In vitro digestion for assessing micronutrient bioavailability: the importance of digestion duration

Static digestion in vitro is a commonly used technique for investigating micronutrient availability which allows the nutrients or foods of interest to be exposed to conditions that simulate those found within the stomach and small intestine. The activity of these digestive enzymes throughout their respective simulated digestion phases has been reported to decline due to the autolytic activity of the proteases and therefore incomplete digestion may result. The degree of protease inactivation under commonly simulated digestion conditions requires further quantification. Pepsin and pancreatic protease activities were assessed throughout a simulated digestion protocol in vitro over multiple time points using stop-rate spectroscopy. The protease activity of both pepsin and pancreatin decreased significantly during their respective digestion phases. Results suggest that gastric and intestinal proteases are destroyed or inactivated during their respective digestive phase. For this reason, prolonged digestion protocols may require protease supplementation throughout digestion to correctly simulate physiological conditions.