Oil-in-Water Emulsions Stabilized by Sodium Caseinate or Whey Protein Isolate as influenced by Glycerol Monostearate

Competitive adsorption between glycerol monostearate (GMS) and whey protein isolate (WPI) or sodium caseinate was studied in oil-in-water emulsions (20 wt % soya oil, deionized water, pH 7). Addition of GMS resulted in partial displacement of WPI or sodium caseinate from the emulsion interface. SDS-PAGE showed that GMS altered the adsorbed layer composition in sodium caseinate stabilized emulsions containing < 1.0 wt % protein. Predominance of β-casein at the interface in the absence of surfactant was reduced in the presence of GMS. The distribution of α-lactalbumin and β-lactoglobulin between the aqueous bulk phase and the fat surface in emulsions stabilized with WPI was independent of the concentration of added protein or surfactant.     

Effects of pH and Ethanol on the Kinetics of Destabilisation of Oil-in-Water Emulsions Containing Milk Proteins

Soya oil-in-water emulsions (1: 4, v/v) were prepared using sodium caseinate or β-lactoglobulin (5 or 10 g litre⁻¹) as the surfactant. The kinetics of aggregation induced by pH changes or the addition of ethanol were measured by light scattering in diluted systems either under shear or quiescent conditions. Under shear, emulsions containing caseinate were stable between pH 3 and 3·5 and also at pH⩾5·3, while those formed with β-lactoglobulin were stable below pH 4 as well as at pH⩾5·6. Under quiescent conditions, the emulsions were also destabilised but at a much slower rate. The destabilisation process generally showed an initial lag period which depended on the pH, followed by an explosive growth stage. In aqueous ethanol (50: 50, v/v), under both shear and quiescent conditions, fresh emulsions formed with β-lactoglobulin were only slightly destabilised. Ageing the emulsions for 24 h, however, increased their susceptibility, and destabilisation was achieved at concentrations of ethanol as low as 30: 70, (v/v). Emulsions formed with caseinate on the other hand were destabilised at ethanol concentrations ⩾40: 60 (v/v), with time-courses showing a high initial slope, followed by a final stage at which particle size remained constant. Shear had a relatively small effect on these reactions. The kinetics of pH-induced aggregation in both sets of emulsions could be explained by orthokinetic flocculation while the ethanol-induced association in caseinate emulsions appeared to be a result of Ostwald ripening.     

Formation and Coalescence Stability of Emulsions Stabilized by Different Milk Proteins

The volume fraction of oil emulsified, surface area, droplet diameter, and coalescence rates of emulsions stabilized by different milk proteins were studied at protein concentrations of 0.25, 0.5, 1.0, and 2.0% (w/w); pH 4, 5, and 7; and ionic strengths 0.1 and 0.2. The emulsion activity index (EAI) and coalescence stability generally increased with increasing protein solubility and hydrophobicity. The volume fraction of oil emulsified decreased with increasing ionic strength. Coalescence stability correlated with droplet diameter for emulsions stabilized by α-lactalbumin, β-lactoglobulin, and sodium caseinate (r²=0.96). With the exception of β-lactoglobulin-stabilized emulsions, coalescence stability was largely unaffected by pH.     

Creaming Stability of Oil-in-Water Emulsions Formed with Sodium and Calcium Caseinates

Creaming stability of emulsions formed with calcium caseinate, determined after storage of emulsions at 20 °C for 24 h, increased gradually with an increase in protein concentration from 0.5% to 2.0%; further increases in caseinate concentration had much less effect. In contrast, the creaming stability of sodium caseinate emulsions showed a decreased with an increase in protein concentration from 0.5% to 3.0%. Confocal laser micrographs of emulsions formed with >2% sodium caseinate showed extensive flocculation of oil droplets with the appearance of a network structure. However, emulsions formed with calcium caseinate or emulsions formed with low concentrations of sodium caseinate (0.5% and 1.0%) were homogenous with no sign of flocculation.     

Interactions at the interface between hydrophobic and hydrophilic emulsifiers: Polyglycerol polyricinoleate (PGPR) and milk proteins,studied by drop shape tensiometry

The equilibrium interfacial tension and dilational elasticity at the soy oil–water interface were studied in the presence of a lipophilic emulsifier, polyglycerol polyricinoleate (PGPR), in the continuous oil phase, and dairy proteins, β-lactoglobulin (β-lg) or sodium caseinate, in the aqueous phase using drop shape tensiometry. The interfacial tension decreased with increasing PGPR concentration, and was <2 mN m⁻¹ at PGPR concentrations beyond 1%. The presence of proteins in the water phase, β-lg and sodium caseinate, further reduced the interfacial tension. Even at the low concentrations (0.008%) tested, PGPR dominated the interfacial elasticity, which was only slightly affected by the addition of elevated levels of protein. While in the presence of β-lg (0.1%) in isolation, the system showed a high interfacial elasticity, the addition of PGPR lowered the elasticity, suggesting that PGPR interfered with protein–protein interactions at the interface, or caused displacement of β-lg. Interfacial elasticity at the oil–water interface showed little dependence on dilation or frequency of the sinusoidal oscillation, when the interface was dominated by PGPR.     

The dominating effect of ionic strength on the heat-induced denaturation and aggregation of β-lactoglobulin in simulated milk ultrafiltrate

The denaturation/aggregation behaviour of heated (78 °C, 10 min) β-lactoglobulin (1%, w/w) was examined as a function of heating pH (5.0–7.0), in the presence of different salts. Heating β-lactoglobulin in the presence of calcium (5 mm) significantly increased the level of aggregated protein at most heating pH values, compared to heating in water or sodium chloride (100 mm). Heating β-lactoglobulin in the presence of calcium (5 mm) and phosphate (5 mm), resulted in similar denaturation levels in the pH range 5.0–5.8 as in the presence of calcium (5 mm) alone but reduced denaturation in the pH range 6.0–7.0, probably due to the formation of insoluble calcium phosphate. The addition of NaCl (100 mm) counteracted the aggregation promoting properties of the calcium and calcium/phosphate systems. Heating β-lg in a simulated milk ultrafiltrate solution was similar to heating in NaCl alone. This suggested that Ca²⁺ effects alone may not explain the heat-induced denaturation/aggregation behaviour of β-lactoglobulin in milk whey systems.     

Sodium Caseinate and Skim Milk Gels Formed by Incubation with Microbial Transglutaminase

Several suspensions and emulsions containing commercial sodium caseinate or skim milk were gelatinized by Ca²⁺-independent microbial transglutaminase treatment. The characteristics of the gels were largely affected by the enzyme concentrations employed. For caseinate gels generally the higher enzyme concentration gave steep decreases in breaking strength, strain and cohesiveness of the gels. The creep tests on emulsified gels prepared to two different enzyme concentrations showed that the gel made with a higher enzyme concentration was the more viscoelastic. For skim milk gels, the enzyme treatment in higher concentration caused substantial increase of the breaking and hardness while the strain and cohesiveness had little or no changes.     

Effect of iron chelates on oil–water interface,stabilized by milk proteins: The role of phosphate groups and pH. Prediction of iron transfer from aqueous phase toward fat globule surface by changes of interfacial properties

Iron incorporated into food systems induces oxidation and precipitation. The consequences are reduced bioavailability and a functional modification of other food components such as proteins. The iron-chelates such as ferrous bisglycinate represent a possibility to avoid side effects, since the iron is protected. The aim of this study is to investigate the effects of iron-chelates compounds on the properties of an oil/water interface stabilized by caseinate or β-lacotoglobulin, under environmental conditions at 20 °C. Analyses were performed using dynamic drop tensiometry during 5000 s. The aqueous bulk phase is an imidazole/acetate buffer (0.1 M), containing 0.4 × 10⁻⁶ M protein, and 0.2 × 10^-6 9 M iron-chelates compounds. The results indicate that, under neutral conditions, the addition of some irons salts (NaFe-EDTA or Fe-bisglycinate) do not change the structure of the interface stabilized by a protein containing no phosphate groups (β-lactoglobulin). In the case of caseinate, NaFe-EDTA addition increases the lowering rate of surface tension at pH 6.5. On the contrary, the lowering rate of surface tension with caseinate is inhibited by Fe-bisglycinate at pH 6.5. Such an effect is not observed with β-lactoglobulin. The low transfer of irons ions from the bulk to the interface stabilized by β-lactoglobulin is confirmed by zetameter and FTIR measurements. These results indicate an effective strategy to follow for controlling the physical and chemical stability of an emulsion stabilized with proteins.     

Butternut and beetroot pectins: Characterization and functional properties

The physicochemical characteristics and functional properties of butternut (Cucumis moschata Duch. ex Poiret) and beetroot (Beta vulgaris L. var. conditiva) pectins obtained by enzymatic extraction from by-products of vegetable processing have been evaluated. The molecular mass distribution was determined using Gel Permeation Chromatography using light scattering, refractive index and UV detectors and the samples were found to be highly heterogeneous and polydisperse. M_w values of 136,000 and 1,309,000 g/mol were determined for butternut and beetroot pectins respectively. Butternut pectin had a high degree of methyl esterification. In the presence of high concentrations of sugar and at low pH, this pectin did not form gels but instead produced viscous solutions. Solutions showed pseudoplastic flow behaviour with a shear thinning index of 0.68 as determined from the Power law model. Beetroot pectin had a low degree of methyl esterification and formed gels with addition of Ca²⁺ at concentrations of 10 mg/g pectin or higher. The maximum value of the storage modulus was obtained at a Ca²⁺/GalA ratio of 0.25. The thermal stability of gels suggested that hydrogen bond interactions prevailed in the absence of Ca²⁺, whereas electrostatic junction zones increasingly developed between pectin chains as the calcium concentration increased. Aqueous solutions of butternut and beetroot pectins significantly reduced surface tension and both samples were able to form stable oil-in-water emulsions. It was found that protein and/or polyphenol – rich fractions present in the pectins adsorbed at the oil–water interface and were responsible for the emulsification properties.     

Gelation of β-Lactoglobulin: Effects of Sodium Chloride and Calcium Chloride on the Rheological and Structural Properties of Gels

Heating β-lactoglobulin solutions at pH 8 causes an increase in viscosity, but self-supporting gels were not formed unless salts, such as sodium chloride or calcium chloride, were added. The rheological and textural properties and gel strength were markedly affected by salt concentration. Thus, gels of maximum compressive strength were obtained with sodium chloride and calcium chloride at concentrations of 200 and 10mM, respectively. Increasing the concentration of sodium chloride resulted in the formation of soft gels which released water easily. Calcium chloride strengthened β-lactoglobulin gels by forming crossbridges. However, above 10 mM it tended to enhance coagulation rather than gelation. This was confirmed by election microscopy of the gel matrix.     

STABILITY OF OIL-IN-WATER EMULSIONS. 2. Effects of Oil Phase Volume, Stability Test, Viscosity, Type of Oil and Protein Additive

SUMMARY –Stability of oil-in-water emulsions stabilized in sodium caseinate, gelatin and soy sodium proteinate was found to be increased by either an increase in the aqueous phase protein concentration (0.5–2.5%) or oil phase volume (20–50%). Both factors were significantly interrelated. Emulsions stabilized by soy sodium proteinate were generally higher in stability as compared to those stabilized by gelatin or sodium caseinate. With emulsions containing gelatin, greater stability occurred when the stability testing temperature was increased from 37–70°C and when the time interval was decreased from 24 hr to 90 min. Maximum relative viscosities of emulsions stabilized by gelatin and sodium caseinate were 2.0 and 2.5, respectively. Emulsions stabilized by soy sodium proteinate were quite viscous, with relative viscosity from 1.5–30 depending on both protein concentration and oil phase volume. Interchanging the emulsified oil among corn, soybean, safflower and peanut oils did not alter emulsion stability when examined at three concentrations of soy sodium proteinate. Changing the oil to olive oil significantly increased emulsion stability at each soy sodium proteinate level with oil phase volumes of 30, 40 and 50%.     

Effect of calcium chloride addition on ice cream structure and quality

The influence of calcium fortification by the addition of calcium chloride on quality parameters of ice cream based on physical properties was investigated, as was the effect of κ-carrageenan at modifying the effects of this calcium fortification. Four ice cream mixes of conventional composition, with added κ-carrageenan (0 or 0.025%) and added calcium chloride (0 or 4.4 g L⁻¹ = 40 mM of added Ca²⁺), were prepared. Modulated temperature-differential scanning calorimetry was used to investigate the effect of calcium chloride on the nucleation temperature, enthalpy of melting, and freezing point depression. The protein composition of 15.4% (wt/wt) reconstituted skim milk powder solutions with or without 4.4 g L⁻¹ added CaCl₂ and in the supernatant after ultracentrifugation was determined. Fat particle size distributions in ice cream were characterized by light scattering. Ice crystal sizes before and after temperature cycling were determined by cold-stage light microscopy. The results demonstrated that the addition of calcium chloride led to a substantial increase in ice crystal sizes and in fat partial coalescence, which were exacerbated by the addition of κ-carrageenan. These results can be explained by the interaction between Ca²⁺ ions and casein micelles, rather than any effects on freezing point depression. The calcium ions led to a more compact micelle, less serum β-casein, and high fat destabilization, all of which would be expected to reduce macromolecular structure and volume occupancy in the unfrozen phase, which led to increased rates of ice recrystallization.     

Interfacial composition and stability of emulsions made with mixtures of commercial sodium caseinate and whey protein concentrate

The interfacial composition and the stability of oil-in-water emulsion droplets (30% soya oil, pH 7.0) made with mixtures of sodium caseinate and whey protein concentrate (WPC) (1:1 by protein weight) at various total protein concentrations were examined. The average volume-surface diameter (d₃₂) and the total surface protein concentration of emulsion droplets were similar to those of emulsions made with both sodium caseinate alone and WPC alone. Whey proteins were adsorbed in preference to caseins at low protein concentrations (<3%), whereas caseins were adsorbed in preference to whey proteins at high protein concentrations. The creaming stability of the emulsions decreased markedly as the total protein concentration of the system was increased above 2% (sodium caseinate >1%). This was attributed to depletion flocculation caused by the sodium caseinate in these emulsions. Whey proteins did not retard this instability in the emulsions made with mixtures of sodium caseinate and WPC.     

Foaming behaviour of EDTA-treated α-lactalbumin

The foaming properties of partially denatured α-lactalbumin was investigated. The partially denatured state was produced by removing bound Ca²⁺ by treatment with ethylenediaminetetraacetic acid (EDTA) at pH 8.0 and 25°C. Surface tension measurements showed that partially denatured α-lactalbumin unfolds easily at liquid interfaces compared with the native protein. The results of foam volume and stability measurements were consistent with the results of surface tension measurements. In the presence of EDTA a considerable amount of foam was obtained at low concentrations, such as 0.1 mg/ml, and the foam stability was improved. This indicates the importance of the protein structure on the adsorption of molecules at liquid interfaces. The presence of Ca²⁺ also resulted in an increase in the foamability and foam stability of α-lactalbumin compared with native protein, due to the saturation of the surface charges. This shows the binding affinity of protein to Ca²⁺. The investigation of the effect of Ca²⁺ on the surface behaviour of β-lactoglobulin, another whey protein, also showed an improvement in the foaming properties of protein.     

Study of commodity properties of food emulsions Part 1. The effect of dispersion medium composition on kinetic stability of O/W emulsions containing proteins,acidic polysaccharides and calcium salts

The effect of concentrations of acidic polysaccharide and calcium ions on the kinetic stability, viscosity and dispersity of protein-containing O/W emulsions is studied. Variation of kinetic stability of the emulsions studied is independent of dispersion composition. In a wide range of calcium acetate concentrations a correlation is observed between kinetic stability and viscosity of emulsions at sodium alginate concentration in the dispersion medium ≤ 0.3%. The transition zone between liquid solution and gel is widened in the presence of sodium caseinate. Maximum kinetic stability is reached at calcium acetate: sodium alginate concentrations of 1.0-1.2 and ～ 6.0, corresponding to optimum conditions for formation of homogeneous crosslinked structures of calcium alginate and calcium caseinate.     

Interfacial Composition of Sodium Caseinate Emulsions

Oil-in-water emulsions stabilized by sodium caseinate were prepared and diluted with water or solutions of α_sl- or β-casein. The emulsions were aged for 24 hr and the composition of the aqueous phase (and hence of the surface) was determined using Fast Protein Liquid Chromatography. There was no distinct preference for either α_sl- or β-casein at the surface during homogenization, but on aging, β-casein replaced some, but not all, of the surface α_sl-casein. The exchange was stoichiometric, unless the initial surface concentration was low, in which case extra adsorption occurred to achieve a protein load of - 1.2 mg m^-2. Build up of multilayers was not observed.     

Sodium caseinate-stabilized fat globules inhibition of the rennet-induced gelation of casein micelles studied by Diffusing Wave Spectroscopy

Sodium caseinate (NaCas)-stabilized oil-in-water emulsions were added to skim milk and the rennet-induced aggregation was observed in situ using light scattering and dynamic oscillatory rheology. The gelation of the recombined milk was greatly inhibited by the addition of the oil droplets, at volume fractions >0.025. The development of the turbidity parameter, 1/l*, and the apparent hydrodynamic radius during renneting were determined using diffusing wave spectroscopy. Although the recombined milk samples contained two scattering particles, namely, casein micelles and fat globules, the latter overwhelmingly contributed to the overall light-scattering signal. This made possible to follow the behaviour of NaCas-stabilized fat globules during the gelation process. The enzymatic reaction associated with the hydrolysis of micellar κ-casein was not significantly affected by the presence of the NaCas-stabilized fat globules. However, the emulsion droplets impeded the aggregation of rennet-altered casein micelles preventing the formation of a gel network. The inability of renneted casein micelles to develop a gel network can be attributed in part to an altered equilibrium between soluble and micellar calcium phosphate, caused by the association of soluble Ca²⁺ with casein molecules, but mostly can be attributed to the effect of non-adsorbed caseins on the surface of the casein micelles.     

Heat Stability of Oil-in-Water Emulsions Containing Milk Proteins: Effect of Ionic Strength and pH

Emulsions (20 wt% soybean oil; 2 wt% protein) made with caseinate at pH 7 and with whey protein isolate (WPI) at pH 7 and 3 were stable to heating at 90 and 121°C. WPI emulsions destabilized at pH values between 3.5 and 4.0. In the presence of KCI (12.5–200 mM), large particles were formed in WPI emulsions at pH 3 and the emulsions were viscous. At pH 7, moderate concentrations of KCI decreased the heat stability and gels were formed. KCI had less effect on WPI emulsions made at pH 3. Combining the emulsions with caseinate allowed some control of the heat-induced gelation.     

Influence of xanthan gum on the formation and stability of sodium caseinate oil-in-water emulsions

Studies have been made of the changes in droplet sizes, surface coverage and creaming stability of emulsions formed with 30% (w/w) soya oil, and aqueous solution containing 1 or 3% (w/w) sodium caseinate and varying concentrations of xanthan gum. Addition of xanthan prior to homogenization had no significant effect on average emulsion droplet size and surface protein concentration in all emulsions studied. However, addition of low levels of xanthan (≤0.2 wt%) caused flocculation of droplets that resulted in a large decrease in creaming stability and visual phase separation. At higher xanthan concentrations, the creaming stability improved, apparently due to the formation of network of flocculated droplets. It was found that emulsions formed with 3% sodium caseinate in the absence of xanthan showed extensive flocculation that resulted in very low creaming stability. The presence of xanthan in these emulsions increased the creaming stability, although the emulsion droplets were still flocculated. It appears that creaming stability of emulsions made with mixtures of sodium caseinate and xanthan was more closely related to the structure and rheology of the emulsion itself rather than to the rheology of the aqueous phase.     

Effect of added calcium chloride on the physicochemical and rheological properties of aqueous mixtures of sodium caseinate/sodium alginate and respective oil-in-water emulsions

Changes induced by addition of calcium chloride in particle size distribution and electrokinetic potential were determined in sodium caseinate/sodium alginate mixtures dissolved in water or acetate buffer at ambient temperature. Rheological properties of aqueous mixtures and respective oil-in-water emulsions (30% oil w/w) were evaluated using a low-stress rheometer. Stability and particle diameter of emulsions were measured. Caseinate and alginate solutions were negatively charged and showed negative electrokinetic potential; however values of mixtures were between those of the values for the individual hydrocolloids. When calcium ions were added the electrokinetic potential diminished while the negative charge was preserved. Aqueous mixtures of caseinate and alginate showed average particles size between of those of caseinate or alginate samples. We observed low viscosity values and Newtonian behavior for both caseinate (1 and 2%) and alginate (0.1%). Addition of 5 mM CaCl₂ to alginate solutions induced shear-thinning behavior as well as the development of viscoelasticity. Both the viscosity and the elastic modulus of these polysaccharide solutions were attenuated by the presence of protein or dispersed oil in mixtures or emulsions, respectively. High average particle diameter of emulsions prepared was obtained (close to 10 μm), however, stability of emulsions was possible only with the addition of CaCl₂ to the mixtures, in both water and acetate buffer. In these cases elastic behavior predominated to viscosity in the formation of emulsions, confirming the prevalence of aqueous phase rheology on emulsions.