Emulsification Properties of Garlic Aqueous Extract: Effect of Heat Treatment and pH Modification.

Despite the broad research available in the literature dealing with garlic health benefits, little information is found regarding the functional properties of garlic components. The aim of this study was to evaluate the emulsification properties of garlic water-soluble compounds (GWSC), encompassing proteins, saponins, and carbohydrates, after heat treatment (10 min at 95 °C) or pH adjustments (2.5, 3.5, and 7.8). After the various treatments, the extracts were used as such or filtrated (0.45 µm), and 10% soybean oil-in-water emulsions were prepared using low (0.48%) or high (6.55% wt/wt) extract concentrations. Results showed that whereas at low GWSC concentrations, both heating and acidifying resulted in the formation of bigger oil droplet sizes (i.e., from d32 = 0.36 µm using unmodified extract to d32 = 7-22 µm at pH 2.5 with or without extract filtration), the effects were opposite at the highest GWSC concentration. In the latter, heat treatment clearly reduced the droplet size as observed from the micrographs as well as the degree of creaming, though the occurrence of depletion and/or bridging flocculation was still strong. The acidification of the extract at this high GWSC concentration significantly reduced the droplet size, as observed from the micrographs; however, a strong flocculation was observed. Removal of protein aggregates, and possibly also saponin micelles, from the extract resulted in an obvious increase in emulsion droplet size. This research brings valuable insights on this study and utilisation of novel natural food emulsifiers from plant sources.

Cholesterol stabilization of phospholipid vesicles against bile-induced solubilization.

Sphingomyelin (SM) and cholesterol complex to form functional liquid-ordered (Lo) domains. It has been suggested that the detergent resistance of these domains plays a key role during gastrointestinal digestion of the milk fat globule membrane (MFGM), which is rich in both SM and cholesterol. Small-angle X-ray scattering was employed to determine the structural alterations that occur when milk sphingomyelin (MSM)/cholesterol, egg sphingomyelin (ESM)/cholesterol, soy phosphatidylcholine (SPC)/cholesterol, and milk fat globule membrane (MFGM) phospholipid/cholesterol model bilayer systems were incubated with bovine bile under physiological conditions. The persistence of diffraction peaks was indicative of multilamellar vesicles of MSM with cholesterol concentrations > 20 % mol, and also for ESM with or without cholesterol. The complexation of ESM with cholesterol is therefore capable of inhibiting the resulting vesicles from disruption by bile at lower cholesterol concentrations than MSM/cholesterol. After subtraction of background scattering by large aggregates in the bile, a Guinier fitting was used to determine changes in the radii of gyration (Rgs) over time for the biliary mixed micelles after mixing the vesicle dispersions with bile. Swelling of the micelles by phospholipid solubilization from vesicles was a function of cholesterol concentration, with less swelling of the micelles occurring as the cholesterol concentration was increased. With 40% mol cholesterol, the Rgs of the bile micelles mixed with MSM/cholesterol, ESM/cholesterol, and MFGM phospholipid/cholesterol were equal to the control (PIPES buffer + bovine bile), indicating negligible swelling of the biliary mixed micelles.

Efficacy information influences intention to take COVID-19 vaccine.

OBJECTIVES: A successful response to the COVID-19 pandemic requires achieving high levels of vaccine uptake. We tested whether directly contrasting the high efficacy of COVID-19 vaccines with the lower efficacy of the annual flu vaccine would increase intentions to take a COVID-19 vaccine. DESIGN: A pre-registered online study of 481 participants compared four information conditions: (1) no information; (2) COVID-19 Vaccine Information Only; and COVID-19 Vaccine Information combined with flu vaccine information suggesting either (3) 60% efficacy or (4) 40% efficacy; we measured COVID-19 and flu vaccine intentions along with several other vaccine-related variables. METHODS: The Prolific platform was used to recruit 481 UK participants (64% female; aged between 18 and 85 years) who had been pre-screened to have intermediate levels of vaccine hesitancy. After reading a short text (~200 words) about COVID-19 vaccines, participants were asked about their vaccination intentions. RESULTS: Providing information about the safety and efficacy of the new COVID-19 vaccines resulted in vaccination intentions that were, on average, 0.39 standard deviations (SDs) higher than those in the no information condition; providing the same COVID vaccine efficacy information in the context of information about flu vaccine efficacy resulted in a further significant increase in vaccination intentions that were 0.68 SD higher than those in the no information condition. This positive contrast effect for the COVID-19 vaccine was not associated with reduced flu vaccine intentions. CONCLUSIONS: Vaccination intentions can be strengthened through a simple messaging intervention that utilizes context effects to increase perceived response efficacy.

The effect of emulsifier type on the secondary crystallisation of monoacylglycerol and triacylglycerols in model dairy emulsions.

Dairy emulsions contain an intrinsically heterogeneous lipid phase, whose components undergo crystallisation in a manner that is critical to dairy product formulation, storage, and sensory perception. Further complexity is engendered by the diverse array of interfacially-active molecules naturally present within the serum of dairy systems, and those that are added for specific formulation purposes, all of which interact at the lipid-serum interface and modify the impact of lipid crystals on dairy emulsion stability. The work described in this article addresses this complexity, with a specific focus on the impact of temperature cycling and the effect of emulsifier type on the formation and persistence of lipid crystals at lipid-solution interfaces. Profile analysis tensiometry experiments were performed using single droplets of the low melting fraction of dairy lipids, in the presence and absence of emulsifiers (Tween 80 and whey protein isolate, WPI) and during the temperature cycling, to study the formation of monoacylglycerol (MAG) crystals at the lipid-solution interface. Companion experiments on the same lipid systems, and at the same cooling and heating rates, were undertaken with synchrotron small angle X-ray scattering, to specifically analyse the effect of emulsifier type on the formation of triacylglycerol (TAG) crystals at the lipid-solution interface of a model dairy emulsion. These two complementary techniques have revealed that Tween 80 molecules delay MAG and TAG crystal formation by lowering the temperature at which the crystallisation occurs during two cooling cycles. WPI molecules delay the crystallisation of MAGs and TAGs during the first cooling cycle, while MAG crystals form without delay during the second cooling cycle at the same temperature as MAG crystals in an emulsifier free system. The crystallisation of TAGs is inhibited during the second cooling cycle. The observed differences in crystallisation behaviour at the interface upon temperature cycling can provide further insight into the impact of emulsifiers on the long-term stability of emulsion-based dairy systems during storage.

Salmon food matrix influences digestion and bioavailability of long-chain omega-3 polyunsaturated fatty acids.

The natural structure of whole food plays an important role in the physiological impact of bioactive compounds present within the food, also known as the "matrix effect". Long-chain omega-3 polyunsaturated fatty acids (LCn-3PUFAs) are one example of a food-derived nutrient, mostly found in fish, that is believed to be influenced by the food matrix. However, most previous studies have compared only the long-term bioavailability of fish versus fish oil and have used commercial sources of fish oil. The present study aimed to investigate whether fish (salmon) matrix influences the transit of LCn-3PUFAs during in vitro digestion and affects bioavailability in healthy females. Meals containing intact salmon (intact structure), minced salmon (some structure) and defatted salmon + oil (no structure) with identical macronutrient compositions were developed. Healthy female participants (n = 13) consumed the meals in a postprandial crossover study and blood was collected at regular time points for 6 h post meal consumption. In parallel, in vitro digestion of the meals was performed using a human gastric simulator (HGS) and digesta samples were collected at regular time points for 6 h. Results: showed that plasma concentration of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) were significantly higher after participants consumed intact salmon compared to the other meals (covariate analysis p < 0.001). The in vitro digestion results showed defatted salmon + oil meal had a faster decrease in pH and faster fat emptying from the HGS than the other two meals. The defatted salmon + oil meal more closely followed fat emptying of a homogeneous unstructured meal, whereas the other meals exhibited phase separation with a delay in fat emptying. Conclusion: The fish matrix (salmon) plays an important role in the bioaccessibility and bioavailability of EPA and DHA. The differences are partly explained by fat digestion and emptying from the stomach. This study suggests that the natural structure of fish has a functional effect on the absorption and bioavailability of fish oil.

Cholesterol-phospholipid interactions resist the detergent effect of bovine bile.

Sphingomyelin (SM) and cholesterol complexation gives rise to detergent-resistant liquid-ordered domains. The persistence of these domains and subsequent mixed micelle formation was examined in the presence of bile under physiological digestive in vitro conditions for vesicles comprising either SM/cholesterol, porcine brain phosphatidylcholine (BPC)/cholesterol, or soy phosphatidylcholine (SPC)/cholesterol bilayers, the latter two systems having no liquid-ordered domains. Micellization of these digested phospholipid multilamellar vesicle systems was confirmed by transmission electron microscopy. Bovine bile was found to consist of large multilamellar sheets which subsumed phospholipid vesicles to form aggregated superstructures. Budding off from these superstructures were vesicle-to-micelle transition intermediates: unilamellar vesicles and cylindrical micelles. The presence of cholesterol (60/40 phospholipid/cholesterol mol/mol) delayed the initial rapid onset of digestion, but not for BPC and SPC vesicle systems. Acyl chain order/disorder before and after vesicle-to-micelle transition of all three phospholipid/cholesterol systems was examined using Raman spectroscopy. The addition of bovine bile to both PC/cholesterol vesicle systems reduced the overall ratio of acyl chain disorder to order. In SM/cholesterol vesicles with ≤ 20% mol cholesterol, only the lateral inter-acyl chain packing was reduced, whereas for SM/cholesterol vesicles with ≥ 30% mol cholesterol, a higher proportion of gauche-to-trans isomerization was apparent, demonstrating that SM/cholesterol complexes modify the acyl chain structure of micelles.

Soy Protein Pressed Gels: Gelation Mechanism Affects the In Vitro Proteolysis and Bioaccessibility of Added Phenolic Acids.

In this study, a model system of firm tofu (pressed gel) was prepared to study how the coagulation mechanism-acidification with glucono δ-lactone (GDL) or coagulation with magnesium sulphate (MgSO4)-affected the physical properties of the gels along with their in vitro proteolysis (or extent of proteolysis). The two types of gels were also fortified with 3.5 mM protocatechuic (PCA) and coumaric acid (CMA) to test whether they can be used as bioactive delivery systems. Texture analysis showed that all MgSO4-induced gels (fortified and control) had a higher hydration capacity and a weaker texture than the GDL-induced gels (p < 0.05). MgSO4 gels had almost double proteolysis percentages throughout the in vitro digestion and showed a significantly higher amino acid bioaccessibility than the GDL gels (essential amino acid bioaccessibility of 56% versus 31%; p < 0.05). Lastly, both gel matrices showed a similar phenolic acid release profile, on a percentage basis (~80% for PCA and ~100% for CMA). However, GDL gels delivered significantly higher masses of bioactives under simulated intestinal conditions because they could retain more of the bioactives in the gel after pressing. It was concluded that the coagulation mechanism affects both the macro- and microstructure of the soy protein pressed gels and as a result their protein digestibility. Both pressed gel matrices are promising delivery systems for bioactive phenolic acids.

A standardised semi-dynamic in vitro digestion method suitable for food - an international consensus.

The link between food and human health is increasingly a topic of interest. One avenue of study has been to assess food disintegration and interactions within the gastrointestinal tract. In vitro digestion models have been widely used to overcome the constrictions associated with in vivo methodology. The COST Action INFOGEST developed an international, harmonised protocol for static simulation of digestion in the upper gastrointestinal tract of adults. This protocol is widely used; however, it is restricted to providing end-point assessment without considering the possible structural changes. On the other hand, there are dynamic models that provide more physiologically relevant data but are expensive and difficult to access. There is a gap between these models. The method outlined in this article provides an intermediate model; it builds upon the harmonised static model and now includes crucial kinetic aspects associated with the gastric phase of digestion, including gradual acidification, fluid and enzyme secretion and emptying. This paper provides guidance and standardised recommendations of a physiologically relevant semi-dynamic in vitro simulation of upper gastrointestinal tract digestion, with particular focus on the gastric phase. Adaptations of this model have already been used to provide kinetic data on nutrient digestion and structural changes during the gastric phase that impact on nutrient absorption. Moreover, it provides a simple tool that can be used in a wide range of laboratories.

Aggregation and coalescence of partially crystalline emulsion drops investigated using optical tweezers.

The solid content of viscoelastic emulsion drops is known to affect their propensity for aggregation and their subsequent coalescence behaviour, where the balance between the drive to reduce surface tension and the straining of an internal viscoelastic network is able to create a plethora of stable partially-coalesced states. The latter has previously been elegantly demonstrated in synthetic systems, generated using oil containing different phase volumes of added solids, with micro-pipette experiments carried out on emulsion drops of several tens of microns in size. Herein we carry out experiments in the same spirit but aided by optical tweezers (OT) and using smaller micron-sized emulsion drops generated from milk fat. Given the size dependence of Brownian fluctuations and Laplace pressure the experimental investigation of these smaller drops is not necessarily a trivial extension of the previous work. The solid content of initially separated drops is controlled using a temperature-cycling regime in the sample preparation protocol, and subsequently the propensity for drops to remain joined or not after being brought into contact was examined. Aggregated pairs of drops were then subjected to an increase in temperature, either locally using a high-powered laser, or more globally using a custom-made Peltier temperature-controller. By heating to different degrees, the amount of fat crystals in the drops was able to be controlled, with progressively more compact partially-coalesced states, and eventually complete coalescence generated as the solid content was reduced. While in contrast to previous studies, the emulsion studied here was quite different in size and nature, and the solid content was controlled using temperature, the same underlying physics was nevertheless observed.

Food matrix and co-presence of turmeric compounds influence bioavailability of curcumin in healthy humans.

The natural food-derived compound curcumin (from turmeric root) is known for its anti-inflammatory and anti-oxidant effects. However, due to its poor solubility when consumed in isolation, it is poorly bioavailable. In this crossover study we compared the bioavailability of curcumin from a meal containing either curcumin powder, turmeric powder or grated fresh turmeric root, all containing 400 mg of curcumin, along with mashed potatoes and cream. Healthy male participants consumed the meals following overnight fasting, and postprandial blood samples were taken to measure plasma curcuminoids (curcumin, dimethylcurcumin (DMC) and bisdimethylcurcumin (BDMC)). All plasma curcumin values refer to total curcumin (sum of free and conjugated curcumin). The meals were also analysed using confocal laser scanning microscopy to determine the location of curcuminoids. Both of the turmeric meals produced significantly higher amounts (p < 0.05) of plasma curcuminoids at 1-3 hours after the meal was consumed, as compared to the curcumin powder. Plasma curcumin Cmax was 4.9 ng ml-1 95% CI (confidence interval) [2.2, 7.5] for the fresh turmeric meal, 8.4 ng ml-1 95% CI [4.4, 12.48] for the turmeric powder meal and 0.19 ng ml-1 95% [-0.08, 0.47] for the curcumin powder meal. Plasma DMC and BDMC were significantly higher (p < 0.05) following the turmeric powder meal, compared with the fresh turmeric meal and the curcumin powder meal. Microscopy images showed that the curcuminoid particles were mostly confined within curcuminoid cells in the fresh turmeric meal. They were unconfined but in clusters in the turmeric powder meal, while the curcuminoid particles appeared smaller in the curcumin powder meal. Conclusion: curcumin bioavailability is enhanced when consumed as fresh or powdered turmeric, which could be due to the co-presence of other turmeric compounds and/or a turmeric matrix effect.

INFOGEST static in vitro simulation of gastrointestinal food digestion.

Developing a mechanistic understanding of the impact of food structure and composition on human health has increasingly involved simulating digestion in the upper gastrointestinal tract. These simulations have used a wide range of different conditions that often have very little physiological relevance, and this impedes the meaningful comparison of results. The standardized protocol presented here is based on an international consensus developed by the COST INFOGEST network. The method is designed to be used with standard laboratory equipment and requires limited experience to encourage a wide range of researchers to adopt it. It is a static digestion method that uses constant ratios of meal to digestive fluids and a constant pH for each step of digestion. This makes the method simple to use but not suitable for simulating digestion kinetics. Using this method, food samples are subjected to sequential oral, gastric and intestinal digestion while parameters such as electrolytes, enzymes, bile, dilution, pH and time of digestion are based on available physiological data. This amended and improved digestion method (INFOGEST 2.0) avoids challenges associated with the original method, such as the inclusion of the oral phase and the use of gastric lipase. The method can be used to assess the endpoints resulting from digestion of foods by analyzing the digestion products (e.g., peptides/amino acids, fatty acids, simple sugars) and evaluating the release of micronutrients from the food matrix. The whole protocol can be completed in ~7 d, including ~5 d required for the determination of enzyme activities.

Fat Sensation: Fatty Acid Taste and Olfaction Sensitivity and the Link with Disinhibited Eating Behaviour.

Perception of fat taste, aroma, and texture are proposed to influence food preferences, thus shaping dietary intake and eating behaviour and consequently long-term health. In this study, we investigated associations between fatty acid taste, olfaction, mouthfeel of fat, dietary intake, eating behaviour, and body mass index (BMI). Fifty women attended three sessions to assess oleic acid taste and olfaction thresholds, the olfactory threshold for n-butanol and subjective mouthfeel ratings of custard samples. Dietary intake and eating behaviour were evaluated using a Food Frequency and Three-Factor Eating Questionnaire, respectively. Binomial regression analysis was used to model fat taste and olfaction data. Taste and olfactory detection for oleic acid were positively correlated (r = 0.325; p < 0.02). Oleic acid taste hypersensitive women had significantly increased n-butanol olfactory sensitivity (p < 0.03). The eating behaviour disinhibition and BMI were higher in women who were hyposensitive to oleic acid taste (p < 0.05). Dietary intake of nuts, nut spreads, and seeds were significantly correlated with high olfactory sensitivity to oleic acid (p < 0.01). These findings demonstrate a clear link between fatty acid taste sensitivity and olfaction and suggest that fat taste perception is associated with specific characteristics of eating behaviour and body composition.

The influence of emulsion structure on the Maillard reaction of ghee.

Food systems, such as cream and butter, have an emulsion or emulsion-like structure. When these food emulsions are heated to high temperatures to make products such as ghee, the Maillard reaction forms a range of volatile flavour compounds. The objective of this paper was to unravel the specific influence of emulsion structure on the Maillard reaction pathways that occur during the cooking of ghee using model systems. Switching the dispersed phase from oil to water provided a means of altering the ratios of volatile compounds produced in the cooked samples. The oil-in-water emulsion generated a volatile compound profile similar to that of the fat containing two phase model matrix, whereas the water-in-oil emulsion produced a different ratio of these compounds. The ability to generate different volatile compound profiles through the use of inverted emulsion structures could point to a new avenue for control of the Maillard reaction in high temperature food systems.

The role of the Maillard reaction in the formation of flavour compounds in dairy products--not only a deleterious reaction but also a rich source of flavour compounds.

Dairy products are heated both during processing and by consumers during food preparation; consumers place a high level of importance on flavour when assessing product acceptability. Of particular importance to the flavour of heated dairy products is the highly complex network of Maillard reactions. Much focus has been placed on the undesirable flavours generated through the Maillard reaction and how to minimise the formation of these flavours. However, beneficial flavours can also be formed by the Maillard reaction; dairy products, such as ghee, are formed by heating and are characterised by the unique flavour generated by this chemistry. This review looks at the Maillard reaction as a source of beneficial flavours for cooked dairy products and the application of models to the study of flavour formation in food systems. Models are typically used to study complex reactions in a simplified way; however, they are not always applicable to food systems.

The application of computer color matching techniques to the matching of target colors in a food substrate: a first step in the development of foods with customized appearance.

UNLABELLED: A predictive color matching model based on the colorimetric technique was developed and used to calculate the concentrations of primary food dyes needed in a model food substrate to match a set of standard tile colors. This research is the first stage in the development of novel three-dimensional (3D) foods in which color images or designs can be rapidly reproduced in 3D form. Absorption coefficients were derived for each dye, from a concentration series in the model substrate, a microwave-baked cake. When used in a linear, additive blending model these coefficients were able to predict cake color from selected dye blends to within 3 ΔE*(ab,10) color difference units, or within the limit of a visually acceptable match. Absorption coefficients were converted to pseudo X10, Y10, and Z10 tri-stimulus values (X10(P), Y10(P), Z10(P)) for colorimetric matching. The Allen algorithm was used to calculate dye concentrations to match the X10(P), Y10(P), and Z10(P) values of each tile color. Several recipes for each color were computed with the tile specular component included or excluded, and tested in the cake. Some tile colors proved out-of-gamut, limited by legal dye concentrations; these were scaled to within legal range. Actual differences suggest reasonable visual matches could be achieved for within-gamut tile colors. The Allen algorithm, with appropriate adjustments of concentration outputs, could provide a sufficiently rapid and accurate calculation tool for 3D color food printing. PRACTICAL APPLICATION: The predictive color matching approach shows potential for use in a novel embodiment of 3D food printing in which a color image or design could be rendered within a food matrix through the selective blending of primary dyes to reproduce each color element. The on-demand nature of this food application requires rapid color outputs which could be provided by the color matching technique, currently used in nonfood industries, rather than by empirical food industry methods.

Impact of oil type on nanoemulsion formation and Ostwald ripening stability.

The formation of stable transparent nanoemulsions poses two challenges: the ability to initially create an emulsion where the entire droplet size distribution is below 80 nm, and the subsequent stabilization of this emulsion against Ostwald ripening. The physical properties of the oil phase and the nature of the surfactant layer were found to have a considerable impact on nanoemulsion formation and stabilization. Nanoemulsions made with high viscosity oils, such as long chain triglycerides (LCT), were considerably larger ( D = 120 nm) than nanoemulsions prepared with low viscosity oils such as hexadecane ( D = 80 nm). The optimization of surfactant architecture, and differential viscosity eta D/eta C, has led to the formation of remarkably small nanoemulsions. With average sizes below 40 nm they are some of the smallest homogenized emulsions ever reported. What is more remarkable is that LCT nanoemulsions do not undergo Ostwald ripening and are physically stable for over 3 months. Ostwald ripening is prevented by the large molar volume of long chain triglyceride oils, which makes them insoluble in water thus providing a kinetic barrier to Ostwald ripening. Examination of the Ostwald ripening of mixed oil nanoemulsions found that the entropy gain associated with oil demixing provided a thermodynamic barrier to Ostwald ripening. Not only are the nanoemulsions created in this work some of the smallest reported, but they are also thermodynamically stable to Ostwald ripening when at least 50% of the oil phase is an insoluble triglyceride.

Emulsions stabilised by food colloid particles: role of particle adsorption and wettability at the liquid interface.

We study the effect of the particle wettability on the preferred type of emulsion stabilised solely by food colloid particles. We present results obtained with the recently developed gel trapping technique (GTT) for characterisation of wettability and surface structuring of individual food colloid particles adsorbed at air-water and oil-water interfaces. This method allows us to replicate a particle monolayer onto the surface of polydimethylsiloxane (PDMS) without altering the position of the particles. By observing the polymer surface with scanning electron microscopy (SEM), we are able to determine the contact angle of the individual particles at the initial liquid interface. We demonstrate that the GTT can be applied to fat crystal particles, calcium carbonate particles coated with stearic acid and spray-dried soy protein/calcium phosphate particles at air-water and oil-water interfaces. Subsequently, we prepare emulsions of decane and water stabilised by the same food colloid particles and correlate the wettability data obtained for these particles to the preferred type of emulsions they stabilise.

Stability and rheology of emulsions containing sodium caseinate: combined effects of ionic calcium and alcohol.

We have investigated the combined effect of ionic calcium and ethanol on the visual creaming behavior and rheology of sodium caseinate-stabilized emulsions (4 wt% protein, 30 vol% oil, pH 6.8, mean droplet diameter 0.4 microm). A range of ionic calcium concentrations, expressed as a calcium/caseinate molar ratio R, was adjusted prior to homogenization and varying concentrations of ethanol were added shortly after homogenization. A stability map was produced on the basis of visual creaming behavior over a minimum period of 8 h for different calcium/caseinate/ethanol emulsion compositions. A single narrow stable (noncreaming) region was identified, indicating limited cooperation between calcium ions and ethanol. The shear-thinning behavior of the caseinate-stabilized emulsions is typical of systems undergoing depletion flocculation. Addition of calcium ions and/or ethanol was found to lead to a pronounced reduction in viscosity and the onset of Newtonian flow. The state of aggregation was correlated with emulsion microstructure from confocal laser scanning microscopy. Time-dependent rheology (18 h) with a density-matched oil phase (1-bromohexadecane) revealed that the visually stable emulsions were time-independent low-viscosity fluids. Surface coverage data showed that increasing amounts of caseinate were associated with the oil-water interface with increasing R and ethanol content. A decrease in free calcium ions in the aqueous phase with moderate increases in R and ethanol content was observed, which is consistent with greater calcium-caseinate binding (aggregation). Ostwald ripening occurred at the high-ethanol emulsion compositions that were stable to depletion flocculation. While the coarsening rate was low, this can account for the cream plug formation observed during gravity creaming experiments. The caseinate emulsion with no ionic calcium or ethanol exhibits depletion flocculation from excess nonadsorbed caseinate submicelles. Addition of calcium ions reduces the submicelle number density via specific calcium-binding in the aqueous phase (fewer, larger calcium-caseinate aggregates) and at the droplet surface (increased surface coverage). Nonspecific ethanol-induced (calcium-dependent) caseinate submicelle aggregation in the bulk phase and on the droplet surface (increased surface coverage) culminates in a reduction in the number density of caseinate submicelles. A narrow window of inhibition of depletion flocculation occurs in systems containing both calcium ions and ethanol, both species combining to aggregate the protein and so reduce the density of free submicelles.