Cereal Chem 66:112-116   |  VIEW ARTICLE

Evaluation of the Precision of High-Performance Liquid Chromatography for Wheat Cultivar Identification.

M. G. Scanlon, H. D. Sapirstein, and W. Bushuk. Copyright 1989 by the American Association of Cereal Chemists, Inc. 

The reproducibility of computer-derived reversed-phase high-performance liquid chromatography (RP- HPLC) quantitation parameters following prolonged use of a single commercially available column was studied. Using a standardized experimental procedure, more than 65 chromatograms were evaluated based on gliadin extracts from bulk-ground meal and composite grinds of four kernels of the bread wheat cultivar Neepawa. Statistical results are reported for a set of more than 30 chromatogram peaks with respect to the precision of retention times, computed peak areas and heights, and percentage peak areas and heights. Whereas chromatographic resolution was relatively constant over time, prolonged column use significantly retarded peak retention times especially for early eluting components. Results show that without appropriate normalization, peak retention times lack sufficient long-term precision in order to obtain reliable results for cultivar identification and other RP-HPLC comparative analysis applications.