Journal of Functional Foods (Jul 2021)
INFOGEST inter-laboratory recommendations for assaying gastric and pancreatic lipases activities prior to in vitro digestion studies
- 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,
- Christine Heerup,
- 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,
- Anette Mullertz,
- 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
Affiliations
- Myriam M.L. Grundy
- PEGASE, INRAE, Institut Agro, 35590 Saint Gilles, France; Corresponding author at: PEGASE, INRAE, Institut Agro, 35590 Saint Gilles, France.
- Evan Abrahamse
- Danone Nutricia Research, 3584CT Utrecht, the Netherlands; Laboratory of Food Chemistry, Wageningen University, the Netherlands
- Annette Almgren
- Food and Nutrition Science, Department of Biology and Biological Engineering, Chalmers University of Technology, Kemivägen 10, SE-41296 Göteborg, Sweden
- Marie Alminger
- Food and Nutrition Science, Department of Biology and Biological Engineering, Chalmers University of Technology, Kemivägen 10, SE-41296 Göteborg, Sweden
- Ana Andres
- Research Institute of Food Engineering for Development, Universitat Politècnica de València, Camino de Vera s/n, 46022 Valencia, Spain
- Renata M.C. Ariëns
- Wageningen Food & Biobased Research, Wageningen University and Research, 6708 WG Wageningen, the Netherlands
- Shanna Bastiaan-Net
- Wageningen Food & Biobased Research, Wageningen University and Research, 6708 WG Wageningen, the Netherlands
- Claire Bourlieu-Lacanal
- STLO Science et Technologie du Lait et de l'oeuf, INRAE – Institut Agro, 35000 Rennes, France; INRAE, Montpellier SupAgro, CIRAD, Université de Montpellier, IATE, 2 place Pierre-Viala, bâtiment 33, F-34060 Montpellier, France
- André Brodkorb
- Department of Food Chemistry and Technology, Teagasc Food Research Centre, Moorepark, Fermoy, County Cork P61 C996, Ireland
- Maria R. Bronze
- iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2781-901 Oeiras, Portugal; Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República, 2780-157 Oeiras, Portugal; iMED, Faculdade de Farmácia da Universidade de Lisboa, 1649-003 Lisboa, Portugal
- Irene Comi
- Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences (NMBU), P.O. Box 5003, NO-1432 Ås, Norway
- Leslie Couëdelo
- ITERG, Université de Bordeaux, F-33076 Bordeaux, France
- Didier Dupont
- STLO Science et Technologie du Lait et de l'oeuf, INRAE – Institut Agro, 35000 Rennes, France
- Annie Durand
- CarMeN, U1060 INSERM INRA 1397 CARMEN, Hôpital Lyon Sud - Secteur 2, Bâtiment CENS ELI-2D, 165 Chemin du Grand Revoyet, F – 69310 Pierre Benite, France
- Sedef N. El
- Department of Food Engineering, Ege University, 35100 Izmir, Turkey
- Tara Grauwet
- Laboratory of Food Technology, Leuven Food Science and Nutrition Research Center (LFoRCe), Department of Microbial and Molecular Systems (M2S), KU Leuven, Kasteelpark Arenberg 22 box 2457, 3001 Heverlee, Belgium
- Christine Heerup
- Bioneer:FARMA, Department of Pharmacy, Faculty of Health and Medical Sciences, University of Copenhagen, 2100 Copenhagen, Denmark
- Ana Heredia
- Research Institute of Food Engineering for Development, Universitat Politècnica de València, Camino de Vera s/n, 46022 Valencia, Spain
- Marcos R. Infantes Garcia
- Laboratory of Food Technology, Leuven Food Science and Nutrition Research Center (LFoRCe), Department of Microbial and Molecular Systems (M2S), KU Leuven, Kasteelpark Arenberg 22 box 2457, 3001 Heverlee, Belgium
- Christian Jungnickel
- Department of Colloid and Lipid Science, Gdańsk University of Technology, 11/12 Gabriela Narutowicza Street 80-233 Gdańsk, Poland
- Ilona E. Kłosowska-Chomiczewska
- Department of Colloid and Lipid Science, Gdańsk University of Technology, 11/12 Gabriela Narutowicza Street 80-233 Gdańsk, Poland
- Marion Létisse
- CarMeN, U1060 INSERM INRA 1397 CARMEN, Hôpital Lyon Sud - Secteur 2, Bâtiment CENS ELI-2D, 165 Chemin du Grand Revoyet, F – 69310 Pierre Benite, France
- Adam Macierzanka
- Department of Colloid and Lipid Science, Gdańsk University of Technology, 11/12 Gabriela Narutowicza Street 80-233 Gdańsk, Poland
- Alan R. Mackie
- School of Food Science and Nutrition, University of Leeds, Leeds LS2 9JT, UK
- David J. McClements
- Biopolymers and Colloids Laboratory, Department of Food Science, University of Massachusetts, Amherst, MA 01003, USA
- Olivia Menard
- STLO Science et Technologie du Lait et de l'oeuf, INRAE – Institut Agro, 35000 Rennes, France
- Anne Meynier
- UR1268 BIA (Biopolymères Interactions Assemblages), INRAE, F-44300 Nantes, France
- Marie-Caroline Michalski
- CarMeN, U1060 INSERM INRA 1397 CARMEN, Hôpital Lyon Sud - Secteur 2, Bâtiment CENS ELI-2D, 165 Chemin du Grand Revoyet, F – 69310 Pierre Benite, France
- Ana-Isabel Mulet-Cabero
- Department of Food Chemistry and Technology, Teagasc Food Research Centre, Moorepark, Fermoy, County Cork P61 C996, Ireland; Food Innovation and Health, Quadram Institute Bioscience, Norwich Research Park, Colney, Norwich NR4 7UA, UK
- Anette Mullertz
- Bioneer:FARMA, Department of Pharmacy, Faculty of Health and Medical Sciences, University of Copenhagen, 2100 Copenhagen, Denmark
- Francina M. Payeras Perelló
- Inovation & Development Centre, Fresenius Kabi, 61352 Bad Homburg, Germany
- Irene Peinado
- Research Institute of Food Engineering for Development, Universitat Politècnica de València, Camino de Vera s/n, 46022 Valencia, Spain
- Mélina Robert
- INRAE, Montpellier SupAgro, CIRAD, Université de Montpellier, IATE, 2 place Pierre-Viala, bâtiment 33, F-34060 Montpellier, France
- Sébastien Secouard
- Inovation & Development Centre, Fresenius Kabi, 61352 Bad Homburg, Germany
- Ana T. Serra
- iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2781-901 Oeiras, Portugal; Instituto de Tecnologia Química e Biológica António Xavier, Universidade Nova de Lisboa, Av. da República, 2780-157 Oeiras, Portugal
- Sandra D. Silva
- iBET, Instituto de Biologia Experimental e Tecnológica, Apartado 12, 2781-901 Oeiras, Portugal
- Gabriel Thomassen
- Laboratory of Food Chemistry, Wageningen University, the Netherlands
- Cecilia Tullberg
- Food and Nutrition Science, Department of Biology and Biological Engineering, Chalmers University of Technology, Kemivägen 10, SE-41296 Göteborg, Sweden
- Ingrid Undeland
- Food and Nutrition Science, Department of Biology and Biological Engineering, Chalmers University of Technology, Kemivägen 10, SE-41296 Göteborg, Sweden
- Carole Vaysse
- ITERG, Université de Bordeaux, F-33076 Bordeaux, France
- Gerd E. Vegarud
- Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences (NMBU), P.O. Box 5003, NO-1432 Ås, Norway
- Sarah H.E. Verkempinck
- Laboratory of Food Technology, Leuven Food Science and Nutrition Research Center (LFoRCe), Department of Microbial and Molecular Systems (M2S), KU Leuven, Kasteelpark Arenberg 22 box 2457, 3001 Heverlee, Belgium
- Michelle Viau
- UR1268 BIA (Biopolymères Interactions Assemblages), INRAE, F-44300 Nantes, France
- Mostafa Zahir
- Food Quality and Design, Wageningen University and Research Centre, 6708 WG Wageningen, the Netherlands
- Ruojie Zhang
- Biopolymers and Colloids Laboratory, Department of Food Science, University of Massachusetts, Amherst, MA 01003, USA
- Frédéric Carrière
- Aix-Marseille Université, CNRS, UMR 7282, 31 chemin Joseph Aiguier, F-13402 Marseille Cedex 9, France
- Journal volume & issue
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Vol. 82
p. 104497
Abstract
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.
