A comparative study of heat and high pressure induced gels of whey and egg albumen proteins and their binary mixtures

Large deformation rheological studies of either egg albumen or whey protein isolate (15% protein w/w) gels induced by heating at 90 °C for 30 min were compared to those induced by a range high pressures (400–800 MPa for 20 min). Gels made by heating indicated higher gel strength and Young's modulus values for whey protein from pressures of 400–600 MPa for 20 min but similar values at 650–800 MPa. In contrast, egg albumen showed no gelation below 500 MPa for 20 min, but there was an increase in both gel strength and Young's modulus with increasing pressure, although values remained lower than those of the heat-induced gels. A mixture of 10:5 whey/egg albumen showed the highest gel strength and Young's modulus for both heated and high pressure-treated (400–600 MPa) gels, although, the heated mixture had the highest values. Electron micrographs indicated that high pressure-treated gels had a porous aggregated network for egg albumen while whey proteins showed a continuous fine stranded network. The heated mixtures of whey:egg albumen (7.5:7.5) showed large dense aggregates whereas high pressure-treated mixtures produced smaller aggregates. Raman spectroscopy of both heated and high pressure-treated whey and egg albumen (15% w/w in D₂O pD7) and their binary mixtures (7.5:7.5, protein w/w) indicated changes in β-sheet structures in the Amide 111′ region (980–990 cm⁻¹); however, peak intensity was reduced for high pressure-treated samples. β-Sheet structure (1238–1240 cm⁻¹) present in heated whey was absent in high pressure-treated whey and in egg albumen. Involvement of hydrophobic regions was reflected by changes in the CH (1350 cm⁻¹) and CH₂ (1450 cm⁻¹) bending vibrations. In addition to the Trp residues at 760 cm⁻¹, there were broad peaks at 874–880 cm⁻¹ and tyrosine 1207 cm⁻¹ in the high pressure-treated samples. Disulphide bands (500–540 cm⁻¹) in heated whey and egg albumen proteins showed higher peak intensities compared to high pressure-treated samples. Differences in the experimental and theoretical spectra indicated changes in the hydrophobic regions, tyrosine (1207 cm⁻¹) and tryptophan (880 cm⁻¹) and CH₂ bending in high pressure-treated samples, whereas heated samples indicated marked changes in β-sheet structures (987 and 1238 cm⁻¹) as well as hydrophobic regions CH (1350 cm⁻¹) and CH₂ (1450 cm⁻¹) bending vibrations.     

Glycosylation and expanded utility of a modified whey protein ingredient via carbohydrate conjugation at low pH

Whey protein, at one time considered a by-product of the cheese-making process, is now commonly used in foods for its thickening and emulsifying properties. Currently, approximately 30% of these proteinaceous resources remain under-utilized. Previously, an acidified, thermally treated whey protein concentrate (mWPC) was developed to produce a cold-set thickening ingredient. Mass spectroscopy revealed an approximate 2.5-fold decrease in the lactosylation of β-lactoglobulin in mWPC starting materials compared with commercial whey protein concentrates, manufactured at a higher pH. Potentially, this should increase the number of reactive sites that remain available for carbohydrate attachment. With this study, the formation of glycoprotein complexes was demonstrated between the mWPC ingredient and lactose, naturally occurring in mWPC powders, or between mWPC protein components with dextran (35 to 45 and 100 to 200 kDa) materials at low pH. In fact, additional dry heating of mWPC powders showed a 3-fold increase in the amount of lactosylated β-lactoglobulin. Evidence of Maillard reactivity was suggested using colorimetry, o-phthaldialdehyde assays, and sodium dodecyl sulfate PAGE followed by glycoprotein staining. Resultant glycoprotein dispersions exhibited altered functionality, in which case steady shear and small amplitude oscillatory rheology parameters were shown to be dependent on the specific reducing sugar present. Furthermore, the emulsion stability of mWPC-dextran fractions was 2 to 3 times greater than either mWPC or commercial WPC dispersions based on creaming index values. The water-holding capacity of all test samples decreased with additional heating steps; however, mWPC-dextran powders still retained nearly 6 times their weight of water. Scanning electron microscopy revealed that mWPC-dextran conjugates formed a porous network that differed significantly from the dense network observed with mWPC samples. This porosity likely affected both the rheological and water-binding properties of mWPC-dextran complexes. Taken together, these results suggest that the functionality of mWPC ingredients can be enhanced by conjugation with carbohydrate materials at low pH, especially with regard to improving the emulsifying attributes.     

Rheological characterizations of texturized whey protein concentrate-based powders produced by reactive supercritical fluid extrusion

A powder blend comprising (by weight) 94% whey protein concentrate (WPC80), 6% pre-gelatinized corn starch, 0.6% CaCl₂, and 0.6% NaCl was texturized using a supercritical fluid extrusion (SCFX) process. The blend was extruded at 90 °C in a pH range of 2.89 to 8.16 with 1% (db) supercritical carbon dioxide (SC-CO₂) and 60% moisture content. The texturized WPC-based (TWPC) samples were dried, grounded into powder, reconstituted in water, and evaluated using a range of rheological studies. Most TWPC samples exhibited shear thinning behavior and their mechanical spectra were typical of weak gel characteristics. The TWPC produced under extremely acidic condition of pH 2.89 with SC-CO₂ yielded the highest η^* (10,049 Pa s) and G′ (9,885 Pa) compared to the unprocessed WPC (η^* = 0.083 Pa s and G’ = 0.036 Pa). The SCFX process rendered WPC into a product with cold-setting gel characteristics that may be suitable for use as a food texturizer over a wide range of temperatures.     

Ferrous bisglycinate content and release in W1/O/W2 multiple emulsions stabilized by protein–polysaccharide complexes

Ferrous bisglycinate aqueous solution was entrapped in the inner phase (W₁) of water-in-oil-in-water (W₁/O/W₂) multiple emulsions. The primary ferrous bisglycinate aqueous solution-in-mineral oil (W₁/O) emulsion contained 15% (w/w) ferrous bisglycinate, had a dispersed phase mass fraction of 0.5, and was stabilized with a mixture of Grindsted PGPR 90:Panodan SDK (6:4 ratio) with a total emulsifiers concentration of 5% (w/w). This primary emulsion was re-emulsified in order to prepare W₁/O/W₂ multiple emulsions, with a dispersed mass fraction of 0.2, and stabilized using protein (whey protein concentrate (WPC)):polysaccharide (gum arabic (GA) or mesquite gum (MG) or low methoxyl pectin (LMP)) complexes (2:1 ratio) in the W₂ aqueous phase. The W₁/O/W₂ multiple emulsion stabilized with WPC:MG (5% w/w total biopolymers concentration) provided smaller droplet sizes (2.05 μm), lower rate of droplet coalescence (7.09 × 10⁻⁷ s⁻¹), better protection against ferrous bisglycinate oxidation (29.75% Fe³⁺) and slower rate of ferrous bisglycinate release from W₁ to W₂ (K_H = 0.69 mg mL⁻¹ min^−0.5 in the first 24 h and 0.07 mg mL⁻¹ min^−0.5 for the next 19 days of storage time). Better encapsulation efficiencies, enhanced protection against oxidation and slower release rates of ferrous bisglycinate were achieved as the molecular weight of the polysaccharide making up protein:polysaccharide complex was higher. Thus, the factor that probably affected most the overall functionality of multiple emulsions was the thickness of the complex adsorbed around the multiple emulsion oil droplets. These thicknesses determined indirectly by measuring the z-average diameter of the complexes, and that of the WPC:MG (529.4 nm) was the largest.     

Effects of spray drying conditions and the addition of surfactants on the foaming properties of a whey protein concentrate

Whey is the main waste by-product from dairy industry and at the same time is the major source of globular proteins. These proteins are concentrated mainly through spray drying, but high temperatures affect the foaming properties of globular protein. The addition of surfactants can have a protective role against thermal effects. The aim of this work was to optimize the spray-drying condition and surfactant concentration to obtain a whey protein concentrate (WPC) to be used in hot beverages according to the industry criteria for foaming stability. Three temperatures and three surfactant concentrations were applied, and the optimization was conducted using response surface analysis. Sensory analysis was applied to the WPC obtained under optimal conditions. The results showed that the foaming stability according to industrial criteria was attained when the spray drying was performed at 210 °C with surfactant concentration of 1.50 g/100 g. This resulted in foaming capacity of 3.80 mL, moisture content of 1.82 g/100 g and apparent density of 0.181 g/cm³. The sensory analysis suggested that aroma was related to dairy, cooked and whey and taste was related to sweet and dairy notes. In conclusion, temperature and surfactant concentration played an important role in the foaming capacity and stability of WPC.     

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.     

Optimisation, by response surface methodology, of degree of hydrolysis and antioxidant and ACE-inhibitory activities of whey protein hydrolysates obtained with cardoon extract

The hydrolysis of bovine whey protein concentrate (WPC), α-lactalbumin (α-La) and caseinomacropeptide (CMP), by aqueous extracts of Cynara cardunculus, was optimized using response surface methodology. Degree of hydrolysis (DH), angiotensin-converting enzyme (ACE)-inhibitory activity and antioxidant activity were used as objective functions, and hydrolysis time and enzyme/substrate ratio as manipulated parameters. The model was statistically appropriate to describe ACE-inhibitory activity of hydrolysates from WPC and α-La, but not from CMP. Maximum DH was 18% and 9%, for WPC and α-La, respectively. 50% ACE-inhibition was produced by 105.4 (total fraction) and 25.6 μg mL⁻¹ (<3 kDa fraction) for WPC, and 47.6 (total fraction) and 22.5 μg mL⁻¹ (<3 kDa fraction) for α-La. Major peptides of fractions exhibiting ACE-inhibition were sequenced. The antioxidant activities of WPC and α-La were 0.96 ± 0.08 and 1.12 ± 0.13 μmol trolox equivalent per mg hydrolysed protein, respectively.     

Stability of oil‐in‐water emulsions as influenced by thermal treatment of whey protein dispersions or emulsions

Properties of whey protein concentrate stabilised emulsions were modified by protein and emulsion heat treatment (60–90 °C). All liquid emulsions were flocculated and the particle sizes showed bimodal size distributions. The state and surface properties of proteins and coexisting protein/aggregates in the system strongly determined the stability of heat‐modified whey protein concentrate stabilised emulsions. The whey protein particles of 122–342 nm that formed on protein heating enhanced the stability of highly concentrated emulsions. These particles stabilised protein‐heated emulsions in the way that is typical for Pickering emulsions. The emulsions heated at 80 and 90 °C gelled due to the aggregation of the protein‐coated oil droplets.     

The Effect of Feed Solids Concentration and Inlet Temperature on the Flavor of Spray Dried Whey Protein Concentrate

Previous research has demonstrated that unit operations in whey protein manufacture promote off‐flavor production in whey protein. The objective of this study was to determine the effects of feed solids concentration in liquid retentate and spray drier inlet temperature on the flavor of dried whey protein concentrate (WPC). Cheddar cheese whey was manufactured, fat‐separated, pasteurized, bleached (250 ppm hydrogen peroxide), and ultrafiltered (UF) to obtain WPC80 retentate (25% solids, wt/wt). The liquid retentate was then diluted with deionized water to the following solids concentrations: 25%, 18%, and 10%. Each of the treatments was then spray dried at the following temperatures: 180 °C, 200 °C, and 220 °C. The experiment was replicated 3 times. Flavor of the WPC80 was evaluated by sensory and instrumental analyses. Particle size and surface free fat were also analyzed. Both main effects (solids concentration and inlet temperature) and interactions were investigated. WPC80 spray dried at 10% feed solids concentration had increased surface free fat, increased intensities of overall aroma, cabbage and cardboard flavors and increased concentrations of pentanal, hexanal, heptanal, decanal, (E)2‐decenal, DMTS, DMDS, and 2,4‐decadienal (P < 0.05) compared to WPC80 spray dried at 25% feed solids. Product spray dried at lower inlet temperature also had increased surface free fat and increased intensity of cardboard flavor and increased concentrations of pentanal, (Z)4‐heptenal, nonanal, decanal, 2,4‐nonadienal, 2,4‐decadienal, and 2‐ and 3‐methyl butanal (P < 0.05) compared to product spray dried at higher inlet temperature. Particle size was higher for powders from increased feed solids concentration and increased inlet temperature (P < 0.05). An increase in feed solids concentration in the liquid retentate and inlet temperature within the parameters evaluated decreased off‐flavor intensity in the resulting WPC80.     

Evaluation of commercially available, wide-pore ultrafiltration membranes for production of α-lactalbumin-enriched whey protein concentrate

Commercially available, wide-pore ultrafiltration membranes were evaluated for production of α-lactalbumin (α-LA)-enriched whey protein concentrate (WPC). In this study microfiltration was used to produce a prepurified feed that was devoid of casein fines, lipid materials, and aggregated proteins. This prepurified feed was subsequently subjected to a wide-pore ultrafiltration process that produced an α-LA-enriched fraction in the permeate. We evaluated the performance of 3 membrane types and a range of transmembrane pressures. We determined that the optimal process used a polyvinylidene fluoride membrane (molecular weight cut-off of 50 kDa) operated at transmembrane pressure (TMP) of 207 kPa. This membrane type and operating pressure resulted in α-LA purity of 0.63, α-LA:β-LG ratio of 1.41, α-LA yield of 21.27%, and overall flux of 49.46 L/m²·h. The manufacturing cost of the process for a hypothetical plant indicated that α-LA-enriched WPC 80 (i.e., with 80% protein) could be produced at $17.92/kg when the price of whey was considered as an input cost. This price came down to $16.46/kg when the price of whey was not considered as an input cost. The results of this study indicate that production of a commercially viable α-LA-enriched WPC is possible and the process developed can be used to meet worldwide demand for α-LA-enriched whey protein.     

Effect of bleaching whey on sensory and functional properties of 80% whey protein concentrate

Whey is a highly functional food that has found widespread use in a variety of food and beverage applications. A large amount of the whey proteins produced in the United States is derived from annatto-colored Cheddar cheese. Color from annatto is undesirable in whey and must be bleached. The objective of this study was to compare 2 commercially approved bleaching agents, benzoyl peroxide (BP) and hydrogen peroxide (HP), and their effects on the flavor and functionality of 80% whey protein concentrate (WPC80). Colored and uncolored liquid wheys were bleached with BP or HP, and then ultrafiltered, diafiltered, and spray-dried; WPC80 from unbleached colored and uncolored Cheddar whey were manufactured as controls. All treatments were manufactured in triplicate. The WPC80 were then assessed by sensory, instrumental, functionality, color, and proximate analysis techniques. The HP-bleached WPC80 were higher in lipid oxidation compounds (specifically hexanal, heptanal, octanal, nonanal, decanal, dimethyl disulfide, and 1-octen-3-one) and had higher fatty and cardboard flavors compared with the other unbleached and BP-bleached WPC80. The WPC80 bleached with BP had lower norbixin concentrations compared with WPC80 bleached with HP. The WPC powders differed in Hunter color values (L, a, b), with bleached powders being more white, less red, and less yellow than unbleached powders. Bleaching with BP under the conditions used in this study resulted in larger reductions in yellowness of the powders made from whey with annatto color than did bleaching with HP. Functionality testing demonstrated that whey bleached with HP treatments had more soluble protein after 10 min of heating at 90°C at pH 4.6 and pH 7 than the no-bleach and BP treatments, regardless of additional color. Overall, HP bleaching caused more lipid oxidation products and subsequent off-flavors compared with BP bleaching. However, heat stability of WPC80 was enhanced by HP bleaching compared with control or BP-bleached WPC80.     

Prebiotic fibre‐incorporated whey protein crisps processed by supercritical fluid extrusion

Prebiotic soluble fibre (fructooligosaccharides)‐incorporated whey protein crisps were produced by low‐shear supercritical fluid extrusion (SCFX), which utilises supercritical CO₂ as an expansion agent instead of steam. Protein crisps with desirable qualities were obtained with a formulation containing 8% prebiotic fibre and 60% whey protein concentrate (WPC‐80), which gave the final product with a protein content of 49.6% (w/w). A maximum of 70% WPC‐80 and 8% prebiotic fibre could be incorporated to produce expanded protein crisps; however, increasing WPC‐80 from 50% to 70% decreased the end‐product expansion ratio from 3.1 to 1.2 and increased the product hardness and piece density from 1.3 to 2.8 kN and 0.63 to 0.75 g mL^?1, respectively. Addition of 8% prebiotic fibre did not affect the textural qualities of final products. The process produced an expanded protein matrix with unique internal microstructure of uniformly distributed closed cells. Amino acid analysis indicated that 90% of the total lysine and 92% of the total essential amino acids were retained after SCFX processing and oven‐drying, indicating the preservation of protein nutritional quality during the process.     

Transglutaminase Catalysis of Modified Whey Protein Dispersions

ABSTRACT: Transglutaminase (TGase) cross-linking reactions were accomplished using a heat-modified whey protein concentrate (mWPC) substrate after pH adjustment to 8. Based on earlier reports, the degree of lactosylation with respect to β-lactoglobulin was lower in mWPC dispersions than measured in commercial whey concentrate (cWPC) protein solutions. In this study, a higher concentration of free sulfhydryl groups was detected in soluble supernatant fractions. Both factors potentially impact the availability of reactive lysine/glutaminyl residues required for TGase reactivity. The addition of 10 mM dithiothreitol (DTT) to the substrate mix, CBZ-glutaminyl glycine and hydroxylamine, revealed a 3.6-fold increase in TGase activity, likely due in part to maintenance of the catalytic cysteine residue in a reduced state. Furthermore, inclusion of DTT to mWPC dispersions significantly raised the apparent viscosity, independently of enzyme modification, while the rate of polymerization increased 2-fold based on OPA assay measurements. Limited cross-linking slightly increased the apparent viscosity, whereas extensive coupling lowered these values compared to equivalent nonenzyme-treated mWPC samples. Carbohydrate-staining revealed formation of glyco-polymers due to covalent linkages between glucosamine and mWPC proteins after TGase processing. Again, the apparent viscosity decreased after extensive enzymatic modification. Larger particles, sized 11.28 μm, were observed in the structural matrix of TGase-mWPC-fixed samples compared to 8 μm particles in control mWPC samples as viewed in scanning electron micrographs. Ultimately, the functional characteristics of TGase-mWPC ingredients may be custom-designed to deliver alternative functional attributes by adjusting the experimental reaction conditions under which catalysis is achieved. Practical Application: Taken together, these results suggest that unique TGase-mWPC and/or TGase-mWPC-glucosamine ingredients may be designed to provide novel, value-added, polymeric/glyco-polymeric protein products that afford added benefit for the milk industry.     

Spray-drying microencapsulation and oxidative stability of conjugated linoleic acid

Conjugated linoleic acid (CLA) as the free acid was microencapsulated using whey protein concentrate (WPC) as a wall material. An emulsion of CLA was prepared using an emulsifier formulated with a 1:4 (w/w) ratio of a 30% WPC solution and was spray-dried with a laboratory unit. Microcapsules were stored at 35 and 45 °C at different water activities. Oxidation was monitored by measuring the CLA concentration, and peroxide, anisidine and total oxidation values. The encapsulation efficiency was 89.60% with a surface oil concentration of 1.77 g/100 g of sample. Microcapsules stored at a_w=0.743–0.898 had very good stability against oxidation for at least 60 days; therefore WPC is considered as an effective microencapsulating agent.     

Influence of storage, heat treatment, and solids composition on the bleaching of whey with hydrogen peroxide

The residual annatto colorant in liquid whey is bleached to provide a desired neutral color in dried whey ingredients. This study evaluated the influence of starter culture, whey solids and composition, and spray drying on bleaching efficacy. Cheddar cheese whey with annatto was manufactured with starter culture or by addition of lactic acid and rennet. Pasteurized fat-separated whey was ultrafiltered (retentate) and spray dried to 34% whey protein concentrate (WPC34). Aliquots were bleached at 60 °C for 1 h (hydrogen peroxide, 250 ppm), before pasteurization, after pasteurization, after storage at 3 °C and after freezing at -20 °C. Aliquots of retentate were bleached analogously immediately and after storage at 3 or -20 °C. Freshly spray dried WPC34 was rehydrated to 9% (w/w) solids and bleached. In a final experiment, pasteurized fat-separated whey was ultrafiltered and spray dried to WPC34 and WPC80. The WPC34 and WPC80 retentates were diluted to 7 or 9% solids (w/w) and bleached at 50 °C for 1 h. Freshly spray-dried WPC34 and WPC80 were rehydrated to 9 or 12% solids and bleached. Bleaching efficacy was measured by extraction and quantification of norbixin. Each experiment was replicated 3 times. Starter culture, fat separation, or pasteurization did not impact bleaching efficacy (P > 0.05) while cold or frozen storage decreased bleaching efficacy (P < 0.05). Bleaching efficacy of 80% (w/w) protein liquid retentate was higher than liquid whey or 34% (w/w) protein liquid retentate (P < 0.05). Processing steps, particularly holding times and solids composition, influence bleaching efficacy of whey. PRACTICAL APPLICATION: Optimization of whey bleaching conditions is important to reduce the negative effects of bleaching on the flavor of dried whey ingredients. This study established that liquid storage and whey composition are critical processing points that influence bleaching efficacy.     

Effect of incorporating whey protein concentrate into stevia‐sweetened Kulfi on physicochemical and sensory properties

In 50% sugar replaced with 0.05% stevia‐added Kulfi, whey protein concentrate (WPC) at 0, 2, 3 and 4% levels were separately incorporated. Increase in WPC level resulted in significant (P < 0.05) decrease in freezing point, melting rate, hardness and moisture percentage and significant (P < 0.05) increase in specific gravity, protein percentage and total calorie content in the product. Among 0, 2, 3 and 4% WPC‐added Kulfi, 3% WPC‐added Kulfi was adjudged as best by a panel of judges. Above 3% WPC addition, the product was very soft and possessed undesirable whey flavour.     

Short communication: Sensitive detection of norbixin in dried dairy ingredients at concentrations of less than 1 part per billion

Norbixin is the water-soluble carotenoid in annatto extracts used in the cheese industry to color Cheddar cheese. The purpose of norbixin is to provide cheese color, but norbixin is also present in the whey stream and contaminates dried dairy ingredients. Regulatory restrictions dictate that norbixin cannot be present in dairy ingredients destined for infant formula or ingredients entering different international markets. Thus, there is a need for the detection and quantification of norbixin at very low levels in dried dairy ingredients to confirm its absence. A rapid method for norbixin evaluation exists, but it does not have the sensitivity required to confirm norbixin absence at very low levels in compliance with existing regulations. The current method has a limit of detection of 2.7 μg/kg and a limit of quantification of 3.5 μg/kg. The purpose of this study was to develop a method to extract and concentrate norbixin for quantification in dried dairy ingredients below 1 μg/kg (1 ppb). A reverse-phase solid-phase extraction column step was applied in the new method to concentrate and quantify norbixin from liquid and dried WPC80 (whey protein concentrate with 80% protein), WPC34 (WPC, 34% protein), permeate, and lactose. Samples were evaluated by both methods for comparison. The established method was able to quantify norbixin in whey proteins and permeates (9.39 μg/kg to 2.35 mg/kg) but was unable to detect norbixin in suspect powdered lactose samples. The newly developed method had similar performance to the established method for whey proteins and permeates but was also able to detect norbixin in powdered lactose samples. The proposed method had a >90% recovery in lactose samples and a limit of detection of 28 ppt (ng/kg) and a limit of quantification of 94 ppt (ng/kg). The developed method provides detection and quantification of norbixin for dairy ingredients that have a concentration of <1 ppb.     

Interactions between food components in ice cream. Part 1: unfrozen emulsions

Ice cream was manufactured on a pilot plant and the structure of the emulsion was estimated in terms of droplet size distribution and protein composition of the aqueous phase after homogenization (two stages: 19 MPa + 3 MPa, 70°C) and after ageing ( 18 h, 4°C). Four different factors were studied: the nature of the milk protein [skim milk powder (SMP), skim milk replacer (SMR) or whey protein concentrate ( WPC) ], the nature of the emulsifier (saturated monoglycerides or Sugin Fl50, which is apolysorbate 80-based emulsifier) and its concentration (0.17–0.67% w/w for Sugin F150; 0.20–0.54% wlw for saturated monoglycerides), and the amount of butter oil (8–12% wlw). Freshly homogenized mixes containing either SMP or an SMR were stable during the ageing stage, irrespective of the nature and the concentration of the emulsifier. WPC-based mixes, however, were destabilized after homogenization: this destabilization was found to be flocculation only, which shows that whey proteins are efficient against coalescence. The quantity of adsorbed protein per surface unit was systematically higher for SMP mix than for both SMR and WPC. After the ageing stage, the structure of the mixes containing monoglycerides or WPC + polysorbate 80 remained unchanged. However, polysorbate 80 used in combination with both SMP and SMR led to a destabilization of the mix during the ageing stage: this destabilization was found to depend upon the mass/surface ratio of polysorbate 80 to butteroil.