INFOGEST inter-laboratory recommendations for assaying gastric and pancreatic lipases activities prior to in vitro digestion studies

In vitro digestion studies often use animal digestive enzyme extracts as substitutes of human gastric and pancreatic secretions. Pancreatin from porcine origin is thus commonly used to provide relevant enzymes such proteases, amylase lipase. Rabbit (RGE) have been recently introduced lipase in addition pepsin. Before preparing simulated with targeted activities described protocols, it important determine the preparations using validated methods. The purpose this inter-laboratory study within INFOGEST network was test repeatability reproducibility assays pH-stat technique for measuring lipases various sources. Twenty-one laboratories having different devices received same protocol identical batches RGE two pancreatin Lipase were performed tributyrin a substrate three amounts (50, 100 200 µg) each preparation. results individual satisfactory coefficients variation (CVs) ranging 4 8% regardless amount tested. However, variability high (CV > 15%) compared existing standards bioanalytical assays. We identified weighted contributions several parameters associated equipment (e.g. reaction vessel volume shape, stirring mode rate, burette automated delivery sodium hydroxide). Based on this, we established recommendations improving technique. Defining accurate complete how correctly quantify activity levels gateway promising comparison data obtained following protocol.