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ARS Home » Plains Area » Fargo, North Dakota » Edward T. Schafer Agricultural Research Center » Cereal Crops Research » Research » Publications at this Location » Publication #235527
Title: Size Exclusion HPLC of Protein Using a Narrow-Bore Column for Evaluation of Bread-Making Quality of Hard Spring Wheat Flours

Author
item Ohm, Jae-Bom
item Hareland, Gary
item SIMSEK, SENAY - North Dakota State University
item Seabourn, Bradford - Brad

Submitted to: Cereal Chemistry
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 6/1/2009
Publication Date: 7/1/2009
Citation: Ohm, J., Hareland, G.A., Simsek, S., Seabourn, B.W. 2009. Size Exclusion HPLC of Protein Using a Narrow-Bore Column for Evaluation of Bread-Making Quality of Hard Spring Wheat Flours. Cereal Chemistry. 86(4) 463-469

Interpretive Summary: This research was performed to improve analyses of unreduced proteins in flour by size exclusion HPLC (SE-HPLC) and subsequent evaluation of bread-making quality of hard spring wheat flours. Total protein extracts and sodium dodecyl sulfate (SDS) buffer extractable and unextractable proteins were analyzed by SE-HPLC using a narrow bore column. A narrow bore column separated proteins in 10 mins at a flow rate of 0.5 mL/min with similar resolution to a regular column which took 30 min. SE-HPLC data obtained from a narrow bore column showed comparable or superior repeatability and prediction capability of flour bread-making characteristics when compared with those of a regular column. Chromatogram data of total protein extracts were calculated using those of SDS extractable and unextractable proteins. The calculated chromatogram data showed superior repeatability and correlations with quality characteristics to those of actual total protein extracts, indicating that they could be used for quality evaluation without additional extraction of total proteins and HPLC analyses. The improvements including employment of a narrow bore column in SE-HPLC and calculation of SE-HPLC profile of total proteins provide enhancement of rapid quality evaluation and decreased consumption of hazardous organic solvents.

Technical Abstract: The objective of this study was to investigate if 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 bread-making quality of hard spring wheat flours. Total protein extracts and sodium dodecyl sulfate (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 bread-making characteristics when compared with those of a regular column. AA values of total protein extracts were calculated by adding AA values of SDS extractable and unextractable proteins. The calculated AA values showed superior repeatability and correlations with quality characteristics to those of actual total protein extracts, indicating that they could be used for quality evaluation without additional extraction of total proteins and HPLC analyses. The improvements including employment of an NBC in SE-HPLC and calculation of SE-HPLC profile of total proteins provide enhancement of rapid quality evaluation and decreased consumption of hazardous organic solvents.