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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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References

  1. WHO. Global Health Observatory—World Health Statistics 172 pp (WHO, Geneva, 2013).

  2. Sullivan, L. M. et al. Gastric digestion of α-lactalbumin in adult human subjects using capsule endoscopy and nasogastric tube sampling. Br. J. Nutr. 112, 638–646 (2014).

    Article  CAS  PubMed  Google Scholar 

  3. Boutrou, R. et al. Sequential release of milk protein–derived bioactive peptides in the jejunum in healthy humans. Am. J. Clin. Nutr. 97, 1314–1323 (2013).

    Article  CAS  PubMed  Google Scholar 

  4. Mackie, A. R., Rafiee, H., Malcolm, P., Salt, L. & van Aken, G. Specific food structures supress appetite through reduced gastric emptying rate. Am. J. Physiol. Gastrointest. Liver Physiol. 304, G1038–G1043 (2013).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Koziolek, M. et al. Intragastric pH and pressure profiles after intake of the high-caloric, high-fat meal as used for food effect studies. J. Control. Release 220, 71–78 (2015).

    Article  CAS  PubMed  Google Scholar 

  6. Minekus, M., Marteau, P., Havenaar, R. & Huis In’t Veld, J. H. J. A multicompartmental dynamic computer-controlled model simulating the stomach and small intestine. Altern. Lab. Anim. 23, 197–209 (1995).

    Google Scholar 

  7. Wickham, M., Faulks, R. & Mills, C. In vitro digestion methods for assessing the effect of food structure on allergen breakdown. Mol. Nutr. Food Res. 53, 952–958 (2009).

    Article  CAS  PubMed  Google Scholar 

  8. Ménard, O. et al. Validation of a new in vitro dynamic system to simulate infant digestion. Food Chem. 145, 1039–1045 (2014).

    Article  PubMed  CAS  Google Scholar 

  9. Molly, K., Woestyne, M. V. & Verstraete, W. Development of a 5-step multi-chamber reactor as a simulation of the human intestinal microbial ecosystem. Appl. Microbiol. Biotechnol. 39, 254–258 (1993).

    Article  CAS  PubMed  Google Scholar 

  10. Kong, F. & Singh, R. P. A human gastric simulator (HGS) to study food digestion in human stomach. J. Food Sci. 75, E627–E635 (2010).

    Article  CAS  PubMed  Google Scholar 

  11. Dupont, D. et al. Can dynamic in vitro digestion systems mimic the physiological reality? Crit. Rev. Food Sci. Nutr. 1-17, https://doi.org/10.1080/10408398.2017.1421900 (2018).

  12. Kaukonen, A. M., Boyd, B. J., Charman, W. N. & Porter, C. J. Drug solubilization behavior during in vitro digestion of suspension formulations of poorly water-soluble drugs in triglyceride lipids. Pharm. Res. 21, 254–260 (2004).

    Article  CAS  PubMed  Google Scholar 

  13. Maldonado-Valderrama, J., Gunning, A. P., Wilde, P. J. & Morris, V. J. In vitro gastric digestion of interfacial protein structures: visualisation by AFM. Soft Matter 6, 4908–4915 (2010).

    Article  CAS  Google Scholar 

  14. Boisen, S. & Fernández, J. A. Prediction of the total tract digestibility of energy in feedstuffs and pig diets by in vitro analyses. Anim. Feed Sci. Technol. 68, 277–286 (1997).

    Article  Google Scholar 

  15. Bohn, T. et al. Correlation between in vitro and in vivo data on food digestion. What can we predict with static in vitro digestion models? Crit. Rev. Food Sci. Nutr. 58, 2239–2261 (2017).

  16. Sanchón, J. et al. Protein degradation and peptide release from milk proteins in human jejunum. Comparison with in vitro gastrointestinal simulation. Food Chem. 239, 486–494 (2018).

    Article  PubMed  CAS  Google Scholar 

  17. The United States Pharmacopeial Convention Inc. The United States Pharmacopeia 26, The National Formulary 21 (USP 26/NF21) (The United States Pharmacopeial Convention Inc., Rockville, MD, 2003).

    Google Scholar 

  18. McCarthy, C. A. et al. In vitro dissolution models for the prediction of in vivo performance of an oral mesoporous silica formulation. J. Control. Release 250, 86–95 (2017).

    Article  CAS  PubMed  Google Scholar 

  19. Griffin, B. T. et al. Comparison of in vitro tests at various levels of complexity for the prediction of in vivo performance of lipid-based formulations: case studies with fenofibrate. Eur. J. Pharm. Biopharm. 86, 427–437 (2014).

    Article  CAS  PubMed  Google Scholar 

  20. Oomen, A. G. et al. Development of an in vitro digestion model for estimating the bioaccessibility of soil contaminants. Arch. Environ. Contam. Toxicol. 44, 0281–0287 (2003).

    Article  CAS  Google Scholar 

  21. Versantvoort, C. H. M., Oomen, A. G., Van de Kamp, E., Rompelberg, C. J. M. & Sips, A. J. A. M. Applicability of an in vitro digestion model in assessing the bioaccessibility of mycotoxins from food. Food Chem. Toxicol. 43, 31–40 (2005).

    Article  CAS  PubMed  Google Scholar 

  22. Wragg, J. et al. Inter-Laboratory Trial of a Unified Bioaccessibility Testing Procedure. Chemical & Biological Hazards Programme. Open Report OR/07/027 (2009); http://nora.nerc.ac.uk/id/eprint/7491/1/OR07027.pdf

  23. Dressman, J. B. et al. Upper gastrointestinal (GI) pH in young, healthy men and women. Pharm. Res. 7, 756–761 (1990).

    Article  CAS  PubMed  Google Scholar 

  24. Lentner, C. Geigy Scientific Tables. Vol. 1, Units of Measurement, Body Fluids, Composition of the Body, Nutrition 8th edn (Ciba-Geigy, Basel, Switzerland, 1981). .

  25. Hur, S. J., Lim, B. O., Decker, E. A. & McClements, D. J. In vitro human digestion models for food applications. Food Chem. 125, 1–12 (2011).

    Article  CAS  Google Scholar 

  26. Dupont, D. et al. An international network for improving health properties of food by sharing our knowledge on the digestive process. Food Digestion 2, 23–25 (2011).

    Article  Google Scholar 

  27. Minekus, M. et al. A standardised static in vitro digestion method suitable for food—an international consensus. Food Funct. 5, 1113–1124 (2014).

    Article  CAS  PubMed  Google Scholar 

  28. Egger, L. et al. The harmonized INFOGEST in vitro digestion method: from knowledge to action. Food Res. Int. 88, 217–225 (2016).

    Article  CAS  Google Scholar 

  29. Egger, L. et al. Physiological comparability of the harmonized INFOGEST in vitro digestion method to in vivo pig digestion. Food Res. Int. 102, 567–574 (2017).

    Article  CAS  PubMed  Google Scholar 

  30. Hempel, J. et al. Ultrastructural deposition forms and bioaccessibility of carotenoids and carotenoid esters from goji berries (Lycium barbarum L.). Food Chem. 218, 525–533 (2017).

    Article  CAS  PubMed  Google Scholar 

  31. Rodrigues, D. B., Mariutti, L. R. B. & Mercadante, A. Z. An in vitro digestion method adapted for carotenoids and carotenoid esters: moving forward towards standardization. Food Funct. 7, 4992–5001 (2016).

    Article  CAS  PubMed  Google Scholar 

  32. Bot, F. et al. The effect of pulsed electric fields on carotenoids bioaccessibility: the role of tomato matrix. Food Chem. 240, 415–421 (2018).

    Article  CAS  PubMed  Google Scholar 

  33. Gomez-Mascaraque, L. G., Perez-Masia, R., Gonzalez-Barrio, R., Periago, M. J. & Lopez-Rubio, A. Potential of microencapsulation through emulsion-electrospraying to improve the bioaccesibility of beta-carotene. Food Hydrocoll. 73, 1–12 (2017).

    Article  CAS  Google Scholar 

  34. Davidov-Pardo, G., Perez-Ciordia, S., Marin-Arroyo, M. R. & McClements, D. J. Improving resveratrol bioaccessibility using biopolymer nanoparticles and complexes: impact of protein–carbohydrate maillard conjugation. J. Agric. Food Chem. 63, 3915–3923 (2015).

    Article  CAS  PubMed  Google Scholar 

  35. Ferreira-Lazarte, A. et al. Study on the digestion of milk with prebiotic carbohydrates in a simulated gastrointestinal model. J. Funct. Foods 33, 149–154 (2017).

    Article  CAS  Google Scholar 

  36. El, S. N. et al. In vitro digestibility of goat milk and kefir with a new standardised static digestion method (INFOGEST cost action) and bioactivities of the resultant peptides. Food Funct. 6, 2322–2330 (2015).

    Article  CAS  Google Scholar 

  37. Wang, B., Timilsena, Y. P., Blanch, E. & Adhikari, B. Mild thermal treatment and in-vitro digestion of three forms of bovine lactoferrin: effects on functional properties. Int. Dairy J. 64, 22–30 (2017).

    Article  CAS  Google Scholar 

  38. Naegeli, H. et al. Guidance on allergenicity assessment of genetically modified plants. EFSA J. 15, e04862 (2017).

  39. Mamone, G. et al. Tracking the fate of pasta (T. durum semolina) immunogenic proteins by in vitro simulated digestion. J. Agric. Food. Chem. 63, 2660–2667 (2015).

    Article  CAS  PubMed  Google Scholar 

  40. Korte, R., Bracker, J. & Brockmeyer, J. Gastrointestinal digestion of hazelnut allergens on molecular level: elucidation of degradation kinetics and resistant immunoactive peptides using mass spectrometry. Mol. Nutr. Food Res. 61, 1700130 (2017).

  41. Di Stasio, L. et al. Peanut digestome: identification of digestion resistant IgE binding peptides. Food Chem. Toxicol. 107, 88–98 (2017).

    Article  PubMed  CAS  Google Scholar 

  42. Mat, D. J. L., Le Feunteun, S., Michon, C. & Souchon, I. In vitro digestion of foods using pH-stat and the INFOGEST protocol: Impact of matrix structure on digestion kinetics of macronutrients, proteins and lipids. Food Res. Int. 88, Part B. 226–233 (2016).

  43. Floury, J. et al. Exploring the breakdown of dairy protein gels during in vitro gastric digestion using time-lapse synchrotron deep-UV fluorescence microscopy. Food Chem. 239, 898–910 (2018).

    Article  CAS  PubMed  Google Scholar 

  44. Sarkar, A. et al. In vitro digestion of Pickering emulsions stabilized by soft whey protein microgel particles: influence of thermal treatment. Soft Matter 12, 3558–3569 (2016).

    Article  CAS  PubMed  Google Scholar 

  45. Fernandez-Avila, C., Arranz, E., Guri, A., Trujillo, A. & Corredig, M. Vegetable protein isolate-stabilized emulsions for enhanced delivery of conjugated linoleic acid in Caco-2 cells. Food Hydrocoll. 55, 144–154 (2016).

    Article  CAS  Google Scholar 

  46. Yang, J., Primo, C., Elbaz-Younes, I. & Hirschi, K. D. Bioavailability of transgenic microRNAs in genetically modified plants. Genes Nutr. 12, 17 (2017).

  47. Aschoff, J. K. et al. Bioavailability of beta-cryptoxanthin is greater from pasteurized orange juice than from fresh oranges—a randomized cross-over study. Mol. Nutr. Food Res. 59, 1896–1904 (2015).

    Article  CAS  PubMed  Google Scholar 

  48. Garrett, D. A., Failla, M. L. & Sarama, R. J. Development of an in vitro digestion method to assess carotenoid bioavailability from meals. J. Agric. Food Chem. 47, 4301–4309 (1999).

    Article  CAS  PubMed  Google Scholar 

  49. Mulet-Cabero, A. I., Rigby, N. M., Brodkorb, A. & Mackie, A. R. Dairy food structures influence the rates of nutrient digestion through different in vitro gastric behaviour. Food Hydrocoll. 67, 63–73 (2017).

    Article  CAS  Google Scholar 

  50. Mulet-Cabero, A. I., Mackie, A., Wilde, P., Fenelon, M. A. & Brodkorb, A. Structural mechanism and kinetics of in vitro gastric digestion are affected by process-induced changes in bovine milk. Food Hydrocoll. 86, 172–183 (2019).

    Article  CAS  Google Scholar 

  51. Roura, E. et al. Critical review evaluating the pig as a model for human nutritional physiology. Nutr. Res. Rev. 29, 60–90 (2016).

    Article  CAS  PubMed  Google Scholar 

  52. Le Huërou-Luron, I. et al. A mixture of milk and vegetable lipids in infant formula changes gut digestion, mucosal immunity and microbiota composition in neonatal piglets. Eur. J. Nutr. 57, 463–476 (2018).

    Article  PubMed  CAS  Google Scholar 

  53. Barbé, F. et al. The heat treatment and the gelation are strong determinants of the kinetics of milk proteins digestion and of the peripheral availability of amino acids. Food Chem. 136, 1203–1212 (2013).

    Article  PubMed  CAS  Google Scholar 

  54. Evenepoel, P. et al. Digestibility of cooked and raw egg protein in humans as assessed by stable isotope techniques. J. Nutr. 128, 1716–1722 (1998).

    Article  CAS  PubMed  Google Scholar 

  55. Normén, L. et al. Phytosterol and phytostanol esters are effectively hydrolysed in the gut and do not affect fat digestion in ileostomy subjects. Eur. J. Nutr. 45, 165–170 (2006).

    Article  PubMed  CAS  Google Scholar 

  56. Edwards, C. H. et al. Manipulation of starch bioaccessibility in wheat endosperm to regulate starch digestion, postprandial glycemia, insulinemia, and gut hormone responses: a randomized controlled trial in healthy ileostomy participants. Am. J. Clin. Nutr. 102, 791–800 (2015).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  57. Bakala N’Goma, J. C., Amara, S., Dridi, K., Jannin, V. & Carriere, F. Understanding the lipid-digestion processes in the GI tract before designing lipid-based drug-delivery systems. Ther. Deliv. 3, 105–124 (2012).

    Article  PubMed  CAS  Google Scholar 

  58. Gargouri, Y. et al. Importance of human gastric lipase for intestinal lipolysis: an in vitro study. Biochim. Biophys. Acta 879, 419–423 (1986).

    Article  CAS  PubMed  Google Scholar 

  59. Ville, E., Carriere, F., Renou, C. & Laugier, R. Physiological study of pH stability and sensitivity to pepsin of human gastric lipase. Digestion 65, 73–81 (2002).

    Article  CAS  PubMed  Google Scholar 

  60. Carrière, F., Barrowman, J. A., Verger, R. & Laugier, R. Secretion and contribution to lipolysis of gastric and pancreatic lipases during a test meal in humans. Gastroenterology 105, 876–888 (1993).

    Article  PubMed  Google Scholar 

  61. Sams, L., Paume, J., Giallo, J. & Carriere, F. Relevant pH and lipase for in vitro models of gastric digestion. Food Funct. 7, 30–45 (2016).

    Article  CAS  PubMed  Google Scholar 

  62. Carrière, F. et al. The specific activities of human digestive lipases measured from the in vivo and in vitro lipolysis of test meals. Gastroenterology 119, 949–960 (2000).

    Article  PubMed  Google Scholar 

  63. Bakala-N’Goma, J. C. et al. Toward the establishment of standardized in vitro tests for lipid-based formulations. 5. Lipolysis of representative formulations by gastric lipase. Pharm. Res. 32, 1279–1287 (2015).

    Article  PubMed  CAS  Google Scholar 

  64. Capolino, P. et al. In vitro gastrointestinal lipolysis: replacement of human digestive lipases by a combination of rabbit gastric and porcine pancreatic extracts. Food Digestion 2, 43–51 (2011).

    Article  CAS  Google Scholar 

  65. Moreau, H., Gargouri, Y., Lecat, D., Junien, J.-L. & Verger, R. Screening of preduodenal lipases in several mammals. Biochim. Biophys. Acta 959, 247–252 (1988).

    Article  CAS  PubMed  Google Scholar 

  66. De Caro, J., Ferrato, F., Verger, R. & de Caro, A. Purification and molecular characterization of lamb pregastric lipase. Biochim. Biophys. Acta 1252, 321–329 (1995).

    Article  PubMed  Google Scholar 

  67. Sams, L. et al. Characterization of pepsin from rabbit gastric extract, its action on β-casein and the effects of lipids on proteolysis. Food Funct. 9, 5975–5988 (2018).

    Article  CAS  PubMed  Google Scholar 

  68. Rich, D. H. et al. Inhibition of aspartic proteases by pepstatin and 3-methylstatine derivatives of pepstatin. Evidence for collected-substrate enzyme inhibition. Biochemistry 24, 3165–3173 (1985).

  69. Williams, H. D. et al. Toward the establishment of standardized in vitro tests for lipid-based formulations, part 1: method parameterization and comparison of in vitro digestion profiles across a range of representative formulations. J. Pharm. Sci. 101, 3360–3380 (2012).

  70. Mat, D. J. L., Cattenoz, T., Souchon, I., Michon, C. & Le Feunteun, S. Monitoring protein hydrolysis by pepsin using pH-stat: in vitro gastric digestions in static and dynamic pH conditions. Food Chem. 239, 268–275 (2018).

    Article  CAS  PubMed  Google Scholar 

  71. Gargouri, Y. et al. Kinetic assay of human gastric lipase on short- and long-chain triacylglycerol emulsions. Gastroenterology 91, 919–925 (1986).

    Article  CAS  PubMed  Google Scholar 

  72. Moreau, H., Gargouri, Y., Lecat, D., Junien, J.-L. & Verger, R. Purification, characterization and kinetic properties of the rabbit gastric lipase. Biochim. Biophys. Acta 960, 286–293 (1988).

    Article  CAS  PubMed  Google Scholar 

  73. Ménard, O. et al. A first step towards a consensus static in vitro model for simulating full-term infant digestion. Food Chem. 240, 338–345 (2018).

    Article  PubMed  CAS  Google Scholar 

  74. Lecomte, M. et al. Milk polar lipids affect in vitro digestive lipolysis and postprandial lipid metabolism in mice. J. Nutr. 145, 1770–1777 (2015).

    Article  CAS  PubMed  Google Scholar 

  75. Grundy, M. M. L. et al. The impact of oat structure and beta-glucan on in vitro lipid digestion. J. Funct. Foods 38, 378–388 (2017).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  76. Salvia-Trujillo, L. et al. Lipid digestion, micelle formation and carotenoid bioaccessibility kinetics: influence of emulsion droplet size. Food Chem. 229, 653–662 (2017).

    Article  CAS  PubMed  Google Scholar 

  77. Bligh, E. G. & Dyer, W. J. A rapid method of total lipid extraction and purification. Can. J. Biochem. Physiol. 37, 911–917 (1959).

    Article  CAS  PubMed  Google Scholar 

  78. Cavalier, J.-F. et al. Validation of lipolysis product extraction from aqueous/biological samples, separation and quantification by thin-layer chromatography with flame ionization detection analysis using O-cholesteryl ethylene glycol as a new internal standard. J. Chromatogr. A 1216, 6543–6548 (2009).

    Article  CAS  PubMed  Google Scholar 

  79. Carriere, F. et al. Purification and biochemical characterization of dog gastric lipase. FEBS J. 202, 75–83 (1991).

    CAS  Google Scholar 

  80. Bourlieu, C. et al. The structure of infant formulas impacts their lipolysis, proteolysis and disintegration during in vitro gastric digestion. Food Chem. 182, 224–235 (2015).

    Article  CAS  PubMed  Google Scholar 

  81. Chatzidaki, M. D., Mateos-Diaz, E., Leal-Calderon, F., Xenakis, A. & Carriere, F. Water-in-oil microemulsions versus emulsions as carriers of hydroxytyrosol: an in vitro gastrointestinal lipolysis study using the pHstat technique. Food Funct. 7, 2258–2269 (2016).

    Article  CAS  PubMed  Google Scholar 

  82. Tyssandier, V. et al. Processing of vegetable-borne carotenoids in the human stomach and duodenum. Am. J. Physiol. Gastrointest. Liver Physiol. 284, G913–G923 (2003).

    Article  CAS  PubMed  Google Scholar 

  83. Reboul, E. et al. Bioaccessibility of carotenoids and vitamin E from their main dietary sources. J. Agric. Food Chem. 54, 8749–8755 (2006).

    Article  CAS  PubMed  Google Scholar 

  84. Biehler, E., Kaulmann, A., Hoffmann, L., Krause, E. & Bohn, T. Dietary and host-related factors influencing carotenoid bioaccessibility from spinach (Spinacia oleracea). Food Chem. 125, 1328–1334 (2011).

    Article  CAS  Google Scholar 

  85. Boon, C. S., McClements, D. J., Weiss, J. & Decker, E. A. Factors influencing the chemical stability of carotenoids in foods. Crit. Rev. Food Sci. Nutr. 50, 515–532 (2010).

    Article  CAS  PubMed  Google Scholar 

  86. Jorgensen, E. M., Marin, A. B. & Kennedy, J. A. Analysis of the oxidative degradation of proanthocyanidins under basic conditions. J. Agric. Food. Chem. 52, 2292–2296 (2004).

    Article  CAS  PubMed  Google Scholar 

  87. Talcott, S. T. & Howard, L. R. Phenolic autoxidation is responsible for color degradation in processed carrot puree. J. Agric. Food. Chem. 47, 2109–2115 (1999).

    Article  CAS  PubMed  Google Scholar 

  88. Bermúdez-Soto, M. J., Tomás-Barberán, F. A. & García-Conesa, M. T. Stability of polyphenols in chokeberry (Aronia melanocarpa) subjected to in vitro gastric and pancreatic digestion. Food Chem. 102, 865–874 (2007).

    Article  CAS  Google Scholar 

  89. Alminger, M. et al. In vitro models for studying secondary plant metabolite digestion and bioaccessibility. Compr. Rev. Food Sci. Food Saf. 13, 413–436 (2014).

    Article  CAS  PubMed  Google Scholar 

  90. Bohn, T. et al. Mind the gap-deficits in our knowledge of aspects impacting the bioavailability of phytochemicals and their metabolites—a position paper focusing on carotenoids and polyphenols. Mol. Nutr. Food Res. 59, 1307–1323 (2015).

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  91. Amiri, M. & Naim, H. Y. Characterization of mucosal disaccharidases from human intestine. Nutrients 9, 1106 (2017).

  92. Bouayed, J., Deusser, H., Hoffmann, L. & Bohn, T. Bioaccessible and dialysable polyphenols in selected apple varieties following in vitro digestion vs. their native patterns. Food Chem. 131, 1466–1472 (2012).

    Article  CAS  Google Scholar 

  93. Coates, E. M. et al. Colon-available raspberry polyphenols exhibit anti-cancer effects on in vitro models of colon cancer. J. Carcinog. 6, 4 (2007).

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  94. Figueira, I. et al. Blood–brain barrier transport and neuroprotective potential of blackberry-digested polyphenols: an in vitro study. Eur. J. Nutr. https://doi.org/10.1007/s00394-017-1576-y (2017)

  95. Garcia, G. et al. Bioaccessible (poly)phenol metabolites from raspberry protect neural cells from oxidative stress and attenuate microglia activation. Food Chem. 215, 274–283 (2017).

    Article  CAS  PubMed  Google Scholar 

  96. Bohn, T. Bioactivity of carotenoids—chasms of knowledge. Int. J. Vitam. Nutr. Res. 10, 1–5 (2016).

    Google Scholar 

  97. Levi, C. S. et al. Extending in vitro digestion models to specific human populations: Perspectives, practical tools and bio-relevant information. Trends Food Sci. Technol. 60, 52–63 (2017).

    Article  CAS  Google Scholar 

  98. Picariello, G. et al. Peptides surviving the simulated gastrointestinal digestion of milk proteins: Biological and toxicological implications. J. Chromatogr. B Analyt. Technol. Biomed. Life Sci. 878, 295–308 (2010).

    Article  CAS  PubMed  Google Scholar 

  99. Garcia-Campayo, V., Han, S., Vercauteren, R. & Franck, A. Digestion of food ingredients and food using an in vitro model integrating intestinal mucosal enzymes. Food Nutr. Sci. 9, 711–734 (2018).

    CAS  Google Scholar 

  100. Picariello, G., Ferranti, P. & Addeo, F. Use of brush border membrane vesicles to simulate the human intestinal digestion. Food Res. Int. 88, Part B. 327–335 (2016).

  101. Cheeseman, C. I. & O’Neill, D. Isolation of intestinal brush-border membranes. Curr. Protoc. Cell Biol. 30, 3.21.1–3.21.10 (2006).

  102. Lin, X. J. & Wright, A. J. Pectin and gastric pH interactively affect DHA-rich emulsion in vitro digestion microstructure, digestibility and bioaccessibility. Food Hydrocoll. 76, 49–59 (2018).

    Article  CAS  Google Scholar 

  103. Lorieau, L. et al. Impact of the dairy product structure and protein nature on the proteolysis and amino acid bioaccessiblity during in vitro digestion. Food Hydrocoll. 82, 399–411 (2018).

    Article  CAS  Google Scholar 

  104. Macierzanka, A., Sancho, A., Mills, E. N. C., Rigby, N. & Mackie, A. Emulsification alters simulated gastrointestinal proteolysis of β-casein and β-lactoglobulin. Soft Matter 5, 538–550 (2009).

    Article  CAS  Google Scholar 

  105. Carriere, F. et al. Inhibition of gastrointestinal lipolysis by Orlistat during digestion of test meals in healthy volunteers. Am. J. Physiol. Gastrointest. Liver Physiol. 281, G16–G28 (2001).

  106. Edwards, C. H., Maillot, M., Parker, R. & Warren, F. J. A comparison of the kinetics of in vitro starch digestion in smooth and wrinkled peas by porcine pancreatic alpha-amylase. Food Chem. 244, 386–393 (2018).

  107. Villemejane, C. et al. In vitro digestion of short-dough biscuits enriched in proteins and/or fibres using a multi-compartmental and dynamic system (2): protein and starch hydrolyses. Food Chem. 190, 164–172 (2016).

    Article  CAS  PubMed  Google Scholar 

  108. Romano, A. et al. Characterisation, in vitro digestibility and expected glycemic index of commercial starches as uncooked ingredients. J. Food Sci. Technol. 53, 4126–4134 (2016).

  109. Bustos, M. C., Vignola, M. B., Perez, G. T. & Leon, A. E. In vitro digestion kinetics and bioaccessibility of starch in cereal food products. J. Cereal Sci. 77, 243–250 (2017).

    Article  CAS  Google Scholar 

  110. Liu, J. Y. et al. Cellular uptake and trans-enterocyte transport of phenolics bound to vinegar melanoidins. J. Funct. Foods 37, 632–640 (2017).

    Article  CAS  Google Scholar 

  111. Corte-Real, J., Richling, E., Hoffmann, L. & Bohn, T. Selective factors governing in vitro beta-carotene bioaccessibility: negative influence of low filtration cutoffs and alterations by emulsifiers and food matrices. Nutr. Res. 34, 1101–1110 (2014).

    Article  CAS  PubMed  Google Scholar 

  112. Hidalgo, A. et al. Bioactive compounds and antioxidant properties of pseudocereals-enriched water biscuits and their in vitro digestates. Food Chem. 240, 799–807 (2018).

    Article  CAS  PubMed  Google Scholar 

  113. Eratte, D., Dowling, K., Barrow, C. J. & Adhikari, B. P. In-vitro digestion of probiotic bacteria and omega-3 oil co-microencapsulated in whey protein isolate-gum Arabic complex coacervates. Food Chem. 227, 129–136 (2017).

    Article  CAS  PubMed  Google Scholar 

  114. Bottari, B. et al. Characterization of the peptide fraction from digested Parmigiano Reggiano cheese and its effect on growth of lactobacilli and bifidobacteria. Int. J. Food Microbiol. 255, 32–41 (2017).

    Article  CAS  PubMed  Google Scholar 

  115. Sanchez-Moya, T. et al. In vitro modulation of gut microbiota by whey protein to preserve intestinal health. Food Funct. 8, 3053–3063 (2017).

    Article  CAS  PubMed  Google Scholar 

Download references

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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