Size-Exclusion HPLC of Protein Using a Narrow-Bore Column for Evaluation of Breadmaking Quality of Hard Spring Wheat Flours

The objective of this study was to investigate whether a narrow-bore column (NBC) (300 x 4.5 mm, i.d.) improved analyses of unreduced proteins in flour by size-exclusion HPLC (SE-HPLC) and subsequent evaluation of breadmaking quality of hard spring wheat flours. Total protein extracts and SDS buffer extractable and unextractable proteins were analyzed by SE-HPLC. NBC separated proteins in 10 min at a flow rate of 0.5 mL/min with similar resolution to a regular column (300 x 7.8 mm, i.d.) which took 30 min. SE-HPLC absorbance area (AA) data obtained from an NBC showed comparable or superior repeatability and correlations with flour breadmaking characteristics when compared with those of a regular column. AA values of total protein that were calculated by adding AA values of SDS extractable and unextractable proteins showed greater repeatability and correlations with quality characteristics than those of actual total protein extracts. The improvements including employment of an NBC in SE-HPLC provide enhancement of rapid quality evaluation and decreased consumption of hazardous organic solvents.