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Cereal Chem 69:371-378   |  VIEW ARTICLE

Reversed-Phase High-Performance Liquid Chromatographic Analysis of Wheat Proteins Using a New, Highly Stable Column.

B. A. Marchylo, D. W. Hatcher, J. E. Kruger, and J. J. Kirkland. Copyright 1992 by the American Association of Cereal Chemists, Inc. 

A new class of sterically protected monofunctional-silane bonded phase columns (C8 and CN) was evaluated to determine resolution capabilities and long-term stability in the reversed-phase high- performance liquid chromatography (RP-HPLC) separation of wheat storage proteins. Up to 450 separations were performed using the high temperature (50 C) and low pH (less than 3) conditions required to resolve gliadins and glutenins. Superior resolution of these proteins was achieved with the new wide- pore, sterically protected columns as compared to that from conventional silica-based RP-HPLC columns. Statistical analysis of retention times and quantified peak areas also indicated improved stability and reproducibility as compared to that from conventional RP-HPLC columns. Column-to-column variability also appeared minimal. Analysis of storage proteins (gliadins or gliadins and glutenins) of different wheat varieties and high molecular weight glutenin subunits exemplified the suitability of these columns for varietal identification and the study of quality-related wheat storage proteins.

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