Time maters: Exploring the dynamics of bioactive compounds content, bioaccessibility and antioxidant activity during Lupinus angustifolius germination.

Germination is a process that enhances the content of health-promoting secondary metabolites. However, the bioaccessibility of these compounds depends on their stability and solubility throughout the gastrointestinal tract. The study aimed to explore how germination time influences the content and bioaccessibility of γ-aminobutyric acid and polyphenols and antioxidant capacity of lupin (Lupinus angustifolius L.) sprouts during simulated gastrointestinal digestion. Gamma-aminobutyric acid showed a decrease following gastrointestinal digestion (GID) whereas phenolic acids and flavonoids exhibited bioaccessibilities of up to 82.56 and 114.20%, respectively. Although the digestion process affected the profile of phenolic acids and flavonoids, certain isoflavonoids identified in 7-day sprouts (G7) showed resistance to GID. Germination not only favored antioxidant activity but also resulted in germinated samples exhibiting greater antioxidant properties than ungerminated counter parts after GID. Intestinal digests from G7 did not show cytotoxicity in RAW 264.7 macrophages, and notably, they showed an outstanding ability to inhibit the production of reactive oxygen species. This suggests potential benefit in mitigating oxidative stress. These findings contribute to understand the dynamic interplay between bioprocessing and digestion in modulating the bioaccessibility of bioactive compounds in lupin, thereby impacting health.

Probiotic Encapsulation: Bead Design Improves Bacterial Performance during In Vitro Digestion.

The stability and release properties of all bioactive capsules are strongly related to the composition of the wall material. This study aimed to evaluate the effect of the wall materials during the encapsulation process by ionotropic gelation on the viability of Lactobacillus fermentum K73, a lactic acid bacterium that has hypocholesterolemia probiotic potential. A response surface methodology experimental design was performed to improve bacterial survival during the synthesis process and under simulated gastrointestinal conditions by tuning the wall material composition (gelatin 25% w/v, sweet whey 8% v/v, and sodium alginate 1.5% w/v). An optimal mixture formulation determined that the optimal mixture must contain a volume ratio of 0.39/0.61 v/v sweet whey and sodium alginate, respectively, without gelatin, with a final bacterial concentration of 9.20 log10 CFU/mL. The mean particle diameter was 1.6 ± 0.2 mm, and the experimental encapsulation yield was 95 ± 3%. The INFOGEST model was used to evaluate the survival of probiotic beads in gastrointestinal tract conditions. Upon exposure to in the vitro conditions of oral, gastric, and intestinal phases, the encapsulated cells of L. fermentum decreased only by 0.32, 0.48, and 1.53 log10 CFU/mL, respectively, by employing the optimized formulation, thereby improving the survival of probiotic bacteria during both the encapsulation process and under gastrointestinal conditions compared to free cells. Beads were characterized using SEM and ATR-FTIR techniques.

A Simple Validated Method for the Estimation of Pepsin Activity in Microtiter Array for the INFOGEST Protocol.

The INFOGEST protocol has been widely used as a static in-vitro simulation of gastrointestinal food digestion for bioaccessibility assessments on bioactive compounds. The standardization of the activity of several enzymes, such as pepsin, via UV-spectrophotometry of digested hemoglobin at 280 nm is a key step in the protocol. Standardization is a crucial stage since it is necessary to determine the quantity of enzyme to be added to the sample for digestion. However, this method is yet to be analytically validated; it requires quartz cuvettes and large volumes of samples and is time-consuming. Thus, we reviewed and adapted a well-known colorimetric method in microplates array by using the Folin-Ciocalteu reagent, and this study is the first to report for miniaturization of this method, the advantages of which include its automation, ease of use, the low volume of samples required, the minimal use of reagents, and speed. This method was compared to the traditional UV method, and the comparison results show no statistical difference between the inter day means for each group (p > 0.05). The proposed method was validated, showing high reproducibility (8% as inter-day CV) and statistically comparable results with the traditional UV spectrophotometric method.

Compartmentalization of lutein in simple and double emulsions containing protein nanoparticles: Effects on stability and bioaccessibility.

Delivery systems designed through protein stabilized emulsions are promising for incorporating carotenoids in different products. Nevertheless, the versatility in structures of such systems raises questions regarding the effect of the bioactive compound localization on their bio-efficacy, in particular for double emulsions. In this context, the aims of this study were to determine the impact of the localization of lutein in different water/oil/water double emulsions versus a single oil/water emulsion on the stability and in vitro bioaccessibility of lutein, a lipophilic carotenoid. The inner aqueous phase, which contained whey protein isolate (WPI) nanoparticles obtained by desolvation, was emulsified in sunflower oil stabilized by polyglycerol polyricinoleate (PGPR). The primary emulsion was then emulsified in a continuous aqueous phase containing whey protein isolate (WPI) and xanthan gum, the latter to increase the viscosity of the outer phase and delay creaming. Lutein was incorporated using different strategies: (1) lutein entrapped by WPI nanoparticles within the inner water phase of a double emulsion (W-L/O/W); (2) lutein incorporated into the oil phase of the double emulsion (W/O-L/W); (3) lutein incorporated in the oil phase of a single emulsion (O-L/W). All systems contained similar whey protein concentrations, as well as all other stabilizers. W-L/O/W sample showed the lowest lutein stability against light exposure during storage, and the highest lutein bioaccessibility after in vitro digestion, for freshly made samples. Furthermore, the in vitro bioaccessibility of lutein incorporated into the single emulsion was considerably lower than those observed for the double emulsions. The results reinforce the importance of designing appropriate structures for delivering improved stability and bioaccessibility of bioactive compounds.

Bioaccessibility of phenolic compounds using the standardized INFOGEST protocol: A narrative review.

The INFOGEST protocol creation was a watershed for phenolic bioaccessibility studies. Because of this important initiative to standardize bioaccessibility studies, data comparisons between different laboratories are now expedited. It has been eight years since the INFOGEST protocol creation, and three from the latest update. However, the current status in terms of phenolic bioaccessibility and how far different laboratories are from reaching a consensus are still unrevealed. In this sense, this narrative review considered an evaluation of different studies that applied the INFOGEST protocol to investigate the bioaccessibility of phenolic compounds. The central objective was to compile the main findings and consensus and to identify possible gaps and future opportunities. This approach intends to further facilitate the use of this protocol by professionals in the field of food science and technology and related areas, generating a reflection on the actual level of standardization of the method. Despite the differences in phenolic compounds from diverse food matrices, and their peculiar behavior, some trends could be elucidated, in terms of phenolic release, stability, and/or transformation upon in vivo digestion. In contrast, there was no general consensus regarding sample preparation, how to report results and the form to calculate bioaccessibility, making it difficult to compare different studies. There is still a long road to effectively standardize the results obtained for phenolic bioaccessibility using the INFOGEST protocol, which is also an opportunity in terms of food analysis that can impact the food industry, especially for the development of nutraceuticals and functional foods.

A guide for the evaluation of in vitro bioaccessibility of carotenoids.

Ongoing efforts to improve the nutritional content of carotenoids in food sources to solve global health problems must also consider bioaccessibility and bioavailability of carotenoids. Carotenoid bioaccessibility encompasses carotenoid release from the food matrix, solubilization in lipid emulsion droplets, and transfer to mixed micelles for further absorption into the intestine. Bioavailability refers to the fraction of carotenoids which enters into circulation. To understand the mechanisms controlling bioaccessibility, in vitro digestion methods have been developed and are a useful approach to overcome some of the disadvantages associated with in vivo protocols. The INFOGEST protocol comprises three phases (oral, gastric and intestinal) to simulate adult human digestion of foods, as an international consensus for the conditions of static in vitro digestion. The last steps in any digestion protocol involve isolation of the target food compounds and subsequent analysis, steps not included in the INFOGEST protocol because they may differ depending on specific physical-chemical characteristics and quantification analysis of the target analyte. In this chapter, we describe the INFOGEST protocol which has been adapted for the analysis of carotenoid bioaccessibility, including determination and calculation of enzyme activities according to INFOGEST guidelines, removal of undigested material, collection of mixed micelles containing carotenoids, extraction, storage and subsequent identification and quantification by liquid chromatography analysis of both free and acylated carotenoids.

Characterization and the impact of in vitro simulated digestion on the stability and bioaccessibility of carotenoids and their esters in two Pouteria lucuma varieties.

Lucuma is a starchy orange-yellow fruit native to the Andean region. It is widely consumed in Latin America and has been recently adapted to the agronomical characteristics of the south region of Spain. However, its carotenoid profile has never been reported. The aim of this study was to characterize the carotenoid and carotenoid ester composition of lucuma pulps (var. Molina and Beltran) and assess their bioaccessibility with an in vitro simulated gastrointestinal digestion according to the INFOGEST® methodology. The carotenoid profile in lucuma pulps revealed a high qualitative diversity composed of 33 compounds, corresponding to 9 free xanthophylls, 9 hydrocarbon carotenes and 15 xanthophyll esters. (13Z)-violaxanthin, (all-E)-violaxanthin and (all-E)-antheraxanthin were the most abundant carotenoids in lucuma fruits and were naturally present as xanthophyll esters: (all-E)-antheraxanthin 3-O-palmitate, (all-E)-violaxanthin laurate and (all-E)-violaxanthin palmitate. Carotenoids were stable during in vitro digestion; however, their release from the food matrix was limited which contributed to their low bioaccessibility.

Comparison of Two Static in Vitro Digestion Methods for Screening the Bioaccessibility of Carotenoids in Fruits, Vegetables, and Animal Products.

In vitro digestion methods are routinely used to assess the bioaccessibility of carotenoids and other dietary lipophilic compounds. Here, we compared the recovery of carotenoids and their efficiency of micellarization in digested fruits, vegetables, egg yolk, and salmon and also in mixed-vegetable salads with and without either egg yolk or salmon using the static INFOGEST method22 and the procedure of Failla et al.16 Carotenoid stability during the simulated digestion was ≥70%. The efficiencies of the partitioning of carotenoids into mixed micelles were similar when individual plant foods and salad meals were digested using the two static methods. Furthermore, the addition of cooked egg or salmon to vegetable salads increased the bioaccessibility of some carotenoids. Our findings showed that the two methods of in vitro digestion generated similar estimates of carotenoid retention and bioaccessibility for diverse foods.