Preliminary study into the factors modulating β-carotene micelle formation in dispersions using an in vitro digestion model

β-Carotene is an active compound associated with prevention of heart disease, cancer and cataracts. For absorption in vivo, β-carotene must be incorporated in mixed micelles. Micelle formulation varies widely and depends on various factors. The aim of this study was to identify and study the main factors governing the bioaccessibility of β-carotene incorporated into dispersions, using an in vitro digestion model. β-Carotene dispersions were prepared by high-pressure homogenization or by combining emulsification and evaporation. The average particle sizes of the dispersions obtained ranged from 45 to 18315 nm. Results show that the concentration of β-carotene, bile extract and pancreatic lipase, pH, and the particle size of the dispersions significantly affected the transfer of β-carotene from dispersions into micelles. The transfer of β-carotene was inversely related to the particle size and the concentration of bile extract and was highest at pH 6 and 0.4 mg/mL pancreatic lipase. Bile salt played different roles depending on the particle sizes of the dispersion. When the mean diameter of β-carotene particle was below 100 nm, the addition of bile extract and pancreatic lipase did not significantly affect bioaccessibility of β-carotene passing through in vitro digestion model. At larger particle sizes, the transfer efficiency of β-carotene increased with bile extract concentration. The outcomes suggest that there is potential to improve the bioavailability of β-carotene by micronizing lipid droplets.     

Relation between in vitro lipid digestion and β-carotene bioaccessibility in β-carotene-enriched emulsions with different concentrations of l-α-phosphatidylcholine

In view of consumer health, it is desirable to promote the bioaccessibility of lipid-soluble compounds like carotenoids, while limiting the lipid intake. The objective of this work was to examine the relation between in vitro lipid digestion and β-carotene bioaccessibility of carrot-based model food emulsions containing water, 5% olive oil enriched with β-carotene (from carrots) and different concentrations (1–2–3–4%) of l-α-phosphatidylcholine (PHC), as an emulsifier. The lipid digestion (hydrolysis of triacylglycerols (TAGS) and incorporation of free fatty acids (FFAs) and monoacylglycerols (MAGs) in the micelles) and the β-carotene bioaccessibility (incorporation of β-carotene in the micelles) were studied after an in vitro digestion procedure wherein the stomach phase was mimicked for 2.0 h (37 °C) and the small intestinal phase was mimicked for 1.0 h, 1.5 h and 2.0 h (37 °C) (both end-over-end rotations). As a consequence, not only the influence of the emulsifier concentration, but also the influence of the duration in the small intestinal phase was investigated in this study. The oil droplet size distributions of the emulsions at different stages of digestion were shown to be dependent on the phosphatidylcholine concentration, but independent on the duration in the small intestinal phase (1.0 h–2.0 h). Furthermore, all TAGs were already hydrolysed into FFAs and MAGs after 1.0 h small intestinal phase and the incorporation of FFAs and MAGs into micelles seemed to reach a maximum for all emulsions (approximately 26.5%), independent on the phoshpatidylcholine concentration and thus on the particle size distributions. Finally, the β-carotene bioaccessibility increased with increasing phosphatidylcholine concentration, ranging from 33.2% to 79.8% for a 1% and 4% PHC emulsion respectively. No significant differences in β-carotene bioaccessibility were however noticed for the different durations in the small intestinal phase tested. In conclusion, a higher phosphatidylcholine concentration in emulsions leads to higher β-carotene bioaccessibility while the incorporation of lipids into micelles did not increase.     

Lycopene and β-carotene transfer to oil and micellar phases during in vitro digestion of tomato and red carrot based-fractions

The natural structural barriers (i.e. chromoplast substructure and cell wall) in conjunction with the carotenoid hydrophobicity were investigated as factors that may play a role governing the carotenoid transfer efficiency from the tomato and red carrot matrices into the oil and micellar phases during digestion. In addition, the effect of thermal processing on the carotenoid transfer during digestion was studied. While the carotenoid transfer efficiency from the matrix into the oil phase was found to be highly determined by the level of bio-encapsulation, the carotenoid hydrophobicity, being lower for lycopene than for β-carotene, was the main factor influencing the transfer from the oil into the micelles. Overall, the incorporation into the micellar fraction represents the critical step for carotenoid bioaccessibility. Moreover, the effect of thermal treatments on these systems depended on carotenoid species, being negative for lycopene and positive for β-carotene.     

Physicochemical characteristics and in vitro bile acid binding and starch digestion of β-glucans extracted from different varieties of Jeju barley

Physicochemical properties of six different varieties of barley and their β-glucans were evaluated along with in vitro bile acid binding and starch digestibility for health beneficial effects. β-Glucan concentrations in less-hulled, beer, black, waxy-naked, naked, and blue barley were 3.44, 3.46, 6.08, 6.75, 6.45, and 5.91%, respectively. Viscosity of waxy-naked barley flour was the highest. While the yield of β-glucan from waxy-naked barley after extraction was 95.49%, less-hulled barley was 70.09%. As the increase of β-glucan purification, in vitro bile acid binding was increased when compared with cholestyramine and cellulose. In vitro starch digestibility of barley flour and the mixture of potato starch with β-glucan were increased by heat and β-glucan concentration. Estimated glycemic index (GI) calculated based on in vitro starch digestibility was decreased by increasing β-glucan. These results suggest that the physicochemical properties of barley were dependent on the variety of barley and especially β-glucan was involved.     

Stability of α-tocopherol, γ-tocopherol and β-carotene during ripening of pasta-filata cheese made from raw and pasteurised milk with different vitamin contents

Ripening stability of α-tocopherol, γ-tocopherol and β-carotene in cheese was evaluated in relation to different milk vitamin content and to the use of either pasteurised or raw milk. Milk from two farms with different management systems was used to obtain different vitamin content. Milk was divided into two parts, of which only one was pasteurised. Four blocks of cheese were made from each batch and ripened for 0, 15, 30, or 60 days at 14–16 °C. There was a notable variation in cheese vitamin levels, with the differences in milk vitamin content due to farm management having the highest impact. Pasteurisation had no effect on cheese vitamin content. Cheese γ-tocopherol and β-carotene content decreased after 30 and 60 days, respectively, whereas α-tocopherol content remained stable. γ-Tocopherol appeared to be the most efficient antioxidant in cheese, followed by β-carotene. Vitamin stability was not influenced by milk vitamin content or pasteurisation.     

Structure and antioxidant activity of Maillard reaction products from α-lactalbumin and β-lactoglobulin with ribose in an aqueous model system

Maillard reaction products (MRPs) were prepared from aqueous model mixtures containing 3% (w/w) ribose and 3% (w/w) of the dairy proteins α-lactalbumin (α-LA) or β-lactoglobulin (β-LG), heated at 95 °C, for up to 5 h. The pH of MRPs decreased significantly during heat treatment of α-LA-Ribose and β-LG-Ribose mixtures from 8.4 to 5.3. The amino group content in MRPs, derived from the α-LA-Ribose and β-LG-Ribose model system, was decreased noticeably during the first hour and did not change thereafter. The loss of free ribose in MRPs was higher for β-LG-Ribose than for α-LA-Ribose. During the Maillard reaction, the concentration of native and non-native α-LA, or β-LG, decreased and the formation of aggregates was observed. Fluorescence intensity of the β-LG-Ribose MRPs reached maximum within 1 h, compared to 2 h for α-LA-Ribose MRPs. Meanwhile, modification of the UV/vis absorption spectra for α-LA and β-LG was mainly due to a condensation reaction with ribose. Dynamic light scattering showed a significant increase in the particle size of the MRPs. Size exclusion chromatography of MRPs revealed the production of both high and low molecular weight material. Electrophoresis of MRPs indicated polymerization of α-LA and β-LG monomers via inter-molecular disulfide bridge, but also via other covelant bonds. MRPs from α-LA-Ribose and β-LG-Ribose exhibited increased antioxidant activities, therefore theses MRPs may be used as natural antioxidants in food products.     

A chemical study of β-carotene oxidation by ozone in an organic model system and the identification of the resulting products

Carotenoids are present in many foods. Due to their polyenic chains, they undergo oxidation reactions which may give several compounds. Ozone, a powerful antimicrobial agent, is applied in the food industry due to its high reactivity and penetrability. This work presents a chemical study of the degradation of β-carotene in solutions, under the influence of ozone. The experiments were carried out at ozone concentrations ranging from 0.8 to 2.5 ppm and the β-carotene solutions were sampled and analysed from zero to seven hours of reaction. The oxidation products were collected in C18 cartridges coated with dinitrophenylhydrazine and the hydrazones formed were analysed by LC-MS. The oxidation reaction was found to follow a zero order kinetic model and the β-carotene decay ranged between 17.2% and 99.8%. Fourteen oxidation products were tentatively identified, amongst them eight which had not been cited yet in the literature as oxidation products of β-carotene.     

Effect of 3 lactobacilli on immunoregulation and intestinal microbiota in a β-lactoglobulin–induced allergic mouse model

Milk is one of the earliest and most common allergen sources in the world, with β-lactoglobulin representing a major allergen protein. Numerous studies have reported that probiotics exert antiallergic and anti-inflammatory effects. Here, we examined the effects of 3 strains of Lactobacillus on immunomodulatory functions, intestinal barrier functions, and intestinal microbiota through a β-lactoglobulin–induced allergic mouse model. We found that the oral administration of Lactobacillus plantarum ZDY2013 and Lactobacillus rhamnosus GG suppressed allergic response, attenuating serum IgE and relieving anaphylaxis symptoms. The 3 strains of Lactobacillus could induce T helper (Th) 1 or T regulatory cells to differentiate to inhibit the Th2-biased response for regulating Th1/Th2 immune balance. Furthermore, L. plantarum ZDY2013 and L. rhamnosus GG enhanced intestinal barrier function through the regulation of tight junction. We also found that L. plantarum ZDY2013 and L. plantarum WLPL04 could regulate alterations in intestinal microbiota caused by allergies. In particular, Rikenella, Ruminiclostridium, and Lachnospiraceae UCG-006 were considerably reduced after treatment with L. plantarum ZDY2013 and L. plantarum WLPL04. These results suggested that 3 Lactobacillus strains may serve as an effective tool for the treatment of food allergies by regulating immune and gut microbiota.     

Simultaneous and confirmative detection of multi-residues of β2-agonists and β-blockers in urine using LC-MS/MS/MS coupled with β-receptor molecular imprinted polymer SPE clean-up

A liquid chromatography–linear ion-trap spectrometry (LC-MS³) method using β-receptor molecular-imprinted polymer (MIP) solid-phase extraction (SPE) as clean-up was developed to determine simultaneously and confirmatively residues of 25 β₂-agonists and 21 β-blockers in urine samples. Urine samples were subjected to enzymatic hydrolysis by β-glucoronidase/arylsulphatase, and then extracted with perchloric acid. Sample clean-up was performed using β-receptor MIP SPE. A Supelco Ascentis^® express Rp-Amide column was used to separate the analytes, and MS³ detection used an electrospray ionisation source in positive-ion mode. Recovery studies were carried out using blank urine samples fortified with the 46 analytes at the levels of 0.5, 1.0 and 2.0 μg l^–1. Recoveries were obtained ranging from 60.1% to 109.9% with relative standard deviations (RSDs, n = 7) from 0.5% to 19.4%. The limits of detection (LODs) and limits of quantitation (LOQs) of the 46 analytes in urine were 0.02–0.18 and 0.05–0.60 μg l^–1, respectively. As a result of the selective clean-up by MIP SPE and MS³ detection of the target drugs, the sensitivity and accuracy of the present method was high enough for monitoring β₂-agonist and β-blocker residues in urine samples. Satisfactory results were obtained in the process of the determination of positive urine samples.     

Water dispersibility of the β-carotene source and its effect on the physical,thermal, and in vitro release properties of an inclusion complex

There is a need to understand the effect of in vitro digestion on the release of β-carotene (BC) from inclusion complexes (IC) prepared with water-dispersible (WDBC) or hexane-soluble (HSBC) sources. IC were prepared by co-precipitating WDBC or HSBC with β-cyclodextrin and characterised using imaging, spectral (FTIR) and thermal techniques. Two solvent systems (absolute ethanol, AE and 4:3 ethanol–hexane, EH) were employed to compare extractability of BC during simulated co-digestion with rice as food matrix. WDBC-IC had greater yield (89% vs. 44%) but lower encapsulation efficiency (30% vs. 89%) than HSBC-IC. Imaging, FTIR and thermal data confirm inclusion complex formation. In vitro WDBC release extracted with AE decreased uniformly and was unaffected by rice, whereas HSBC extracted with EH exhibited gradual release towards intestinal digestion but decreased with rice. Fabricated microcapsules with suitable forms of BC may be applied for food fortification purposes.     

Encapsulation of β-carotene in 2-hydroxypropyl-β-cyclodextrin/carrageenan/soy protein using a modified spray drying process

The objective of this study was to produce ternary complex encapsulates to improve solubility and bioaccessibility of the β-carotene (β-c). A modified spray drying process was used to produce encapsulated β-c particles with 2-hydroxypropyl-β-cyclodextrin (HβCD), carrageenan (CRG), and soy protein (SP). Physicochemical properties, dissolution behaviour, and in vitro gastrointestinal (GI) stability of the encapsulate were studied. The optimised ternary complex (β-c/HβCD/CRG/SP) particles had a spherical shape with a particle size of 726.1 nm, polydispersity index (PDI) of 0.41, the zeta potential of −45 mV, and higher retention of β-c with 84% encapsulation efficiency. Differential scanning calorimetry (DSC) analysis revealed that encapsulates are in the amorphous state and had excellent solubility. The bioaccessibility of β-c was found to be significantly higher for β-c/HβCD/CRG/SP (78%), compared to β-c/HβCD (34%) and β-c/HβCD/CRG (62%), in the in vitro study. This study concludes that the HβCD/CRG/SP ternary complex facilitates a potential delivery system for hydrophobic bioactive compounds with improved solubility and bioaccessibility.     

Comparison of volatiles in raw and boiled potatoes using a mild extraction technique combined with GC odour profiling and GC–MS

Aromas of raw and boiled potatoes were compared using a mild extraction technique, ensuring preservation of the very labile composition of potato aroma during analysis. The extracts were evaluated by GC–MS and GC-sniffing using a panel of four judges. A total of 29 compounds were identified by GC–MS in raw potatoes and 25 in boiled. Twenty compounds were found in both raw and boiled potatoes, but most often in very different concentrations. During GC-sniffing, 33 odours were detected in boiled, and 27 in raw potatoes. Eight odours corresponded to compounds identified by GC–MS. Another four odours could be tentatively identified. It is concluded that the change in aroma during boiling of potatoes depends on compounds from lipid oxidation as well as compounds from other types of reactions, for instance the Strecker-degradation.     

Effect of oral administration of β-glucans derived from Aureobasidium pullulans SM-2001 in model mice and rat with atopic dermatitis-like phenotypes

Food Science and Biotechnology - In this study, we investigated the anti-atopic dermatitis (AD) activity of β-glucans derived from Aureobasidium pullulans SM-2001 (βGdAP). βGdAP was...     

Short communication: Inhibition of angiotensin 1-converting enzyme by peptides derived from variants of bovine β-casein upon apical exposure to a Caco-2 cell monolayer

This study investigated the consequence of genetically contingent amino acid substitutions in bovine β-casein (CN) genetic variants A¹, A², B, and I on the structure and bioactive potential of peptides following in vitro digestion. The β-CN variants were digested in vitro using pepsin and pancreatin, and a peptide profile was obtained by liquid chromatography tandem mass spectrometry, revealing among others, the β-casomorphin precursor peptides VYPFPGPIHN and VYPFPGPIPN, derived from variant A¹/B and from A²/I, respectively. These 2 peptides were synthesized and assessed for angiotensin 1-converting enzyme (ACE) inhibitory capacity before and after incubation with a monolayer of Caco-2 intestinal cells. The VYPFPGPIHN was a stronger ACE inhibitor than VYPFPGPIPN, with the concentration needed to reach half-maximal inhibition (IC₅₀) of 123 ± 14.2 μM versus 656 ± 7.6 μM. Exposure to a Caco-2 intestinal cell monolayer did not affect ACE inhibition by VYPFPGPIHN, but resulted in an almost 2-fold increase in inhibition by VYPFPGPIPN after incubation. Subsequent tandem mass spectrometric analysis identified the truncated peptide VYPFPGPIP, suggesting hydrolysis by a cell membrane associated peptidase. Thus, genetic variation in bovine β-CN results in the generation of peptides that differ in bioactivity, and are differently affected by intestinal brush border peptidases.     

Monitoring and identification of bacteria associated with safety concerns in the manufacture of São Jorge, a Portuguese traditional cheese from raw cow's milk

The aim of this study was to assess the hygienic quality of raw milk used in the manufacture of S?o Jorge, a Protected Denomination of Origin Portuguese semihard cheese, as well as to ascertain the sanitary conditions prevailing during its processing. Viable counts of Enterobacteriaceae and Micrococcaceae were accordingly obtained, pertaining to 21 independent batches (including samples of raw milk, curd, and cheeses after 1, 3, and 4 months of ripening), from 7 dairy farms. Standard plate counts (log CFU per milliliter or per gram) ranged from 6.1 to 8.6 in raw milk, whereas they ranged from 7.0 to 8.0 in 4-month-old cheeses. Viable counts of Enterobacteriaceae ranged between 5.9 and 7.0 in raw milk and between 0.0 and 1.3 in 4-month-old cheeses. Species identified within this family encompassed Klebsiella oxytoca, Klebsiella cloacae, Klebsiella pneumoniae, Enterobacter sakazakii, and Escherichia coli; Klebsiella ornithinolytica, Klebsiella terrigena, and Serratia odorifera were detected only in raw milk. No Salmonella whatsoever could be detected in any of the samples. Viable counts of Micrococcaceae ranged between 4.7 and 5.9 and between 1.3 and 3.3 in raw milk and 4-month-old cheeses, respectively. Species identified within this family encompassed Staphylococcus sciuri, Staphylococcus epidermidis, Staphylococcus saprophyticus (which was found mainly in ripened cheeses), and Staphylococcus aureus (which was not detected in 4-month-old cheeses). Accompanying physicochemical analyses included determination of moisture, salt, and pH. Statistical analyses revealed a negative correlation between salt content and viable numbers of Enterobacteriaceae in cheese, whereas in the case of Micrococcaceae, a more negative correlation was found between viable numbers and moisture content than between viable numbers and pH. The results of our study indicate, in general, poor milk handling conditions in all farms, given that the indicators total mesophile and Enterobacteriaceae counts were high, between 100- and 1,000-fold those enforced by international standards pertaining to the matrices in question. However, by the time of regular consumption (i.e., after 4 months of ripening), S?o Jorge cheeses exhibit low levels of contamination by Enterobacteriaceae and S. aureus, as well as absence of Salmonella.     

Physicochemical properties, α-amylase and α-glucosidase inhibitory effects of the polysaccharide from leaves of Morus alba L. under simulated gastro-intestinal digestion and its fermentation capability in vitro by human gut microbiota

The investigation aimed at determining the impact of sequential simulated digestion on the physicochemical properties and digestive enzymes inhibitory effects of the polysaccharides fraction (MLP-2) of Morus alba L. leaves as well as its in vitro fermentation behaviours. After artificial salivary, gastric and intestinal digestions, the chemical components and microstructure of MLP-2 were altered with significantly (P < 0.05) decreased molecular weight. The α-amylase and α-glucosidase inhibitory activities of MLP-2 were significantly (P < 0.05) improved throughout simulated digestion. MLP-2I, the intestinal digested fraction of MLP-2, could significantly (P < 0.05) decrease the pH value of fermented culture and increase the short-chain fatty acids (SCFA) concentrations, especially acetic, propionic and butyric acids. In conclusion, MLP-2 could be gradually degraded under simulated digestion with altered physicochemical properties and enhanced α-amylase and α-glucosidase inhibitory effects, and further utilised by human gut microbiota to decrease pH value and promote SCFA production.