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BACKGROUND: A colorimetric method for the quantification of hydrogen sulfide (H2S) produced in microbial fermentations was developed using lead gelled alginate microparticles packed in glass columns. The formation of a lead sulfide complex, between H2S and lead ion (Pb2+) immobilized on the microparticles, allowed simple and accurate quantification by colorimetry. RESULTS: The microparticle-loaded columns were calibrated and showed significant analytical sensitivity. The calibration curve of the system showed a correlation coefficient (r2) of 0.995 and a detection limit of 1.29 ± 0.02 µg L-1. The application of the columns in laboratory wine fermentations was able to detect variations in H2S production from 10.6 to 23.5 µg L-1 by increasing the sugar content in the medium, and from 10.6 to 3.2 µg L-1 with decreasing nitrogen content in the medium. CONCLUSION: Validation of the proposed method was carried out by determining H2S in a vinic fermentation model, the results of which were compared with those obtained using a reference chemical method. The data obtained showed no statistically significant differences between the two methods, confirming the reliability and accuracy of the developed system. © 2024 Society of Chemical Industry.

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A common challenge in hydrogel-based delivery systems is the premature release of low molecular weight encapsulates through diffusion or swelling and reduced cell viability caused by the low pH in gastric conditions. A second biopolymer, such as chitosan, can be incorporated to overcome this. Chitosan is usually associated with colonic drug delivery systems. We intended to formulate chitosan-coated pectin beads for use in delaying premature release of the encapsulate under gastric conditions but allowing release through disintegration under intestinal conditions. The latter is of utmost importance in delivering most functional food ingredients. Therefore, this study investigated the impact of formulation and process conditions on the size, sphericity, and dissolution behavior of chitosan-coated hydrogel beads prepared by interfacial coacervation. The size and sphericity of the beads depend on the formulation and range from approximately 3 to 5 mm and 0.82 to 0.95, respectively. Process conditions during electro-dripping may be modulated to tailor bead size. Depending on the voltage, bead size ranged from 1.5 to 4 mm. Confocal laser scanning microscopy and scanning electron microscopy confirmed chitosan shell formation around the pectin bead. Chitosan-coated beads maintained their size and shape in simulated gastric fluid but experienced structural damage in simulated intestinal fluid. Therefore, they represent a novel delivery system for functional food ingredients.

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Alginate encapsulates loaded with clove essential oil (CEO) were prepared by ionic gelation, with subsequent freeze-drying. The objective of the present work was to develop a product with the ability to protect CEO against its easy volatility and oxidation. The following techniques were used to characterize the formulations: eugenol release, degree of swelling, GC/MS, TGA/DSC, and SEM. The alginate solution (1.0%) containing different concentrations of CEO (LF1: 1.0%; LF2: 0.5%; LF3: 0.1%) was dropped into a 3.0% CaCl2 solution. After lyophilization, the encapsulated samples were wrinkled and rigid, with high encapsulation power (LF3: 76.9% ± 0.5). Three chemical components were identified: eugenol (the major one), caryophyllene, and humulene. The antioxidant power (LF1: DPPH IC50 18.1 µg mL-1) was consistent with the phenol content (LF1: 172.2 mg GAE g-1). The encapsulated ones were thermally stable, as shown by analysis of FTIR peaks, eugenol molecular structure was kept unaltered. The degree of swelling was 19.2% (PBS). The release of eugenol (92.5%) in the PBS solution was faster than in the acidic medium. It was concluded that the low-cost technology used allows the maintenance of the content and characteristics of CEO in the three concentrations tested, offering a basis for further research with essential oil encapsulates.

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In situ forming hydrogels are promising for biomedical applications, especially in drug delivery. The precursor solution can be injected at the target site, where it undergoes a sol-gel transition to afford a hydrogel. In this sense, the most significant characteristic of these hydrogels is fast gelation behavior after injection. This study describes an all-polysaccharide, rapidly in situ-forming hydrogel composed of carboxymethyl chitosan (CMCHT) and hydroxyethyl cellulose functionalized with aldehyde groups (HEC-Ald). The HEC-Ald was synthesized through acetal functionalization, followed by acid deprotection. This innovative approach avoids cleavage of pyran rings, as is inherent in the periodate oxidation approach, which is the most common method currently employed for adding aldehyde groups to polysaccharides. The resulting hydrogel exhibited fast stress relaxation, self-healing properties, and pH sensitivity, which allowed it to control the release of an encapsulated model drug in response to the medium pH. Based on the collected data, the HEC-Ald/CMCHT hydrogels show promise as pH-sensitive drug carriers.

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The effect of the cold-set and heat-set gelling mechanism of whey protein isolate on bigel production was assessed. For this purpose, hydrogel phase was produced with whey protein isolated (10 % w/v) and for oleogel sunflower oil and glycerol monostearate (7.5 % w/v) were used. Bigels were produced by hot emulsification of different hydrogel:oleogel ratios (from 90:10 up to 10:90). For cold-set bigels (CSB) NaCl (200 mM) was added to the aqueous phase prior to the emulsification and the emulsion was cooled to promote the 3D network formation. On the other hand, heat-set bigels (HSB) were produced by heating the emulsion (80 °C, 60 min). Bigels were evaluated through microscopy, FTIR, thermal and texture analyzes. Results showed that depending on the hydrogel:oleogel ratio and gelling mechanism different structures organization were obtained. CSB were more organized, showing that the rate of gelation was the mechanism responsible for the structure. However, for HSB the heat treatment destabilized the emulsion and disorganized structures were observed for high oleogel content. FTIR corroborates the visual observation and showed that the arrangement was purely physical. In addition, the structural arrangement led to different mechanical properties. In general, HSB produced gels with rubber-like behavior, higher elasticity modulus and the presence of a breaking point. In contrast, CSB behaves as squeezing gel, with no breaking point and lower values of elasticity modulus. Moreover, for O/W bigels the dispersed oleogel particles disrupted the WPI network decreasing the gel strength in comparison to pure hydrogels. However, for systems where oleogel was the continuous phase, the gel strength was recovered due to the metastable and dynamic character of these systems. Thus, results showed that the gelling mechanism of the protein exerted an effect on the physical properties of bigels. In addition, the mechanical properties also can be modulated according to the bigel composition, allowing its application in products with different sensorial characteristics.

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Alginate is a biopolymer widely used on delivery systems when bioactive protection at acidic pH is required, while chitosan can enhance mucoadhesion and controlled release at alkaline pHs. In this work, alginate ionotropic gelation and electrostatic complexation to chitosan were evaluated concomitantly or in a two-step approach to improve the delivery properties of systems in different pHs. The effect of pH on alginate gelation and chitosan interactions were also evaluated. Alginate microspheres were prepared by ionotropic gelation in CaCl2 at different pH values (2.5 and 6.0) by extrusion. Complexation with chitosan was carried out during alginate ionotropic gelation (one-step approach) or after alginate gel formation (two-step approach). Alginate microparticles without chitosan showed larger pores and lower mechanical strength. Extruded microspheres at pH 6.0 were more stable to pH and showed smaller pores than the formed at pH 2.5. One-step production retained a large amount of bioactive at pH 7.0 and resulted in lower release at the pH of intestinal digestion. The two-step approach retained less amount of bioactive but confer more protection to the pH of the stomach phase and higher release in pH of the intestinal phase than one-step samples. These results indicate that the formation of alginate gels by ionotropic gelation followed by the complexation with chitosan (in two-step) is promising for the transport and delivery of bioactives into intestinal conditions, whereas the ionotropic gelation concomitantly to electrostatic complexation (one-step approach) is indicated to the delivery of bioactives into lower pH environments.

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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.

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Nanotechnology has emerged as a possible solution to improve phytochemicals' limitations. The objective of the present study was to encapsulate beetroot extract (BR Ext) within a chitosan (CS)-based nanogel (NG) designed via ionic crosslinking with tripolyphosphate (TPP) for betanin (Bet) delivery, mainly in the ophthalmic environment. BR Ext is rich in betanin (Bet) according to thin layer chromatography (TLC), UV-visible spectroscopy, and HPLC analysis. NG presented a monodisperse profile with a size of 166 ± 6 nm and low polydispersity (0.30 ± 0.03). ζ potential (ζ-Pot) of +28 ± 1 is indicative of a colloidally stable system. BR Ext encapsulation efficiency (EE) was 45 ± 3%. TEM, with the respective 3D-surface plots and AFM, showed spherical-elliptical-shaped NG. The BR Ext release profile was biphasic with a burst release followed by slow and sustained phase over 12 h. Mucoadhesion assay demonstrated interactions between NG with mucin. Moreover, NG provided photoprotection and pH stability to BR Ext. FRAP and ABTS assays confirmed that BR Ext maintained antioxidant activity into NG. Furthermore, in vitro assays using human retinal cells displayed absence of cytotoxicity as well as an efficient protection against injury agents (LPS and H2O2). NGs are a promising platform for BR Ext encapsulation, exerting controlled release for ophthalmological use.

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Phenolic compounds that are present in pineapple by-products offer many health benefits to the consumer; however, they are unstable to many environmental factors. For this reason, encapsulation is ideal for preserving their beneficial effects. In this work, extracts were obtained by the combined method of solid-state fermentation with Rhizopus oryzae and ultrasound. After this process, the encapsulation process was performed by ionotropic gelation using corn starch, sodium alginate, and Weissella confusa exopolysaccharide as wall material. The encapsulates produced presented a moisture content between 7.10 and 10.45% (w.b), a solubility of 53.06 ± 0.54%, and a wettability of 31.46 ± 2.02 s. The total phenolic content (TPC), antioxidant capacity of DPPH, and ABTS of the encapsulates were also determined, finding 232.55 ± 2.07 mg GAE/g d.m for TPC, 45.64 ± 0.9 µm Trolox/mg GAE for DPPH, and 51.69 ± 1.08 µm Trolox/mg GAE for ABTS. Additionally, ultrahigh performance liquid chromatography (UHPLC) analysis allowed us to identify and quantify six bioactive compounds: rosmarinic acid, caffeic acid, p-coumaric acid, ferulic acid, gallic acid, and quercetin. According to the above, using ionotropic gelation, it was possible to obtain microencapsulates containing bioactive compounds from pineapple peel extracts, which may have applications in the development of functional foods.

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Studies show that yerba mate (Ilex paraguariensis) has high antioxidant capacity occasioned by its high contents of total phenolic compounds. Microencapsulation, specifically ionic gelation, since it does not use heating during process, is considered as an alternative for preserving and applying the extract. The purpose of this study was to evaluate general characteristics and stability of hydroalcoholic extract of yerba mate, conduct the extract microencapsulation by ionic gelation followed by microparticle fluidized bed drying. The extract was evaluated for color stability, total phenolic compounds, and antioxidant activity for nine weeks and at three temperatures (5, 15, and 25 °C). From the extract, a double emulsion (W/O/W), generation of microparticles (ionic gelation by dripping), and fluidized bed drying were conducted. The extract had 32912.55 mg GAE/100 g of phenolic compounds and 2379.49 µmol TE/g of antioxidant activity. The main compound observed was chlorogenic acid (5-CQA) with 0.35 ± 0.01 g/100 mL. In the stability study, the temperature was observed to influence in phenolic compounds reduction, as well as in total color difference of the extract. Double emulsion has shown to be stable and appropriate for use. The values of microparticles total phenolic compounds and antioxidant activity were 423.18 ± 8.60 mg GAE/100 g and 21.17 ± 0.24 µmol TE/g, respectively. After drying, the moisture of microparticles was reduced from 79.2% to 19%. The extract had high total phenolic compound content and high antioxidant activity. Storage at the lowest temperature (5 °C) assured better preservation of extract total phenolic compounds. The dried microparticles showed content of total phenolic compounds and antioxidant activity with potential for commercialization and future application in food matrices.

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In this work, extracellular colored metabolites obtained from the filamentous fungi Talaromyces australis and Penicillium murcianum, isolated in the Andean-Patagonian native forests of Chile, were studied as prospect compounds to increase the sustainability of cosmetic products. The chemical and antioxidant properties of these natural pigments were characterized and strategies for their microencapsulation were also studied. UHPLC/MS-MS analyses indicated that the predominant metabolites detected in the cultures of P. murcianum were monascin (m/z = 411.15) and monashexenone (m/z = 319.10), while athrorosin H (m/z = 458.20) and damnacanthal (m/z = 281.05) were detected in cultures of T. australis. ORAC tests revealed that P. murcianum's metabolites had the greatest antioxidant properties with values higher than 2000 µmol of trolox equivalents/g. The fungal metabolites were successfully microencapsulated by ionic gelation into structures made of 1.3% sodium alginate, 0.2% chitosan, and 0.07% hyaluronic acid. The microencapsulation process generated structures of 543.57 ± 0.13 µm of mean diameter (d50) with an efficiency of 30% for P. murcianum, and 329.59 ± 0.15 µm of mean diameter (d50) and 40% efficiency, for T. australis. The chemical and biological characterization show the biotechnological potential of these fungal species to obtain pigments with antioxidant activity that could be useful in the cosmetic industry. The encapsulation process enables the production of easy-to-handle dry powder from the fungal metabolites, which could be potentially marketed as a functional cosmetic ingredient. KEY POINTS: • The predominant fungal pigments were of azaphilone and anthraquinoid classes. • The fungal pigments showed high antioxidant activity by ORAC assay. • Fungal pigment microcapsules obtained by ionic gelation were characterized.

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This study aimed to develop a functional strawberry pulp containing the combination of Lactobacillus casei and bioactive compounds from red onion peel extract into the microparticles formulations to improve bacteria survival during storage and product consumption. To achieve this goal, the addition of different concentrations of red onion peel extract added to the microparticles was evaluated: 5, 20 and 40 %. Microparticles were morphologically characterized and the encapsulation efficiency of the bioactive compounds were evaluated. The physicochemical and microbiological characteristics of the fruit pulp were within the required standards, regardless of the formulation evaluated. As for the pulp added from the microparticles, their physicochemical and microbiological features and probiotic survival under simulated gastrointestinal conditions and storage were analyzed; the size of the microparticles ranged from 136.00 to 305.00 µm. The encapsulation efficiency of both, probiotics and compounds was satisfactory over the different treatments. Indeed, the results pointed out values in the range from 77.77 to 92.11 % for probiotic bacteria; from28.88 to 50.18 % for reducing compounds; 35.72 to 69.01 % for flavonoids; and 25.39 to 60.00 % for total monomeric anthocyanins. The formulations of alginate microparticles and alginate +5 % extract had the best results of L. casei probiotic viability in strawberry pulp under simulated gastrointestinal conditions and during storage at -18 °C for 60 days. In conclusion, red onion peel extract at low concentrations can help the survival of the probiotic L. casei under different conditions.

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Encapsulation is a process in which a base material is encapsulated in a wall material that can protect it against external factors and/or improve its bioavailability. Among the different encapsulation techniques, ionic gelation stands out as being useful for thermolabile compounds. The aim of this work was to encapsulate Saccharomyces boulardii by ionic gelation using agavins (A) and whey protein (WP) as wall materials and to evaluate the morphostructural changes that occur during in vitro gastrointestinal digestion. Encapsulations at different levels of A and WP were analyzed using microscopic, spectroscopic and thermal techniques. Encapsulation efficiency and cell viability were evaluated. S. boulardii encapsulated at 5% A: 3.75% WP (AWB6) showed 88.5% cell survival after the simulated gastrointestinal digestion; the bead showed a significantly different microstructure from the controls. The mixture of A and WP increased in the survival of S. boulardii respect to those encapsulated with alginate, A or WP alone. The binary material mixture simultaneously allowed a controlled release of S. boulardii by mostly diffusive Fickian mechanisms and swelling. The cell-release time was found to control the increment of the Damköhler number when A and WP were substrates for S. boulardii, in this way allowing greater protection against gastrointestinal conditions.

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In this work, purified pectins from Araçá fruits (Psidium cattleianum Sabine) were obtained and characterized after partial demethylation. On each prepared sample, the carboxylic yield was obtained by titration, the degree of methylation (DM) by 1H-NMR, and the molecular weight distribution by steric exclusion chromatography (SEC). Then, the gelation ability in the presence of calcium counterions was investigated and related to DM (59-0%); the pectin concentration (2-10 g L-1); and the CaCl2 concentration (0.1-1 mol L-1) used for dialysis. The critical pectin concentration for homogeneous gelation was above 2 g L-1 when formed against 1 mol L-1 CaCl2. The elastic modulus (G') increased with pectin concentration following the relationship G'~C2.8 in agreement with rigid physical gel network predictions. The purified samples APP and APP-A with DM ≥ 40% in the same conditions released heterogeneous systems formed of large aggregates. Gels formed against lower concentrations of CaCl2 down to 0.1 mol L-1 had a higher degree of swelling, indicating electrostatic repulsions between charged chains, thus, counterbalancing the Ca2+ cross-linkage. Compression/traction experiments demonstrated that an irreversible change in the gel structure occurred during small compression with an enhancement of the G' modulus.

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The accumulation of chitin waste from the seafood industry is a serious environmental problem. However, this residue can be degraded by chitinases and its subproducts, such as chitosan, economically exploited. In this study, a chitinase producer bacteria, identified as Paenibacillus illinoisensis, was isolated from the Brazilian coastal city of Terra de Areia - Rio Grande Do Sul (RS) and was immobilized within alginate beads to evaluate its chitinase production. The alginate beads containing cells presented an average size of 4 mm, 99% of immobilization efficiency and increased the enzymatic activity in 40.71% compared to the free cells. The biomass during enzymatic production increased 62.01% and the total cells leaked from the alginate beads corresponded to 6.46% after 96 h. Immobilized cells were reused in a sequential batch system and remained stable for production for up to four 96-h cycles, decreasing only 21.04% of the initial activity at the end of the fourth cycle. Therefore, the methodology used for cell immobilization resulted in adequate beads to maintain cell viability during the enzymatic production, increasing enzymatic activity, showing low cell leakage from the support and appropriate recyclable capacity.

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Two phases are generally recognized in the enzymatic coagulation of milk: hydrolysis and aggregation, although nowadays more and more researchers consider the non-enzymatic phase to actually be a stage of gel formation made up of two sub-stages: micellar aggregation and hardening of the three-dimensional network of para-κ-casein. To evaluate this controversy, the main descriptive models have been reviewed. Most of them can only model micellar aggregation, without modeling the hardening stage. Some are not generalizable enough. However, more recent models have been proposed, applicable to a wide range of conditions, which could differentiate both substages. Manufacturing quality enzymatic cheeses in a cost-effective and consistent manner requires effective control of coagulation, which implies studying the non-enzymatic sub-stages of coagulation separately, as numerous studies require specific measurement methods for each of them. Some authors have recently reviewed the micellar aggregation models, but without differentiating it from hardening. Therefore, a review of the proposed models is necessary, as coagulation cannot be controlled without knowing its mechanisms and the stages that constitute it.

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Biopolymer beads can be used as carrier and encapsulation system for a wide variety of materials in food, medical, pharmaceutical, cosmetics, agricultural, and environmental applications. Beads of low acyl gellan gum (0.4-1.2% w/w) were produced using extrusion technique (dripping) followed by an ionotropic gelation step with calcium or potassium chloride. In this methodology, gel formation is accomplished by cations diffusion at room temperature and, as a consequence, different structure and gel properties could be obtained. Gellan beads were subjected to uniaxial compression measurements. The force-displacement curves showed that the occurrence of structural failure under tested conditions depended on beads formulation and was only observed at polysaccharide concentration above 0.8% (w/w). Maximum force or force at failure was mainly dependent on the type (monovalent or divalent cation) and salt concentration. Moreover, at fixed salt amounts, higher values of maximum force were reached using a concentration of 1% (w/w) gellan. Young modulus, determined using Hertz approach, showed values between 445 and 840 kPa depending on polysaccharide concentration and salt type added. Mechanical properties are critical features of gel beads and can define their suitability for a specific application. Therefore, the results obtained, mainly intrinsic properties such as Young modulus, could be a tool for comparing and choosing polysaccharides for specific uses.

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Isomaltulose is a potential substitute for sucrose, with a high stability and prebiotic potential, for wide use in candies and soft drinks. This sugar is obtained from sucrose through enzymatic conversion using microbial glucosyltransferases. This work aimed to optimize a matrix to immobilize glucosyltransferase producing Erwinia sp. D12 cells using a sequential experimental strategy. The cell mass of Erwinia sp. D12 obtained in a bioreactor was immobilized in beads formed by ionic gelation. The conversion of sucrose into isomaltulose using the beads was performed in batch and continuous processes, and the isomaltulose was recovered through crystallization. The stability of isomaltulose was assessed in beverages of different pH values, and its prebiotic potential was verified with the growth of probiotic microorganisms. The optimized matrix composed of alginate (2.0% w/v), CaCl2 (2.0% w/v), gelatin (2.0% w/v), and transglutaminase (0.2% w/v) showed the highest mean of produced isomaltulose (199.82 g/L) after four batches. In addition, high stability during the continuous process resulted in an isomaltulose production above of 230 g/L for up to 72 h. The produced isomaltulose was more stable than sucrose in lemon soft drink and orange and grape energy drinks after 30 days of storage; and promoted the growth of Bifidobacterium animalis and Lactobacillus lactis. In conclusion, the production of isomaltulose by Erwinia sp. D12 cells immobilized using optimized conditions is recommended, due to its high conversion capacity, high stability, and prebiotic potential of crystals obtained.

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Caseinomacropeptide (CMP) is derived from the chymosin cleavage of κ-casein during cheese production. This study developed gels from CMPs, which were isolated by different ultrafiltration systems, and whey protein isolate (WPI), and studied their rheological and ultrastructural characteristics. The 30% WPI gel showed high elastic modulus (G') values and stronger structure than the other samples with CMP. Another gel, with 50% protein, 30% WPI and 20% CMP sample isolated from the 30 kDa retentate, had a weaker structure and lower G' value. The third gel, with 30% WPI and 20% CMP sample from the 5 kDa retentate derived from the 30 kDa retentate, presented intermediate structural strength. Despite the increase in protein concentration from the addition of CMP, there was a decrease in the strength of the gel network. Different CMP isolation processes also contributed to differences in the microscopic analysis of gel structures with the same protein content.

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Oleaginous microorganisms, including the fungus Umbelopsis isabellina, have emerged as a biotechnological alternative to obtain polyunsaturated fatty acid-rich oils, which are strongly linked to energy purposes (biofuel) than the food industry. Considering the composition of microbial oil and its use by the food industry, it is necessary to investigate strategies that increase its lipid stability. Ergo, this pioneering study aimed to microencapsulate the oil produced by Umbelopsis isabellina and evaluate its oxidative stability throughout the storage period against factors such as temperature and luminosity. The microbial oil was microencapsulated through the external ionic gelation technique, producing an encapsulation efficiency of 80% and proving to be a suitable method because it maintained oil composition. Combining microencapsulation and refrigerated storage led to the best effects on storage time, increasing the evaluated lipid stability through the peroxide values and conjugated diene formation. Moreover, saturated and monounsaturated fatty acid content increased, and polyunsaturated fatty acid content decreased during storage for both the free and microencapsulated oil, regardless of storage temperature, although microencapsulation reduced the changes. The results primarily demonstrate how microencapsulation prolongs the oxidative stability and unsaturated fatty acid content of the microbial oil by reducing its reactions to external environmental factors, thus facilitating its use in the food industry.

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