In vitro macronutrient digestibility and mineral bioaccessibility of lentil-based pasta: The influence of cellular intactness.

Recently, pulse ingredients with (partial) cellular intactness are put forward as promising innovative food ingredients with slowed macronutrient digestibility. This study compared cooking quality and nutrient (starch, protein, and mineral) digestibility/bioaccessibility of lentil-based pasta prepared from 100% raw-milled flour, and by substituting 30% of the formulation by isolated cotyledon cell powder or whole precooked powder. Formulation had little effect on cooking properties. Both amylolysis and proteolysis were significantly slowed by incorporating cellular ingredients: towards the end of simulated digestion, amylolysis was lowered by 16-25%, while differences in proteolysis became small. Cellular ingredient incorporation slightly decreased Zn and Mg but did not affect Ca and Fe bioaccessibility, overall yielding a low mineral bioaccessibility comparable to cooked whole pulses. To conclude, lentil-based pasta substituted with cellular ingredients showed improved nutritional properties (i.e., high in digestible protein and slowed amylolysis), with perspectives for the development of different innovative foods with targeted nutritional properties.

Effect of manufacturing conditions on in vitro starch and protein digestibility of (cellular) lentil-based ingredients.

(Cellular) pulse powders are being proposed as ingredients for different foods. However, the effect of manufacturing conditions on the properties of those powders remained unknown. Therefore, this study investigated the effect of specific manufacturing conditions (cooking time, application of cell isolation, and drying method) on the composition, microstructure, and in vitro starch and protein digestibility of lentil powders. Next to powders consisting of isolated cotyledon cells (ICC), this study proposes the production of precooked whole lentil powders (WL), without a cellular isolation step. In a model food system (heat-treated suspension), starch and protein digestion were significantly attenuated for both WL and ICC compared to raw-milled lentil flour. The applied cooking time determined macronutrient digestibility in the powders by (i) affecting the susceptibility of ICC to in vitro digestion, and (ii) determining the microstructural properties of WL. Freeze-dried ICC powder showed a stronger attenuation of amylolysis compared air-dried ICC. This study showed that WL powders have an important potential as innovative food ingredients higher in fiber but lower in starch compared to ICC.

Studying semi-dynamic digestion kinetics of food: Establishing a computer-controlled multireactor approach.

In this work, a multireactor system to study digestion (MuReDi) kinetics is introduced. For this, a custom-made automated system with four independent syringe pumps (BioXplorer 100, H.E.L Group) was acquired. This system consists of multiple, small-scale reactors allowing to study digestion as a function of time and thus to determine digestion kinetics. The different digestion conditions used in the oral, gastric, and small intestinal phase were based on the digestion protocols published by the INFOGEST consortium. We showed that the minimum working volume of a reactor is 30 mL. Besides, repeatability of the digestion kinetics was shown for two food systems: a liquid Ensure® Plus Vanilla drink, and a solid, cooked lentil sample. When comparing static digestion kinetics with semi-dynamic ones, a significantly different digestion pattern was observed. In the static case, a relatively fast hydrolysis rate was observed until a clear plateau was reached. Oppositely, for the semi-dynamic case, a delayed start of the hydrolysis process was noticed. In the gastric phase, this was explained by the decreasing pH and the large pH dependency of pepsin activity. In the small intestine, the lag phase was relatively shorter, yet clearly present. Here we related it to the gradual enzyme (and bile salt) secretion that had to diffuse towards the substrate before hydrolysis could start. Generally, this work showed that the MuReDi system could be used to perform a semi-dynamic digestion approach which largely impacted the overall digestion kinetics. This is important to consider in future in vitro food digestion simulation work to come closer to physiologically relevant digestion kinetics.

Digestion kinetics of lipids and proteins in plant-based shakes: Impact of processing conditions and resulting structural properties.

In this work, plant-based shakes were prepared (5% oil, 6% protein, 1% lecithin, 88% water) (w/w) using two processing techniques (i) only mixing versus (ii) mixing followed by high pressure homogenisation, as well as two processing sequences (i) adding all ingredients together versus (ii) stepwise addition of ingredients. Shakes only mixed consisted of large, irregular particles (1-100 µm). Eventually, this resulted in a relatively low lipid and protein digestion extent after 2 h of gastric pre-digestion (9% and < 1%, respectively). In contrast, shakes that were subjected to high pressure homogenisation displayed small, homogeneous particles (<10 µm). Besides, lipids and proteins were digested to a high extent in the stomach (40% and 10%, respectively). The small intestinal digestion kinetics indicated a significant impact of proteins on lipid digestion kineticsbutno significant effect of lipids on protein digestion kinetics. The results highlighted the relevance of food processing on macronutrient (micro)structure and further gastrointestinal functionality.

Enzymatic and chemical conversions taking place during in vitro gastric lipid digestion: The effect of emulsion droplet size behavior.

This investigation reports the effect of droplet size behavior on the overall lipolysis profile and molecular lipolysis mechanisms under in vitro gastric conditions. O/W emulsions (5% triolein, 1% sodium taurodeoxycholate) with different initial droplet sizes (fine: 0.58 µm; medium: 1.82 µm; and large: 4.00 µm) were subjected to static in vitro digestion. For the first time, multiple lipolysis products including diolein and monoolein regioisomers were quantified within a single HPLC run. An inverse relation was found between the droplet size and the initial rate and final extent of lipolysis based on the digested triolein. Furthermore, a mechanistic gastric lipolysis model was established based on a reaction scheme including enzymatic and chemical isomerization conversions. The estimated rate of the sn-1/3 hydrolysis was around two- to thirty-fold faster compared to the rates of sn-2 cleavage and isomerization, respectively. These findings resulted in a profound insight in in vitro gastric molecular lipolysis mechanisms.

Pectin influences the kinetics of in vitro lipid digestion in oil-in-water emulsions.

Oil-in-water emulsions were prepared with 5% (w/v) carrot-enriched olive oil and stabilized with Tween 80 (TW), phosphatidylcholine (PC), citrus pectin (CP) or a combination of these emulsifiers. Additionally, the methylesterification degree (DM) of citrus pectin was modified, resulting in three different studied pectin structures: CP82, CP38 and CP10. All initial emulsions presented small initial oil droplet sizes and were submitted to an in vitro simulated gastric and small intestinal phase. The latter was executed in a kinetic way to determine the time dependency of the lipolysis reaction, micelle formation and carotenoid bioaccessibility. The results showed that the pectin DM mainly influenced the reaction rate constants, while the emulsifier (combination) determined the extent of lipolysis and carotenoid bioaccessibility. Moreover, a direct relation was observed between the lipolysis reaction and bioaccessibility extent. The presented study showed that targeted emulsion design can be used to tailor lipid digestion kinetics.

Emulsion stability during gastrointestinal conditions effects lipid digestion kinetics.

Oil-in-water emulsions were prepared with carrot- or tomato-enriched olive oil (5%w/v) and stabilized with Tween80 or sucrose esters (0.5%w/v) with different hydrophilic-lipophilic balance (8; 11 or 16). All emulsions had similar initial oil droplet sizes and were submitted to simulated gastrointestinal conditions using a kinetic digestion procedure. Sucrose esters induced an unstable system after gastric conditions leading to coalesced oil droplets, while Tween80 emulsions remained stable. Emulsion particle sizes at the end of the gastric phase were directly associated with the lipolysis kinetics during the intestinal phase. Moreover, a direct relationship was observed between lipolysis and carotenoid micellarisation for all emulsions, and depended mainly on the surfactant structure used. Tween80 emulsions led to a higher lipolysis extent (53-57%) and carotenoid bioaccessibility (17-42%) compared to sucrose ester emulsions (33-52% and 9-27%, respectively). These findings show the importance of the emulsifier structure and emulsion stability during gastrointestinal conditions in modulating lipolysis kinetics.

Lipid digestion, micelle formation and carotenoid bioaccessibility kinetics: Influence of emulsion droplet size.

Carotenoid-enriched oil-in-water emulsions with different droplet sizes (small: d43 0.72µm; medium: d43 1.9µm; large: d43 15.1µm) were subjected to simulated gastrointestinal conditions. The kinetics of lipolysis, micelle formation and carotenoid bioaccessibility were monitored during the intestinal phase. The rates of all three processes increased with decreasing droplet size. The large droplet size emulsion contained undigested oil at the end of digestion, whereas an almost complete hydrolysis was observed for the other two emulsions. The sub-micron emulsion presented a higher conversion of MAGs to FFAs during digestion, which led to a higher concentration of FFAs in the mixed micelles. The incorporation of carotenoids into mixed micelles occurred faster and reached a higher final value for the small droplet size emulsion, leading to final carotenoids bioaccessibility values of around 70%. This work provides valuable information for developing in silico models to simulate the lipid digestibility and carotenoid bioaccessibility.