ARS | Publication request: Size exclusion HPLC of protein using a micro-bore column for evaluation of bread-making quality of hard spring wheat flours

Submitted to: American Association of Cereal Chemists Meetings
Publication Type: Abstract
Publication Acceptance Date: June 25, 2008
Publication Date: July 25, 2008
Citation: Ohm, J., Hareland, G.A., Simsek, S., Seabourn, B.W. 2008. Size exclusion HPLC of protein using a micro-bore column for evaluation of bread-making quality of hard spring wheat flours. American Association of Cereal Chemists Meetings. Cereal Foods World 53(4): 72A

Technical Abstract: Variations in quantity and quality of wheat flour protein have significant associations with flour quality characteristics and development of rapid analytical method to analyze molecular weight distributions of flour protein is highly expected to enhance characterization of protein for flour quality evaluation specifically when sample set is large. The objective of this experiment was to investigate if employment of a micro-bore column (MBC) (300 x 4.5 mm i.d.) could improve analysis of unreduced proteins in flour by size exclusion HPCL (SE-HPLC) and subsequent evaluation of flour quality. Total protein was extracted with sonication from 33 hard spring wheat flours and separated by SE-HPLC with measurement of absorbance at 214 nm. MBC separated proteins in 10 min at a flow rate of 0.5 mL/min and in 8 min at 0.7 mL/min, while a regular column (300 x 7.8 mm i.d.) took 30 min at a flow rate of 0.5 mL/min. Flour protein content showed significant (P<0.001) correlation coefficients (r) of 0.82, 0.91, and 0.57 with total absorbance areas (AA) using the regular column and MBC at flow rates of 0.5 and 0.7 ml/min, respectively. Bread loaf volume also showed higher r values with total AA using the MBC at a flow rate of 0.5 mL/min (r=0.73) versus the regular column (r=0.61). Correlation analysis was also performed between flour quality characteristics and AA values over SE-HPLC retention time, and AA values from MBC at a flow rate of 0.5 mL/min showed a higher or similar range of r values with quality characteristics when compared with the regular column. Molecular weight distribution of flour protein could be analyzed more rapidly by SE-HPLC at 0.5 mL/min when using a MBC, and r values indicate that it could enhance determination of protein functionality for flour quality evaluation as well as decreased consumption of organic solvents.