Physicochemical Properties of Whey Protein-Stabilized Emulsions as affected by Heating and Ionic Strength

Corn oil-in-water emulsions (19.6 wt%; d₃₂～ 0.6 μm) stabilized by 2 wt% whey protein isolate (WPI) were prepared with a range of pH (3–7) and salt concentrations (0–100 mM NaCl). These emulsions were heated between 30 and 90°C and their particle size distribution, rheological properties and susceptibility to creaming measured. Emulsions had a paste-like texture around the isoelectric point of WPI (～φ 5) at all temperatures, but tended to remain fluid-like at pH >6 or <4. Heating caused flocculation in pH 7 emulsions between 70 and 80°C (especially at high salt concentrations), but had little effect on pH 3 emulsions. Flocculation increased emulsion viscosity and creaming. Results were interpreted in terms of colloidal interactions between droplets.     

Physical Properties of Whey Protein Stabilized Emulsions as Related to pH and NaCl

Corn oil-in-water emulsions (20 wt%, d₃₂～ 0.6 μm) stabilized by 2 wt% whey protein isolate were prepared with a range of pH (3–7) and salt concentrations (0–100 mM NaCl), and particle size, rheology and creaming were measured at 30°C. Appreciable droplet flocculation occurred near the isoelectric point of whey protein (pH 4–6), especially at higher NaCl concentrations. Droplet flocculation increased emulsion viscosity and decreased stability to creaming. Results are related to the influence of environmental conditions on electrostatic and other interactions between droplets.     

The effect of chitosan on the properties of emulsions stabilized by whey proteins

The influence of the cationic amino polysaccharide chitosan content (0–0.5%) on particle size distribution, creaming stability, apparent viscosity, and microstructure of oil-in-water emulsions (40% of rapeseed oil) containing whey protein isolate (WPI) (4%) at pH 3 was investigated. The emulsifying properties, apparent viscosity and phase separation behaviour of aqueous WPI/chitosan mixture at pH 3 were also studied. The interface tension data showed that WPI/chitosan mixture had a slightly higher emulsifying activity than had whey protein alone. An increase in chitosan content resulted in a decreased average particle size, higher viscosity and increased creaming stability of emulsions. The microstructure analysis indicated that increasing concentration of chitosan resulted in the formation of a flocculated droplet network. This behaviour of acidic model emulsions containing WPI and chitosan was explained by a flocculation phenomenon.     

Mangosteen Oil-In-Water Emulsions: Rheology, Creaming, and Microstructural Characteristics during Storage

The rheological properties and physical stability of mangosteen (Garcinia mangostana L.) extract in oil-in-water (MIO/W) emulsions were investigated. Rheological study on the emulsions exhibited Newtonian flow behavior. The 20?wt.% emulsion showed higher apparent viscosity than 10?wt.% MIO/W sample. The effects of salt (NaCl) concentration (0, 50, 100, and 200?mM) and heat treatment (70?°C) on the stability of the emulsions were also examined. Heat (70?°C)- and NaCl (100 and 200?mM)-treated emulsions showed creaming and droplet aggregation on storage for a period of 60?days. The 10?wt.% MIO/W emulsions stored at 4?°C showed a homogeneous distribution of oil droplets with good stability to creaming and viscosity independent of shear stress (i.e., a Newtonian liquid).     

Rheology and stability of whey protein stabilized emulsions with high CaCl2 concentrations

The influence of pH and CaC1₂ on the rheology and physical stability of emulsions stabilized by whey protein isolate (WPI) has been studied. The particle size, creaming index and shear viscosity of 10 wt% soy bean oil-in-water emulsions (d=0.55 μm) were measured with varying pH (3, 5 and 7) and CaC1₂ concentration (0–150 mM). In the absence of CaCl₂ extensive droplet aggregation occurred around the isoelectic point of the whey proteins (4<pH<6) because of their low electrical charge. In the presence of CaC1₂, extensive droplet aggregation, viscosity enhancement and creaming instability occurred at pH 7 for CaC1₂>3 mM. These effects were much less pronounced in emulsions at pH 3 even at 150 mM CaC1₂. Droplet aggregation, creaming and viscosity of emulsions at pH 5 were fairly independent of CaC1₂ concentration. Droplet aggregation was induced by CaC1₂ probably because of the reduction in electrostatic repulsion between droplets. Re-stabilization of oil-in-water emulsions at high CaC1₂ concentrations was not observed in this study.     

Interactions between Whey Protein Isolate and Soy Protein Fractions at Oil–Water Interfaces: Effects of Heat and Concentration of Protein in the Aqueous Phase

ABSTRACT:  The 2 main storage proteins of soy—glycinin (11S) and β-conglycinin (7S)—exhibit unique behaviors during processing, such as gelling, emulsifying, or foaming. The objective of this work was to observe the interactions between soy protein isolates enriched in 7S or 11S and whey protein isolate (WPI) in oil–water emulsion systems. Soy oil emulsion droplets were stabilized by either soy proteins (7S or 11S rich fractions) or whey proteins, and then whey proteins or soy proteins were added to the aqueous phase. Although the emulsifying behavior of these proteins has been studied separately, the effect of the presence of mixed protein systems at interfaces on the bulk properties of the emulsions has yet to be characterized. The particle size distribution and viscosity of the emulsions were measured before and after heating at 80 and 90 °C for 10 min. In addition, SDS-PAGE electrophoresis was carried out to determine if protein adsorption or exchanges at the interface occurred after heating. When WPI was added to soy protein emulsions, gelling occurred with heat treatment at WPI concentrations >2.5%. In addition, whey proteins were found adsorbed at the oil–water interface together with 7S or 11S proteins. When 7S or 11S fractions were added to WPI-stabilized emulsions, no gelation occurred at concentrations up to 2.5% soy protein. In this case also, 7S or 11S formed complexes at the interface with whey proteins during heating.     

Factors influencing the production of o/w emulsions stabilized by β-lactoglobulin–pectin membranes

A primary emulsion was prepared by homogenizing 10 wt% corn oil with 90 wt% aqueous β-lactoglobulin solution (0.5 wt% β-lg, pH 3 or 7) using a two-stage high-pressure valve homogenizer. This emulsion was mixed with aqueous pectin (citrus, 59% DE) stock solution (2 wt%, pH 3 or 7) and NaCl solution to yield secondary emulsions with 5 wt% corn oil, 0.225 wt% β-lactoglobulin, 0.2 wt% pectin and 0 or 100 mM NaCl. The final pH of the emulsions was then adjusted (3–8). Primary and secondary emulsions were ultrasonically treated (30 s, 20 kHz, 40% amplitude) to disrupt any flocculated droplets. Secondary emulsions were more stable than primary emulsions at intermediate pHs. Secondary emulsions prepared at pH 7 had smaller particle diameters (0.35 to 6 μm) than those prepared at pH 3 (0.42 to 18 μm) across the whole pH range studied, and also had smaller diameters than the primary emulsions (0.35 to 14 μm). Ultrasound treatment reduced the particle diameter of both primary and secondary emulsions and lowered the rate of creaming. The presence of NaCl screened the charges and thus the electrostatic interaction between biopolymer molecules and primary emulsion droplets. Secondary emulsions were more stable to the presence of 100 mM NaCl at low pHs (3–4) than primary emulsions. This study shows that stable emulsions can be prepared by engineering their interfacial membranes using the electrostatic interaction of natural biopolymers, especially at intermediate pHs where proteins normally fail to function.     

Rheology and Oxidative Stability of Whey Protein Isolate-Stabilized Menhaden Oil-in-Water Emulsions as a Function of Heat Treatment

ABSTRACT:  Menhaden oil-in-water emulsions (20%, v/v) were stabilized by 2 wt% whey protein isolate (WPI) with 0.2 wt% xanthan gum (XG) in the presence of 10 mM CaCl₂ and 200 μM EDTA at pH 7. Droplet size, lipid oxidation, and rheological properties of the emulsions were investigated as a function of heating temperature and time. During heating, droplet size reached a maximum at 70 °C and then decreased at 90 °C, which can be attributed to both heating effect on increased hydrophobic attractions and the influence of CaCl₂ on decreased electrostatic repulsions. Combination of effects of EDTA and heat treatment contributed to oxidative stability of the heated emulsions. The rheological data indicate that the WPI/XG-stabilized emulsions undergo a state transition from being viscous like to an elastic like upon substantial thermal treatment. Heating below 70 °C or for less than 10 min at 70 °C favors droplet aggregation while heating at 90 °C or for 15 min or longer at 70 °C facilitates WPI adsorption and rearrangement. WPI adsorption leads to the formation of protein network around the droplet surface, which promotes oxidative stability of menhaden oil. Heating also aggravates thermodynamic incompatibility between XG and WPI, which contributes to droplet aggregation and the accumulation of more WPI around the droplet surfaces as well.     

Effect of pH and Ionic Strength on the Physicochemical Properties of Coconut Milk Emulsions

ABSTRACT:  Coconut milk (16% to 17% fat, 1.8% to 2% protein) was extracted from coconut ( Cocos nucifera L.) endosperm and diluted in buffer to produce natural oil-in-water emulsions (10 wt% oil). The effect of pH (3 to 7) and NaCl (0 to 200 mM) on the properties and stability, namely, mean particle size, ζ-potential, viscosity, microstructure, and creaming stability, of the natural coconut milk emulsions was investigated. At pH values close to the isoelectric point (IEP) of the coconut proteins (pH 3.5 to 4) and in the absence of NaCl, coconut milk flocculated, but did not coalesce. Flocculation corresponded to low surface charges and was accompanied by an increase in emulsion viscosity. Adding up to 200 mM NaCl to those flocculated emulsions did not change the apparent degree of flocculation. Coconut milk emulsion at pH 6 was negatively charged and not flocculated. Upon addition of salt, the ζ-potential decreased from –16 to –6 mV (at 200 mM NaCl) but this was not sufficient to induce flocculation in coconut milk emulsions. At low pH (< IEP), the positively charged droplets of coconut milk emulsions only flocculated when the NaCl concentration exceeded 50 mM, as the ζ-potential approached zero.     

Comparison of Gum Arabic, Modified Starch, and Whey Protein Isolate as Emulsifiers: Influence of pH, CaCl2 and Temperature

ABSTRACT: The particle size and zeta potential of model beverage emulsions (0.01 wt% soybean oil-in-water emulsions, d ≅ 1 mm) stabilized by gum arabic, modified starch, or whey protein isolate (WPI) were studied with varying pH (3 to 9), CaCl₂ concentration (0 to 25 mM), and temperature (30 °C to 90 °C). Temperature, pH, CaCl₂ strongly influenced emulsions stabilized by WPI because its stabilizing mechanism was mainly electrostatic repulsion, but not those stabilized by gum arabic or modified starch because their stabilizing modes of action were mainly steric repulsion. This study may have important implications for the application of WPI as an emulsifier in beverage emulsions.     

Physical and Oxidative Stabilities of O/W Emulsions Formed with Rice Dreg Protein Hydrolysate: Effect of Xanthan Gum Rheology

The shortening of shelf-life of food emulsions is frequently due to poor creaming and lipid oxidation stability. The lipid oxidation of O/W emulsions can be inhibited by rice dreg protein hydrolysate (RDPH); however, emulsions were stabilized by Tween-20. Polysaccharides can control the rheology and network structure of the aqueous continuous phase by increasing viscosity and yield stress, hence retarding phase separation and gravity-induced creaming, especially for xanthan gum. The objective of this research was to evaluate whether emulsions formed with 2 wt% RDPH and stabilized by xanthan gum (0–0.5 wt%) could produce 20 % (v/v) soybean oil-in-water emulsions that had good physical and oxidative stability. The degree of flocculation of droplets as a function of xanthan gum concentration was assessed by the microstructure, rheology, and the creaming index of emulsions. Addition of xanthan gum prior to homogenization had no significant effect on the mean droplet diameter in all emulsions studied. Increase in xanthan gum concentration led to the increase in creaming stability of emulsions, due to an increase in viscosity of the continuous phase and/or the formation of a droplet network with a yield stress, as well as the enhanced steric and electrostatic repulsion between the droplets. Lipid oxidation of the emulsions was significantly inhibited at xanthan gum concentrations of 0.12 wt% or above with RDPH, which could due to the fact that xanthan gum increases the viscosity of the aqueous phase and hindered the diffusion of oxidants to the oil droplet surface area, synergistic effect between RDPH and xanthan gum to suppress oil peroxidation, and metal ion chelation capability of xanthan gum. Thus, stable protein hydrolyzates-type emulsions could be obtained with increasing concentration of xanthan gum.     

Influence of biopolymer emulsifier type on formation and stability of rice bran oil-in-water emulsions: whey protein, gum arabic, and modified starch

Rice bran oil (RBO) is used in foods, cosmetics, and pharmaceuticals due to its desirable health, flavor, and functional attributes. We investigated the effects of biopolymer emulsifier type and environmental stresses on the stability of RBO emulsions. Oil-in-water emulsions (5% RBO, 10 mM citrate buffer) stabilized by whey protein isolate (WPI), gum arabic (GA), or modified starch (MS) were prepared using high-pressure homogenization. The new MS used had a higher number of octenyl succinic anhydride (OSA) groups per starch molecule than conventional MS. The droplet diameters produced by WPI and MS were considerably smaller (d < 300 nm) than those produced by GA (d > 1000 nm). The influence of pH (3 to 8), ionic strength (0 to 500 mM NaCl), and thermal treatment (30 to 90 °C) on the physical stability of the emulsions was examined. Extensive droplet aggregation occurred in WPI-stabilized emulsions around their isoelectric point (4 < pH < 6), at high salt (> 200 mM, pH 7), and at high temperatures (>70 °C, pH 7, 150 mM NaCl), which was attributed to changes in electrostatic and hydrophobic interactions between droplets. There was little effect of pH, ionic strength, and temperature on emulsions stabilized by GA or MS, which was attributed to strong steric stabilization. In summary: WPI produced small droplets at low concentrations, but they had poor stability to environmental stress; GA produced large droplets and needed high concentrations, but they had good stability to stress; new MS produced small droplets at low concentrations, with good stability to stress. Practical Application: This study showed that stable rice bran oil-in-water emulsions can be formed using biopolymer emulsifiers. These emulsions could be used to incorporate RBO into a wide range of food products. We compared the relative performance of whey protein, GA, and a new MS at forming and stabilizing the emulsions. The new OSA MS was capable of forming small stable droplets at relatively low concentrations.     

Heat stability of oil-in-water emulsions formed with intact or hydrolysed whey proteins: influence of polysaccharides

The heat stability of emulsions (4 wt% corn oil) formed with whey protein isolate (WPI) or extensively hydrolysed whey protein (WPH) products and containing xanthan gum or guar gum was examined after a retort treatment at 121 °C for 16 min. At neutral pH and low ionic strength, emulsions stabilized with both 0.5 and 4 wt% WPI (intact whey protein) were stable against retorting. The amount of β-lactoglobulin (β-lg) at the droplet surface increased during retorting, especially in the emulsion containing 4 wt% protein, whereas the amount of adsorbed α-lactalbumin (α-la) decreased markedly. Addition of xanthan gum or guar gum caused depletion flocculation of the emulsion droplets, but this flocculation did not lead to their aggregation during heating. In contrast, the droplet size of emulsions formed with WPH increased during heat treatment, indicating that coalescence had occurred. The coalescence during heating was enhanced considerably with increasing concentration of polysaccharide in the emulsions, up to 0.12% and 0.2% for xanthan gum and guar gum, respectively; whey peptides in the WPH emulsions formed weaker and looser, mobile interfacial structures than those formed with intact whey proteins. Consequently, the lack of electrostatic and steric repulsion resulted in the coalescence of flocculated droplets during retort treatment. At higher levels of xanthan gum or guar gum addition, the extent of coalescence decreased gradually, apparently because of the high viscosity of the aqueous phase.     

Stability of citral in protein- and gum arabic-stabilized oil-in-water emulsions

Citral is a major flavor component of citrus oils that can undergo chemical degradation leading to loss of aroma and formation of off-flavors. Engineering the interface of emulsion droplets with emulsifiers that inhibit chemical reactions could provide a novel technique to stabilize citral. The objective of this study was to determine if citral was more stable in emulsions stabilized with whey protein isolate (WPI) than gum arabic (GA). Degradation of citral was equal to or less in GA- than WPI-stabilized emulsion at pH 3.0 and 7.0. However, formation of the citral oxidation product, p-cymene was greater in the GA- than WPI-stabilized emulsion at pH 3.0 and 7.0. Emulsions stabilized by WPI had a better creaming stability than those stabilized by GA because the protein emulsifier was able to produce smaller lipid droplets during homogenization. These data suggest that WPI was able to inhibit the oxidative deterioration of citral in oil-in-water emulsions. The ability of WPI to decrease oxidative reactions could be due to the formation of a cationic emulsion droplet interface at pH 3.0 which can repel prooxidative metals and/or the ability of amino acids in WPI to scavenge free radical and chelate prooxidative metals.     

Influence of EDTA and citrate on thermal stability of whey protein stabilized oil-in-water emulsions containing calcium chloride

The influence of calcium ions and chelating agents on the thermal stability of model nutritional beverages was examined. Oil-in-water emulsions (6.94% (w/v) soybean oil, 0.35% (w/v) WPI, 0.02% (w/v) sodium azide, 20 mM Tris buffer, 0–10 mM CaCl₂, and 0–40 mM EDTA or citrate, pH 7.0) were stored at temperatures between 30 and 120 °C for 15 min. The particle size, particle charge, creaming stability, rheology, and free-calcium concentration of the emulsions were then measured. In the absence of chelating agents, appreciable droplet aggregation occurred in emulsions held at temperatures from 80 to 120 °C, which led to increased emulsion particle diameter, shear-thinning behavior, apparent viscosity, and creaming instability. Addition of chelating agents to the emulsions prior to heating decreased, but did not prevent, droplet aggregation in the emulsions. EDTA was more effective than citrate in decreasing droplet aggregation. Heat treatment increased the amount of chelating agents required to prevent droplet aggregation in the emulsions. Free-calcium concentration and droplet surface potential was independent of heat-treatment temperature, indicating that the performance of the chelating agents in binding calcium ions was not affected by the heat treatment. It was suggested that increased hydrophobic attractive interactions between the droplets occurred during heating, which induced droplet aggregation.     

Impact of whey protein isolates and concentrates on the formation of protein nanoparticles‐stabilised Pickering emulsions

Whey protein nanoparticles (NPs) were prepared by heat‐induced method. The influences of whey protein isolates (WPIs) and concentrates (WPCs) on the formation of NPs were first investigated. Then Pickering emulsions were produced by protein NPs and their properties were evaluated. After heat treatment, WPC NPs showed larger particle size, higher stability against NaCl, lower negative charge and contact angle between air and water. Dispersions of WPC NPs appeared as higher turbidity and viscosity than those of WPI NPs. The interfacial tension of WPC NPs (~7.9 mN/m at 3 wt% NPs) was greatly lower than that of WPI NPs (~12.1 mN/m at 3 wt% NPs). WPC NPs‐stabilised emulsions had smaller particle size and were more homogeneous than WPI NPs‐stabilised emulsions. WPC NPs‐stabilised emulsions had higher stability against NaCl, pH and coalescence during storage.     

Comparison of properties of oil-in-water emulsions stabilized by coconut cream proteins with those stabilized by whey protein isolate

Coconut cream protein (CCP) fractions were isolated from coconuts using two different isolation procedures: isoelectric precipitation (CCP1-fraction) and freeze–thaw treatment (CCP2-fraction). The ability of these protein fractions to form and stabilize oil-in-water emulsions was compared with that of whey protein isolate (WPI). Protein solubility was a minimum at ∼pH 4, 4.5 and 5 for CCP1, CCP2, and WPI, respectively, and decreased with increasing salt concentration (0–200 mM NaCl) for the coconut proteins. All of the proteins studied were surface active, but WPI was more surface active than the two coconut cream proteins. The two coconut cream proteins were used to prepare 10 wt% corn oil-in-water emulsions (pH 6.2, 5 mM phosphate buffer). CCP2 emulsions had smaller mean droplet diameters (d₃₂ ≈ 2 μm) than CCP1 emulsions (d₃₂ ≈ 5 μm). Corn oil-in-water emulsions (10 wt%) stabilized by 0.2 wt% CCP2 and WPI were prepared with different pH values (3–8), salt concentrations (0–500 mM NaCl) and thermal treatments (50–90 °C for 30 min). Considerable droplet flocculation occurred in the emulsions near the isoelectric point of the proteins: CCP2 (pH ∼ 4.3); WPI (pH ∼ 4.8). Emulsions with monomodal particle size distributions, small mean droplet diameters, and good creaming stability could be produced at pH 7 for WPI, but CCP2 produced bimodal distributions at this pH. The CCP2 and WPI emulsions remained relatively stable to droplet aggregation and creaming at NaCl concentrations ⩽50 and ⩽100 mM, respectively. In the absence of salt, both CCP2 and WPI emulsions were quite stable to thermal treatments (50–90 °C for 30 min).     

Effect of CaCl2 and KCl on Physiochemical Properties of Model Nutritional Beverages Based on Whey Protein Stabilized Oil-in-Water Emulsions

ABSTRACT: Calcium chloride (0 to 10 mM) and potassium chloride (0 to 600 mM) were added into model nutritional beverage emulsions containing 7% (w/w) soybean oil droplets and 0.35% (w/w) whey protein isolate (pH 6.7). The particle size, surface charge, viscosity, and creaming stability of the emulsions then were measured. The surface charge decreased with increasing mineral ion concentration. The particle size, viscosity, and creaming instability of the emulsions increased appreciably above critical CaCl₂ (3 mM) and KCl (200 mM) concentrations because of droplet flocculation. The origin of this effect was attributed to reduction of the electrostatic repulsion between droplets due to electrostatic screening and ion binding. CaCl₂ promoted emulsion instability more efficiently than KCl because Ca²⁺ ions are more effective at reducing electrostatic repulsion than K⁺ ions.     

Flocculation of Whey Protein Stabilized Emulsions as Influenced by Dextran Sulfate and Electrolyte

The creaming stability and viscosity of oil-in-water emulsions stabilized by whey protein isolate were monitored as functions of dextran sulfate (DS) and electrolyte (NaCl) concentration. At a specific DS concentration (the critical flocculation concentration, CFC), the droplets became flocculated, which promoted creaming. Addition of electrolyte caused an increase in CFC. At NaCl concentrations <0.5 wt%, addition of electrolyte decreased emulsion viscosity, but at concentrations >0.5 wt% it caused an increase in viscosity due to increased flocculation. The results were due to the influence of electrostatic screening on the effective volume of DS molecules and colloidal interactions between droplets.     

Disulfide-mediated Polymerization Reactions and Physical Properties of Heated WPI-stabilized Emulsions

Heating a 19 wt% corn oil-in-water emulsion stabilized by 1 wt% whey protein isolate from 30 to 70°C and then cooling to 25°C for at least 15 hr, brought about minimal changes in droplet aggregation, apparent viscosity and susceptibility to creaming. At 75°C, droplet aggregation occurred but this decreased on heating to 90°C. The apparent viscosity and susceptibility of droplets to creaming increased as the degree of droplet aggregation increased. Inclusion of the sulfhydryl blocking agent N-ethylmaleimide to inhibit thiol/disulfide interchange reactions did not affect droplet aggregation but resulted in higher apparent viscosity values and susceptibility to creaming at 85 and 90°C and not at lower temperatures. The results suggest that droplet aggregation results from noncovalent interactions between unfolded protein molecules adsorbed on different droplets and that the interactions are strengthened by disulfide bonds.