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Background: The complexity of food structure is such as to hinder its inclusion in mathematical models predicting food properties and transformations, although a considerable impulse is being determined by using artificial intelligence. As a matter of fact, food definition currently neglects the structural description, even in those fields for which structure is demonstrated to have a decisive role, such as nutrition. Scope and approach: This review aims to analyse the current knowledge about the structure of foods and its potential use to numerically define the sensory and nutritional quality, as well as the stability properties. Starting from this information, a possible methodology is explored to build, even in an automated way, mathematical models for simulating and predicting the properties of food. A model pipeline has been proposed and applied to pasta, in particular exploiting the description of the structural changes occurring upon cooking. Key findings and conclusions: Foods may be designed in silico, based on automated pipelines for direct extraction of information on rheological and sensory properties as derived from structure images and from data on the dynamic state of the water. The ultimate goal of these approaches is to make more limited use of expensive and time-consuming experiments on physically prepared foods to get to use digital twins of foods designed in the laboratory.

Wheat consumption can represent one of the nutritional factors involved in the onset of diabetes. We specifically investigated the potential diabetogenic effects of Hammurabi, a T. monococcum wheat cultivar, in non-obese diabetic (NOD) mice and analysed the levels of resistant starch in pasta manufactured with Hammurabi after in vitro gastroduodenal digestion. NOD mice were fed with Hammurabi, bread wheat or rice flour to evaluate diabetes incidence and insulitis score. An enzymatic method was applied to compare the content of resistant starch in Hammurabi pasta and durum wheat pasta (control). In NOD mice, the Hammurabi-based diet significantly delayed diabetes onset (p = 0.0042) and reduced insulitis score compared to rice or wheat-based diet. Furthermore, the resistant starch value following in vitro digestion of Hammurabi pasta was significantly higher (4.08%) than that of durum wheat pasta (2.28%). Taken together, these results highlighted the potential positive effects of the Hammurabi-based diet on diabetes incidence.

The increasing interest of consumers for food with enhanced nutritional characteristics has spurred industry to look for unconventional raw materials for novel food design. Pasta is a staple food consumed all over the world, and therefore represents an ideal matrix for the enrichment with functional ingredients. Despite durum wheat semolina is considered the best raw material for Italian pasta making, due to its physicochemical, rheological and sensory properties, nutritionally enriched pasta is becoming quite popular among specific segments of consumers. Nowadays there is a wide choice of ingredients available to modify and improve the nutritional profile of durum wheat semolina pasta. Among them dietary fibres, legume flours, rice, corn, emmer, cricket flour among others. The novel ingredients and process also allow the production of high quality gluten-free pasta for sensitive and celiac consumers categories. The use of alternative ingredients in pasta formulation results in interesting modifications in quality as well as nutritional and sensory characteristics.

Osteosarcoma is a bone cancer characterized by the production of osteoid tissue and immature bone from mesenchymal cells. Osteosarcoma mainly affects long bones (femur is most frequently site) and occur in children and young adults with greater incidence. Here, we investigated the role accomplished by polydatin, a natural antioxidative compound, in promoting osteogenic differentiation alone or after radiation therapy on osteosarcoma cells. In vitro, polydatin significantly induced cell cycle arrest in S-phase and enhanced bone alkaline phosphatase activity. Moreover, the differentiation process was paralleled by the activation of Wnt-β-catenin pathway. In combination with radiotherapy, the pretreatment with polydatin promoted a radiosensitizing effect on osteosarcoma cancer cells as demonstrated by the upregulation of osteogenic markers and reduced clonogenic survival of tumor cells. Additionally, we analyzed, by mass spectrometry, the secretion of sphingolipid, ceramides, and their metabolites in osteosarcoma cells treated with polydatin. Overall, our results demonstrate that polydatin, through the secretion of sphingolipids and ceramide, induced osteogenic differentiation, alone and in the presence of ionizing therapy. Future investigations are needed to validate the use of polydatin in clinical practice as a potentiating agent of radiotherapy-induced anticancer effects.

Aim of this work was to evaluate the impact of bread enrichment with different concentrations of lentil flour (LF) (0 g/100g, 10 g/100g and 20 g/100g) on some physico-chemical, structural, and nutritional features of bread. The addition of LF in formulation affected its properties, leading to a darker appearance and to a more compact structure (e.g., higher density and lower gas bubble area fraction). Besides, it led to a significant rise in the protein, free glucose, and rapidly digestible starch content as well as in the expected glycemic index. Concerning the investigation of nutritional properties, starch and protein digestibility has been evaluated by the combination of different digestion models. Each type of bread was subjected to an in vivo oral phase followed by a semi-dynamic in vitro gastrointestinal digestion. During the oral and the gastric phases of digestion, bread with the greatest LF content (20 g/100g) exhibited an extensive starch digestibility and a partial protein digestibility compared to the other samples, due to its higher protein content. In contrast, during the intestinal phase, control (0 g/100g) showed a stronger starch digestibility compared with the LF samples, while no differences could be observed in terms of protein digestibility investigated by SDS-PAGE., This work has received financial support from project PID2019-107663RB-I00 from the Spanish Ministry of Science and Innovation (MICINN).

Every year, the coffee supply chain produces tons of waste materials from the processing of the coffee bean. Coffee silverskin (CS) is the only waste product from the coffee bean roasting phases but as roasting is a key step in obtaining the coffee beverage, there are tons of CS collected and thrown away by coffee roasting industries. This study is based on the characterization of this integument, with a view to its use in nutritional, nutraceutical and industrial fields. From the results obtained, silverskin can be used in the food sector for its nutritional profile with a good amount of protein (18.9%) and total dietary fiber (34.7%) andits low-fat content (3.0%). Food safety was also determined by evaluating the low levels of contaminants such as ochratoxin A (OTA) and acrylamide (AA), with values < 0.1 mg/kg and

Alternative sources of edible proteins are required to feed the world's growing population, such as Moringa oleifera leaves, a protein source with a balanced amino acid composition. Since Moringa leaf proteins is a novel food in the EU and UK, an assessment of their potential allergenicity of is required. Proteins from Moringa leaf powder were characterised using traditional proteomic approaches. The proteins identified were evaluated for their allergenic potential using in-silico tools. The main proteins identified belonged to photosynthetic and metabolic pathways. In-silico analysis of the leaf proteome identified moritides as potential allergens by homology with a latex allergen implicated in fruit-latex syndrome. This analysis also identified a nsLTP, a major panallergen in food. The presence of these putative allergens was confirmed by de-novo sequencing. Our study allowed identification of putative allergens, Morintides and nsLTP. Further in-vitro and in-vivo investigations are required to confirm their allergenic potential. Proteins from Moringa leaf powder were characterised using traditional proteomic approaches. The proteins identified were evaluated for their allergenic potential using in-silico tools. The main proteins identified belonged to photosynthetic and metabolic pathways. In-silico analysis of the leaf proteome identified moritides as potential allergens by homology with a latex allergen implicated in fruit-latex syndrome. This analysis also identified a nsLTP, a major panallergen in food. The presence of these putative allergens was confirmed by de-novo sequencing. Our study allowed identification of putative allergens, Morintides and nsLTP. Further in-vitro and in-vivo investigations are required to confirm their allergenic potential.

Digestomics can be defined as the systematic characterization of the biomolecular complement produced by food digestion. Protein digestomics, that essentially corresponds to tracking the fate of dietary proteins in human body, assumes special relevance because food-derived peptides are virtually capable of inducing positive (health-promoting) or negative (allergy, adverse reactions) effects. In general, food digestion generates extremely heterogeneous mixtures, which include free amino acids and variously sized (poly) peptides, in addition to products from other macro- and micronutrients. Modern proteomics and peptidomics have enabled the comprehensive study of complex digestomes. On the other hand, several gaps still need to be bridged to define kinetics and dynamics of food-derived peptides, also as a result of food structure, in order to ultimately assess the effects of dietary proteins on human health. This chapter surveys the recent advances in the comprehension of mechanisms of food degradation, highlighting research trends and still unaddressed issues.

Wheat consumption can represent one of the nutritional factors involved in the onset of diabetes. We specifically investigated the potential diabetogenic effects of Hammurabi, a

Heat-stable peptidases released in refrigerated raw milk by psychrotrophic bacteria are responsible for UHT milk gelation. K-casein-derived caseinomacropeptides, identified by mass spectrometry, were constantly detected in gelled milk by capillary electrophoresis. Strains of Pseudomonas fluorescens, Ps. poae and Chryseobacterium joostei, selected among aprX-positive strains from raw milk, were incubated in milk up to 6 days at 4 degrees C before sterilization (98 degrees C/4 min). Samples were then stored at 25 or 40 degrees C, visually observed for gelation, and analysed for presence of caseinomacropeptides throughout 90 days of storage. Depending on cold pre-incubation time, caseinomacropeptides accumulated well before gelation onset in milk stored at 25 degrees C. Caseinomacropeptides were successively degraded, especially in milk stored at 40 degrees C, due to extensive proteolysis, and an abundant sediment developed instead of a gel. The caseinomacropeptides are here presented as an early indicator of UHT milk gelation and a mechanism explaining this phenomenon is proposed.

Analyzing an in vitro gastroduodenal digest of whey proteins by high-performance liquid chromatography (HPLC) coupled to high-resolution/high-sensitivity tandem mass spectrometry (MS/MS), we sought to evaluate if state-of-art peptidomics provide comprehensive peptide coverage of food "digestomes". A multitude of small-sized peptides derived from both a-lactalbumin and beta-lactoglobulin as well as disulfide cross-linked hetero-oligomers remained unassigned, even when the digests were compared before and after S-S reduction. The precipitation with 12% trichloroacetic acid demonstrated the occurrence of large-sized polypeptides that escaped the bioinformatic identification. The analysis of a HPLC-MS/MS run with different proteomic search engines generated dissimilar peptide subsets, thus emphasizing the 'demand of refined searching algorithms. Although the MS/MS fragmentation of monocharged ions with exclusion of non-peptide-interfering compounds enlarged the inventory of short peptides, the overall picture of the "digestome" was still incomplete. These findings raise relevant implications for the identification of possible food-derived bioactive peptides or allergenic determinants.

In these last years, consumers'choices are being directed towards healthier food and beverages with an increasing demand for functional products. In this study, we investigated the sensory and biochemical properties of novel drinks based on tomato juice. To this purpose, different blends were formulated mixing fresh tomato juice with other fruit juices and nectars in different proportions and then assayed to investigate their sensory (panel test), compositional and biochemical characteristics. Our results indicated that it is possible to formulate tasty drinks based on tomato juice with improved nutritional properties. The combinations of red fruits/tomato (60/40 v/v) and red fruits/orange/tomato (40/30/30 v/v/v) showed a sugar content lower than those of different soft drinks on the market including energy drinks, and suitable lycopene levels as well. Interestingly, the blended red fruits/orange/tomato had a greater number of polyphenols and vitamin C, a softer tomato flavour and high sensory appreciation. High pasteurization (90ºC, 7 min), performed to increase storability, did not significantly affect sensory and biochemical properties of drinks. These achievements may be useful to modulate tomato flavour release and consumer acceptability of novel drinks based on tomato juice.

The addition of frozen curd (FC) during the production process of “Mozzarella di Bufala Campana”, an Italian cheese with Protected Designation of Origin (PDO), is a common fraud not involving modifications of the chemical composition in the final product. Its detection cannot thus be easily obtained by common analytical methods, which are targeted at changes in concentrations of diagnostic chemical species. In this work, the possibility of spotting this fraud by focusing on the modifications of the supramolecular structure of the food matrix, detected by time domain nuclear magnetic resonance (TD-NMR) experiments, was investigated. Cheese samples were manufactured in triplicate, according to the PDO disciplinary of production, except for using variable amounts of FC (i.e., 0, 15, 30, and 50% w/w). Relaxation data were analysed through different approaches: (i) Discrete multi-exponential fitting, (ii) continuous Laplace inverse fitting, and (iii) chemometrics approach. The strategy that lead to best detection results was the chemometrics analysis of raw Carr-Purcell-Meiboom-Gill (CPMG) decays, allowing to discriminate between compliant and adulterated samples, with as low as 15% of FC addition. The strategy is based on the use of machine learning for projection on latent structures of raw CPMG data and classification tasks for fraud detection, using quadratic discriminant analysis. By coupling TD-NMR raw decays with machine learning, this work opens the way to set up a system for detecting common food frauds modifying the matrix structure, for which no official authentication methods are yet available.

Donkey's milk (DM) has been extensively investigated as a valuable substitute of breast milk, often suitable to manage cow's milk protein allergy in infants. DM exhibits potent inhibitory properties against numerous microbial species. Although oligosaccharides and lipids might contribute to the antimicrobial potential, the current inventory of proteins is not able to justify the low count of microorganisms generally observed in DM. The shotgun proteomic analysis of fractionated DM disclosed a set of 94 gene products, 41% of which have documented antimicrobial activity or are involved in transferring the passive immunity to the donkey offspring. The concerted action of lysozyme, lactoferrin, immunoglobulins provides the molecular basis for part of the DM antibacterial potential. The pH -4.6 insoluble fraction contained significant levels of L-amino acid oxidase, identified with 11 unique peptides matching the horse homologue gene product. This enzyme catalyses the oxidative deamination of amino acids into ketoacids, producing ammonia and H2O2. κ-casein, likely occurring as a fully O-glycosylated protein, may concur to inhibit the adhesion of pathogenic microorganisms, along with other glycoproteins. Proteomics supports the alimentary use of DM not only as a substitute of human milk in early infancy, but also for growing children, convalescent, elderly people and general population. Significance Donkey's milk (DM) is acquiring increasing popularity because it is a suitable substitute of the human milk, when breastfeeding is not possible and infants suffer from cow's milk allergy. DM is characterized by a much lower microbial load compared to ruminants' milk. This feature has been traditionally attributed to the high content of lysozyme. DM exhibits potent activity against a broad range of bacteria, viruses and fungi, suggesting that other protein components can be responsible of the antimicrobial potential. The gel-free proteomic analysis of pH 4.6-insoluble and soluble (whey) fractions demonstrated that DM contains a large number of gene products involved in antimicrobial mechanisms and in transferring passive immunity to the donkey offspring. DM contains relatively high levels of L-amino acid oxidase that catalyses the oxidative deamination of amino acid substrates into ketoacids, with production of ammonia and H2O2. In combination with lysozyme, lactoferrin and immunoglobulins, the presence of L-amino acid oxidase provides the molecular basis of the antibacterial potential observed for DM. Considered the low microbial load, DM can be sanitated at mild conditions, thereby preserving many of the native nutritional traits. Thus, DM can be considered a safe and nutritionally valid alimentary resource for growing children, convalescent, elderly people and general population. Data of this study represent the largest inventory of proteins identified in Equidae milk, so far.

Wheat, an essential ingredient for several bakery preparations, is also responsible for gluten-related diseases in sensitive subjects. The effect of the N fertilization rate (80 vs 160 kg N ha−1) on gluten protein expression profile has been evaluated considering two soft wheats (landrace and modern) and one tritordeum cultivar (cv), grown in the same experimental field in North Italy. The proteins of refined flour were characterized through advanced proteomic approaches, including chromatography (RP-HPLC) and electrophoresis. A static model system was used to simulate in vitro digestion and the digestome peptides were examined by mass spectrometry and in silico approaches, to investigate the celiac and allergenic sequences. The CD-toxic epitopes in the digested samples were quantified by means of a R5 ELISA assay. The N fertilization rate increased the grain protein content, but it did not lead to any difference in gluten composition, with exception of glu/glia ratio in the modern wheat cv. Moreover, the gluten composition and the occurrence of toxic/allergenic epitopes varied to a great extent, according mostly to the genotype. A lower immunoreactivity, determined using R5 ELISA, was detected for the digested tritordeum flours than for the landrace (-51%) or modern (-58%) cvs, while no significant difference was observed for the N rates between each genotype. In silico analysis showed that tritordeum has fewer CD epitopes belonging to the ω-gliadins and a lower LMW-GS than the landrace or modern cv. Tritordeum presented fewer α-gliadin allergenic epitopes than the modern wheat cv. The lower frequency of celiac epitopes in tritordeum, compared to the old and the modern wheat, is probably due to the absence of a D genome.

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The use of lentil flour in bakery (bread, cake, crackers), extruded (pasta, snacks) and other products (dressings, soups, dairy and meat products) is gaining attention of food technologists and of industry as well as popularity among consumers due to an excellent and balanced nutritional composition. This research interest has extended our knowledge of nutritional and functional properties (solubility, emulsification, gelation, foaming) of lentil flour, which, in turn, has disclosed its technological potential for preparation of high-quality foods (gluten free bakery, yogurt and meat products). However, addition of lentil flour may introduce technological problems and novel allergens. This review covers the use of lentil flour in food preparations, with focus on protein and starch digestibility and on the challenges to design lentil-based products with improved quality.

Egg proteins are among the major food allergens. Very often, the same pasta-making plants are used for industrial production of egg-based pasta (EBP) and semolina-only pasta (SP), so that residual egg proteins may be present in SP. This calls for defining the amount of semolina pasta that should be discarded when switching production lines. In this study, the egg proteins content was measured in pasta samples taken at various times after switching production lines from EBP to SP Both long and short pasta shapes were sampled before and after a drying step. Protocols meant to circumvent the difficulties associated with detecting egg proteins in a complex matrix after processing were set up for using commercial ELISA kits to monitor the disappearance of egg proteins from the products. The use of both denaturants and disulphide reductants to solubilise egg proteins was found to be mandatory, as verified by ovalbumin detection by ELISA and by using mass spectrometry to assess residual egg white lysozyme. Appropriate sample preparation protocols were used to monitor the progressive disappearance of egg proteins in the products when shifting production lines in an industrial pasta plant, providing a basis for credible, reliable, and consistent self-control procedures. For lines with a production capacity of 2200-2400 kg h-1, the amount of material to be discarded to ensure that products meet the strictest analytical requirements has been found to be around 2000-3000 kg (for long pasta) and 3000-4000 kg (for short pasta).

The aim of this study was to evaluate the dynamics of proteolysis during dough fermentation started with different lactic acid bacteria species, through the identification of intermediate and small-sized peptides generated during fermentation. Single-strain cultures of Levilactobacillus brevis, Fructilactobacillus sanfranciscensis, Companilactobacillus alimentarius, and Leuconostoc pseudomesenteroides were assayed as sourdough starters. Assays were carried out at lab-scale for 48 h of fermentation, using both unstarted and yeast-leavened dough as controls. Physicochemical and microbiological analyses were combined with peptidomic and proteomic profiling, identifying several hundreds of peptides mainly released from the water-soluble wheat proteins, including β-amylase, triticin, and serpins. Both α- and γ-gliadins were hydrolyzed, though only at the N-terminal domain, while the central protein region – encrypting celiac disease epitopes– remained unaffected. The bacterial-mediated consumption of sugars and the concomitant hydrolysis of starch degrading β-amylase could underlie improved digestibility and several nutritionally beneficial effects of sourdough baked products.

In this work, we explored the “deep” seed peanut proteome by using both two dimensional electrophoresis (2- DE)-based analysis run under reducing and non-reducing condition (protein-centric) and LC-MS/MS gel-free proteomic (peptide-centric). The former approach allowed to identify high molecular weight disulfide-linked Ara h 1 and Ara h 3 heteroligomers and Ara h 1 homoligomers linked through covalent bonds other than disulfides. The occurrence of these protein complexes revealed natural interactions between Ara(s) subunits with a possible involvement in the allergenic potential of peanut. The second approach, also referred to as shot-gun proteomics, allowed the identification of 149 gene products, including low-abundance proteins escaped the 2-DE detection. Interestingly, we identified 60 proteins never catalogued previously. The complementary exploitation of two proteomic approaches enabled the access to new relevant information about the complexity of the peanut proteome, with special emphasis to the complement of allergens (allergome).

Hemp (Cannabis sativa L.), traditionally cultivated for industrial use and harvested for fibers and seeds, has raised much interest as a sustainable crop in the last years. Recently, hemp seeds and derived oil have started to be used in a variety of food products. Hemp-based food products are considered less allergenic than those from other edible seeds, although this statement has never been experimentally verified. In this study high purity grade hemp flour (HF) and hemp protein isolate (HPI) were obtained through a fast and cheap process starting from defatted hemp cakes, a residue of hempseed oil extraction. HPI resulted enriched at nearly 86% protein, mainly constituted by the storage protein edestin (accounting for 70% total protein). In vitro protein digestibility was determined using a static model of gastrointestinal digestion (GID), which included a final step with purified brush border membrane (BBM) enzyme preparations. HF and HPI showed a high degree of digestibility. The survival of potential bioactive and/or allergenic peptide sequences in digests was investigated by peptidomic analysis. Only a limited number of sequences survived GID. Among them, fragments from 12 seed proteins. These fragments were precursors of sequences with potential bioactive peptides, which might justify the bioactivity of HPI hydrolysates, reported in previous studies. More importantly, all known hemp allergens, including the major thaumatin-like protein and LTP, were entirely eliminated by the HPI production process, neither fragments of the proteins were present after GID. These data support the use of HPI as an ingredient for hypoallergenic foods.

The demand for sustainably produced proteins is increasing with the world population and is prompting a dietary shift toward plant sourced proteins. Vegetable proteins have lower digestibility and biological value compared to animal derived counterparts. We explored sprouting of chickpea seeds as a strategy for improving digestibility. Protein evolution associated with by the sprouting process was assessed by proteomics. The sprouting induced breakdown of seed storage proteins and doubled the release of free alpha-amino nitrogen in sprouted chickpea flour. During sprouting, several enzymes involved in plant development were newly expressed. An ex vivo model of gastroduodenal and jejunal digestion was applied to assess the bioaccessibility of the protein digests. Proteins from chickpea sprouts showed a greater susceptibility to digestion with a 10% increase in alpha amino nitrogen. Peptides with potential immunoreactivity or bioactivity were catalogued in both digested chickpea sprouts and seeds using an in-silico approach. Peptides belonging to the non-specific transfer proteins, which are allergens in pulses, and peptides belonging to an IgE-binding hemagglutinin protein could only be identified in the digested chickpea sprouts. The observation collected paved the way to immune-based evaluations to assess the effect of germination on the allergenic potential.

Kashk is a fermented dairy product typical of the Middle East, traditionally produced with sour milk and/or dairy waste. The kashk water-soluble peptide fraction was characterized at the molecular level by liquid chromatography-mass spectrometry and its antibacterial and skin healing activity was evaluated. Antibacterial assays showed a significant antibacterial activity against clinical isolates of Staphylococcus aureus (S. aureus) from patients with atopic dermatitis, inhibiting bacterial growth by approximately 45% (500μg/mL). Skin repair activity was evaluated on keratinocytes through scratch tests showing accelerated wound closure in vitro in the presence of TNF-α, by approximately 44% (500μg/mL), compared to control cells. Furthermore, based on the MTT assay, the kashk peptide fraction did not show toxicity on keratinocytes. The results suggested that the peptide kashk extract may be useful in skin care for patients with atopic dermatitis.

The field of food analytical chemistry has greatly evolved over the last years, moving from the descriptive view of raw food composition into the modern instrumental approaches known as the foodomics. The development and optimization of these integrated approaches, based on the use of high resolution techniques for tracing the chemical fingerprint of a food, is arriving to define molecular parameters for the qualification, authentication, and safety of the products. With these new tools, omics are solving some of the new issues of food quality and traceability, including the development of analytical methods to ensure the origin of food, as well as the discovery of biomarkers to identify potential food safety problems. Here we review the main methods used in foodomics from its birth until now, highlighting the use of high throughput instrumentation. This includes the computational problems they generate, but also the significant prospects for development of knowledge that are able to open in important areas such as food technology and biotechnology.

Unravelling the fate of food in the gastrointestinal tract is essential to better understand the health effects of the food and to fight against diet-related pathologies such as cardiovascular diseases and type-2 diabetes. Digestion is the process that transforms food into nutrients. The first step of digestion occurs in the mouth, where mastication transforms solid and semi-solid foods into particles while mixing with saliva allows bolus formation and initiates digestion of carbohydrates.Then the bolus is transferred into the stomach, where acid conditions and specific enzymes (pepsin, gastric lipase) start hydrolyzing macronutrients like proteins and triglycerides. The next step occurs in the small intestine, where other digestive enzymes further degrade macronutrients allowing their absorption. In the small intestine, proteins are hydrolyzed by trypsin, chymotrypsin, elastase, carboxypeptidase etc, lipids by pancreatic lipases, and carbohydrates by pancreatic amylase. Small intestinal digestion is completed by the enzymes of the brush border membrane that release macronutrients, which can be absorbed by enterocytes to reach the bloodstream. Undigested material, fiber for example, reaches the large intestine where it is further metabolized by the intestinal microbiota. Investigating food digestion requires the use of models and a myriad of in vitro (static and dynamic), animal and human models have been described in the literature with the objectives of understanding the fate of food in the gastrointestinal tract. In particular, static in vitro digestion simulations are extremely popular because they are very easy to use and do not require sophisticated equipment. They have been shown to be adapted for screening large series of similar samples in identical conditions or to understand interactions at the molecular scale (Bohn et al.,2017). However, they are too simple to study more complex phenomena and the kinetics of food digestion for which dynamic in vitro models are more appropriate (Dupont et al., 2018). There was a high heterogeneity between the different static in vitro digestion models that were used by the research groups all around the world. Models were differing in the pH used in the different phases (gastric and intestinal), their duration, the digestive enzyme/substrate ratio, etc For that reason, comparing results obtained from one study to another was impossible and there was a crucial need for a harmonized method that could be used by everyone allowing comparison between studies. This was one of the main objectives of the INFOGEST COST Action.