Stability and bioaccessibility of micronutrients and phytochemicals present in processed leek and Brussels sprouts during static in vitro digestion.

Vegetables are frequently processed before consumption. However, vegetable functionalization continues beyond ingestion as the human digestive tract exposes vegetable products to various conditions (e.g. elevated temperature, pH alterations, enzymes, electrolytes, mechanical disintegration) which can affect the stability of micronutrients and phytochemicals. Besides the extent to which these compounds withstand the challenges posed by digestive conditions, it is equally important to consider their accessibility for potential absorption by the body. Therefore, this study investigated the impact of static in vitro digestion on the stability (i.e. concentration) and bioaccessibility of vitamin C, vitamin K1, glucosinolates, S-alk(en)yl-l-cysteine sulfoxides (ACSOs) and carotenoids in Brussels sprouts (Brassica oleracea var. gemmifera) and leek (Allium ampeloprasum var. porrum). Water-soluble compounds, glucosinolates and ACSOs, remained stable during digestion while vitamin C decreased by >48%. However, all water-soluble compounds were completely bioaccessible. Lipid-soluble compounds were also stable during digestion but were only bioaccessible for 26-81%.

Pea protein extraction method impacts the protein (micro)structural organisation and in vitro digestion kinetics.

There is increasing interest in including pulse proteins into food products due to their nutrient-rich and sustainable character. However, little is known regarding the consequences of different extraction approaches on the pulse protein structure and the subsequent protein (micro)structural organization and protein digestion kinetics. Therefore, three green pea protein extracts were created: (i) cooking followed by cotyledon cell isolation, (ii) alkaline extraction followed by isoelectric precipitation, or (iii) salt extraction, and compared to the original pea flour as well as to sodium caseinate. The results showed that encapsulated, denatured protein inside pea cotyledon cells presented the (s)lowest digestion, while accessible and more native protein (e.g., pea flour, pea protein salt extract) presented much faster and higher digestion. Moreover, the alkali extracted pea protein was denatured to some extent, significantly lowering in vitro digestion kinetics. In the second part, three different in vitro approaches were applied to digest the salt extracted pea protein. Semi-dynamic gastric digestion approaches simulate in vivo conditions more closely which especially impacted the rate of digestion.

Impact of refrigerated storage on (bio)chemical conversions of health-related compounds in pretreated, pasteurized Brussels sprouts and leek.

Vegetable processing often consists of multiple processing steps. Research mostly focused on the impact of individual processing steps on individual health-related compounds. However, there is a need for more holistic approaches to understand the overall impact of the processing chain on the health potential of vegetables. Therefore, this work studied the impact of pretreatment (relatively intact versus pureed vegetable systems), pasteurization and subsequent refrigerated storage (kinetic evaluation) on multiple health-related compounds (vitamin C, vitamin K1, carotenoids, glucosinolates and S-alk(en)yl-L-cysteine sulfoxides (ACSOs)) in Brussels sprouts and leek. It could be shown that differences introduced by different types of pretreatment were not nullified during pasteurization and refrigerated storage. Clearly, enzymatic conversions controlled during pretreatment resulted in different health-related compound profiles still observable after pasteurization. Moreover, about -42% and -100% relative concentration differences of ACSOs and dehydroascorbic acid, respectively, were detected immediately after pasteurization, while glucosinolates concentrations decreased by about 47% during refrigerated storage. All other compounds were stable during pasteurization and refrigerated storage.

From static to semi-dynamic in vitro digestion conditions relevant for the older population: starch and protein digestion of cooked lentils.

In the context of adequately feeding the rising older population, lentils have an important potential as sources of (plant-based) protein as well as slowly digestible bio-encapsulated starch and fibre. This study evaluated in vitro digestion of protein and starch in lentils under conditions representing the gastrointestinal tract of older adults. Both static and semi-dynamic simulations were applied to analyze the effect of specific gastrointestinal conditions (healthy versus older adult) on macronutrient digestion patterns. Gastric proteolysis was strongly dependent on applied gastric pH (gradient), leading to a lower extent of protein hydrolysis for simulations relevant for older adults. Fewer and smaller (lower degree of polymerization, DP) bioaccessible peptides were formed during gastric proteolysis under older adult compared to healthy adult conditions. These differences, developed during the in vitro gastric phase, were compensated during small intestinal digestion, yielding similar final proteolysis levels regardless of the applied simulation conditions. In contrast, in the presence of saliva, amylolysis was generally accelerated under older adult conditions. Moreover, the current work highlighted the importance of considering saliva (or salivary amylase) incorporation in simulations where the applied gastric pH (gradient) allows salivary amylase activity. Under both healthy and older adult conditions, in vitro starch hydrolysis bio-encapsulated in cotyledon cells of cooked lentils was attenuated, compared to a white bread reference.

Pre-duodenal lipid digestion of emulsions: Relevance, colloidal aspects and mechanistic insight.

The digestion of lipids in the human body has several health and nutritional implications. Lipid digestion is an interfacial phenomenon meaning that water-soluble lipases need to first adsorb to the oil-water interface before enzymatic conversions can start. The digestion of lipids mainly occurs on colloidal structures dispersed in water, such as oil-in-water (o/w) emulsions, which can be designed during food formulation/processing or structured during digestion. From a food design perspective, different in vitro studies have demonstrated that the kinetics of lipid digestion can be influenced by emulsion properties. However, most of these studies have been performed with pancreatic enzymes to simulate lipolysis in the small intestine. Only few studies have dealt with lipid digestion in the gastric phase and its subsequent impact on intestinal lipolysis. In this aspect, this review compiles information on the physiological aspects of gastric lipid digestion. In addition, it deals with colloidal and interfacial aspects starting from emulsion design factors and how they evolve during in vitro digestion. Finally, molecular mechanisms describing gastric lipolysis are discussed.

How Cooking Time Affects In Vitro Starch and Protein Digestibility of Whole Cooked Lentil Seeds versus Isolated Cotyledon Cells.

Lentils are sustainable sources of bioencapsulated macronutrients, meaning physical barriers hinder the permeation of digestive enzymes into cotyledon cells, slowing down macronutrient digestion. While lentils are typically consumed as cooked seeds, insights into the effect of cooking time on microstructural and related digestive properties are lacking. Therefore, the effect of cooking time (15, 30, or 60 min) on in vitro amylolysis and proteolysis kinetics of lentil seeds (CL) and an important microstructural fraction, i.e., cotyledon cells isolated thereof (ICC), were studied. For ICC, cooking time had no significant effect on amylolysis kinetics, while small but significant differences in proteolysis were observed (p < 0.05). In contrast, cooking time importantly affected the microstructure obtained upon the mechanical disintegration of whole lentils, resulting in significantly different digestion kinetics. Upon long cooking times (60 min), digestion kinetics approached those of ICC since mechanical disintegration yielded a high fraction of individual cotyledon cells (67 g/100 g dry matter). However, cooked lentils with a short cooking time (15 min) showed significantly slower amylolysis with a lower final extent (~30%), due to the presence of more cell clusters upon disintegration. In conclusion, cooking time can be used to obtain distinct microstructures and digestive functionalities with perspectives for household and industrial preparation.

Importance of adapted digestion conditions to simulate in vitro lipid digestion of broilers in different life stages.

In vitro digestion studies demonstrate large potential to gain more and quicker insights into the underlying mechanisms of feed additives, allowing the optimization of feed design. Unfortunately, current in vitro digestion models relevant for broiler chickens lack sufficient description in terms of protocols and standardisation used. Furthermore, no distinction is made between the different life phases of these animals (starter, grower, and finisher). Hence, our research aimed to establish adapted in vitro digestion conditions, corresponding to the 3 life phases in broilers, with specific focus on lipid digestion. The effect of 3 different bile salt concentrations of 2, 10, and 20 mM, and 3 different lipase activities of 5, 20, and 100 U/mL, on in vitro lipid digestion kinetics were evaluated using a full factorial design. These values were selected to represent starter, grower, and finisher birds, respectively. Our findings showed that the extent of lipid digestion was mainly influenced by lipase activity. The rate of lipid digestion was affected by an interplay between bile salt concentration and lipase activity, due to possible lipase inhibition at certain bile salt concentrations. Overall, this work resulted in 3 in vitro lipid digestion models representative for starter, grower, and finisher birds. In conclusion, this research showed the impact of adapted in vitro digestion conditions on lipid digestion kinetics and thus the need for these conditions relevant for each life phase of broilers.

Heat and Light Stability of Pumpkin-Based Carotenoids in a Photosensitive Food: A Carotenoid-Coloured Beverage.

This study aimed to evaluate carotenoid degradation kinetics in a beverage coloured with pumpkin juice concentrate during storage at dark and illuminated conditions at four temperatures (10, 20, 35 and 45 °C). Carotenoids were quantified by HPLC-DAD, and kinetic parameters for carotenoid degradation were estimated by one-step nonlinear regression analysis. During dark storage, degradation kinetics was modelled by fractional conversion (all-trans-ß-carotene) and zero-order equations (all-trans-antheraxanthin, all-trans-lutein, all-trans-violaxanthin and all-trans-neoxanthin). Storage of samples in a climatic chamber with intense light intensity (1875-3000 lux) accelerated the carotenoid losses. At illuminated conditions, degradation followed a first-order (all-trans-lutein, all-trans-violaxanthin and all-trans-neoxanthin) and fractional conversion model (all-trans-ß-carotene and all-trans-antheraxanthin). Carotenoid degradation followed an Arrhenius temperature-dependency, with Ea values lower than 50 kJ/mol. Degradation was shown to be mainly by oxidative reactions. Packaging under minimal oxygen conditions, use of antioxidants (e.g., ascorbic acid), and proper choice of light sources at retail shelves may be considered to optimize the pigment retention in a carotenoid-coloured beverage during storage.

Impact of processing on the production of a carotenoid-rich Cucurbita maxima cv. Hokkaido pumpkin juice.

Pumpkin juice with high carotenoid content can be attractive natural alternative for artificial food colourants. We evaluated the impact of processing treatments on aqueous carotenoid extraction from pumpkins, aiming to enhance carotenoid transfer into the juice fraction. Crushed whole pumpkins were processed by high pressure homogenization (HPH) for mechanical cell disruption, by enzymatic treatment for cell wall polysaccharide degradation or by a pulsed electric field (PEF) treatment for cell membrane electroporation. Processed purees were separated into juice and pomace and carotenoids were quantified by HPLC-DAD. Whereas only 54-60% of the carotenoids in non-processed puree was transferred into the juice, HPH- and enzyme-assisted processing of purees significantly increased juice yields and total soluble solids, and consequently, carotenoid concentrations in these juices up to 90-98% and 72-90%, respectively. No significant improvement was observed for PEF-treated samples. Results obtained can be industrially useful in producing natural colouring plant concentrates as clean-label ingredients.

Modified Rhamnogalacturonan-Rich Apple Pectin-Derived Structures: The Relation between Their Structural Characteristics and Emulsifying and Emulsion-Stabilizing Properties.

In the context of the increasing interest in natural food ingredients, the emulsifying and emulsion-stabilizing properties of three rhamnogalacturonan-rich apple pectin-derived samples were assessed by evaluating a range of physicochemical properties. An apple pectin (AP74) was structurally modified by a ß-eliminative reaction to obtain a RG-I-rich pectin sample (AP-RG). Subsequent acid hydrolysis of AP-RG led to the generation of pectin material with partially removed side chains (in particular arabinose depleted) (AP-RG-hydrolyzed), thus exhibiting differences in rhamnose, arabinose, and galactose in comparison to AP-RG. All samples exhibited surface activity to some extent, especially under acidic conditions (pH 2.5). Furthermore, the viscosity of the samples was assessed in relation to their emulsion-stabilizing properties. In a stability study, it was observed that the non-degraded AP74 sample at pH 2.5 exhibited the best performance among all the apple pectin-derived samples evaluated. This emulsion presented relatively small oil droplets upon emulsion production and was less prone to creaming than the emulsions stabilized by the (lower molecular weight) RG-I-rich materials. The AP-RG and AP-RG-hydrolyzed samples presented a slightly better emulsion stability at pH 6.0 than at pH 2.5. Yet, neither pectin sample was considered having good emulsifying and emulsion-stabilizing properties, indicated by the presence of coalesced and flocculated oil droplets.

Kinetic Modeling of In Vitro Small Intestinal Lipid Digestion as Affected by the Emulsion Interfacial Composition and Gastric Prelipolysis.

This research evaluated the impact of the emulsion interfacial composition on in vitro small intestinal lipolysis kinetics with the inclusion of rabbit gastric lipase resulting in a gastric prelipolysis step. O/w emulsions contained 5% triolein (w/w) and 1% (w/w) of the following emulsifiers: sodium taurodeoxycholate, citrus pectin, soy protein isolate, soy lecithin, and tween 80. Emulsions were subjected to static in vitro digestion and diverse lipolysis species quantified via a HPLC-charged aerosol detector. Single-response modeling indicated that the kinetics of lipolysis in the small intestinal phase were impacted by the emulsion particle size at the beginning of this phase. Multiresponse modeling permitted the elucidation of the lipolysis mechanism under in vitro conditions. The final reaction scheme included enzymatic and chemical conversions. The modeling strategies used in this research allowed to gain more insights into the kinetics and mechanism of in vitro lipid digestion.

In vitro ß-Carotene Bioaccessibility and Lipid Digestion in Emulsions: Influence of Pectin Type and Degree of Methyl-Esterification.

Citrus pectin (CP) and sugar beet pectin (SBP) were demethoxylated and fully characterized in terms of pectin properties in order to investigate the influence of the pectin degree of methyl-esterification (DM) and the pectin type on the in vitro ß-carotene bioaccessibility and lipid digestion in emulsions. For the CP based emulsions containing ß-carotene enriched oil, water and pectin, the ß-carotene bioaccessibility, and lipid digestion were higher in the emulsions with pectin with a higher DM (57%; "CP57 emulsion") compared to the emulsions with pectin with a lower DM (30%; "CP30 emulsion") showing that the DM plays an important role. In contrast, in SBP-based emulsions, nor ß-carotene bioaccessibility nor lipid digestion were dependent on pectin DM. Probably here, other pectin properties are more important factors. It was observed that ß-carotene bioaccessibility and lipid digestion were lower in the CP30 emulsion in comparison with the CP57, SBP32, and SBP58 emulsions. However, the ß-carotene bioaccessibility of CP57 emulsion was similar to that of the SBP emulsions, whereas the lipid digestion was not. It seems that pectin type and pectin DM (in case of CP) are determining which components can be incorporated into micelles. Because carotenoids and lipids have different structures and polarities, their incorporation may be different. This knowledge can be used to engineer targeted (digestive) functionalities in food products. If both high ß-carotene bioaccessibility and high lipid digestion are targeted, SBP emulsions are the best options. The CP57 emulsion can be chosen if high ß-carotene bioaccessibility but lower lipid digestion is desired.