Production of O/W Emulsions Containing Astaxanthin by Repeated Premix Membrane Emulsification

ABSTRACT: Oil-in-Water (O/W) emulsions are mainly produced by application of high mechanical stress or by membrane emulsification processes at low pressures. Advantages such as narrower droplet size distribution and lower operating costs make membrane emulsification processes more suitable for producing astaxanthin-loaded O/W emulsions. The characteristics of 1 of these membrane emulsification methods, called repeated premix membrane emulsification, are studied in this work. In this emulsification process, a pre-emulsion is repeatedly pushed through a hydrophilic or hydrophobic membrane. In this research, ahydrophilic membrane was used because the objective was to obtain an O/W emulsion. Pre-emulsions were produced by dispersing palm oil containing dissolved astaxanthin in water. The oil droplets were stabilized with a combination of 2 emulsifiers. Each O /W emulsion passed the membrane 3 times underpressures and disperse phase fractions of 5 to 15 bar and from 10 wt% to 40 wt%, respectively. To investigate the production of O/W emulsions by repeated premix membrane emulsification, mean Sauter diameters and fluxes were measured. To find the optimal ranges of pressure and dispersed phase fraction, a "2-level factorial design with central composite and stars points" experimental design was applied.     

Viscosity change in oil/water food emulsions prepared using a membrane emulsification system

This paper reports viscosity measurements of oil/water (O/W) monodispersed emulsions of different droplet diameters obtained in a membrane emulsification system. Hydrophilic microporous glass membranes of different pore diameters were used to prepare O/W emulsions. The results showed that the droplet diameter of the emulsions varied with the average pore diameter of the membrane. The average droplet diameter was found to be about five times greater than the average membrane pore diameter. A correlation was found for the relationship between the average droplet diameter and the emulsion viscosity. As the dispersed droplet size became smaller, the total surface area of the droplets increased. Therefore, the emulsion viscosity and the relative viscosity increased. Few studies have reported the viscosity of O/W emulsions with droplet diameter of 5 μm or more and an oil phase concentration of 10 vol% or less. In the present study a correlation between the droplet diameter and the emulsion viscosity was statistically established. ©     

碱热改性米渣谷蛋白对W/O/W型双重乳液稳定性的影响

目的：实现米渣谷蛋白在乳浊体系中的应用。方法：选取碱热改性米渣谷蛋白和span80,采用一步乳化法制备W/O/W型双重乳液,并考察蛋白浓度对双重乳液稳定性的影响。结果：当蛋白质量分数从0.5%升高至2.5%时,乳液大粒径峰消失,显微结构中液滴的双重结构增强,表观黏度及黏弹性提高,离心稳定性和贮藏稳定性增强。当蛋白质量分数为2.5%时,离心后乳清析出指数从37.21%降至10.56%,分层时间从6 h延长至96 h。蛋白质与span80形成复合膜共同稳定油水界面,形成中间态液滴,当界面蛋白足以形成刚性界面膜时,液滴从中间态转为稳定的双重结构;当蛋白质量分数为3.0%时,双重乳液发生絮凝使大粒径峰重新出现,稳定性下降,离心后乳清析出指数为16.48%,制备后96 h左右分层,过剩的蛋白质一部分参与内相液滴的构建,另一部分单独形成O/W型液滴吸附于大体积液滴外侧。结论：一步乳化法下,蛋白质量分数为2.5%时,可制得稳定双重乳液。     

Influence of Emulsification Technique and Wall Composition on Physicochemical Properties and Oxidative Stability of Fish Oil Microcapsules Produced by Spray Drying

Encapsulation of fish oil is an effective way to protect it against oxidation and masking its fishy odor. One of the possible ways to produce fish oil microcapsules is to produce an oil-in-water (O/W) emulsion followed by spray drying. This study compares the production of the O/W emulsion by mechanical homogenization (rotor–stator) with membrane emulsification and examines the effect of the type and amount of wall material added before drying. The membrane emulsification process selected for the emulsion production is premix membrane emulsification (ME), which consists of the production of a coarse emulsion by mechanical means followed by droplet breakup when the coarse emulsion is forced through a membrane. The emulsions produced had an oil load of 10 and 20 % and were stabilized using whey protein (isolate and hydrolyzate at 1 or 10 %) and sodium caseinate with concentrations of 2 and 10 %. Regarding the material used to build the microcapsule wall, whey protein, maltodextrin, or combinations of them were used at three different oil/wall ratios (1:1, 1:2, 1:3). The results clearly show that premix ME is a suitable technology for producing O/W emulsions stabilized with proteins, which have a smaller droplet size and are more monodisperse than those produced by rotor–stator emulsification. However, protein concentrations of 10 % are required to reduce the droplet size down to 2–3 μm. Small and monodisperse emulsions have been found to produce microcapsules with lower surface oil content, which increases oil encapsulation efficiency and presents lower levels of oxidation during storage at 30 °C. Of all the possible combinations studied, the one with the highest oil encapsulation efficiency is the production of a 20 % O/W emulsion stabilized with 10 % sodium caseinate followed by the addition of 50 % maltodextrin and drying.     

枯草芽孢杆菌对大豆蛋白-磷脂复合乳液在体外消化中稳定性的影响

为探讨枯草芽孢杆菌在消化过程中对大豆蛋白-磷脂复合乳液稳定性的影响。本试验设计胃肠消化模型,通过对加菌与不加菌的O/W型和W/O型乳液的乳化活性指数（Emulsifying activity index,EAI）、乳化稳定指数（Emulsification stability index,ESI）、浊度、粒径、Zeta电位检测和显微观察,考察乳液是否出现分离、絮凝、聚集上浮等现象。结果表明,在胃消化阶段,O/W型和W/O型乳液EAI、ESI、粒径、电位都明显减小,显微观察显示液滴密度减小,体积增大;加入枯草芽孢杆菌的乳液与不加菌乳液相比,各指标减小的幅度更大。在小肠消化阶段,O/W型和W/O型乳液EAI和ESI整体呈上升趋势,而粒径和电位则明显下降,另外,两者的显微观察显示液滴密度减小,体积也减小。与不加菌的相比,加入枯草芽孢杆菌后乳液EAI、ESI、粒径显著降低,电位显著升高（P<0.05）。综合上述结果,枯草芽孢杆菌促进了大豆蛋白-磷脂复合乳液在消化过程中的破乳现象。     

Fish Oil Microcapsules from O/W Emulsions Produced by Premix Membrane Emulsification

Fish oil microcapsules were prepared by combining a low-energy emulsification method (premix membrane emulsification) with spray drying. Oil-in-water (O/W) emulsions were prepared using a two-step emulsification method that used a rotor–stator homogenizer followed by membrane emulsification. The influence of the emulsification method (mechanical stirring or membrane emulsification), the emulsification conditions (membrane and emulsifier type), and the amount of wall material on the physicochemical characteristics of the microcapsules was studied. The results show that the emulsification method and the type and amount of emulsifier and wall material affect the final amount of encapsulated oil. Microcapsules produced by membrane emulsification and stabilized with 2 % Tween-20 or 10 % whey protein presented the highest values (higher than 50 %) of oil encapsulation efficiency (OEE). It has been found that the OEE increases when decreasing the droplet size of the emulsions as well as with the increase of the amount of wall material employed during drying. Morphology analysis showed that the microcapsules obtained from O/W emulsions produced by premix membrane emulsification were rounder in shape, without visible cracks on the surface and no vacuoles on the inside. Oxidation stability tests performed on some selected samples indicate that the microcapsules with higher stability are the ones produced with a higher amount of wall material and have less surface oil.     

Alginate‐Based Emulsion Template Containing High Oil Loading Stabilized by Nonionic Surfactants

Oil‐in‐water (O/W) emulsion‐gel systems containing high oil payloads are of increasing interest for food applications because of the reduction in encapsulation cost, consumption frequency or volume of food products. This study shows a facile approach to prepare stable alginate‐based O/W emulsions at high oil loading using a mixture of nonionic surfactants (Tween 80 and Span 20) as a template to form gelled‐emulsions. The synergistic effects of alginate and surfactants on the O/W emulsion properties were evaluated in terms of oil droplet size and emulsion stability. At 2% (w/v) of alginate and 1% (w/v) of surfactants, the size distribution of oil droplets was narrow and monomodal, even at an oil loading of 70% (v/v). The emulsions formed were stable against phase separation. The oil droplet size could be further reduced to below 1 μm using a high‐shear homogenizer. The emulsions formed could be easily molded and gelled into solids of different shapes via ionic gelation. The findings of this study create possible avenues for applications in food industries.     

Stability and Oil Migration of Oil‐in‐Water Emulsions Emulsified by Phase‐Separating Biopolymer Mixtures

Oil‐in‐water (O/W) emulsions with varying concentration of oil phase, medium‐chain triglyceride (MCT), were prepared using phase‐separating gum arabic (GA)/sugar beet pectin (SBP) mixture as an emulsifier. Stability of the emulsions including emulsion phase separation, droplet size change, and oil migration were investigated by means of visual observation, droplet size analysis, oil partition analysis, backscattering of light, and interfacial tension measurement. It was found that in the emulsions prepared with 4.0% GA/1.0% SBP, when the concentration of MCT was greater than 2.0%, emulsion phase separation was not observed and the emulsions were stable with droplet size unchanged during storage. This result proves the emulsification ability of phase‐separating biopolymer mixtures and their potential usage as emulsifiers to prepare O/W emulsion. However, when the concentration of MCT was equal or less than 2.0%, emulsion phase separation occurred after preparation resulting in an upper SBP‐rich phase and a lower GA‐rich phase. The droplet size increased in the upper phase whereas decreased slightly in the lower phase with time, compared to the freshly prepared emulsions. During storage, the oil droplets exhibited a complex migration process: first moving to the SBP‐rich phase, then to the GA‐rich phase and finally gathering at the interface between the two phases. The mechanisms of the emulsion stability and oil migration in the phase‐separated emulsions were discussed.     

Effects of water-soluble soybean polysaccharide on rheological properties,stability and lipid digestibility of oil-in-water emulsion during in vitro gastrointestinal digestion

Water-soluble soybean polysaccharide (SSPS) is a naturally occurring emulsifier. SSPS was used as the sole emulsifier to stabilize an oil-in-water (O/W) emulsion. The effects were investigated of different SSPS concentrations (3–20% (w/w)) on the lipid digestibility, rheological properties and stability of O/W emulsions during in vitro digestion model. The droplet size of the emulsions tended to increase during the oral phase because the emulsions were unstable and droplets coalesced, except with a SSPS concentration of 20% (w/w). The presence of SSPS markedly reduced the free fatty acid (FFA) content after its stabilized O/W emulsion passed through in vitro gastrointestinal digestion. The amount of FFA significantly decreased as the concentration of SSPS increased due to SSPS stabilization film on oil droplet surface and high viscous system. SSPS may be an attractive alternative ingredient to control the lipid digestibility of emulsions for various food products.     

The characterization of the semi-solid W/O/W emulsions with low concentrations of the primary polymeric emulsifier

Semi-solid multiple W/O/W emulsions with low concentrations (0.8, 1.6 and 2.4% w/w) of lipophilic polymeric primary emulsifier PEG-30-dipolyhydroxystearate (PDHS) have been formulated. Both emulsions, primary and multiple, were prepared with high content of inner phase (Phi1 = Phi2 = 0.8). All the formulations differ only in the lipophilic emulsifier concentration. Evaluating several parameters such as macroscopic and microscopic aspect, droplet size, accelerated stability under centrifugation and flow and oscillatory rheological behaviour, assessed the multiple systems. It is possible to formulate the semi-solid W/O/W multiple emulsions with low concentrations of PDHS as the primary emulsifier. It appeared that the highest long-term stable multiple emulsion with the lowest droplet size, the highest apparent viscosity and highest elastic characteristic, was the sample with the highest concentration (2.4% w/w) of the primary emulsifier.     

Synergistic effects of polyglycerol ester of polyricinoleic acid and sodium caseinate on the stabilisation of water–oil–water emulsions

The inherent thermodynamic instability of water–oil–water (W/O/W) emulsions has restrictions for their application in food systems. The objective of this study was to develop a food grade W/O/W emulsions with high yield and stability using minimal concentrations of surfactants. Emulsions were prepared using soybean oil, polyglycerol ester of polyricinoleic acid (PGPR) alone or in combination with sodium caseinate (NaCN) as emulsifier(s) for primary water-in-oil (W/O) emulsions and NaCN as the sole emulsifier for secondary W/O/W emulsions. Increasing the concentration of PGPR (0.5–8%w/v) had no effect on the droplet sizes of the resulting W/O/W emulsions. However, significant increases in droplet sizes of W/O/W emulsions were observed when the concentration of NaCN in external phase was reduced from 0.5 to 0.03% (w/v) (p<0.05). Percentage yields of emulsions (using a water-soluble dye) improved when PGPR concentration in the inner phase was increased from 0.5 to 8% (w/v). A stable W/O/W emulsion with a yield >90% could be prepared with 4% (w/v) PGPR alone as primary hydrophobic emulsifier and 0.5% (w/v) NaCN as external hydrophilic emulsifier. The concentration of PGPR in the inner phase could be reduced to 2% (w/v) without affecting the yield and stability of the W/O/W emulsion by partially replacing PGPR with 0.5% (w/v) NaCN, which was added to the aqueous phase of the primary W/O emulsion. The results indicate that a possible synergistic effect may exist between PGPR and NaCN, thus allowing formulation of double emulsions with reduced surfactant concentration.     

米糠蛋白O/W及W/O/W乳液制备及界面稳定性

为对比不同米糠蛋白质量浓度下O/W及W/O/W乳液的稳定性,以米糠蛋白作为基料,采用双乳化法制备O/W及W/O/W乳液,考察不同米糠蛋白质量浓度下乳液的微观形态和稳定性并探究其界面稳定机理。结果表明：W/O/W乳液的贮存稳定性显著优于O/W乳液;与相同蛋白含量的O/W乳液相比,W/O/W乳液的黏度显著提高;当米糠蛋白质量浓度为0.4 g/100 mL时,W/O/W乳液的稳定性较O/W乳液提高了1 倍以上;乳液内部包裹更多的W/O液滴,W/O/W乳液的粒径较大;而此时静电斥力也较大,起到稳定乳液的目的。同时,米糠蛋白质量浓度不小于0.4 g/100 mL时,O/W及W/O/W乳液中蛋白质的吸附率较高,达到78%以上。本研究为天然米糠蛋白质在食品级乳液中的开发提供参考,为粮食副产物的综合利用提供了新思路。     

Preparation of Monodisperse Food-Grade Oleuropein-Loaded W/O/W Emulsions Using Microchannel Emulsification and Evaluation of Their Storage Stability

W/O/W emulsion is an emerging system in developing new functional and low-calorie food products. The aim of this study is to produce food-grade monodisperse water-in-oil-in-water (W/O/W) emulsions loaded with a hydrophilic bioactive oleuropein. W/O/W emulsions were prepared via high-pressure homogenization and subsequent microchannel (MC) emulsification. The internal aqueous phase was a 5-mM sodium phosphate buffer containing d(+)-glucose (5 wt.%) and oleuropein (0.1–0.7 wt.%). The oil phase consisted of soybean oil and tetraglycerin monolaurate condensed ricinoleic acid esters (TGCR; 3–8 wt.%). The external aqueous phase was a 5-mM sodium phosphate buffer containing d(+)-glucose (5 wt.%) and decaglycerol monolaurate (1 wt.%). Oleuropein-loaded submicron W/O emulsions with average droplet diameters as small as 0.15 μm and monomodal droplet size distributions were prepared by high-pressure homogenization when applying high TGCR concentrations of 5–8 wt.% and low oleuropein concentrations of 0.1–0.3 wt.%. Monodisperse oleuropein-loaded W/O/W emulsions with average W/O droplet diameters of around 27 μm and coefficients of variation of below 5 % were successfully prepared when using a silicon MC array plate with wide channels of 5-μm depth and 18-μm width. The monodisperse W/O/W emulsions prepared at high TGCR concentrations and low oleuropein concentrations were the most stable during 40 days of storage. The adsorption behavior of oleuropein at the internal aqueous–oil interface was relevant to W/O/W emulsions microstructure and stability. The results are believed to provide useful information for successfully preparing stable monodisperse W/O/W emulsions loaded with hydrophilic functional compounds. The surface activity of the loaded material seems to be a key parameter in optimizing the formulation of W/O/W food emulsion.     

Surface-active solid lipid nanoparticles as Pickering stabilizers for oil-in-water emulsions

Oil-in-water (O/W) emulsions solely stabilized by surface-active solid lipid nanoparticles (SLNs) were developed. The SLNs were generated by quench-cooling hot O/W nanoemulsions consisting of 7.5% glyceryl stearyl citrate (GSC) dispersed in water. Their initial volume-weighted mean particle diameter (～152 nm) and zeta potential (ca.-49 mV) remained unchanged for 24 weeks. O/W emulsions (oil phase volume fraction: 0.2) containing 7.5% (w/w) GSC SLNs in the aqueous phase were kinetically-stable for 12 weeks and did not visually phase-separate over 24 weeks. The O/W emulsions generated with solid-state GSC SLNs had a volume-weighted mean oil droplet diameter of ～459 nm and a zeta potential of ca.-43 mV. Emulsion microstructure evaluated with TEM revealed dispersed oil droplets sparsely covered with adsorbed Pickering-type SLNs as well aggregated SLNs present in the continuous phase. Gradual emulsion destabilization resulted from GSC SLN dissolution during the experimental timeframe. Overall, surface-active SLNs developed via nanoemulsions effectively kinetically stabilized O/W emulsions.     

Temperature and pH as factors influencing droplet size distribution and linear viscoelasticity of O/W emulsions stabilised by soy and gluten proteins

The influence of pH and two post-emulsification treatments (pH modification and thermal cycles) over linear dynamic viscoelasticity and droplet size distribution, DSD, of O/W emulsions (75% oil) stabilized either by soy protein isolate, SPI, or wheat gluten, WG were studied in the present work. Rheological properties and droplet size of fresh emulsions showed an important dependence on pH as a consequence of the role of electrostatic interactions, not being possible to obtain a stable emulsion for pH values close to the protein isoelectric point, pI, (4–5 for SPI and 6 for WG). In order to overcome this inconvenient, an alternative emulsification procedure, basically consisting in a modification of pH after emulsification (indirect emulsification), was successfully developed. Emulsions obtained after this post-emulsification treatment, showed higher elastic (G′) and loss (G″) moduli and also larger oil droplets than fresh emulsions prepared at the same pH. Moreover, the application of upward/downward temperature cycles from 20 to 70 °C to emulsions directly prepared at a pH yielded to significantly higher values of the rheological functions when compared to those found for fresh emulsions. Accordingly, both post-emulsification treatments lead to apparent enhancements in emulsion rheology and microstructure, which is indicative of a good potential to improve long-term emulsion stability.     

阿拉伯树胶-肌原纤维蛋白共建乳状液体系的物理稳定性

研究不同质量分数阿拉伯树胶(arabic gum,AG)对肌原纤维蛋白(myofibrillar protein,MP)为乳化剂的乳化体系稳定性的影响。结果显示,乳化活性和乳化稳定性随着AG质量分数的增加呈先增加后降低的变化趋势。AG质量分数为0.3%时,MP-AG共建乳状液体系表现出最高的物理稳定性,显著地增加了ζ-电势,降低了粒径大小,表现出最低的乳析指数(P0.05)。激光共聚焦显微镜(confocal laser scanning microscopy,CLSM)观察结果表明,与单独以MP为乳化剂的样品相比,添加0.3%AG的乳状液样品液滴颗粒最小,这与粒径大小和分布的结果相一致。通过CLSM进一步观察MP在界面上的吸附行为,结果表明,与未添加AG的乳状液样品相比,添加0.3%AG的MP-AG共建乳状液体系所形成的界面膜更加坚固和致密。总之,AG可以促进蛋白质在油水界面上的吸附作用,提高MP乳化的水包油型乳状液的物理稳定性。     

Effect of chitosan on the stability and properties of modified lecithin stabilized oil-in-water monodisperse emulsion prepared by microchannel emulsification

The effect of chitosan (CHI) on the stability of monodisperse modified lecithin (ML) stabilized soybean oil-in-water (O/W) emulsion was investigated. Monodisperse emulsion droplets with particle size of 24.4 ± 0.7 μm and coefficient of variation below 12% were prepared by microchannel (MC) emulsification using a hydrophilic asymmetric straight-through MC silicon 24 × 24 mm microchip consisting of 23,348 microchannels. The stability of the ML stabilized monodisperse emulsion droplets was investigated as a function of CHI addition at various concentration, pH, ionic strength, thermal treatment and freezing-thawing treatment by means of particle size and ζ-potential measurements as well as microscopic observation. The monodisperse O/W emulsions were diluted with CHI solution at various concentrations to a final droplet concentration of 1 wt% soybean oil, 0.25 wt% ML and 0–0.5 wt% CHI at pH 3. Pronounced droplet aggregation was observed when CHI was present at a concentration range of between 0.01 and 0.04 wt%. Above this concentration range, flocculations were less extensive, indicating some restabilization. ML stabilized emulsions were stable at a wide range of NaCl concentrations (0–1000 mM) and pH (3–8). On the contrary, in the presence of CHI, aggregation of the emulsion droplets was observed when NaCl concentration was above 200 mM and when the pH started to approach the pKa of CHI (i.e. ∼6.2–7.0). Emulsions containing CHI were found to have better stability at high temperature (>70 °C) in comparison to the emulsion stabilized only by ML. With sucrose/sorbitol as cryoprotectant aids, emulsions with the addition of CHI were found to be more resistant to droplet coalescence as compared to those without CHI after freezing at −20 °C for 22 h and thawing at 30 °C for 2 h. The use of CHI may potentially destabilize ML-stabilized O/W emulsions but its stability can be enhanced by selectively choosing the appropriate CHI concentrations and conditions of preparation.     

Emulsifying properties of collagen fibers: Effect of pH,protein concentration and homogenization pressure

The emulsifying properties of collagen fibers were evaluated in oil-in-water (O/W) emulsions produced under different conditions of pH, protein content and type of emulsification device (rotor–stator and high-pressure homogenizer). The stability, microstructure and rheology of the O/W emulsions were measured. The phase separation and droplet size of the emulsions prepared using the rotor–stator device (primary emulsion) decreased with protein concentration and reduction in pH, allowing the production of electrostatically stable emulsions at pH 3.5. In contrast, emulsions at higher pH values (4.5, 5.5 and 7.5) showed a microscopic three-dimensional network responsible for their stability at protein contents higher than 1.0% (w/w). The emulsions at pH 3.5 homogenized by high pressure (up to 100 MPa) showed a decrease in surface mean diameter (d₃₂) with increasing pressure and the number of passes through the homogenizer. These emulsions showed droplets with lower dispersion and d₃₂ between 1.00 and 4.05 μm, six times lower than values observed for primary emulsions. The emulsions presented shear-thinning behavior and lower consistency index and viscosity at higher homogenization pressures. In addition, the emulsions showed a less structured gel-like behavior with increase in homogenization pressure and number of passes, since the pressure disrupted the collagen fiber structure and the oil droplets. The results of this work showed that the collagen fiber has a good potential for use as an emulsifier in the food industry, mainly in acid products.     

Formulation and Stability Characterization of Rutin-Loaded Oil-in-Water Emulsions

In this work, formulation and characterization of oil-in-water (O/W) emulsions loaded with rutin were successfully overhead. We investigated the effect of homogenization pressure on the mean droplet size, droplet size distribution, physical stability, and rutin retention of these emulsions. O/W emulsions with a mean droplet size (d _3,2) of about 150 nm and a span of nearly the unit were formulated by microfluidization at the homogenization pressure 20–150 MPa. The O/W emulsion droplets loaded with rutin were physically stable in terms of variations of d _3,2 and span during 30 days of storage in the dark condition at 4 and 25 °C. The creaming velocity was characterized using centrifugal method showing a relative good shelf life. HPLC analysis demonstrated that 71–85% of initial rutin was retained in the fresh O/W emulsions and declined to 22–35% (w/w) for 30-day storage at 25 °C. Antioxidant activity assays confirmed that rutin-loaded emulsion participated in the antioxidant activity after encapsulation similarly to pure rutin. These results indicate that O/W emulsion systems can function as potential delivery systems to enhance bioavailability to encapsulate liposoluble antioxidant rutin for potential applications in the food industry.     

Factors that influence the coalescence stability of protein-stabilised emulsions,estimated from the proportion of oil extracted by hexane

The coalescence stability of protein-stabilised emulsions was estimated by measuring the degree to which the oil content could be extracted by hexane. The hexane extraction method is an empirical one but it correlates well with both an absolute method, such as increase in droplet size, and with another ‘accelerating’ technique, oil separation by centrifugation. Moreover, the hexane extraction method is capable of measuring coalescence stability over a wide range of instability, whereas the centrifugation method only provides information about the final stages of emulsion instability. Among the proteins studied, caseinates were generally the best stabilisers, especially at pH 6. Soya proteins gave rise to emulsions of minimal stability, whereas whey protein concentrate and blood plasma resulted in emulsions of medium stability. The coalescence instability of the protein-stabilised emulsions, viewed overall, was significantly and positively related to the droplet size, the degree of flocculation and the amount of protein in the membrane. High values of these emulsion parameters were due mainly to frequent recoalescence and bridging during emulsification. To minimise these effects emulsifying conditions creating high protein: surface area ratios should be used, as well as proteins that quickly change their conformation at an interface and have low aggregation numbers in solution.