In vitro digestibility testing
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1 In vitro digestibility testing Improving health properties of food by sharing our knowledge on the digestive process Prof Alan Mackie, University of Leeds Dr. Didier DUPONT, Senior Scientist, INRA, France
2 INFOGEST Chair Didier Dupont - France Vice-chair Alan Mackie - UK Characterization of raw materials and processed food matrices for optimized nutrient bioaccessibility WG1 BFC identification Stability during processing Food multi-scale characterization In vitro, in vivo and in silico models of mammalian gastrointestinal digestion WG2 Digestion models harmonization Comparison in vitro / in vivo Digestion products identification BFC absorption /bioavailability Evaluation of the health effects WG3 Immunomodulatory properties Regulation of appetite and satiety Effect of BFC on human microbiota B. Murray UK F Capozzi Italy A. Brodkorb Ireland I. Recio Spain A. Bordoni Italy Tor Lea Norway 2
3 Food Structure and Nutrient Release Cognitive and sensory enhancement of satiety and enzyme secretion Oral processing: aroma, taste and texture sensing starch hydrolysis and chewing gastric processing: proteolysis, lipolysis, acidification and storage -> fullness gastric processing: shearing, grinding intestinal processing: proteolysis, lipolysis, amylolysis, mixing and absorption intestinal processing: nutrient sensing, hormone secretion hormone secretion: controlling GI motility, enzyme and bile secretion, appetite
4 The objective for Infogest To produce a protocol to simulate human digestion that was: Simple and could be used in any laboratory Based on human physiology Giving results that are: Reproducible Consistent with human data on the same samples
5 Gastric Phase Mix 1:1 with SGF + pepsin (2000 U/mL), ph 3.0, 2h
6 Dissemination Minekus, M., et al. (2014). A standardised static in-vitro digestion method suitable for food an international consensus. Food and Function, 5, Egger, L., et al. (2016). The harmonized INFOGEST in vitro digestion method: From knowledge to action. Food Research International, 88, Part B, Mackie, A. R., & Rigby, N. M. (2015). Infogest Concensus Method. In K. Verhoeckx, P. Cotter, I. López-Expósito, C. Kleiveland, T. Lea, A. R. Mackie, T. Requena, D. Swiatecka & H. J. Wichers (Eds.), The Impact of Food Bioactives on Health In Vitro and Ex Vivo Models: Springer. Youtube: Dropbox folder: dl=0
7 Harmonisation Digestion of skimmed milk powder (SMP)
8 Harmonisation Release of free amino acids from SMP after gastric and intestinal phases of in vitro digestion. HPLC analysis of samples from interlaboratory trials applying the harmonized protocol
9 Pros and cons Pros: Simple to use Has been used in different labs giving the same results The end points seem the same as in vivo (SMP in pigs) Cons: Cannot be used for kinetics Only mimics adult conditions No gastric lipase included
10 Updates Semi-dynamic Dilution in the oral phase to be based on dry weight Inclusion of gastric emptying (based on caloric density), gradual secretion of simulated gastric fluid including acid and enzymes, inclusion of gastric lipase Infant conditions tba Elderly conditions tba
11 Semi-dynamic Gastric emptying 35,00 30,00 25,00 Volume (ml) 20,00 15,00 10,00 5,00 0,00 0,00 22,50 45,00 67,50 90,00 112,50 135,00 157,50 180,00 Time (minutes) Actual gastric volume (ml) Cumulative secretion volume (ml) volume to empty (ml) A 500mL meal is assumed for calculating the emptying rate. volumes are then scaled based on a smaller experimental sample (in this case 20g of food). The caloric density (0.72) gives 360 calories to 2 kcal/min = 180 mins Gastric secretion occurs over the same time.
12 Semi-dynamic Assuming 20g of food with a dry weight of 8g, the oral phase volume = 20+8 = 28g The final volume of gastric secretion = 28g, 10% is put in at the start Gastric lipase(rabbit) is included at 50 U/mL Intestinal digestion is in parallel. In this case 7g is emptied and diluted with 7g of simulated intestinal fluid
13 Pros and cons Pros: More physiological simulation of the gastric phase Can be used to assess kinetics Still based on small volumes and simple apparatus Can be used in many labs Cons: More complicated procedure The emptying may be difficult with some foods (solids) Sourcing a suitable gastric lipase
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