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

Research output: Contribution to journalJournal articleResearchpeer-review

  • Myriam M.L. Grundy
  • Evan Abrahamse
  • Annette Almgren
  • Marie Alminger
  • Ana Andres
  • Renata M.C. Ariëns
  • Shanna Bastiaan-Net
  • Claire Bourlieu-Lacanal
  • André Brodkorb
  • Maria R. Bronze
  • Irene Comi
  • Leslie Couëdelo
  • Didier Dupont
  • Annie Durand
  • Sedef N. El
  • Tara Grauwet
  • Ana Heredia
  • Marcos R. Infantes Garcia
  • Christian Jungnickel
  • Ilona E. Kłosowska-Chomiczewska
  • Marion Létisse
  • Adam Macierzanka
  • Alan R. Mackie
  • David J. McClements
  • Olivia Menard
  • Anne Meynier
  • Marie Caroline Michalski
  • Ana Isabel Mulet-Cabero
  • Francina M. Payeras Perelló
  • Irene Peinado
  • Mélina Robert
  • Sébastien Secouard
  • Ana T. Serra
  • Sandra D. Silva
  • Gabriel Thomassen
  • Cecilia Tullberg
  • Ingrid Undeland
  • Carole Vaysse
  • Gerd E. Vegarud
  • Sarah H.E. Verkempinck
  • Michelle Viau
  • Mostafa Zahir
  • Ruojie Zhang
  • Frédéric Carrière

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

Original languageEnglish
Article number104497
JournalJournal of Functional Foods
Volume82
ISSN1756-4646
DOIs
Publication statusPublished - 2021

Bibliographical note

Publisher Copyright:
© 2021

    Research areas

  • Enzyme activity, INFOGEST, Inhibitor, Lipases, Lipolysis, Titration method

ID: 273634235