Effects of heat on physicochemical properties of whey protein-stabilised emulsions

The effect of heating has been studied for whey protein-stabilised oil-in-water emulsions (25.0% (w/w) soybean oil, 3.0% (w/w) whey protein isolate, pH 7.0). These emulsions were heated between 55 and 95 °C as a function of time and the effect on particle size distribution, adsorbed protein amount, protein conformation and rheological properties was determined. Heating the emulsions as a function of temperature for 25 min resulted in an increase of the mean diameter (d₃₂) and shear viscosity with a maximum at 75 °C. Heating of the emulsions at different temperatures as a function of time in all cases resulted in a curve with a maximum for d₃₂. A maximum increase of d₃₂ was observed after about 45 min at 75 °C and after 6–8 min at 90 °C. Similar trends were observed with viscosity measurements. Confocal scanning laser micrographs showed that after 8 min of heating at 90 °C large, loose aggregates of oil droplets were formed, while after 20 min of heating compact aggregates of two or three emulsion droplets remained. An increase of the adsorbed amount of protein was found with increasing heating temperature. Plateau values were reached after 10 min of heating at 75 °C and after 5 min of heating at 90 °C. Based on these results we concluded that in the whole process of aggregation of whey protein-stabilised emulsions an essential role is played by the non-adsorbed protein fraction, that the kinetics of the aggregation of whey protein-stabilised emulsions follow similar trends as those for heated whey protein solutions and that upon prolonged heating rearrangements take place leading to deaggregation of initially formed large, loose aggregates of emulsion droplets into smaller, more compact ones.     

Effects of milk protein type and pre-heating on physical stability of whipped and frozen emulsions

Various milk protein mixtures were used to manufacture complex emulsions differing by their total protein concentration (1, 2.25 and 3.5%) and by their weight proportion of casein (from 0 to 75%) or whey proteins (WP) (containing from 10 to 80% β-lactoglobulin). Beside those changes in protein concentration and composition, impact of milk protein heat treatment, which was applied before emulsification, was also investigated. The resulting structuration effects on the corresponding emulsions were determined through changes in the particle size distribution and in the amount of adsorbed protein at the fat globule surface. Furthermore, fat destabilisation under a whipping and freezing steps was underlined as functions of the processing parameters (protein concentration and composition, and application or not of an additional heat treatment), and it was discussed in terms of the resulting amount of displaced protein from the oil–water interface.     

Molecular aspects of milk allergens and their role in clinical events

Milk allergy is the most frequent food allergy in childhood. Even though cases of newly developed milk allergy in adulthood are known, this allergy is less frequent in adults since it is normally outgrown by children during the first years of life. One of the reasons why allergy to cow’s milk shows its highest prevalence in children is its early introduction into the diets of babies when breast feeding is not possible. The major allergens are caseins and β-lactoglobulin, but allergies to other minor proteins (immunoglobulins, bovine serum albumin) have also been reported. Milk allergenicity can be reduced by various treatments (mainly hydrolysis), meaning that formulas based on cow’s milk can often be safely fed to children allergic to milk proteins. Cross-reactivity has been described between different mammalian milks and between milk and meat or animal dander. Cross-contamination can result from inadequate cleaning of industrial equipment and constitutes a hidden danger for allergic subjects who unknowingly ingest milk proteins. Figure Involvement (expressed as percentage of total subjects) of the most abundant milk proteins in the sensitization of 80 children allergic to cow’s milk. The upper panel includes all positive responses, even minor ones; data in the lower panel are restricted to the most severe positive responses (see text for details). SPT, skin prick test; CAP, CAP test; IMM, immunoblotting; alpha-LA, α-lactalbumin; beta-LG, β-lactoglobulin; cas, caseins; BSA, bovine serum albumin     

Behaviour of protein-stabilised emulsions under various physiological conditions

Emulsion forms a major part of many processed food formulations. During the past few decades, the physico-chemical properties of oil-in-water emulsions under various food processing conditions have been extensively studied. However, over the recent years, interest has turned to understanding the behaviour of emulsions during consumption, i.e. physiological processing. In general, on ingestion, an emulsion is exposed to a relatively narrow range of physical (e.g. shear and temperature) and biochemical (e.g. dilution, pH, pepsin, pancreatin, mucins and bile salts) environments as it passes through the mouth into the stomach and then the intestines. There is currently limited knowledge of the physico-chemical and structural changes, which an emulsion may undergo when it passes through the physiologically active regime. A better understanding of the gastro-intestinal processing of emulsions would allow manipulation of physico-chemical and interfacial properties to modulate lipid ingestion, improve bioavailability of lipid soluble nutrients and reduce absorption of saturated fats, cholesterol and trans fats.Food emulsions are commonly stabilised by proteins, as they are not only excellent emulsifiers but also provide nutritional benefits to the product. The effects of digestion conditions on interfacial protein structures are complicated because of potential breakdown of these structures by proteolytic enzymes of the gastrointestinal tract. Studies dealing directly with the behaviour of protein-based emulsions under digestion conditions are very limited. This paper provides an overview of the behaviour of oil-in-water emulsions stabilised with globular proteins, namely lactoferrin and β-lactoglobulin. Recent advances in understanding the interactions between interfacial proteins on oil droplets and various physiological materials (e.g. enzymes and bile salts) in in vitro digestion systems are considered. Major emphasis is placed on the recent work carried out in our laboratory at Massey University on the behaviour of milk protein based emulsions (lactoferrin or β-lactoglobulin) during their passage through the gastro-intestinal tract.     

Sulphoxine immobilized onto chitosan microspheres by spray drying: application for metal ions preconcentration by flow injection analysis

A new chelating resin based on chitosan biopolymer modified with 5-sulphonic acid 8-hydroxyquinoline using the spray drying technique for immobilization is proposed. The chelating resin was characterized by thermogravimetric analysis (TGA) and X-ray diffraction (XRD) and surface area by nitrogen sorption. The efficiency of the chelating resin was evaluated by the preconcentration of metal ions Cu(II) and Cd(II) present in aqueous samples in trace amounts. The metal ions were previously enriched in a minicolumn and the concentrations of the analytes were determined on-line by flame atomic absorption spectrometry (FAAS). The maximum retention for Cu(II) occurred in the pH range 8-10, and for Cd(II) at pH 7. The optimum flow rate for sorption was found to be 7.2 ml min⁻¹ for the preconcentration of the metal ions. The analytes gave relative standard deviations (R.S.D.) of 0.7 and 0.6% for solutions containing 20 μg l⁻¹ of Cu(II) and 15 μg l⁻¹ of Cd (II), respectively (n=7). The enrichment factors for Cu(II) and Cd (II) were 19.1 and 13.9, respectively, and the limits of detection (LOD) were 0.2 μg l⁻¹ for Cd(II) and 0.3 μg l⁻¹ for Cu(II), using a preconcentration time of 90 s (n=11). The accuracy of the proposed method was evaluated by the metal ion recovery technique, in the analysis of potable water and water from a lake, with recoveries being between 97.2 and 107.3%.     

Influence of the fat characteristics on the physicochemical behavior of oil-in-water emulsions based on milk proteins-glycerol esters mixtures

Oil-in-water emulsions based on 10% milk protein preparation, 0.3% mono-di-glycerides (MDG) and 8% vegetable oil were prepared for models typifying ice cream formulations. Two MDG (saturated and partially unsaturated) and four fats (oleic oil, hydrogenated and refined coconut oils, refined palm oil) were chosen to investigate the interactions occurring between the oil phase, the MDG and the milk proteins. Influence of temperature (4 °C) and ageing (24 h at 4 °C) was also tested. The emulsions were characterized for protein desorption, particle size distribution and rheological properties. The dynamic surface activity of the milk proteins and the MDG at the oil-water interface was also determined. At 20 °C, emulsions were mostly stabilized by proteins although the protein load at the globule surface strongly depended on the emulsifier and the oil phase natures. A displacement of the proteins adsorbed at the oil droplet interface by the lipid surfactant was a consequence of the temperature decrease and/or ageing step, suggesting a disruption of the interfacial protein interactions. This disruption was more or less marked depending on the physicochemical characteristics of the surfactant and the oil used (amount of crystallized matter, fatty acid chain length and unsaturation degree). In parallel, the variation of the apparent viscosity of the various emulsions upon temperature was well correlated with the solid fat content. On the whole, the results obtained suggested that not only the surfactant molecules, i.e. emulsifiers and proteins, but also the fat used in the emulsion formulation participated in the development of the interface characteristics and rheological properties.     

Fabrication and characterization of hexagonal boron nitride powder by spray drying and calcining-nitriding technology

Hexagonal boron nitride (hBN) powder was fabricated prepared by the spray drying and calcining-nitriding technology. The effects of nitrided temperature on the phases, morphology and particle size distribution of hBN powder, were investigated. The synthesized powders were characterized by X-ray diffraction (XRD), field emission scanning electron microscope (FESEM), Fourier transformed infrared spectrum, ultraviolet-visible (UV-vis) spectrum and photoluminescence (PL) spectrum. UV-vis spectrum revealed that the product had one obvious band gap (4.7 eV) and PL spectrum showed that it had a visible emission at 457 nm (λ_ex=230 nm). FESEM image indicated that the particle size of the synthesized hBN was mainly in the range of 0.5-1.5 μm in diameter, and 50-150 nm in thickness. The high-energy ball-milling process following 900 °C calcining process was very helpful to obtain fully crystallized hBN at lower temperature.     

A differential microcalorimetric study of whey proteins and their behaviour in oil-in-water emulsions

Oil-in-water emulsions (20% soya oil, 1% protein) have been prepared containing lysozyme or isolates of -lactalbumin and β-lactoglobulin from whey protein. The structural characteristics of these proteins adsorbed at an oil/water interface were determined by following their thermal transitions using differential scanning microcalorimetry. Thermograms of the proteins in the adsorbed state differed markedly from the corresponding transitions seen for the proteins in solution. This suggests that the proteins underwent substantial changes in secondary and tertiary structure upon adsorption. In general, for all the proteins studied, a net decrease in the total energy absorbed during denaturation was found when the proteins were in an adsorbed state. Both lysozyme and -lactalbumin were in part “surface denatured”, and they showed a certain degree of reversibility between solution and the adsorbed state. β-Lactoglobulin showed the highest degree of denaturation upon adsorption and the conformational change was irreversible.     

Detection of cow's milk in ewe's or goat's milk by HPLC

Summary High-pressure liquid chromatography (HPLC) was used to detect added cows' milk in goat and ewe's milk. Analysis of the whey proteins enabled detection of mixtures in proportions of less than 1%. The method does not distinguish between goat and ewe's milk.     

Physical properties of diglycerol esters in relation to rheology and stability of protein-stabilised emulsions

The surface activity and lyotropic phase behaviour of concentrated diglycerol-esters of fatty acids with chain length of C14, C16, C18 and C18:1 (cis-oleic acid) are investigated. Diglycerol-esters show a much stronger reduction in the interfacial tension at a low concentration (0.01–0.1%) than corresponding monoglycerides. The diglycerol-esters form lamellar mesophases above their Krafft point, and no other types of mesophases are found in the temperature region examined (0–80°C). The lamellar phases show a limited swelling capacity, corresponding to a water layer thickness of ≈24 Å, which is found when the ratio of diglycerol-ester to water is 60:40, or lower. At high water concentrations (>90%) multi-lamellar liposomes are formed. The diglycerol-monooleate form lamellar phases in water in the temperature region from zero to 80°C. This is in strong contrast to the corresponding glycerol-monooleate, which forms cubic and reversed hexagonal mesophases in water. Oil in water emulsions are stabilised by diglycerol-esters by formation of liquid crystalline interfacial films around the oil droplets, which can be seen in polarised light microscopy. In presence of milk proteins in the aqueous phase the emulsion stability is depending on the protein to emulsifier ratio. At 40°C a mixed interfacial film of diglycerol-monooleate (DIGMO) and protein is present at the oil–water interface, but when cooled to 5°C, the proteins are displaced by DIGMO. This behaviour affects the stability and rheological properties of emulsions stored at low temperatures.     

Analysis of major ovine milk proteins by reversed-phase high-performance liquid chromatography and flow injection analysis with electrospray ionization mass spectrometry

Ovine milk proteins were analyzed both by coupling HPLC and electrospray ionization mass spectrometry (ESI-MS) and by flow injection analysis and ESI-MS detection after separation and collection of fractions from gel permeation chromatography. These methods resolved the four ovine caseins and whey proteins and made it possible to study the complexity of these proteins associated with genetic polymorphism, post-translational changes (phosphorylation and glycosylation) and the presence of multiple forms of proteins. The experimental molecular masses of ewe milk proteins were: 19 373 for κ-casein 3P; 25 616 for _s2-casein 10P; 23 411 for _s1-casein C-8P; 23 750 for β-casein 5P; 18 170 and 18 148 for β-lactoglobulins A and B; 14 152 for -lactalbumin A and 66 322 for serum albumin.     

Physicochemical behavior of oil-in-water emulsions: influence of milk protein mixtures, glycerol ester mixtures and fat characteristics

Different emulsions based on two protein mixtures (skim milk powder (SMP) and functional dairy proteins (FDP)), two mono-di-glyceride mixtures (MDG) (saturated and partially unsaturated), three fats (hydrogenated and refined coconut oils and refined palm oil) were studied to investigate the interactions occurring between the oil phase, low molecular weight emulsifiers and proteins. Immediately following the emulsification process, high diameters of fat globules were obtained in FDP-based systems, relevant of an aggregation phenomenon. At this stage, the fat globule size characteristics were dependent on the emulsifier and fat types present in the formulation. In contrast, SMP-based emulsions were characterized by low proportions of aggregated particles regardless the formulations. Ageing (24 h at 4 °C) promoted disaggregation in FDP formulations, while SMP emulsions were well stabilized. Just after the homogenization step, less proteins were required to stabilize the globule interface in FDP systems as compared to SMP ones. Only with SMP, the amount of protein load at the fat globule surface was influenced by the oil nature and/or by the emulsifier type. A competitive adsorption of caseins, over whey proteins, was demonstrated in the case of FDP. The ageing period promoted a displacement of the proteins adsorbed at the oil droplet interface, suggesting a disruption of the interfacial protein interactions. This disruption was more marked with SMP than with FDP and, in both cases, was more or less influenced by the emulsifier and oil phase natures. The variations of the viscosity and rheological parameters (elastic and viscous moduli) were not dependent on one specific component of the formulation.     

Development of biodegradable co-poly( -lactic/glycolic acid) microspheres for the controlled release of 5-FU by the spray drying method

The water-soluble anti-cancer drug, 5-fluorouracil (5-fluoro-2,4-pyrimidinedione) (5-FU) is encapsulated into biodegradable co-poly ( -lactic/glycolic acid) (PLGA) using the spray drying method for the development of long-lasting controlled release systems. In this study, the effects of both polymeric composition and technological parameters on release profiles of 5-FU were investigated. The degradation of various microspheres was also investigated. The mixture of dichloromethane/chloroform/methanol (1:1:2 v/v) instead of dichloromethane/chloroform (1:1 v/v) resulted in the modification of morphology, while the physical structure of the microsphere varied from a porous PLGA microsphere to a dense PLGA microsphere. The results show that the average diameter was 2 μm and the anti-cancer drug loading of microspheres approached approximately 8% (w/w). In addition, the lactide/glycolide ratio of the polymer is an important parameter for controlling the release profile of the entrapped anticancer drug. Our results indicate that the mixture solvent using the spray drying method was more efficient than emulsification solvent diffusion.     

Influence of sodium dodecyl sulfate on the physicochemical properties of whey protein-stabilized emulsions

The influence of sodium dodecyl sulfate (SDS) on the flocculation of droplets in 20 wt.% soybean oil-in-water emulsions stabilized by whey protein isolate (WPI) was investigated by light scattering, rheology and creaming measurements. The SDS concentrations used were low enough to prevent depletion flocculation by surfactant micelles and extensive protein displacement. In the absence of SDS, emulsions were prone to droplet flocculation near the isoelectric point of the proteins (4<pH<6), but were stable at a higher and lower pH. Flocculation led to an increase in emulsion viscosity, pronounced shear thinning behavior and accelerated creaming. When the surfactant-to-protein molar ratio was increased from 0 to 10, the emulsion instability range shifted to lower pH values due to binding of the negatively charged SDS molecules to the droplets. Our results indicate that the physicochemical properties of protein-stabilized emulsions can be modified by utilizing surfactant–protein interactions.     

Effect of pH on interface composition and on quality of oil-in-water emulsions made with hen egg yolk

Constituents of egg yolk are key ingredients of many food emulsions. They contribute to create an interfacial film between oil and water, which determines largely the characteristics of the emulsions. Food emulsions prepared with yolk are made at various pHs. However, the effect of pH on the adsorption of yolk constituents and on the composition of the interfacial film is not known. The present study deals with the influence of pH (3, 6 and 9), on protein interface concentration and composition, change in interfacial tension, and oil droplet diameter, of emulsions made with yolk. Emulsions were prepared as follows: 0.5% w/v of yolk; oil volume fraction: 0.375, homogenisation rate: 20 000 rpm/2 min. pH 6 provided the best conditions to prepare emulsion with yolk. The average diameter of oil droplets was lower at pH 6 (8.5 μm) than at pH 3 (11.8 μm) and pH 9 (13.5 μm). The interfacial protein concentration was higher at pH 6 (1.7 mg m⁻²) than at pH 3 and pH 9 (0.5 mg m⁻²). At pH 6, all the proteins of yolk, except phosvitin, were adsorbed at the interface and the interfacial tension at steady-state was lower (10 mN m⁻¹) than at pH 3 (15 mN m⁻¹) and pH 9 (30 mN m⁻¹). At pH 3, proteins at the interface are mainly phosvitin, and, at pH 9, some apoproteins of LDL and HDL. The pH modulates the composition of yolk proteins at the interface, mainly by modifying the net charge of the proteins causing their repulsion or dimerisation.     

柴油-生物质油乳化燃料最佳HLB值及理化性质研究

采用超声波乳化法制备柴油-生物质油乳化燃料,并研究了乳化燃料所需乳化剂的最佳亲水亲油平衡（HLB）值以及乳化条件对乳化燃料稳定性的影响,测定了乳化燃料的密度、黏度、闭口闪点、烟点、凝点和总热值等理化性质。结果表明,柴油-生物质油乳化燃料乳化剂的最佳HLB值为5.5~6.2;当乳化温度为50℃~60℃、单位容积输入功为180J/mL~300J/mL时,乳化燃料的稳定性最好;乳化燃料的密度、黏度、闪点和烟点随生物质油比例的增加而增大,而凝点和总热值则随生物质油比例的增加而降低。     

Casein adsorption on the surfaces of oil-in-water emulsions modified by lecithin

The effects of incorporating an additional component, egg-yolk lecithin, on the properties of oil-in-water emulsions stabilized by casein have been studied. The impact of lecithin on the stability of the emulsions was studied using integrated light scattering and the casein-oil-lecithin interaction was studied with photon correlation spectroscopy combined with breakdown of the adsorbed protein layers by proteolysis. Lecithin was found to enhance the stability of the emulsions at low cascin concentrations, below the limiting surface coverage of 1 mg m⁻² of casein which is found in the absence of lecithin. Conversely, small amounts of casein also stabilized flocculating oil-lecithin emulsions. The hydrodynamic thickness of the adsorbed protein layer on the hydrophobic oil surface was modified by the presence of lecithin. When the total surface area occupied by lecithin was less than 10% (5 mg lecithin for 2 ml oil), the thickness of the adsorbed casein layer was not significantly different from that in the absence of phospholipid. At higher concentrations of lecithin, the adsorbed casein layer had a lower minimum value for the layer thickness of 6.5 nm at low casein concentration and an upper plateau value of 8 nm at saturated adsorption, compared to a low limit of 5 nm and a plateau value of 10 nm in the absence of lecithin, demonstrating that the structure of the adsorbed casein layer was changed by the presence of phospholipid.     

蛋白质在羟基磷灰石界面吸附的研究

考察了酪蛋白酸钠(sodium caseinate,SC)和乳清分离蛋白(whey protein isolate,WPI)在表面性质不同的3种羟基磷灰石(hydroxyapatite,HA)颗粒上的界面吸附,分析了蛋白质的分子构型和HA颗粒的表面性质等因素对蛋白质在HA界面吸附的影响,重点讨论了SC和WPI肽链上磷酸化丝氨酸基团(phosphorylated serine residues,Ser-P)的数量和分布对吸附差异的影响.通过傅里叶变换红外光谱和表面电位分析发现SC和WPI无法被比表面积较小的HA颗粒有效吸附,但是在有效吸附面积较高的球状纳米HA和棒状微米HA上能够被吸附.Ser-P的存在使得SC在HA界面的吸附量更高、吸附能力更强.Ser-P数量和分布的不同则导致了SC中不同的蛋白组分在HA界面的竞争性吸附:β-酪蛋白在2μmHA界面始终存在优先吸附性;当纳米HA的浓度低于15 mg/mL时,纳米HA界面会优先吸附αs-酪蛋白.     

Competitive adsorption between egg yolk lipoproteins and whey proteins on oil-in-water interfaces

Adsorption characteristics of mixtures of egg yolk lipoproteins and whey protein isolate (WPI) were studied in emulsions (20% oil, v/v 0.5% protein, w/v pH 7.0) made with pure triolein or n-tetradecane. Emulsions stabilized by granule lipoproteins (GLP) or low-density lipoproteins (LDL) had smaller particle sizes than emulsions stabilized by WPI. In protein mixtures containing egg yolk lipoproteins and WPI, there was a decrease in particle size with increased concentration of the yolk lipoproteins. The reduction in particle size of emulsions was greater when WPI was mixed with LDL than with GLP, for both n-tetradecane and triolein. Emulsions made with triolein had smaller particle sizes than those made with n-tetradecane, irrespective of the type or ratio of lipoproteins used. Therefore, the protein concentration per unit area of the interface was greater for emulsions containing n-tetradecane than for triolein. In displacement experiments, emulsions made with only WPI were mixed with 0.1 and 0.5% GLP or LDL for a given period of time and the relative concentrations of β-lactoglobulin and -lactalbumin determined. Displacement of β-lactoglobulin by LDL increased with time and was greater in emulsions made with n-tetradecane than with triolein. However, displacement of β-lactoglobulin by GLP was greater in emulsions made with triolein than with n-tetradecane. -lactalbumin was completely displaced from the interface within 1 min of addition of either 0.5% GLP or LDL, whereas addition of 0.1% GLP or LDL resulted only in a partial displacement. The results suggest that egg yolk lipoproteins are more surface active than WPI and that LDL penetrates the n-tetradecane–water interface more than GLP, while GLP penetrates the triolein–water interface more than LDL.     

粉煤灰物化性质对单质汞吸附性能的影响

在固定床实验台上考察了三种不同来源粉煤灰对单质汞的吸附性能,采用X射线荧光光谱仪、X射线光电子能谱仪、激光粒度分析仪、扫描电子显微镜等对不同粉煤灰的物化性质进行了表征,并探讨了物化性质对汞吸附性能的影响。结果表明,粉煤灰对汞的吸附包括物理吸附和化学吸附;粉煤灰中未燃尽碳是影响粉煤灰汞吸附性能的重要因素之一,其中,表面C元素与Ti、Si等元素(M)相互作用形成的"C-M"化学键促进了单质汞的氧化,同时粉煤灰中含有的无机化合物Fe₂O₃等也对单质汞的氧化有促进作用;增大比表面积和减小孔径,及适中的颗粒粒径均有利于提高粉煤灰单质汞吸附性能。