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INFOGEST static in vitro simulation of gastrointestinal food digestion

Nature Protocols volume 14, pages 991–1014 (2019)Cite this article

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Abstract

Developing a mechanistic understanding of the impact of food structure and composition on human health has increasingly involved simulating digestion in the upper gastrointestinal tract. These simulations have used a wide range of different conditions that often have very little physiological relevance, and this impedes the meaningful comparison of results. The standardized protocol presented here is based on an international consensus developed by the COST INFOGEST network. The method is designed to be used with standard laboratory equipment and requires limited experience to encourage a wide range of researchers to adopt it. It is a static digestion method that uses constant ratios of meal to digestive fluids and a constant pH for each step of digestion. This makes the method simple to use but not suitable for simulating digestion kinetics. Using this method, food samples are subjected to sequential oral, gastric and intestinal digestion while parameters such as electrolytes, enzymes, bile, dilution, pH and time of digestion are based on available physiological data. This amended and improved digestion method (INFOGEST 2.0) avoids challenges associated with the original method, such as the inclusion of the oral phase and the use of gastric lipase. The method can be used to assess the endpoints resulting from digestion of foods by analyzing the digestion products (e.g., peptides/amino acids, fatty acids, simple sugars) and evaluating the release of micronutrients from the food matrix. The whole protocol can be completed in ~7 d, including ~5 d required for the determination of enzyme activities.

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Fig. 1: Flow diagram of the INFOGEST 2.0 digestion method.
Fig. 2: Protein separation by gel electrophoresis of in vitro–digested skim milk powder.
Fig. 3: Comparison of in vitro–digested skim milk powder peptide patterns of κ-casein with in vivo (pig) digestion.

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Acknowledgements

We acknowledge COST action FA1005 INFOGEST26 (http://www.cost-infogest.eu/) for providing funding for travel, meetings and conferences (2011–2015). We acknowledge the French National Institute for Agricultural Research (INRA, http://www.inra.fr) for its continuous support of the INFOGEST network by organizing and co-funding the International Conference on Food Digestion and workgroup meetings. We thank A. G. F. Lopes (Universidade de Lisboa, Portugal) and V. S. N. Mishra (Teagasc Food Research Centre, Moorepark, Ireland) for their help in the final preparation of the videos. We also acknowledge the many other researchers, mostly associated with the above COST action and subsequent events, who have contributed to the discussion of digestion parameters.

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Authors and Affiliations

  1. Teagasc Food Research Centre, Moorepark, Fermoy, Ireland

    André Brodkorb

  2. Agroscope, Bern, Switzerland

    Lotti Egger & Reto Portmann

  3. Department of Biology and Biological Engineering, Chalmers University of Technology, Gothenburg, Sweden

    Marie Alminger

  4. National Institute of Health Doutor Ricardo Jorge, Lisbon and CESAM, University of Aveiro, Aveiro, Portugal

    Paula Alvito, Ricardo Assunção & Carla Martins

  5. Nofima AS, Ås, Norway

    Simon Ballance & Bente Kirkhus

  6. Luxembourg Institute of Health, Strassen, Luxembourg

    Torsten Bohn

  7. INRA/Montpellier SupAgro, Montpellier, France

    Claire Bourlieu-Lacanal

  8. STLO, INRA, Agrocampus Ouest, Rennes, France

    Rachel Boutrou, Didier Dupont, Steven Le Feunteun & Olivia Ménard

  9. Aix-Marseille, CNRS, UMR7281 Bioénergétique et Ingénierie des Protéines, Marseille, France

    Frédéric Carrière

  10. Estación Experimental del Zaidin, Consejo Superior de Investigaciones Científicas (CSIC), Granada, Spain

    Alfonso Clemente

  11. Department of Food Science, Aarhus University, Tjele, Denmark

    Milena Corredig

  12. SQPOV, INRA, Avignon, France

    Claire Dufour

  13. Quadram Institute Bioscience, Norwich, UK

    Cathrina Edwards

  14. Riddet Institute, Massey University, Palmerston North, New Zealand

    Matt Golding

  15. Department of Food Engineering, Faculty of Engineering, Ege University, Izmir, Turkey

    Sibel Karakaya

  16. Israel Institute of Technology, Technion City, Haifa, Israel

    Uri Lesmes

  17. Faculty of Chemistry, Gdansk University of Technology, Gdansk, Poland

    Adam Macierzanka

  18. School of Food Science & Nutrition, University of Leeds, Leeds, UK

    Alan R. Mackie

  19. BIA, INRA, Nantes, France

    Sébastien Marze

  20. Chenoweth Lab, Department of Food Science, University of Massachusetts, Amherst, MA, USA

    David Julian McClements

  21. Triskelion, Zeist, the Netherlands

    Mans Minekus

  22. Instituto de Biologia Experimental e Tecnológica, Oeiras, Portugal

    Cláudia N. Santos

  23. Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Oeiras, Portugal

    Cláudia N. Santos

  24. GMPA, AgroParisTech, INRA, Université Paris-Saclay, Thiverval-Grignon, France

    Isabelle Souchon

  25. Departments of Biological and Agricultural Engineering, and of Food Science and Technology, University of California, Davis, Davis, CA, USA

    R. Paul Singh

  26. Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, Aas, Norway

    Gerd E. Vegarud

  27. Reacta Biotech Limited, The Langley Building, Manchester, UK

    Martin S. J. Wickham

  28. Ernst Moritz Arndt University of Greifswald, Greifswald, Germany

    Werner Weitschies

  29. Instituto de Investigación en Ciencias de la Alimentación (CIAL, CSIC-UAM), Madrid, Spain

    Isidra Recio

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  1. André Brodkorb
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Contributions

A.B., L.E. and I.R. wrote the manuscript. M.A., S.B., T.B., F.C., A.C., D.D., C.D., C.E., S.L.F., U.L., A.M., A.R.M., O.M., M.M., R.P., C.N.S. and I.S. contributed to the writing of the manuscript. A.B., L.E., M.A., P.A., S.B., T.B., C.B.-L, R.B., F.C., A.C., M.C., D.D., C.D., C.E., M.G., S.K., B.K., S.L.F., U.L., A.M., A.R.M., S.M., O.M., M.M., R.P., C.N.S., I.S., G.E.V., M.S.J.W., W.W. and I.R. contributed to the definition of digestion parameters. R.P. wrote the online tools. R.A. and C.M. prepared the videos. M.G., D.J.M. and R.P.S. contributed to the manuscript by critical revision of the digestion parameters.

Corresponding author

Correspondence to André Brodkorb.

Ethics declarations

Competing interests

Rabbit lipase from rabbit gastric extract is available commercially from Lipolytech, a start-up company founded by a researcher who had previously worked in F. Carrière’s (coauthor of this paper) group. The F. Carrière laboratory, a joint unit of the Centre National de la Recherche Scientifique (CNRS) and Aix Marseille University (AMU), has a research collaboration contract with Lipolytech (CNRS reference no. 163451; signed on 30 June 2017). However, the coauthor F. Carrière does not financially benefit from this contract and, as an employee of CNRS and civil servant of the French state, is not allowed to have private consulting activity for a company contracting with his own laboratory.

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Related links

Key references using this protocol

Egger, L. et al. Food Res. Int. 88, 217–225 (2016): https://doi.org/10.1016/j.foodres.2015.12.006

Egger, L. et al. Food Res. Int. 102, 567–574 (2017): https://doi.org/10.1016/j.foodres.2017.09.047

Sanchón, J. et al. Food Chem. 239, 486–494 (2018): https://doi.org/10.1016/j.foodchem.2017.06.134

Integrated supplementary information

Supplementary Figure 1 Oral bolus hydration in vivo.

Bolus hydration (g of saliva/g of foods) in vivo just before swallowing, for various foods based on published data116–123. Supplementary Methods (enzyme activity assays) were adapted from Minekus et al.28 under a Creative Commons Attribution 3.0 license (https://creativecommons.org/licenses/by/3.0/legalcode).

Supplementary information

Supplementary Text and Figures

Supplementary Figure 1 and Supplementary Methods

Supplementary Data 1

Excel spreadsheets for calculating the enzyme activities of all digestive enzymes.

Supplementary Data 2

Excel spreadsheets for calculating all volumes of simulated digestive fluids, enzyme and bile solutions on the basis of the initial amount of digested food.

Supplementary Video 1

INFOGEST 2.0 digestion procedure part 1.

Supplementary Video 2

INFOGEST 2.0 digestion procedure part 2.

Supplementary Video 3

Amylase activity assay.

Supplementary Video 4

Pepsin activity assay.

Supplementary Video 5

Lipase activity assay (both gastric and pancreatic).

Supplementary Video 6

Trypsin activity assay.

Supplementary Video 7

Chymotrypsin activity assay.

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Brodkorb, A., Egger, L., Alminger, M. et al. INFOGEST static in vitro simulation of gastrointestinal food digestion. Nat Protoc 14, 991–1014 (2019). https://doi.org/10.1038/s41596-018-0119-1

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  • DOI: https://doi.org/10.1038/s41596-018-0119-1

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