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Oat β-glucan has been shown to play a positive role in influencing lipid and cholesterol metabolism. However, the mechanisms behind these beneficial effects are not fully understood.The purpose of the current work was to investigate some of the possible mechanisms behind the cholesterol lowering effect of oat β-glucan, and how processing of oat modulates lipolysis. β-Glucan release, and the rate and extent of lipolysis measured in the presence of different sources of oat β-glucan, were investigated during gastrointestinal digestion.Only a fraction of the original β-glucan content was released during digestion. Oat flakes and flour appeared to have a more significant effect on lipolysis than purified β-glucan.These findings show that the positive action of β-glucan is likely to involve complex processes and interactions with the food matrix. This work also highlights the importance of considering the structure and physicochemical properties of foods, and not just the nutrient content.