Evaluation of Pure Bile Salts in Place of Bile Extract in the Standardized INFOGEST Digestion Protocol for Quantification of Sterol Bioaccessibility

Improving standardized in vitro digestion protocols for phytosterols (PSs) is critical for understanding their bioaccessibility (BA) in food products and supplements. In this study, in vitro BA of phytosterol esters (PSEs) and free cholesterol (Ch) was compared under modified digestion conditions. The addition of Ch esterase (CE) to the INFOGEST model containing bovine bile resulted in a 70% increase in PS BA and an 18.5% reduction in Ch micellarization. Relative to the standardized INFOGEST model, substitution of pure bile salts (PBSs) did not significantly change PS BA or Ch micellarization. In the presence of CE, the substitution resulted in a 49.9% reduction in PS BA and a 13% increase in Ch micellarization. The differing results may be due to inhibitory effects of PBSs on the activity of intestinal enzymes, including CE. These results suggest that the INFOGEST model should include Ch esterase and the continued use of bile extract to evaluate PS BA.