Department of Agriculture and does not imply its approval to the exclusion of other products that may also be suitable. ²Graduate research assistant and adjunct professor, respectively, Department of Grain Science and Industry, Kansas State University, Manhattan, KS 66506. ³Research chemist, USDA-ARS, Grain Marketing and Production Research Center, Manhattan, KS 66502.

Gliadins and glutenins from four hard red winter wheat cultivars were separated by a novel two-dimensional (2D) technique. Protein extracts were separated by reversed-phase high performance liquid chromatography as the first dimension with each 30-sec interval collected separately. Those fractions were then separated by free-zone capillary electrophoresis (FZCE) for the second dimension. Data was combined into 2D surface contour plots similar to traditional gel electrophoresis 2D maps. For HPLC, C₈ and C₁₈ columns were used in the first dimension to separate gliadins and glutenins, respectively. Uncoated fused silica capillaries (27 cm × 25 μm, i.d.) were used for the 2D FZCE separations. Differences in the 2D maps of both gliadin and glutenin fractions were found between pairs of both closely related and sister lines that varied in quality.