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The fate of β-glucan (BG) health promoting properties during food production is crucial, but not predictable yet. Therefore, high molar mass BG (hBG) and control BG (cBG) were extracted from two barley varieties, characterized and added to wheat breads at levels of 3% and 6%. Bread quality criteria, carbohydrate contents and BG content and structural properties were determined. Additionally, breads were subjected to an in vitro digestion. The BG content in the chyme, molar mass, molar ratio, viscosity and bile acid retention were determined. The hBG and the cBG decreased loaf volume and increased crumb hardness with increasing BG content. The reduction in BG content during bread making was similar for hBG and cBG, but the molar mass of cBG decreased to a greater extent. As a result, only 10% of cBG entering in vitro digestion were found in the chyme afterwards, while 40% of the ingested hBG were detected. Molar mass reduction was much more severe for cBG compared to hBG. The use of hBG showed higher viscosity and better bile acid retention, indicating cholesterol lowering properties, compared to similar or higher amounts of cBG. These results provide valuable knowledge on the criteria to select BG-rich raw materials for ideal health promoting properties.

Relevance of β-Glucan Molecular Properties on Its Suitability as Health Promoting Bread Ingredient