Carob is the fruit obtained from Ceratonia siliqua L. and it is a source of bioactive compounds that have been linked to several health promoting effects, including lowering blood cholesterol concentration. The objective of this study was to connect the physicochemical changes of carob flour occurring during roasting with its capacity to bind glycoconjugates of bile acids. Carob flour samples were roasted for different times at 150 °C and chemically characterized by measuring the concentrations of tannins and polyphenols. Data showed that carob flour binds high amounts of bile acids: 732.6 μmol of bound bile acid per g of carob flour which is comparable to the 836.2 μmol per g bound by cholestyramine, a known cholesterol lowering drug. The carob flour ability to bind cholesterol decreases up to 40% during roasting. Data suggested that tannins and insoluble components play a major role in binding bile salts, as a result of hydrophobic interactions.