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      A standardised static in vitro digestion method suitable for food - an international consensus.

      Food & function
      Animals, Bile Acids and Salts, metabolism, Consensus, Digestion, physiology, Food, Gastrointestinal Contents, chemistry, Humans, Hydrogen-Ion Concentration, Models, Biological, Models, Theoretical, Pancreatin, Saliva

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          Abstract

          Simulated gastro-intestinal digestion is widely employed in many fields of food and nutritional sciences, as conducting human trials are often costly, resource intensive, and ethically disputable. As a consequence, in vitro alternatives that determine endpoints such as the bioaccessibility of nutrients and non-nutrients or the digestibility of macronutrients (e.g. lipids, proteins and carbohydrates) are used for screening and building new hypotheses. Various digestion models have been proposed, often impeding the possibility to compare results across research teams. For example, a large variety of enzymes from different sources such as of porcine, rabbit or human origin have been used, differing in their activity and characterization. Differences in pH, mineral type, ionic strength and digestion time, which alter enzyme activity and other phenomena, may also considerably alter results. Other parameters such as the presence of phospholipids, individual enzymes such as gastric lipase and digestive emulsifiers vs. their mixtures (e.g. pancreatin and bile salts), and the ratio of food bolus to digestive fluids, have also been discussed at length. In the present consensus paper, within the COST Infogest network, we propose a general standardised and practical static digestion method based on physiologically relevant conditions that can be applied for various endpoints, which may be amended to accommodate further specific requirements. A frameset of parameters including the oral, gastric and small intestinal digestion are outlined and their relevance discussed in relation to available in vivo data and enzymes. This consensus paper will give a detailed protocol and a line-by-line, guidance, recommendations and justifications but also limitation of the proposed model. This harmonised static, in vitro digestion method for food should aid the production of more comparable data in the future.

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          Most cited references74

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          THE ESTIMATION OF PEPSIN, TRYPSIN, PAPAIN, AND CATHEPSIN WITH HEMOGLOBIN

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            In vitro human digestion models for food applications

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              Total phenolics, flavonoids, anthocyanins and antioxidant activity following simulated gastro-intestinal digestion and dialysis of apple varieties: Bioaccessibility and potential uptake.

              In the present study, an in vitro model simulating gastrointestinal (GI) digestion, including dialysability, was adapted to assess free soluble polyphenols from apples (four varieties). Results indicated that polyphenol release was mainly achieved during the gastric phase (ca. 65% of phenolics and flavonoids), with a slight further release (<10%) during intestinal digestion. Anthocyanins present after the gastric phase (1.04-1.14mg/100g) were not detectable following intestinal digestion. Dialysis experiments employing a semipermeable cellulose membrane, presenting a simplified model of the epithelial barrier, showed that free soluble dialysable polyphenols and flavonoids were 55% and 44% of native concentrations, respectively, being approximately 20% and 30% lower than that of the GI digesta. Similar results were found for the antioxidant capacity of dialysable antioxidants, being 57% and 46% lower compared to total antioxidants in fresh apples (FRAP and ABTS test, respectively). It is suggested that some polyphenols are bound to macromolecular compounds that are non-dialysable, that the presented method allowed the study of free soluble polyphenols available for further uptake, and that both chemical extraction and concentrations in final digesta would overestimate polyphenol availability.
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