Plant Protein-Based Delivery Systems: An Emerging Approach for Increasing the Efficacy of Lipophilic Bioactive Compounds.

The development of plant protein-based delivery systems to protect and control lipophilic bioactive compound delivery (such as vitamins, polyphenols, carotenoids, polyunsaturated fatty acids) has increased interest in food, nutraceutical, and pharmaceutical fields. The quite significant ascension of plant proteins from legumes, oil/edible seeds, nuts, tuber, and cereals is motivated by their eco-friendly, sustainable, and healthy profile compared with other sources. However, many challenges need to be overcome before their widespread use as raw material for carriers. Thus, modification approaches have been used to improve their techno-functionality and address their limitations, aiming to produce a new generation of plant-based carriers (hydrogels, emulsions, self-assembled structures, films). This paper addresses the advantages and challenges of using plant proteins and the effects of modification methods on their nutritional quality, bioactivity, and techno-functionalities. Furthermore, we review the recent progress in designing plant protein-based delivery systems, their main applications as carriers for lipophilic bioactive compounds, and the contribution of protein-bioactive compound interactions to the dynamics and structure of delivery systems. Expressive advances have been made in the plant protein area; however, new extraction/purification technologies and protein sources need to be found Their functional properties must also be deeply studied for the rational development of effective delivery platforms.

Films Based on Biopolymers Incorporated with Active Compounds Encapsulated in Emulsions: Properties and Potential Applications-A Review.

The rising consumer demand for safer, healthier, and fresher-like food has led to the emergence of new concepts in food packaging. In addition, the growing concern about environmental issues has increased the search for materials derived from non-petroleum sources and biodegradable options. Thus, active films based on biopolymers loaded with natural active compounds have great potential to be used as food packaging. However, several lipophilic active compounds are difficult to incorporate into aqueous film-forming solutions based on polysaccharides or proteins, and the hydrophilic active compounds require protection against oxidation. One way to incorporate these active compounds into film matrices is to encapsulate them in emulsions, such as microemulsions, nanoemulsions, Pickering emulsions, or double emulsions. However, emulsion characteristics can influence the properties of active films, such as mechanical, barrier, and optical properties. This review addresses the advantages of using emulsions to encapsulate active compounds before their incorporation into biopolymeric matrices, the main characteristics of these emulsions (emulsion type, droplet size, and emulsifier nature), and their influence on active film properties. Furthermore, we review the recent applications of the emulsion-charged active films in food systems.

Role of aqueous phase composition and hydrophilic emulsifier type on the stability of W/O/W emulsions.

Double W1/O/W2 emulsions can act as fat substitutes in food matrices, although synthetic emulsifiers are commonly used due to their inherent instability and susceptibility to coalescence. In order to guarantee the stability of the W/O interface, the synthetic emulsifier polyglycerol polyricinoleate (PGPR - 4.5% w/w) was used. However, the replacement of chemically synthesized ingredients by natural alternatives has been extensively pursued in food applications. In this sense, whey protein isolate (WPI) and sodium caseinate (SC) were used to stabilize the external aqueous phase (W2) of water-in-oil-in-water double emulsions (W1/O/W2), in addition to Tween 80 that was used as a control. The composition of the internal aqueous phase and its effects on the double emulsion were studied by the addition of sodium chloride (0.2% w/w NaCl), gallic acid (0.5% w/w GA) or a GA/NaCl mixture (0.5% / 0.2% w/w). The effect of these different hydrophilic components was evaluated from measurements of droplet size, viscosity, ζ potential, interfacial tension and kinetic stability. SC-stabilized W/O/W emulsions showed better kinetic stability than WPI- and T80-systems. These results can be attributed to the initial droplet size (SC < T80 < WPI) and viscosity of the emulsions (SC < T80 < WPI). SC is a more flexible and unfolded protein that can quickly adsorb and rearrange at the interface, favoring the formation of smaller droplets and limiting the migration of inner water droplets to the outer phase. In addition to smaller droplets, the addition of SC (8% w/w) to the external aqueous phase promoted an increase in the viscosity of bulk systems, which reduced the destabilization rates by creaming and coalescence. All W/O/W systems containing NaCl in the inner aqueous phase presented greater kinetic stability during 7 days of storage. Although the addition GA was less efficient to stabilize double emulsions compared to NaCl, this phenolic compound reduced the interfacial tension, favoring the formation of WPI- and T80-droplets with smaller diameters. However, the use of GA/NaCl blend improved the stability and functionally of W/O/W double emulsions. We concluded that the type of hydrophilic emulsifier, the properties of the inner water droplets and the viscosity of phases influenced the droplet size, viscosity and kinetic stability of double emulsions. This work provides a better understanding of how composition influences the properties of double emulsions and how it can be used to design W/O/W emulsions as fat substitutes in more complex food systems.

Stabilization mechanisms of O/W emulsions by cellulose nanocrystals and sunflower protein.

Oil-in-water (O/W) emulsions stabilized by cellulose nanocrystals (CNC) and/or sunflower proteins (SFP) were produced, aiming to study the effects of each and the mixture of these stabilizers on the interfacial behavior and physicochemical properties of O/W emulsions. The presence of CNC (non-surface activity compound) did not affect SFP solutions' adsorption kinetics since there were no differences in the interfacial tension curves of SFP and mixtures of stabilizers over time. However, either stabilizer provided alone high resistance against droplet coalescence over time (no evidence of oiling-off and no difference in the mean droplet size values), even systems with less viscoelastic interface (2 % CNC). Although droplet coalescence was prevented by steric hindrance and reduction of interfacial tension between the oil-water phases provided by CNC and SFP, respectively, these emulsions were unstable to the creaming phenomenon. Only the mixture of these stabilizers was able to prevent both destabilization mechanisms, initially by adsorption and anchoring of SFP on the interface, followed by adsorption of CNC in the free interface spaces, and finally by the interaction of non-adsorbed CNC particles in the continuous phase, which led to an increase in system viscosity. Thus, based on the results of interfacial properties and emulsions characteristics, we had a better understanding of stabilization mechanisms of O/W emulsions by a food-grade particle and a plant-derived protein.

Interactions of ß-carotene with WPI/Tween 80 mixture and oil phase: Effect on the behavior of O/W emulsions during in vitro digestion.

This study investigated the impact of adding ß-carotene on the structure of fresh O/W emulsions with different oil phase (sunflower oil-LCT or NEOBEE®1053-MCT) and emulsifiers (WPI, Tween 80 - T80 or WPI/T80 mixture). In this sense, the behavior of emulsions through the gastrointestinal tract, the stability and bioaccessibility of ß-carotene were also assessed. The ß-carotene reduced the interfacial tension of the LCT/MCT-water systems. The addition of ß-carotene promoted an increase of viscoelasticity of LCT/MCT-T80 (0.5%WPI/0.5%T80 and 1%T80 w/w) interfaces, but an increase of WPI content reduced the viscoelasticity of interfacial layers (LCT/MCT-1% WPI). These changes in the interface properties influenced the mean droplet size and ζ-potential of the fresh emulsions. LCT systems presented similar bioaccessibility/stability of ß-carotene. However, ß-carotene entrapped within protein-coated MCT droplets was more stable than within T80-MCT systems. Our results show that ß-carotene interacted with other ingredients of emulsions changing their properties and behavior under gastrointestinal tract as well as the stability/bioaccessibility of ß-carotene.

Hybrid oil-in-water emulsions applying wax(lecithin)-based structured oils: Tailoring interface properties.

This study addressed the impact of fruit wax(lecithin)-based oleogels as dispersed phase in formation and stability of oil-in-water emulsions. These hybrid emulsions were prepared above the melting point of the oleogels, using Tween 80 (T80) or whey protein isolate (WPI) as emulsifiers. Both mono- and mixed-component oleogels comprised of fruit wax (FW) or FW + lecithin (FWLEC), respectively, were studied as lipid phases. After hot-homogenization, emulsions were submitted to quiescent cooling and stored over 14 days at 5 or 25 °C, in such temperatures supposed to assist or hinder oleogelation, respectively. Time course promoted a slight decrease in zeta potential only for WPI-stabilized emulsions and particle size distribution was shifted to larger size values, but showing a lesser extent to those stored at 5 °C. The presence of oleogels improved kinetic stability of emulsions compared to liquid oil at both temperatures, disclosing the role of the combined effects of the type of emulsifier and oleogelator(s)-emulsifier interactions. These outcomes are associated with the interfacial activity played by both oleogelators, but mainly lecithin that led to lower values of interfacial tension. In addition FWLEC combined with WPI showed the lowest complex modulus from dilational rheology, which can be related with WPI-LEC complex formation. Overall, results suggest that oleogelators migrated to the O/W interface of dispersed droplets, no longer reflecting oleogel bulk properties and showing a more complex behavior. However, the formation of more complex structures at the interface favored greater stability of the emulsions. Thus, the new perspective of oleogel-inspired fat droplets in hybrid systems can expand the conventional approach of oil structuring to create mixed interfaces tailoring oil-in-water emulsions properties.

Modulating in vitro digestibility of Pickering emulsions stabilized by food-grade polysaccharides particles.

An in vitro digestibility protocol was used to elucidate the role of different emulsifying polysaccharides particles on the lipid digestion rate of oil-in-water Pickering emulsions. Emulsions stabilized by cellulose crystals (CCrys), cellulose nanofibers (CNFs), chitosan particles and a conventional emulsifier (Tween 80) were evaluated concerning microstructure, droplet size, zeta potential and free fatty acids released during digestion. After gastric step, the high positive charge of chitosan-stabilized emulsions favored the droplets disaggregation resulting in a mild effect of bridging flocculation by particles sharing and displacement of the size curve distribution toward lower size. After passing through the intestinal condition, these emulsions presented few droplets and chitosan aggregates with a monomodal size distribution and high mean droplet size (D4,3 = 197 ±â€¯8 µm). On the other hand, Tween 80, CCrys and CNFs were able to inhibit lipid digestion and no changes on mean droplet size were observed following intestinal step. CNFs-stabilized emulsion showed the lowest lipid digestion, whereas the strong adherence of the CCrys particles onto the droplet interface became them resistant to displacement by surface-active components (i.e. bile salts and lipase enzyme). On the other hand, a slow lipid hydrolysis could be observed in chitosan-stabilized emulsions promoted by competition between chitosan aggregates and intestinal fluids by the oil droplet interface. Studying the emulsions stabilized using different polysaccharides particles on gastrointestinal conditions we could elucidate important features for their potential application as control systems of lipid digestion rate, as well as, as delivery systems of lipophilic compounds.

Impact of whey protein/surfactant mixture and oil type on the gastrointestinal fate of emulsions: Ingredient engineering.

The engineering of ingredients emerges as a strategy to design emulsified products aiming to control the lipid hydrolysis. In this context, oil-in-water (O/W) emulsions composed of different oil phases (Sunflower oil - LCT or NEOBEE® 1053 - MCT) and stabilized by whey protein isolate - WPI (1% w/w), Tween 80 - T80 (1% w/w) or varied ratios of WPI/T80 (0.9975%WPI/0.0025%T80; 0.75%WPI/0.25%T80; 0.5%WPI/0.5%T80 w/w) were produced and submitted to simulated gastrointestinal conditions. The lipolysis of LCT was influenced by the fatty acid chain length and emulsifier composition, while only the fatty acid chain length affected the lipolysis of MCT. The emulsions produced with LCT and 1%WPI or 09975%WPI/00025%T80 showed the highest release rate of free fatty acids (FFAs), but similar result was observed for the 0.5%WPI/0.5%T80 system. In the 0.5%WPI/0.5%T80 mixture, WPI and T80 worked together and achieved an improved performance during the gastric (stability similar as 1%T80 emulsion) and small intestinal phases (lipolysis similar as 1%WPI emulsion). The rational selection of ingredients is useful to design emulsions with improved performance as a delivery system since the emulsion structural stability during digestion, the oil type and interaction between lipase-interface had a marked impact on the efficiency of lipid digestion.

Ultrasound stabilization of raw milk: Microbial and enzymatic inactivation, physicochemical properties and kinetic stability.

The aim of this study was to evaluate the effects of non-thermal and thermal high-intensity ultrasound (HIUS) treatment on the microbial and enzymatic inactivation, physicochemical properties, and kinetic stability of the raw milk by applying different energy densities (1, 3, 5, and 7 kJ/mL). Two HIUS treatments were evaluated based on different nominal powers, named HIUS-A and HIUS-B, using 100 W and 475 W, respectively. HIUS-A treatment was non-thermal processing while HIUS-B was a thermal treatment only for the energy densities of 5 and 7 kJ/mL since the final temperature was above 70 °C. The HIUS-B treatment showed to be more efficient. Log reductions up to 3.9 cycles of aerobic mesophilic heterotrophic bacteria (AMHB) were achieved. Significant reductions of the fat globule size, with diameters lower than 1 µm, better color parameters, and kinetic stability during the storage were observed. Also, HIUS-B treatment inactivated the alkaline phosphatase and lactoperoxidase. The HIUS-B treatment at 3 kJ/mL worked below 57 °C being considered a border temperature since it did not cause unwanted physicochemical effects. Furthermore, a microbial inactivation of 1.8 ± 0.1 log cycles of AMHB was observed. A proper inactivation of only the Alkaline phosphatase and a significant reduction of the fat globules sizes, which kept the milk kinetically stable during storage was achieved.

Bioaccessibility of Lipophilic Compounds Vehiculated in Emulsions: Choice of Lipids and Emulsifiers.

Great efforts have been made to design emulsions considering the need to perform an effective encapsulation, protection, vehiculation, and bioaccessibility of lipophilic compounds. This task can be achieved by manipulating the structure of the emulsion based on the choice of the processes and ingredients of the aqueous phase, interface, and lipid matrix. Thus, the main focus of this perspective is to provide insights into the use of ingredient engineering in manipulating/building emulsion structures that enhance lipophilic compound release and bioaccessibility.

Formation and stability of W/O-high internal phase emulsions (HIPEs) and derived O/W emulsions stabilized by PGPR and lecithin.

Water-in-oil high internal phase emulsions (HIPEs) can provide interesting textures that could be used to reduce trans- and/or saturated fat content in food products. On the other hand oil-in-water emulsions can be found in a variety of food and beverages. Moreover, strategies aiming synthetic or semi-synthetic ingredients replacement by natural alternatives for food applications has been pursuit. For these purposes, the effect of partial replacement of PGPR by lecithin on properties of either W/O-HIPEs or O/W emulsions manufactured from the same initial composition but showing different volume fraction of dispersed phase were investigated aiming to understand the behaviour of emulsifiers' mixture in water-oil or oil-water interfaces. Firstly, water-in-oil HIPEs were produced using a rotor-stator device. At fixed total amount of emulsifier (2% w/w), W/O emulsions stabilized with LEC:PGPR ratios of 0.5:1.5 and 1.0:1.0 showed similar droplet size with a better kinetic stability compared to emulsions containing only PGPR. These results indicated good interaction between LEC and PGPR, which was also confirmed by dynamic interfacial tension profile and interfacial dilational rheology. In order to reduce the droplet size of W/O-HIPEs, these emulsions were subsequently subjected to high-pressure homogenization and interestingly phases inversion was observed. Confocal microscopy confirmed the phases inversion attributed to high input of energy leading to the formation of O/W emulsions. Then both W/O-HIPEs and O/W emulsions were investigated regarding LEC:PGPR mixtures as emulsifiers. All W/O-HIPEs showed shear thinning behavior and high viscosity at low shear rate whereas O/W emulsions showed low viscosity and Newtonian behavior. The increase of lecithin content in emulsifier mixture led to more stable O/W emulsions, whereas more stable W/O-HIPEs were produced by lecithin and PGPR mixtures ratio of 0.5:1.5 and 1.0:1.0.

Impact of oil type and WPI/Tween 80 ratio at the oil-water interface: Adsorption, interfacial rheology and emulsion features.

The relationship between the composition and structure of food emulsions was evaluated from the effect of a mixture of emulsifiers Whey protein (WPI) - Tween 80 (T80) and the oil phase features, such as chain length and unsaturation degree (sunflower oil, a long chain triacylglycerol - LCT or NEOBEE® 1053, a medium chain triacylglycerol - MCT). Emulsions with LCT showed higher droplet size than MCT as a consequence of its higher viscosity. All emulsions exhibited shear thinning behavior, but the viscosity was influenced by their interface composition. An occurrence of the destabilization mechanism by creaming was observed in turbidimetric measurements, but no visual phase separation could be observed, indicating a good kinetic stability after a 7-day storage. The initial interfacial tension of the water-LCT or water-MCT oil was about 25 mN/m, but the WPI addition (1% w/w) reduced the initial interfacial tension to approximately 20 mN/m. The increase of T80 concentration led to a decrease of the interfacial tension, reaching a value around 10 mN/m in systems with pure T80. The curves of interfacial tension of systems with LCT or MCT showed differences in the decay rate of tension over time. These differences were attributed to characteristics of the oil phase (hydrophobicity, unsaturation degree, presence of impurities) and the different proportions of each emulsifier within the mixture of emulsifiers. Finally, a higher viscoelastic interface was observed in LCT emulsions, which were mainly stabilized by WPI molecules. Such molecules presented a higher resistance to the displacement due to the competitive adsorption phenomenon, since the LCT is a more hydrophobic oil. On the other hand, the interface with MCT and a higher T80 concentration was less viscoelastic due to an easier displacement of WPI from the interface and the replacement by T80. The results indicate that T80 can be used in combination with WPI to produce emulsions with good stability and lower concentration of synthetic compounds. Lastly, the interfacial layer composition is not only dependent on the WPI-T80 ratio in the bulk phase, but also on the oily phase features. These results provide a potential strategy for designing emulsified foods based on the choice of ingredients and knowledge of the interaction between them.

Coupling of high-intensity ultrasound and mechanical stirring for producing food emulsions at low-energy densities.

In this study, coupling of ultrasound (US) device and rotor-stator (RS), operating at low-energy densities, was studied as an alternative process to individual US and RS to produce modified starch-stabilized oil-in-water emulsions, as well as its potential use to encapsulate eugenol. To this aim, a full factorial design was employed to evaluate the effects of the US nominal power (0, 360 and 720 W) and RS nominal power (0, 150 and 300 W) on the physical properties, encapsulation efficiency and kinetic stability of emulsions produced. Firstly, the action of modified starch and eugenol onto interface oil-water was evaluated. The emulsifier was rapidly adsorbed on the interface water-sunflower oil reducing the interfacial tension from 25 to 16 mN/m, while eugenol did not show surface activity. The increase of energy density, in general, resulted in droplet size reduction, indicating the relevant role of the forces involved in the droplet breakup on emulsion stability. Coupling was more efficient on the droplets breakup producing smaller droplet size with narrower size distribution. While the coupled system work during 5 min for an energy density of 583 J/mL, the corresponding emulsification time for operating singly US and RS were 7.09 min and 17.04 min, respectively. Therefore, the main advantage associate to coupled process is the reduction of processing time to produce an emulsion with better kinetic stability.

Cellulose nanofibers from banana peels as a Pickering emulsifier: High-energy emulsification processes.

Cellulose nanofibers (CNFs) from banana peels was evaluated as promising stabilizer for oil-in-water emulsions. CNFs were treated using ultrasound and high-pressure homogenizer. Changes on the size, crystallinity index and zeta potential of CNFs were associated with the intense effects of cavitation phenomenon and shear forces promoted by mechanical treatments. CNFs-stabilized emulsions were produced under the same process conditions as the particles. Coalescence phenomenon was observed in the emulsions produced using high-pressure homogenizer, whereas droplets flocculation occurred in emulsions processed by ultrasound. In the latter, coalescence stability was associated with effects of cavitation forces acting on the CNFs breakup. Thus, smaller droplets created during the ultrasonication process could be recovered by particles that acted as an effective barrier against droplets coalescence. Our results improved understanding about the relationship between the choice of emulsification process and their effects on the CNFs properties influencing the potential application of CNFs as a food emulsifier.

Rheological properties, microstructure, and encapsulation efficiency of inulin-type dietary fiber-based gelled emulsions at different concentrations.

Gelled emulsion systems offer promising matrices for encapsulating bioactive compounds, enhancing stability, bioavailability, and controlled release. Incorporating inulin-type dietary fibers into emulsion-filled gels can innovate food products. This study explored the impact of inulin concentration (0-15 % w/w) on visual aspect, microstructure, particle size distribution, creaming stability, rheological behavior, and encapsulation efficiency of emulsions and gelled emulsions with clove bud oil rich in eugenol. Regardless of inulin concentration, systems exhibited evenly distributed small oil droplets, ensuring good creaming stability. Emulsions with 10-15 % inulin formed gels upon natural cooling to approximately 30 °C. Viscoelastic properties varied with inulin concentration, attributed to increased polymer chain approximation and mobility. Higher inulin content decreased the transition temperature (66 °C, 56 °C, and 54 °C for 10 %, 12.5 %, and 15 % inulin, respectively). While inulin did not enhance creaming stability, it acted as a physical barrier, improving encapsulation efficiency of eugenol to nearly 100 %. Inulin-based emulsion-filled gels offer potential for functional food development, enriching nutritional value and health benefits.