Abraham Aserin

Protein-Polysaccharide Interactions for Stabilization of Food Emulsions

ABSTRACT Proteins, polysaccharides and their blends, as examples of natural biopolymers, are surface active materials. Biopolymers may be considered as amphiphilic macromolecules that play an essential role in stabilizing food formulations (foams, emulsions and dispersions). Under specific conditions (such as protein-to-polysaccharide ratio, pH, ionic strength, temperature, mixing processing), it has been stated that proteins and polysaccharides form hybrids (complexes) with enhanced functional properties in comparison to the proteins and polysaccharides alone. Different protein-polysaccharide pairs are reviewed with particular attention to the emulsification capability of their mixtures. In the case of uncomplexed blends of biopolymers, competitive adsorption onto hydrophobic surfaces is generally reported. Conversely, electrostatic complexation between oppositely charged proteins and polysaccharides allows better anchoring of the new-formed macro-molecular amphiphile onto oil-water interfaces. Moreover, improved thermal stability and increased resistance to external treatment (high pressure) involved in food processing are obtained. This review presents basic and applied knowledge on protein-polysaccharide interactions in aqueous medium and at the oil-water interface in food emulsion systems. Electrostatic interactions and thermodynamic incompatibility in mixed biopolymer solutions are correlated to the functional properties (rheology, surface hydrophobiciry, emulsification power) of these interesting blends. Basic and industrial selected systems of different families of hydrocolloids (as gum Arabic, galactomannans, pectins) and protein (caseins, whey, soya, gelatin) mixtures are reviewed.

Phase behavior of microemulsions based on food-grade nonionic surfactants: effect of polyols and short-chain alcohols ☆

Abstract The improved water and oil solubilization in the presence of polyols (propylene glycol, PG, and glycerol, Gly) and short-chain alcohol (ethanol) in U-type nonionic W/O and O/W food microemulsions was investigated. The phase behavior of systems based on Tweens (ethoxylated sorbitan esters) was compared with non-food-grade systems based on C18:1E10 (Brij 96v). Short-chain alcohol (ethanol in food-grade systems) together with polyols (glycerol and propylene glycol) when added to a three component system (oil–surfactant–water) induce the formation of both water-in-oil (W/O) and oil-in-water (O/W) microemulsions. Alcohols and polyols destabilize the liquid crystalline phase and extend the isotropic region to higher surfactant concentrations. The total monophasic area, AT, at R(+)-limonene/ethanol of 1/1 (w/w) and aqueous phase of water/PG of 1/1 (w/w), was 73 and 64% of the total area of the phase diagram for Brij 96v and Tween 60, respectively. The transition from a W/O microemulsion into an O/W microemulsion happens gradually, and continuously without any phase separation. The total monophasic area depends also on the type of the oil, on the composition of the polar and apolar phases, and on the nature of the polyol. The results are discussed in terms of BSO equation, spontaneous curvature, H0, film flexibility, κ and κ , surfactant oil and surfactant cosolvent compatibility and the participation of the polyol at the interface. The difference in temperature sensitivity of PG-based microemulsions vs. temperature sensitivity of Gly-based is demonstrated and explained.

Double emulsions stabilized by macromolecular surfactants

Abstract Multiple emulsions are emulsions within emulsions, stabilized traditionally by monomeric emulsifiers both at the inner and outer interface. Double emulsions are thermodynamically unstable and fast coalescence, as well as fast release of markers and drugs, have been the main drawbacks of this technology. Polymeric synthetic emulsifiers, as well as natural macromolecules in combination with monomeric emulsifiers, have recently been studied and evaluated. The review brings new results on the use of amphiphilic proteins such as BSA and casein along with monomeric emulsifiers in relation to improved stability and release properties. Kinetic results are brought to demonstrate the role of BSA as the inner emulsifier and the outer emulsifier. Diffusion controlled mechanisms are suggested. Synthetic block copolymers based on silicon backbones and polyethylene oxide side-chains have also been studied. Double emulsions with relatively small droplets (5 μm) are formed and excellent stability (to storage, sheer and dilution) is obtained. Release data indicates that the newly designed double emulsions have longer shelf-life and very slow rates of release of markers. Such emulsions are good candidates for agricultural formulations.

Enhanced stabilization of cloudy emulsions with gum Arabic and whey protein isolate

Cloudy emulsions are oil-in-water (O/W) emulsions normally prepared as concentrates, further diluted, per request, into the final beverage. The cloudy emulsion provides flavor, color, and cloud (turbidity) to the soft drink. These systems are stabilized by emulsifiers and/or amphiphilic polysaccharides. Cloudy emulsions based on naturally occurring food grade emulsifiers were studied in the present work. Two charged natural biopolymers, whey protein isolate (WPI) and gum Arabic (GA), are interacted in aqueous solution to form charge-charge interactions improving the emulsion stability. The emulsions were high sheared (Microfluidizer) and characterized by particle size distribution analysis (DLS), optical centrifugation (LUMiFuge), optical microscopy observations, and turbidity measurements. Emulsions obtained from 10wt% of 3:1wt. ratio WPI:GA, at pH 7 (10wt% canola oil) show better stability than emulsions stabilized by GA or WPI alone. The droplet sizes were smaller than 1microm and did not grow significantly during 1 month of incubation at 25 degrees C. The D-limonene-based emulsion droplets were larger (> 2microm) than those made with vegetable oils immediately after preparation and underwent significant droplet size increase (coalescence) within 1 month (>8 microm). The emulsion with turbidity suitable as a cloudy emulsion was composed of 3wt% WPI:GA (3:1) and 20wt% canola oil.

Mechanistic considerations on the release of electrolytes from multiple emulsions stabilized by BSA and nonionic surfactants

Abstract The stability of w/o/w emulsions has been significantly improved by using a blend of nonionic surfactant (Span 80) and bovine serum albumin (BSA) as an interfacial complex in the inner phase. Improved stability was obtained by replacing the common nonionic hydrophilic monomeric emulsifier by BSA in the outer phase. Optimum stability and droplet size of double emulsion was achieved with 0.2 wt% BSA+10 wt% Span 80 as emulsifier I and 5.0 wt% Span 80:Tween 80 (2:3) or 0.1 wt% BSA as emulsifier II. Significant slow release of NaCl was obtained. The results were examined in view of Higuchi mechanism and it was found that one can account for the thickness of the inner interfacial complex from the plot of B (diffusion parameter correlated to the fraction of electrolyte release) vs. time. It can be clearly seen that in the inner phase the surfactant and the BSA act synergistically (active interfacial complex), and enhance stability and reduce release. In the outer phase BSA has mainly a stabilizing effect with a limited release retardation effect. Effective diffusion coefficients for each BSA concentration in the inner and outer phase have been calculated and evaluated in view of the diffusion controlled mechanism and the thickness of the viscoelastic film which is formed.

HII mesophase and peptide cell-penetrating enhancers for improved transdermal delivery of sodium diclofenac

This study develops a novel transdermal delivery vehicle for the enhanced delivery of sodium diclofenac (Na-DFC). The system utilizes the advantages of reversed hexagonal lyotropic liquid crystals (H(II)LC), combined with a peptide cell penetration enhancer (CPE), creating together an adaptable system that provides versatile options in the field of transdermal delivery. This enhancer peptide is based on a family of amphipatic peptides that exhibit improved membrane permeability. Franz permeation cell experiments revealed that the peptide enhancer (RALA) improved Na-DFC skin penetration of the liquid crystal 2.2-fold. We studied the structural effects of RALA solubilization on the H(II) mesophase. RALA acts as a chaotropic agent, interfering in the structure of the water, and causes a measurable swelling of the aqueous cylinders by 5A. Small angle X-ray scattering (SAXS) and attenuated total reflectance-Fourier transform infrared (ATR-FTIR) measurements reveal enhanced hydration of the glycerol monooleate (GMO) headgroups and a 6.5% increase in the fraction of non-freezable water resulting from RALA incorporation. RALA caused a gradual increase in the GMO effective headgroup area due to the hydration, leading eventually to a transform of the hexagonal structure towards a lamellar one. Circular dichroism and ATR-FTIR measurements showed a conservation of the peptide structure when incorporated into the H(II) mesophase. The combined H(II)LC-CPE systems can serve as high potential vehicles for a variety of drugs, as they can easily be modified by varying the composition and temperature, according to the required dose and delivery features.

In vitro permeation of diclofenac salts from lyotropic liquid crystalline systems.

In this paper we examined feasible correlations between the structure of different lyotropic mesophases and transdermal administration of three diclofenac derivatives with varying degrees of kosmotropic or chaotropic properties, solubilized within the mesophases. It was found that the most chaotropic derivative of diclofenac diethyl amine (DEA-DFC) interacted with the polar heads of glycerol monooleate (GMO), thus expanding the water-lipid interface of the lamellar and cubic mesophases. This effect was detected by an increase in the lattice parameter of both mesophases, enhanced elastic properties, and increased solid-like response of the systems in the presence of DEA. Potassium diclofenac (K-DFC), a less chaotropic salt, had less pronounced effect on the structural features of the mesophases. Kosmotropic Na+ salt (Na-DFC) had only minor influence on both lamellar and cubic structures. The locus of solubilization of the molecules with the host mesophases was correlated with their delivery. It was suggested that transdermal delivery of kosmotropic Na-DFC was accelerated by the aqueous phase and less constrained by the interaction with monoglyceride. On the other hand, the chaotropic cations (K+ and DEA+), presumably entrapped in the water-lipid interface, interacted with monoglyceride headgroups, which is likely to be the key cause for their sustained administration.

Polyols, High Pressure, and Refractive Indices Equalization for Improved Stability of W/O Emulsions for Food Applications

Mixtures of polyols (glycerol, propylene glycol, glucose) and water were emulsified in oil (isopropyl myristate (IPM), medium chain triglycerides (MCT), long chain triglycerides (LCT), and d-limonene) under elevated pressures and homogenization, in the presence of polyglycerol polyricinoleate (PGPR), glycerol monooleate (GMO), and their mixture as emulsifiers to form water-in-oil emulsions. High pressures was applied to: a) the emulsion, b) the aqueous phase and c) the oil phase in the presence of the emulsifiers (PGPR and GMO). Under optimal pressure (2000 atms) applied to the ready-made emulsion or to the aqueous phase prior to its emulsification, and with optimal composition (30wt% polyol in the aqueous phase and MCT as the oil phase), the aqueous droplets were stable for months and submicron in size (0.1 μm). Moreover, due to equalization of the oil and the aqueous phases refractive indices, the emulsions were almost transparent. Pressure and polyols have synergistic effects on the emulsions stability. During preparation, surface tensions and interfacial tensions were dramatically reduced until an optimal water/polyols ratio was achieved, which allows rupturing of the droplets to submicronal size (0.1 μm) without recoalescence and fast diffusion to the interface. These unique W/O emulsions are suitable for preparing W/O/W double emulsions for sustained release of active materials for food applications.

Lysozyme entrapped within reverse hexagonal mesophases: physical properties and structural behavior.

A model protein (lysozyme) was incorporated into monoolein-based reverse hexagonal (H(II)) mesophase and its structure effects were characterized by small angle X-ray scattering, ATR-FTIR spectroscopy, and rheological measurements. Modifications in molecular organization of the H(II) mesophases as well as the conformational stability of lysozyme (LSZ) as a function of pH and denaturating agent (urea) were clarified. Up to 3 wt.% LSZ can be solubilized into the H(II). The vibration FTIR analysis revealed that LSZ interacted with OH groups of glycerol monooleate (GMO) in the outer interface region, resulting in strong hydrogen bonding between the surfactant and its environment. Simultaneously, the decrease in the hydrogen-bonded carbonyl population of GMO was monitored, indicating dehydration of the monoolein carbonyls. These molecular interactions yielded a minor decrease in the lattice parameter of the systems, as detected by small angle X-ray scattering. Furthermore, LSZ was crystallized within the medium of the hexagonal structures in a single crystal form. The alpha-helix conformation of lysozyme was stabilized at high pH conditions, demonstrating greater helical structure content, compared to D(2)O solution. Moreover, the hexagonal phase decreased the unfavorable alpha-->beta transition in lysozyme, thereby increasing the stability of the protein under chemical denaturation. The rheological behavior of the hexagonal structures varied with the incorporation of LSZ, reflected in stronger elastic properties and pronounced solid-like response of the systems. The hydrogen bonding enhancement in the interface region of the structures was most likely responsible for these phenomena. The results of this study provided valuable information on the use of hexagonal systems as a carrier for incorporation and stabilization of proteins for various applications.

Solubilization of Hydrophobic Guest Molecules in the Monoolein Discontinuous QL Cubic Mesophase and Its Soft Nanoparticles

Hydrophobic bioactive guest molecules were solubilized in the discontinuous cubic mesophase (QL) of monoolein. Their effects on the mesophase structure and thermal behavior, and on the formation of soft nanoparticles upon dispersion of the bulk mesophase were studied. Four additives were analyzed. They were classified into two types based on their presumed location within the lipid bilayer and their influence on the phase behavior and structure. Differential scanning calorimetry (DSC), small-angle X-ray scattering (SAXS), polarized light microscopy, cryogenic-transmission electron microscopy (cryo-TEM), and dynamic light scattering (DLS) were used for the analysis. We found that carbamazepine and cholesterol (type I molecules) likely localize in the hydrophobic domains, but close to the hydrophobic-hydrophilic region. They induce strong perturbation to the mesophase packing by influencing both the order of the lipid acyl chains and interactions between lipid headgroups. This results in significant reduction of the phase transition enthalpy, and phase separation into lamellar and cubic mesophases above the maximum loading capacity. The inclusion of type I molecules in the mesophase also prevents the formation of soft nanoparticles with long-range internal order upon dispersion. In their presence, only vesicles or sponge-like nanoparticles form. Phytosterols and coenzyme Q10 (type II molecules) present only moderate effects. These molecules reside in the hydrophobic domains, where they cannot alter the lipid curvature or transform the QL mesophase into another phase. Therefore, above maximum loading, excess solubilizate precipitates in crystal forms. Moreover, when type II-loaded QL is dispersed, nanoparticles with long-range order and cubic symmetry (i.e., cubosomes) do form. A model for the growth of the ordered nanoparticles was developed from a series of intermediate structures identified by cryo-TEM. It proposes the development of the internal structure by fusion events between bilayer segments.