Double emulsions for iron encapsulation: is a high concentration of lipophilic emulsifier ideal for physical and chemical stability?

BACKGROUND: Worldwide iron deficiency in diets has led to a growing interest in the development of food-compatible encapsulation systems for soluble iron, which are able to prevent iron's undesirable off-taste and pro-oxidant activity. Here, we explore the use of double emulsions for this purpose, and in particular, how the lipophilic emulsifier (polyglycerol polyricinoleate, PGPR) concentration influences the physicochemical stability of water-in-oil-in-water (W1 /O/W2 ) double emulsions containing ferrous sulphate in the inner water droplets. Double emulsions were prepared with sunflower oil containing 10 to 70 g kg-1 PGPR in the oil phase, and were monitored for droplet size distribution, morphology, encapsulation efficiency (EE) and oxidative stability over time. RESULTS: Fresh double emulsions showed an initial EE higher than 88%, but EE decreased upon storage, which occurred particularly fast and to a high extent in the emulsions prepared with low PGPR concentrations. All double emulsions underwent lipid oxidation, in particular those with the highest PGPR concentration, which could be due to the small inner droplet size and thus promoted contact between oil and the internal water phase. CONCLUSION: These results show that a too high PGPR concentration is not needed, and sometimes even adverse, when developing double emulsions as iron encapsulation systems. © 2019 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Food-grade micro-encapsulation systems that may induce satiety via delayed lipolysis: A review.

The increasing prevalence of overweight and obesity requires new, effective prevention and treatment strategies. One approach to reduce energy intake is by developing novel foods with increased satiating properties, which may be accomplished by slowing down lipolysis to deliver substrates to the ileum, thereby enhancing natural gut-brain signaling pathways of satiety that are normally induced by meal intake. To develop slow release food additives, their processing in the gastrointestinal tract has to be understood; therefore, we start from a general description of the digestive system and relate that to in vitro modeling, satiety, and lipolytic mechanisms. The effects of physicochemical lipid composition, encapsulation matrix, and interfacial structure on lipolysis are emphasized. We give an overview of techniques and materials used, and discuss partitioning, which may be a key factor for encapsulation performance. Targeted release capsules that delay lipolysis form a real challenge because of the high efficiency of the digestive system; hardly any proof was found that intact orally ingested lipids can be released in the ileum and thereby induce satiety. We expect that this challenge could be tackled with structured o/w-emulsion-based systems that have some protection against lipase, e.g., by hindering bile salt adsorption and/or delaying lipase diffusion.

Tailored microstructure of colloidal lipid particles for Pickering emulsions with tunable properties.

Sub-micron colloidal lipid particles (CLPs) can successfully be used as Pickering stabilizers in oil-in-water (O/W) emulsions, leading to an enhanced physical stability compared to conventional emulsifier-stabilized emulsions. Varying the lipid solid-liquid ratio leads to particles with distinct nanostructure and morphology, resulting in tunable emulsion stabilization performance. Our CLPs are produced by hot high pressure homogenization of high melting point fats in water, and subsequent cooling to induce lipid crystallization. Lath-like tripalmitin and palm stearin CLPs form jammed, cohesive interfacial layers that prevent relaxation of emulsion droplets, and form a three-dimensional network in the continuous aqueous phase. CLPs consisting of a mixture of solid tripalmitin and liquid tricaprylin are polycrystalline platelet-like particles that form O/W emulsions with spherical and bridged droplets covered by a thin particle layer. Our results present a versatile approach to interfacial design that also opens up new perspectives for development of novel delivery systems for active ingredients.

Unravelling the effect of droplet size on lipid oxidation in O/W emulsions by using microfluidics.

Lipid oxidation in emulsions is hypothesised to increase with decreasing droplet size, as this increases the specific oil-water interfacial area, where lipid oxidation is expected to be initiated. In literature, however, contradictory results have been reported, which can be caused by confounding factors such as the oil droplet polydispersity and the distribution of components between the available phases. In this work, monodisperse surfactant-stabilised emulsions with highly controlled droplet sizes of 4.7, 9.1, and 26 µm were produced by microfluidic emulsification. We show that lipid oxidation increases with decreasing droplet size, which we ascribe to the increased contact area between lipids and continuous phase prooxidants. Besides, a significant amount of oxygen was consumed by oxidation of the surfactant itself (Tween 20), an effect that also increased with decreasing droplet size. These insights substantiate the importance of controlling droplet size for improving the oxidative stability of emulsions.

Physical and Oxidative Stabilization of Oil-In-Water Emulsions by Roasted Coffee Fractions: Interface- and Continuous Phase-Related Effects.

Emulsions fortified with polyunsaturated fatty acids are highly relevant from a nutritional perspective; however, such products are prone to lipid oxidation. In the current work, this is mitigated by the use of natural antioxidants occurring in coffee. Coffee fractions with different molecular weights were extracted from roasted coffee beans. These components were positioned either at the interface or in the continuous phase of emulsions where they contributed to emulsion stability via different pathways. Coffee brew as a whole, and its high-molecular-weight fraction (HMWF), was able to form emulsions with good physical stability and excellent oxidative stability. When added post-homogenization to the continuous phase of dairy protein-stabilized emulsions, all coffee fractions were able to slow down lipid oxidation considerably without altering the physical stability of emulsions, though HMWF was more effective in retarding lipid oxidation than whole coffee brew or low-molecular-weight fraction. This is caused by various effects, such as the antioxidant properties of coffee extracts, the partitioning of components in the emulsions, and the nature of the phenolic compounds. Our research shows that coffee extracts can be used effectively as multifunctional stabilizers in dispersed systems leading to emulsion products with high chemical and physical stability.

A unifying approach to lipid oxidation in emulsions: Modelling and experimental validation.

Lipid oxidation is a longstanding topic within the field of food technology, and is strongly related to loss of product quality and consumer acceptance. Both for bulk oils and emulsions, the chemical phenomena involved in lipid oxidation have been extensively researched, and various reaction pathways have been identified. They are different in bulk oil compared to oil-in-water (O/W) emulsions in which the oil-water interface plays a prominent role. Most probably because of the complexity of the reaction scheme in combination with mass transfer effects, there is no model that describes lipid oxidation in emulsions in a unified fashion, and that is the aim that we have set ourselves to achieve. We use lipid oxidation data previously obtained in O/W emulsions made with 5 different emulsifiers (2 surfactants, and 3 proteins), in well-mixed systems where the oxygen-to-oxidizable lipid ratio is strictly controlled. We use data pertaining to headspace oxygen concentration, and to primary and secondary lipid oxidation products to develop a model based on reaction kinetics, including not only the classical reaction scheme (starting from an unsaturated lipid, LH) but also radical initiation from hydroperoxides, which is thought to be an effect that is overlooked in the classical description of the initiation step. We were able to describe the course of the reactions in these emulsions using the same reaction rate constants for all emulsions, with the exception of the two related to radical-based initiation. In Tween 20- and Tween 80-stabilized emulsions, initiation stems most probably solely from decomposition of hydroperoxides; this implies that lipid oxidation in these emulsions is co-determined by the initial ("pre-existing") hydroperoxide concentration. In protein-stabilized emulsions, on the other hand, lipid radical initiation is probably linked to reactions involving proteins (co-oxidation reactions), whereas initiation through decomposition of hydroperoxides seems less important, if at all. From this, we can conclude that the difference between both types of emulsions with regard to lipid oxidation mechanisms is related to differences in radical initiation. The developed model can serve as a unified basis for understanding lipid oxidation in emulsions, through which additional effects beyond the bare reaction kinetics, such as mass transfer effects, can be identified and used to e.g., quantify antioxidant effects, which is part of follow-up research.

Interfacial protein-protein displacement at fluid interfaces.

Protein blends are used to stabilise many traditional and emerging emulsion products, resulting in complex, non-equilibrated interfacial structures. The interface composition just after emulsification is dependent on the competitive adsorption between proteins. Over time, non-adsorbed proteins are capable of displacing the initially adsorbed ones. Such rearrangements are important to consider, since the integrity of the interfacial film could be compromised after partial displacement, which may result in the physical destabilisation of emulsions. In the present review, we critically describe various experimental techniques to assess the interfacial composition, properties and mechanisms of protein displacement. The type of information that can be obtained from the different techniques is described, from which we comment on their suitability for displacement studies. Comparative studies between model interfaces and emulsions allow for evaluating the impact of minor components and the different fluid dynamics during interface formation. We extensively discuss available mechanistic physical models that describe interfacial properties and the dynamics of complex mixed systems, with a focus on protein in-plane and bulk-interface interactions. The potential of Brownian dynamic simulations to describe the parameters that govern interfacial displacement is also addressed. This review thus provides ample information for characterising the interfacial properties over time in protein blend-stabilised emulsions, based on both experimental and modelling approaches.

Evaluation of oxygen partial pressure, temperature and stripping of antioxidants for accelerated shelf-life testing of oil blends using 1H NMR.

Lipid oxidation compromises the shelf-life of lipid-containing foods, leading to the generation of unpleasant off-flavours. Monitoring lipid oxidation under normal shelf-life conditions can be time-consuming (i.e. weeks or months) and therefore accelerated shelf-life conditions are often applied. However, little is known on their impact on the lipid oxidation mechanisms. In this study, different oxygen partial pressures (PO2; 10 and 21%), temperatures (20, 30 and 40 °C), and the removal of antioxidants through stripping of the oil were tested to accelerate lipid oxidation. Increasing the incubation temperature of stripped oil blends from 30 to 40 °C reduced the onset of lipid oxidation from 4 to 2 weeks, whereas the PO2 had no impact. Surprisingly, at room temperature, an increase in PO2 resulted in a longer onset time (10 weeks under 10% oxygen, 15 weeks under 21% oxygen). We hypothesize that this is due to a shift in (initiation) mechanism. In non-stripped oil, an increase in PO2 from 10 to 21% decreased the onset time from 16 to 10 weeks (40 °C). Temperature elevations and stripping led to a shift towards more trans-trans diene hydroperoxides, as compared to the cis-trans conformation. Additionally, oil stripping led to an increase in oxidized PUFAs with three or more double bonds in which the hydroperoxide group is located between the double bond pattern, instead of on the edge of it. Lastly, it was shown that small additions of LC-PUFAs (0, 0.3, 0.6, 1.2 and 2.3%, w/w) accelerate lipid oxidation, even in relatively stable stripped oils. In conclusion, increased PO2 and slightly elevated temperatures hold fair potential for accelerated shelf-life testing of non-stripped oils with a limited impact on the lipid oxidation mechanisms, whereas stripping significantly changes propagation mechanisms.

Sequential adsorption and interfacial displacement in emulsions stabilized with plant-dairy protein blends.

HYPOTHESIS: Many traditional or emergent emulsion products contain mixtures of proteins, resulting in complex, non-equilibrated interfacial structures. It is expected that protein displacement at oil-water interfaces depends on the sequence in which proteins are introduced during emulsion preparation, and on its initial interfacial composition. EXPERIMENTS: We produced emulsions with whey, pea or a whey-pea protein blend and added extra protein post-emulsification. The surface load was measured indirectly via the continuous phase, or directly via the creamed phase. The interfacial composition was monitored over a three-day period using SDS-PAGE densitometry. We compared these findings with results obtained using an automated drop tensiometer with bulk-phase exchange to highlight the effect of sequential protein adsorption on interfacial tension and dilatational rheology. FINDINGS: Addition of a second protein increased the surface load; especially pea proteins adsorbed to pre-adsorbed whey proteins, leading to thick interfacial layers. The addition of whey proteins to a pea protein- or whey-pea protein blend-stabilized emulsion led to significant displacement of the pea proteins by ß-lactoglobulin. We determined that protein-protein interactions were the driving force for this displacement, rather than a decrease in interfacial tension. These outcomes could be instrumental in defining new strategies for plant-animal protein hybrid products.

Natural particles can armor emulsions against lipid oxidation and coalescence.

Traditional functional ingredients, such as conventional emulsifiers (surfactants, animal-derived proteins), and synthetic antioxidants may become obsolete in the development of clean-label, plant-based, sustainable food emulsions. Previously, we showed that tailor-made antioxidant-loaded particles can yield both physically and oxidatively stable emulsions, and we expected that natural particles with related properties could also show these beneficial effects. Here, we investigated Pickering emulsions prepared with natural plant particulate materials. Particles that showed weak aggregation in acidic aqueous media, indicating a relatively hydrophobic surface, were able to physically stabilize oil-in-water emulsions, through either Pickering stabilization (powders of matcha tea, spinach leaves, and spirulina cake), or an increase in viscosity (pineapple fibers). Matcha tea and spinach leaf particle-stabilized emulsions were highly stable to lipid oxidation, as compared to emulsions stabilized by conventional emulsifiers. Taking this dual particle functionality as a starting point for emulsion design is, in our view, essential to achieve clean-label food emulsions.

Glycation of soy proteins leads to a range of fractions with various supramolecular assemblies and surface activities.

Dry and subsequent wet heating were used to glycate soy proteins with dextran or glucose, followed by fractionation based on size and solubility. Dry heating led to protein glycation (formation of furosine, Nε-(carboxymethyl)-l-lysine, Nε-(carboxyethyl)-l-lysine, and protein-bound carbonyls) and aggregation (increased particle size); while subsequent wet heating induced partial unfolding and de-aggregation. The measurable free amino group content of soy proteins changed from 0.77 to 0.14, then to 0.62 mmol/g upon dry and subsequent wet heating; this non-monotonic evolution is probably due to protein structural changes, and shows that this content should be interpreted with caution as a glycation marker. After both heating steps, the smaller-sized water-soluble fractions showed higher surface activity than the larger insoluble ones, and dextran conjugates exhibited a higher surface activity than their glucose counterparts. We thereby achieved a comprehensive understanding of the properties of various fractions in plant protein fractions, which is essential when targeting applications.

Conformational Changes of Whey and Pea Proteins upon Emulsification Approached by Front-Surface Fluorescence.

Proteins are widely used to stabilize emulsions, and plant proteins have raised increasing interest for this purpose. The interfacial and emulsifying properties of proteins depend largely on their molecular properties. We used fluorescence spectroscopy to characterize the conformation of food proteins from different biological origins (dairy or pea) and transformation processes (commercial or lab-made isolates) in solution and at the oil-water interface. The fourth derivative of fluorescence spectra provided insights in the local environment of tryptophan (Trp) residues and thus in the protein structure. In emulsions, whey proteins adsorbed with their Trp-rich region at the oil-water interface. Proteins in the commercial pea isolate were present as soluble aggregates, and no changes in the local environment of the Trp residues were detected upon emulsification, suggesting that these structures adsorb without conformational changes. The lab-purified pea proteins were less aggregated and a Trp-free region of the vicilin adsorbed at the oil-water interface.

Air-water interfacial behaviour of whey protein and rapeseed oleosome mixtures.

HYPOTHESIS: Plant seeds store lipids in oleosomes, which are storage organelles with a triacylglycerol (TAG) core surrounded by a phospholipid monolayer and proteins. Due to their membrane components, oleosomes have an affinity for the air/oil-water interface. Therefore, it is expected that oleosomes can stabilise interfaces, and also compete with proteins for the air-water interface. EXPERIMENTS: We mixed rapeseed oleosomes with whey protein isolate (WPI), and evaluated their air-water interfacial properties by interfacial rheology and microstructure imaging. To understand the contribution of the oleosome components to the interfacial properties, oleosome membrane components (phospholipids and membrane proteins) or rapeseed lecithin (phospholipids) were also mixed with WPI. FINDINGS: Oleosomes were found to disrupt after adsorption, and formed TAG/phospholipid-rich regions with membrane fragments at the interface, forming a weak and mobile interfacial layer. Mixing oleosomes with WPI resulted in an interface with TAG/phospholipid-rich regions surrounded by whey protein clusters. Membrane components or lecithin mixed with proteins also resulted in an interface where WPI molecules aggregated into small WPI domains, surrounded by a continuous phase of membrane components or phospholipids. We also observed an increase in stiffness of the interfacial layer, due to the presence of oleosome membrane proteins at the interface.

Interrelated Routes between the Maillard Reaction and Lipid Oxidation in Emulsion Systems.

In foods, the Maillard reaction (MR) and lipid oxidation lead to the formation of several molecules through interrelated chemical pathways. MR and lipid oxidation products were investigated in model oil-in-water emulsions consisting of canola oil, water, and Tween 20, a nonionic surfactant, with glucose and phenylalanine. The presence of 1% Tween 20, either in emulsion or as a control surfactant solution, sped up the formation of N-(1-deoxy-d-fructos-1-yl)-phenylalanine and of phenylacetaldehyde. Overall, the formation of MR products was up to sixteen times higher in emulsions than in an aqueous system without a surfactant. The formation of conjugated dienes, total aldehydes, hexanal, and (Z)-2-octenal was reduced down to six times when MR products were present in the emulsion. These results confirm that the formation of MR intermediates is influenced by the reactants' location, and the presence of a discrete nonpolar environment (oil droplets or surfactant micelles) promotes MR volatile formation through Strecker degradation.

Formation of Taste-Active Pyridinium Betaine Derivatives Is Promoted in Thermally Treated Oil-in-Water Emulsions and Alkaline pH.

The oil-water interface can be used as an efficient reaction controller in foods by carrying specific reactants and products in either the hydrophobic or hydrophilic phase. The formation of the taste-active compounds N-(1-carboxyethyl)-6-hydroxymethyl-pyridinium-3-ol inner salt (alapyridaine) and 1-(1-carboxyethyl)-3-hydroxy-pyridinium inner salt is influenced by the presence of a dispersed saturated triglyceride oil phase and by the pH of the aqueous phase. At pH 6.5, the formation of both betaines was 1.24 and 6 times higher in emulsions than in aqueous solution after 4 min at 140 °C. In alkaline emulsions (pH = 9.5, 4 min), the concentrations of alapyridaine and 1-(1-carboxyethyl)-3-hydroxy-pyridinium ion were 6.2 and 3.8 times higher, respectively, than in unbuffered emulsions as a result of the interaction between the polar head group of the surfactant and pyridinium rings. Here, we reported for the first time the effects of multiphase systems on the formation of nonvolatile, taste-active end products.

Pickering particles as interfacial reservoirs of antioxidants.

HYPOTHESIS: Emulsions are common structures encapsulating lipophilic bioactive molecules, both in biological systems and in manufactured products. Protecting these functional molecules from oxidation is essential; Nature excels at doing so by placing antioxidants at the oil-water interface, where oxidative reactions primarily occur. We imagined a novel approach to boost the activity of antioxidants in designer emulsions by employing Pickering particles that act both as physical emulsion stabilizers and as interfacial reservoirs of antioxidants. EXPERIMENTS: α-Tocopherol or carnosic acid, two model lipophilic antioxidants, were entrapped in colloidal lipid particles (CLPs) that were next used to physically stabilize sunflower oil-in-water emulsions ("concept" Pickering emulsions). We first assessed the physical properties and stability of the CLPs and of the Pickering emulsions. We then monitored the oxidative stability of the concept emulsions upon incubation, and compared it to that of control emulsions of similar structure, yet with the antioxidant present in the oil droplet interior. FINDINGS: Both tested antioxidants are largely more effective when loaded within Pickering particles than when solubilized in the oil droplet interior, thus confirming the importance of the interfacial localization of antioxidants. This approach revisits the paradigm for lipid oxidation prevention in emulsions and offers potential for many applications.

Behavior of plant-dairy protein blends at air-water and oil-water interfaces.

Recent work suggests that using blends of dairy and plant proteins could be a promising way to mitigate sustainability and functionality concerns. Many proteins form viscoelastic layers at fluid interfaces and provide physical stabilization to emulsion droplets; yet, the interfacial behavior of animal-plant protein blends is greatly underexplored. In the present work, we considered pea protein isolate (PPI) as a model legume protein, which was blended with well-studied dairy proteins (whey protein isolate (WPI) or sodium caseinate (SC)). We performed dilatational rheology at the air-water and oil-water interface using an automated drop tensiometer to chart the behavior and structure of the interfacial films, and to highlight differences between films made with either blends, or their constituting components only. The rheological response of the blend-stabilized interfaces deviated from what could be expected from averaging those of the individual proteins and depended on the proteins used; e.g. at the air-water interface, the response of the caseinate-pea protein blend was similar to that of PPI only. At the oil-water interface, the PPI and WPI-PPI interfaces gave comparable responses upon deformation and formed less elastic layers compared to the WPI-stabilized interface. Blending SC with PPI gave stronger interfacial layers compared to SC alone, but the layers were less stiff compared to the layers formed with WPI, PPI and WPI-PPI. In general, higher elastic moduli and more rigid interfacial layers were formed at the air-water interface, compared to the oil-water interface, except for PPI.

Oxidative stability of soy proteins: From ground soybeans to structured products.

The production of soy protein-based foods requires multiple-step, intensive processing and storage of soy ingredients, which can increase the product's susceptibility to oxidation. Therefore, we investigated the oxidative stability of soy protein-based products subjected to different relevant conditions or treatments: over storage of soy flours, over fractionation to yield soy protein isolate (SPI), and over subsequent thermomechanical processing to yield a model structured product. Soy flours were stable to lipid and protein oxidation over 250 days storage in chilled or ambient conditions. The fractionation process applied to make SPI did not increase substantially protein carbonylation, but increased surface-exposed hydrophobicity and decreased free thiols, compared to the starting defatted flour. Subsequent processing of hydrated SPI powder at 140 °C further increased protein carbonylation to a high extent. Therefore, we conclude that soy flours can be stable over long storage times, but processing to yield structured foods products promote protein oxidation.

Can we prevent lipid oxidation in emulsions by using fat-based Pickering particles?

Interest has recently been rising in the development of food-compatible Pickering emulsions, i.e., particle-stabilized emulsions, and various biobased particles have been demonstrated as useful for such a purpose. Most of the related work has focused on the physical stability of the emulsions, but whether such particles can be advantageous in terms of chemical stability, and in particular, with regard to lipid oxidation, is largely unexplored. Recently, we found that colloidal lipid particles (CLPs) are efficient Pickering stabilizers, and the objective of the present study was to investigate the oxidative stability of emulsions stabilized with those particles. Three types of sunflower oil-in-water (O/W) emulsions were considered: Pickering emulsions stabilized with colloidal lipid particles (CLPs) made of high melting point (HMP) fat (tripalmitin or palm stearin), adsorbed onto the liquid oil droplets; and, as references, two conventional sodium caseinate-stabilized emulsions, of which one contained only liquid oil, and the other liquid oil mixed with HMP fat as the core of the emulsion droplets. In the presence of iron, the latter oxidized faster than conventional liquid oil and Pickering emulsions, resulting in 2- to 3-fold higher amounts of primary and secondary lipid oxidation products. This may be due to intra-droplet HMP fat pushing oxidizable lipids towards the oil-water interface, which would promote lipid oxidation. This shows that the localization of solid fat in O/W emulsions affects lipid oxidation. We also found that CLP-stabilized Pickering emulsions had similar oxidation rates as conventional sodium caseinate-stabilized emulsions containing only liquid oil. This suggests that the potential of such Pickering particles to prevent lipid oxidation is limited. This could be because diffusion of small pro-oxidant molecules is not hindered by Pickering particles, as they cannot form an interfacial barrier that is structurally homogeneous at such a small scale.

Encapsulation of lipids as emulsion-alginate beads reduces food intake: a randomized placebo-controlled cross-over human trial in overweight adults.

The objective of this study was to investigate the efficacy of lipid emulsions encapsulated in calcium-alginate beads in reducing food intake and appetite sensations. These emulsion-alginate beads were ingested in a yogurt (active) and compared to an equienergetic yogurt containing nonencapsulated nutrients with comparable sensory properties (control) in a randomized placebo-controlled trial with crossover design. Thirty-three healthy overweight volunteers (mean age: 43 years; body mass index: 27.7 kg/m2; 14 male) received the 2 treatments. Test days started with a standardized small breakfast (t = 0) followed by an active or control yogurt (t = 90 minutes). Appetite sensations and gastrointestinal symptoms were monitored prior to and after consumption of the yogurt, and food intake was measured during ad libitum pasta meal consumption (t = 210 minutes). The hypothesis for this study was that delayed release of encapsulated lipids suppresses appetite sensations and reduces food intake. Food intake was significantly reduced with 51 ±â€¯20 kcal (213 ± 84 kJ) (P = .016) after intake of the active yogurt (770 ±â€¯38 kcal (3222 ± 159 kJ)) compared to the control (821 ±â€¯40 kcal (3435 ± 167 kJ)). The approach that we chose is promising to reduce food intake and could contribute to the development of an easy-to-use product for weight management.