Depletion-Induced Flocculation of Concentrated Emulsions Probed by Photon Density Wave Spectroscopy.

Stable, creaming-free oil in water emulsions with high volume fractions of oil (ϕ = 0.05-0.40, density matched to water) and polysorbate 80 as an emulsifier were characterized without dilution by Photon Density Wave spectroscopy measuring light absorption and scattering behavior, the latter serving as the basis for droplet size distribution analysis. The emulsion with ϕ = 0.10 was used to investigate flocculation processes induced by xanthan as a semi-flexible linear nonabsorbing polymer. Different time regimes in the development of the reduced scattering coefficient µs' could be identified. First, a rapid, temperature-dependent change in µs' during the depletion process was observed. Second, the further decrease of µs' follows a power law in analogy to a spinodal demixing behavior, as described by the Cahn-Hilliard theory.

Interfacial structurization between triolein and water from pH and buffer ions.

HYPOTHESIS: Understanding and manipulating the oil/water interface is important across various industries, including food, pharmaceuticals, cosmetics, and detergents. Many of these processes occur under elevated pH conditions in buffer systems, where base-catalyzed hydrolysis of triglyceride ester bonds leads to amphiphilic reaction products such as fatty acids. EXPERIMENTS: Here, pH-triggered alterations of the triolein/water interface are analyzed in the presence of phosphate (PB) and tris(hydroxymethyl)aminomethane (TRIS). Ellipsometry at the liquid/liquid interface, tensiometry, and scanning small angle X-ray scattering are used to study the formation of structures at the oil/water interface. Confocal Raman microscopy, nuclear magnetic resonance spectroscopy, and in silico modeling analyze compositional changes in the interfacial region. FINDINGS: pH and buffer ions were discovered to significantly modify the triglyceride/water interface, contrary to the decane/water control. Decreasing interfacial tensions from 32.4 to 2.2 mN/m upon pH increase from 6.5 to 9.5 is seen with multilamellar interfacial layers forming at pH around 9.0 in the presence of TRIS. Oleic acid from triolein hydrolysis and its further interaction with TRIS is held responsible for this. The new understanding can guide the design of pH- and ion-responsive functional materials and optimize industrial processes involving triglyceride/water interfaces.

Directed Discovery of Tetrapeptide Emulsifiers.

Oil in water emulsions are an important class of soft material that are used in the food, cosmetic, and biomedical industries. These materials are formed through the use of emulsifiers that are able to stabilize oil droplets in water. Historically emulsifiers have been developed from lipids or from large biomolecules such as proteins. However, the ability to use short peptides, which have favorable degradability and toxicity profiles is seen as an attractive alternative. In this work, we demonstrate that it is possible to design emulsifiers from short (tetra) peptides that have tunability (i.e., the surface activity of the emulsion can be tuned according to the peptide primary sequence). This design process is achieved by applying coarse grain molecular dynamics simulation to consecutively reduce the molecular search space from the 83,521 candidates initially considered in the screen to four top ranking candidates that were then studied experimentally. The results of the experimental study correspond well to the predicted results from the computational screening verifying the potential of this screening methodology to be applied to a range of different molecular systems.

Colloidal dynamics of emulsion droplets in mouth.

The interaction of emulsions with the tongue is key to the sensory appeal of food and can potentially be exploited for oral/buccal pharmaceutical delivery. Whilst there is good understanding of the different mucoadhesive forces governing emulsion interaction with the tongue, their relative importance is not well understood. In addition, the physical location of emulsions within the saliva papillae on the tongue is not understood at all. A combination of ex vivo salivary film, and in vivo oral coating experiments were used to determine the importance of different mucoadhesive forces. Mucoadhesion of cationic emulsions was largely driven by electrostatic complexation. SDS-PAGE of the in vivo saliva coating highlighted that mucins were largely responsible for cationic emulsion mucoadhesion. Anionic emulsions were bound via hydrophobic/steric interactions to small salivary proteins typically located away from the mucin anchor points. The physical location and clustering of emulsions relative to the salivary film/papillae was probed via the invention of a fluorescent oral microscope. Cationic emulsions were densely clustered close to the papillae whilst anionic emulsions were suspended in the salivary film above the papillae. Interestingly, non-ionic emulsions were also trapped within the salivary film above the papillae as individual droplets. These findings highlight that whilst electrostatic complexation with saliva is a powerful mucoadhesive force, hydrophobic and steric interactions also act to induce oral retention of emulsions. The differences in physical location and clustering of emulsions within the salivary film hint at the 3D locations of the different salivary proteins driving each mucoadhesive interaction. This novel understanding of emulsion saliva/papillae interactions has potential to aid efficacy of buccal pharmaceutical delivery and the reduction of astringency in plant-based foods.

Fabrication of a Novel Protein Sponge with Dual-Scale Porosity and Mixed Wettability Using a Clean and Versatile Microwave-Based Process.

An open-porous protein sponge with mixed wettability is presented made entirely from whey proteins and with promising applications in biomedicine, pharmaceutical, and food industry. The fabrication relies on an additive-free, clean and scalable process consisting of foaming followed by controlled microwave-convection drying. Volumetric heating throughout the matrix induced by microwaves causes fast expansion and elongation of the foam bubbles, retards crust formation and promotes early protein denaturation. These effects counteract collapse and shrinkage typically encountered in convection drying of foams. The interplay of high protein content, tailored gas incorporation and controlled drying result in a dried structure with dual-scale porosity composed of open macroscopic elongated foam bubbles and microscopic pores in the surrounding solid lamellae induced by water evaporation. Due to the insolubility and mixed wettability of the denatured protein network, polar and non-polar liquids are rapidly absorbed into the interconnected capillary system of the sponge without disintegrating. While non-watery liquids penetrate the pores by capillary suction, water diffuses also into the stiff protein matrix, inducing swelling and softening. Consequently, the water-filled soft sponge can be emptied by compression and re-absorbs any wetting liquid into the free capillary space.

Influence of the stabilizer concentration on the internal liquid crystalline order and the size of oil-loaded monolinolein-based dispersions.

The internal phase of monolinolein-based dispersions loaded with tetradecane or (R)-(+)-limonene was investigated as a function of the stabilizer content by small-angle X-ray scattering. Phase transitions at the colloidal scale were found in some of nanostructured aqueous dispersions by increasing the stabilizer content. For particles containing a bicontinuous cubic phase, a large increase of the stabilizer concentration promoted a liquid crystalline phase transition from the Pn3m to the Im3m cubic symmetry. The coexistence of both phases is observed in an intermediate stabilizer concentration range. For particles with an internal micellar cubic Fd3m symmetry, the internal structure changes in the isotropic fluid L(2) phase. In case of particles with an internal hexagonal phase (H(2) symmetry), the increasing amount of stabilizer did not alter the lattice parameter but decreased the size of the nanostructured domain. Moreover, we showed for hexagonal and emulsified micellar phase particles that the increase of the stabilizer content induced a strong decrease of the mean hydrodynamic size of the particles, allowing producing nanostructured lipid-based liquid crystalline particles down to a radius of 70 nm at the same energy input.

Enzymatic cross-linking of pectin in a high-pressure foaming process.

The enzyme laccase is a copper-containing oxidoreductase with the ability to oxidize a wide range of substrates, such as ferulic acid. Thus, the ferulic acid-containing sugar beet pectin (SBP) can be cross-linked through laccase-mediated oxidation. As cross-linking increases viscosity, it could be applied to stabilize SBP-containing foams. In this study, laccase-mediated cross-linking of SBP was investigated under conditions of a high-pressure foaming process. Shear, presence of CO2, and pressure were simulated in a rheometer equipped with a high-pressure cell. At rest, addition of laccase to SBP solution led to the formation of a stiff gel. Application of shear upon mixing of laccase and SBP solution decreased the storage modulus with increasing shear duration and shear rate. This can be attributed to the formation of a fluid gel. However, when shear was stopped before all available ferulic acid groups were cross-linked, a stronger and more coherent network was formed. Pressure exerted by CO2 did not affect cross-linking. Additionally, this approach was tested in a stirred high-pressure vessel where SBP was foamed through CO2 dissolution under pressure and shear followed by controlled pressure release. While pure SBP foam was highly unstable, addition of laccase decelerated collapse. Highest stability was reached when laccase and SBP were mixed prior to depressurization. At the point of foam formation, the continuous phase was thereby viscous enough to increase foam stability. At the same time, continuation of cross-linking at rest caused gel templating of the foam structure.

Model studies on the release of aroma compounds from structured and nonstructured oil systems using proton-transfer reaction mass spectrometry.

Relative retention, volatility, and temporal release of volatile compounds taken from aldehyde, ester, and alcohol chemical classes were studied at 70 degrees C in model systems using equilibrium static headspace analysis and real time dynamic headspace analysis. These systems were medium-chain triglycerides (MCT), sunflower oil, and two structured systems, i.e., water-in-oil emulsion and L2 phase (water-in-oil microemulsion). Hydrophilic domains of the emulsion type media retained specifically the hydrophilic compounds and alcohols. Four kinetic parameters characterizing the concentration- and time-dependent releases were extracted from the aroma release curves. Most of the kinetic parameter values were higher in structured systems than in oils particularly when using MCT. The oil nature was found to better control the dynamic release profiles than the system structures. The release parameters were well-related (i) to the volatile hydrophobicity as a function of the oil used and (ii) to the retention data in the specific case of the L2 phase due to a specific release behavior of alcohols.

Self-assembly of polar food lipids.

Polar lipids, such as monoglycerides and phospholipids, are amphiphilic molecules commonly used as processing and stabilization aids in the manufacturing of food products. As all amphiphilic molecules (surfactants, emulsifiers) they show self-assembly phenomena when added into water above a certain concentration (the critical aggregation concentration). The variety of self-assembly structures that can be formed by polar food lipids is as rich as it is for synthetic surfactants: micelles (normal and reverse micelles), microemulsions, and liquid crystalline phases can be formulated using food-grade ingredients. In the present work we will first discuss microemulsion and liquid crystalline phase formation from ingredients commonly used in food industry. In the last section we will focus on three different potential application fields, namely (i) solubilization of poorly water soluble ingredients, (ii) controlled release, and (iii) chemical reactivity. We will show how the interfacial area present in self-assembly structures can be used for (i) the delivery of functional molecules, (ii) controlling the release of functional molecules, and (iii) modulating the chemical reactivity between reactive molecules, such as aromas.

A new insight into the formation of odor active carbonyls by thermally-induced degradation of phospholipids in self-assembly structures.

The role of molecular organization in heated aqueous dispersions of egg phosphatidylcholine (PC) and egg phosphatidylethanolamine (PE) was studied with respect to the formation of key odorants. Evidence was found for the crucial role of self-assembly structures adopted by phospholipid molecules on the quantitative composition of volatile constituents. The concentrations of seven aldehydes and one vinyl ketone were determined by isotope dilution assay in heated aqueous dispersions of PC and PE present in various ratios. Addition of PE to PC drastically decreased the amount of (E,E)-2,4-decadienal formed, which cannot be explained by the differences in the fatty acid composition of PC and PE. The free amino group in PE does not explain this phenomenon either, as replacing PE by phosphatidic acid distearylester also reduced the amounts of (E,E)-2,4-decadienal. We suggest that the type of self-assembly structure adopted by phospholipids in water significantly influences the reaction yields. However, the mechanisms leading to the preferred formation of phospholipid-derived odorants in a lamellar phase, as compared to the reversed hexagonal phase, remain unknown.

Five-component food-grade microemulsions: structural characterization by SANS.

In this paper we present the structural characterization of a five-component food-grade microemulsion containing Tween 80, R(+)-limonene, ethanol, glycerol, and water. Our main approach to investigating the microstructure of dense microemulsions, and how it can be influenced by the various components, was to employ small-angle neutron scattering and the new evaluation technique for dense, interacting systems, the Generalized Indirect Fourier Transformation. We started our investigation with the impact of glycerol and ethanol on Tween 80 micelles in water. We found that glycerol increases the aggregation number and withdraws the hydrating agents from the headgroup region of the surfactant, resulting in a higher packing density of molecules in a micelle at slightly increasing size. The same trend holds when the micelles are oil swollen and/or ethanol is present. Ethanol, on the other hand, redistributes mainly between water and the interface-headgroup region of the surfactant. Part of it replaces surfactant molecules in the micelles, which increases the available interface and results in a higher number of micelles with shrinking size. The same trend holds when the micelles are oil swollen and/or glycerol is present in the aqueous phase. We also investigated samples along the dilution of a mixture of surfactant and oil phase (R(+)-limonene and ethanol), which can be diluted with aqueous phase (mixture of water and glycerol) without the occurrence of phase separation. In some samples of this dilution most probably bicontinuous structures are present. To elucidate this point, we also employed dynamic light scattering, viscosity, and conductivity measurements.

Internally self-assembled particles entrapped in thermoreversible hydrogels.

The present study describes the development of thermogelling emulsions by the entrapment of internally self-assembled emulsion droplets (ISAsomes) within a thermoreversible hydrogel made of kappa-carrageenan. The droplets are emulsified mesophases of cubic or hexagonal order, or emulsified micro-emulsions. Above 60 degrees C, the system was fluid and composed of a mixture of internally nanostructured small droplets and polymer chains dispersed in water. Below 60 degrees C, a physical gel with entrapped droplets was formed. A tuning of the temperature in order to switch between the gel and solution state did not affect the particles in terms of size. The thermoreversible behavior of the loaded polymer network and the effects on the internal structure of cubosomes, hexosomes and emulsified micro-emulsions was investigated by SAXS. We showed that the phase borders may be shifted due to the presence of the kappa-carrageenan network, which alter the internal nanostructure of the droplets. This can induce a transformation from emulsified micro-emulsions to micellar cubosomes. In the hexagonal case, the lattice parameters of the hexosomes are slightly modified.

Internally self-assembled thermoreversible gelling emulsions: ISAsomes in methylcellulose, kappa-carrageenan, and mixed hydrogels.

Self-assembled thermo-gelling emulsions were developed by blending internally self-assembled particles (ISAsomes) with thermoreversible polysaccharide hydrogels of methylcellulose (MC), kappa-carrageenan (KC), and their 1:1 mixture. In this way, the hierarchical structure of ISAsome samples was successfully promoted. The gelified polymer network corresponds to the highest level of the hierarchical structure and as such represents the capturing matrix for the medium structural level, i.e., dispersed emulsion particles, which are further internally structured as the lowest level of structure. Utilizing small-angle X-ray scattering, differential scanning calorimetry, dynamic light scattering, and oscillatory rheological experiments in the temperature regime from 20 to 70 degrees C, we were able to show that the ISAsomes stay practically intact during such embedment into a hydrogel matrix retaining its internal self-assembled structure and its functionality. The characteristic sol-gel and gel-sol transition temperatures of the ISAsome-loaded hydrogel samples showed a slight shift in comparison to the unloaded hydrogel samples. Furthermore, we found that MC is actually able to stabilize the ISAsomes at higher temperatures (tests were conducted up to 90 degrees C). Gels made from MC and KC show quite different features in terms of rheology and differential scanning calorimetry. However, the most interesting results were obtained for the ISAsome-loaded MC-KC (1:1) gelifying system, which behaves as a low- and high-temperature gel with a narrow intermediate temperature window where it is a sol. This specific thermal behavior allows for easy temperature tuning of the system's aggregate state as well as the internal self-assembled structure. As such, this system is suggested to be further tested as the potential media for a temperature-controlled burst/sustained release media of various hydrophilic, hydrophobic, or amphiphilic guest functional molecules.

Mixed layers of sodium caseinate + dextran sulfate: influence of order of addition to oil-water interface.

We report on the interfacial properties of electrostatic complexes of protein (sodium caseinate) with a highly sulfated polysaccharide (dextran sulfate). Two routes were investigated for preparation of adsorbed layers at the n-tetradecane-water interface at pH = 6. Bilayers were made by the layer-by-layer deposition technique whereby polysaccharide was added to a previously established protein-stabilized interface. Mixed layers were made by the conventional one-step method in which soluble protein-polysaccharide complexes were adsorbed directly at the interface. Protein + polysaccharide systems gave a slower decay of interfacial tension and stronger dilatational viscoelastic properties than the protein alone, but there was no significant difference in dilatational properties between mixed layers and bilayers. Conversely, shear rheology experiments exhibited significant differences between the two kinds of interfacial layers, with the mixed system giving much stronger interfacial films than the bilayer system, i.e., shear viscosities and moduli at least an order of magnitude higher. The film shear viscoelasticity was further enhanced by acidification of the biopolymer mixture to pH = 2 prior to interface formation. Taken together, these measurements provide insight into the origin of previously reported differences in stability properties of oil-in-water emulsions made by the bilayer and mixed layer approaches. Addition of a proteolytic enzyme (trypsin) to both types of interfaces led to a significant increase in the elastic modulus of the film, suggesting that the enzyme was adsorbed at the interface via complexation with dextran sulfate. Overall, this study has confirmed the potential of shear rheology as a highly sensitive probe of associative electrostatic interactions and interfacial structure in mixed biopolymer layers.

Competition between lipases and monoglycerides at interfaces.

Tensiometry (the pendant drop technique), interfacial shear rheology, and ellipsometry have been used to study the effect of polar lipids that are generated during fat digestion on the behavior of lipases at the oil-water interface. Both Sn-1,3 regiospecific and nonregiospecific lipases have been used, and a noncatalytically active protein, beta-lacloglobulin, has been used as reference in the interfacial shear rheology experiments. The results from the pendant drop measurements and the interfacial rheology studies were in agreement with each other and demonstrated that the Sn-2 monoglyceride, which is one of the lipolysis products generated when a Sn-1,3 regiospecific lipase catalyzes triglyceride hydrolysis, is very interfacially active and efficiently expels the enzyme from the interface. Ellipsometry conducted at the liquid-liquid interface showed that the lipase forms a sublayer in the aqueous phase, just beneath the monoglyceride-covered interface. Sn-1/3 monoglycerides do not behave this way because they are rapidly degraded to fatty acid and glycerol and the fatty acid (or the fatty acid salt) does not have enough interfacial activity to expel the lipase from the interface. Since the lipases present in the gastrointestinal tract are highly Sn-1,3 regiospecific, we believe that the results obtained can be transferred to the in vivo situation. The formation of stable and amphiphilic Sn-2 monoglycerides can be seen as a self-regulatory process for fat digestion.

Influence of Surfactants on Lipase Fat Digestion in a Model Gastro-intestinal System.

In the present study, we use a model gastro-intestinal system to study the influence of different food-grade surface-active molecules (Sn-2 monopalmitin, beta-lactoglobulin, or lysophosphatodylcholine) on lipase activity. The interfacial activity of lipase and surfactants are assessed with the pendant drop technique, a commonly used tensiometry instrument. A mathematical model is adopted which enables quantitative determination of the composition of the water-oil interface as a function of bulk surfactant concentration in the water-oil mixtures. Our results show a decrease in gastric lipolysis when interfacially active molecules are incorporated into a food matrix. However, only the Sn-2 monopalmitin caused a systematic decrease in triglyceride hydrolysis throughout the gastro-intestinal tract. This effect is most likely due to exclusion of both lipase and triglyceride from the water-oil interface together with a probable saturation of the solubilization capacity of bile with monoglycerides. Addition of beta-lactoglobulin or lysophopholipids increased the hydrolysis of fat after the gastric phase. These results can be attributed to an increasing interfacial area with lipase and substrate present at the interface. Otherwise, beta-lactoglobulin, or lysophopholipids reduced fat hydrolysis in the stomach. From the mathematical modeling of the interface composition, we can conclude that Sn-2 monopalmitin can desorb lipase from the interface, which, together with exclusion of substrate from the interface, explains the gradually decreased triglyceride hydrolysis that occurs during the digestion. Our results provide a biophysics approach on lipolysis that can bring new insights into the problem of fat uptake.

Oil-loaded monolinolein-based particles with confined inverse discontinuous cubic structure (Fd3m).

In our recent work, we reported on the effect of varying temperature and solubilizing tetradecane (TC) on the structural transitions observed in dispersed particles based on the monolinolein (MLO)-water-TC system. At a given temperature, the addition of TC induces a transition of the internal structure from the bicontinuous cubic phase, Pn3m, to the reversed hexagonal, H2, and to the isotropic liquid phase (water-in-oil (W/O) microemulsions). Our present study focuses on the discovery of a Fd3m phase (reversed discontinuous micellar cubic), which is formed in the MLO-water-TC system at a specific TC/MLO weight ratio. It is situated between the H2 and the isotropic liquid phase (W/O microemulsion). Remarkably, it is not found in the absence of TC by increasing the temperature. The Fd3m structure was investigated in detail by means of small-angle X-ray scattering (SAXS), and cryogenic transmission electron microscopy (cryo-TEM). The present work proves that the structural transformation in the dispersed particles from H2 (hexosomes) to the W/O microemulsion system (emulsified microemulsion (EME)) is indirect and it occurs gradually via an emulsified intermediate phase. Specifically, in addition to the nanostructured aqueous dispersions described above, we present new TC-loaded aqueous dispersions with a confined intermediate phase, which is a discontinuous micellar cubic phase of the symmetry Fd3m. We denoted this type of emulsified particles as "micellar cubosomes".

Control of the internal structure of MLO-based isasomes by the addition of diglycerol monooleate and soybean phosphatidylcholine.

This work describes the effect of two different surfactants on the internal nanostructure of the kinetically stabilized isasomes (internally self-assembled particles or "somes"), which are a new family of colloidal particles (cubosomes, hexosomes, micellar cubosomes, and emulsified microemulsions, EME). The stabilization of these systems is performed by using the polymeric stabilizer F127. We demonstrate that the internal structure of these oil-free and oil-loaded dispersed particles can be modulated by varying the lipid composition. To achieve this goal, we replaced part of our primary lipid monolinolein (MLO) with diglycerol monooleate (DGMO) or soybean phosphatidylcholine (PC). We found that DGMO has a counter effect to that of tetradecane (TC) and allows us to tune back the self-assembled nanostructure in the TC-loaded dispersions from H2 (hexosomes) to Im3m (cubosomes). Although TC has a higher impact on confined structures than does DGMO, we demonstrate that the addition of DGMO significantly affects the internal structure of the TC-solubilized dispersions and favors the formation of large water channels. PC can also be used to modify the internal structure for MLO-based systems. It is somehow different from DGMO due to the fact that the fully hydrated Pn3m cubic structure in the presence of PC for the TC-free dispersion is preserved after dispersing. The results also indicate that PC is less effective than DGMO for tuning back the TC-loaded internal structure from H2 to cubic phase, in which it makes the confined structure less ordered. In addition, we found that DGMO has a significant effect on the internal structure of isasomes. It increases the water solubilization capacity for dispersed and nondispersed bulk phases. In contrast to the MLO-based dispersions, the present results indicate that F127 plays an important role in the internal structure of these dispersions due to its penetration into the oil-free cubic phase changing the symmetry from Pn3m to Im3m.

Emulsified microemulsions and oil-containing liquid crystalline phases.

Self-assembled nanostructures, such as inverted type mesophases of the cubic or hexagonal geometry or reverse microemulsion phases, can be dispersed using a polymeric stabilizer, such as the PEO-PPO-PEO triblock copolymer Pluronic F127. The particles, which are described in the present study, are based on monolinolein (MLO)-water mixtures. When adding tetradecane (TC) to the MLO-water-F127 system at constant temperature, the internal nanostructure of the kinetically stabilized particles transforms from a Pn3m (cubosomes) to a H2 (hexosomes) and to a water-in-oil (W/O, L2) microemulsion phase (emulsified microemulsion (EME)). To our knowledge, this is the first time that the formation of stable emulsified microemulsion (EME) systems has been described and proven to exist even at room temperature. The same structural transitions can also be induced by increasing temperature at constant tetradecane content. The internal nanostructure of the emulsified particles is probed using small-angle X-ray scattering (SAXS) and cryogenic transmission electron microscopy (cryo-TEM). At each investigated composition and temperature, the internal structure of the dispersions is observed to be identical to the corresponding structure of the nondispersed, fully hydrated bulk phase. This is clear evidence for the fact that the self-assembled inner particle nanostructure is preserved during the dispersion procedure. In addition, the internal structure of the particles is in thermodynamic equilibrium with the surrounding water phase. The internal structure of the dispersed, kinetically stabilized particles is a "real" and stable self-assembled nanostructure. To emphasize this fact, we denoted this new family of colloidal particles (cubosomes, hexosomes, and EMEs) as "ISASOMES" (internally self-assembled particles or "somes").

Effect of time on the interfacial and foaming properties of beta-lactoglobulin/acacia gum electrostatic complexes and coacervates at pH 4.2.

The electrostatic complexation between beta-lactoglobulin and acacia gum was investigated at pH 4.2 and 25 degrees C. The binding isotherm revealed a spontaneous exothermic reaction, leading to a DeltaHobs = -2108 kJ mol(-1) and a saturation protein to polysaccharide weight mixing ratio of 2:1. Soluble electrostatic complexes formed in these conditions were characterized by a hydrodynamic diameter of 119 +/- 0.6 nm and a polydispersity index of 0.097. The effect of time on the interfacial and foaming properties of these soluble complexes was investigated at a concentration of 0.1 wt % at two different times after mixing (4 min, referred as t approximately 0 h and t = 24 h). At t approximately 0 h, the mixture is mainly made of aggregating soluble electrostatic complexes, whereas after 24 h these complexes have already insolubilize to form liquid coacervates. The surface elasticity, viscosity and phase angle obtained at low frequency (0.01 Hz) using oscillating bubble tensiometry revealed higher fluidity and less rigidity in the film formed at t approximately 0 h. This observation was confirmed by diminishing bubble experiments coupled with microscopy of the thin film. It was thicker, more homogeneous and contained more water at t approximately 0 h as compared to t = 24 h (thinner film, less water). This led to very different gas permeability's of Kt approximately 0 h = 0.021 cm s(-1) and Kt=24 h) = 0.449 cm s(-1), respectively. Aqueous foams produced with the beta-lactoglobulin/acacia gum electrostatic complexes or coacervates exhibited very different stability. The former (t approximately 0 h) had a stable volume, combining low drainage rate and mainly air bubble disproportionation as the destabilization mechanism. By contrast, using coacervates aged for 24 h, the foam was significantly less stable, combining fast liquid drainage and air bubble destabilization though fast gas diffusion followed by film rupture and bubble coalescence. The strong effect of time on the air/water interfacial properties of the beta-lactoglobulin/acacia gum electrostatic complexes can be understood by their reorganization at the interface to form a coacervate phase that is more fluid/viscous at t approximately 0 h vs rigid/elastic at t = 24 h.