The effect of chitosan on the properties of emulsions stabilized by whey proteins

The influence of the cationic amino polysaccharide chitosan content (0–0.5%) on particle size distribution, creaming stability, apparent viscosity, and microstructure of oil-in-water emulsions (40% of rapeseed oil) containing whey protein isolate (WPI) (4%) at pH 3 was investigated. The emulsifying properties, apparent viscosity and phase separation behaviour of aqueous WPI/chitosan mixture at pH 3 were also studied. The interface tension data showed that WPI/chitosan mixture had a slightly higher emulsifying activity than had whey protein alone. An increase in chitosan content resulted in a decreased average particle size, higher viscosity and increased creaming stability of emulsions. The microstructure analysis indicated that increasing concentration of chitosan resulted in the formation of a flocculated droplet network. This behaviour of acidic model emulsions containing WPI and chitosan was explained by a flocculation phenomenon.     

Influence of encapsulation of emulsified lipids with chitosan on their in vivo digestibility

The influence of interfacial composition on the in vivo digestibility of emulsified and encapsulated lipids was investigated. An electrostatic layer-by-layer deposition technique was used to prepare soybean oil-in-water emulsions that contained lipid droplets coated by lecithin or by lecithin–chitosan. Thirty six 4-week-old male mice were divided into four groups and fed treatment diets for 4 weeks; atherogenic diets supplemented with (A) non-emulsified fat, without chitosan (control), (B) non-emulsified fat, with chitosan, (C) emulsified fat, without chitosan, or (D) emulsified fat encapsulated by chitosan. There were no differences in body weights, food intake, major organ weights, or fecal fat contents between all treatment groups, where total fat absorption was >90%. The results suggest that encapsulation of lipids by chitosan does not inhibit their in vivo digestibility, even though previous studies indicate that chitosan does inhibit their in vitro digestibility. Consequently, it should be possible to use chitosan to microencapsulate lipids and lipid-soluble components without compromising their bioavailability, although further human studies are needed to confirm this.     

Effects of xanthan gum on physicochemical properties and stability of corn oil-in-water emulsions stabilized by polyoxyethylene (20) sorbitan monooleate

The influence of added xanthan gum on rheological and dispersion characteristics and stability of concentrated (50% w/w) corn oil-in-water emulsions, stabilized with 5% (percentage on oil amount) polyoxyethylene (20) sorbitan monooleate (Tween 80), have been investigated. Emulsion with no xanthan indicated coalescence and poor creaming stability. All emulsions, with and without xanthan, showed shear-thinning flow behavior. Addition of xanthan protected emulsions from coalescence during 15 days of storage. Increase in xanthan concentration led to decrease in droplet average radius and creaming index, and increase in elastic properties of emulsions. Decrease in the emulsions flow behavior indexes, which suggested the extent of non-Newtonian behavior of emulsions, was influenced by increase in xanthan concentration. Above 0.04% of xanthan concentration, G′ and G″ values indicated formation of weak gels. Gel structure existence arises from droplet network association, due to depletion flocculation. Standard deviation of emulsions droplet size mean diameter decreased while concentration of added xanthan increased.     

Improved heat stability by whey protein–surfactant interaction

One of the main changes that occur during heat treatment of milk is whey protein denaturation, which in its turn may lead to protein aggregation and gelation. In this contribution, the effect of lysophospholipids, the main components of lysolecithins, as well as alternative surfactants, on heat-induced whey protein aggregation has been studied. Hereby, attention was paid to the relation between polar lipid molecular structure (e.g. effect of alkyl chain length, effect of polar head group) and heat-stabilising properties. Residual protein determination in the supernatant obtained after centrifugation of heated whey protein solutions learned that whey protein aggregation was at least partly prevented in the presence of surfactants. As the short alkyl chain lysophospholipids were particularly effective heat stabilisers, hydrophilic surfactants seemed to be most effective, which may be ascribed to their higher critical aggregation concentration. Upon more severe heat treatment, protein aggregation was probed either in-situ by oscillatory rheology, or ex-situ by yield rheometry. As some surfactants significantly reduced the gel strength, or even prevented heat-induced gel formation, these experiments corroborated the heat-stabilising effect of hydrophilic surfactants. Nuclear Magnetic Resonance (NMR) enabled a more direct evaluation of the protein–surfactant interaction. A strong hydrophobic interaction between small molecular weight surfactants and whey proteins became obvious from the chemical shift of the surfactant hydrophobic groups in the NMR spectrum and could be quantified by pulsed field gradient NMR (pfg-NMR) diffusiometry. The results indicated that protein–surfactant interaction did not occur upon thermal denaturation, but already took place at room temperature. However, the effect of this interaction became mainly obvious during thermal treatment.     

Influence of chitosan concentration on the stability,microstructure and rheological properties of O/W emulsions formulated with high-oleic sunflower oil and potato protein

Relatively concentrated (40 wt%) O/W emulsions formulated with high-oleic sunflower oil as disperse phase, potato protein isolate as emulsifier and chitosan as stabiliser were prepared by rotor–stator/high-pressure valve/rotor–stator homogenization. The influence of chitosan concentration on the physical stability of emulsions was studied in (0.25–1) wt% range by visual inspection, rheological and microstructural techniques. Steady shear flow curves were sensitive to the occurrence of creaming upon the rise of zero-shear viscosity values. The effect of increasing concentration of chitosan on the zero-shear viscosity turned out to be dependent on emulsion ageing and always resulted in a stepwise increase of the critical shear rate for the onset of shear thinning flow. The critical oscillatory shear stress for the onset of non-linear viscoelastic behaviour was more sensitive than the critical shear rate to detect creaming in emulsions. Mechanical spectra are definitely demonstrated to be the most powerful tool to detect not only creaming but also oil droplet flocculation on account of changes in the plateau relaxation zone. CSLM micrographs supported the interpretation of dynamic viscoelastic results, especially when flocculation as well as coalescence took place. Cryo-SEM micrographs evidenced the formation of increasingly denser protein–polysaccharide networks with chitosan concentration and the fact that the latter governs the microstructure of the emulsion when reaches 1 wt% concentration promoting enhanced physical stability.     

Preparation of nanoparticles from denatured whey protein by pH-cycling treatment

Low temperature cross-linking of denatured whey protein through pH-cycling is proposed to develop nanoparticles with controlled size and properties. Soluble polymers were produced by heating whey protein dispersions at low ionic strength and neutral pH. Nanoparticulation was induced by acidification of diluted polymer dispersions followed by pH neutralization. The effect of aggregation conditions on the physicochemical characteristics and stability of nanoparticles was studied. Nanoparticles with a diameter ranging from 100 to 300 nm were produced depending on the pH of aggregation (5.0, 5.5, 6.0), the added calcium concentration (0, 2.5, 5 mM) and the ageing time at the aggregation pH (0–75 h). The size and the turbidity of nanoparticle dispersions increased with increasing ageing time and calcium concentration. Nanoparticle voluminosity decreased with increasing calcium concentration during pH-cycling, suggesting a more compact and less porous internal structure. The stability of nanoparticles in the presence of different dissociating buffers (EDTA, urea, SDS and DTT) was evaluated and the results showed that whey protein nanoparticles were covalently cross-linked by disulphide bonds.     

Effect of whey protein concentrate addition on the physical properties of homogenized sweetened dairy creams

The influence on their whipping properties of homogenization at first and second stage pressures of 3.5/1.5 MPa and addition of whey protein concentrate (WPC) powder at three different (0.7, 1.4, and 2.1 wt percentage) concentrations to sweetened and homogenized creams was studied. Homogenization of cream significantly decreased maximum overrun and made the foam microstructure less open, while increasing whipping time, cream and foam lightness (Hunter L -value) and apparent viscosity. It also resulted in a less elastic foam structure with an increased drainage. Addition of WPC decreased the amount of maximum overrun, foam drainage and its lightness in parallel with developing a more compact microstructure. It increased the whipping time, apparent viscosity of unwhipped creams and foams, and resulted in a less elastic foam structure. The apparent viscosity of whipped cream with 2.1 wt percentage WPC, however, was lower than that of whipped cream with 1.4 wt percentage WPC, due most probably to the start up of gel formation at 2.1% WPC concentration in sweetened cream when it was sheared. Fresh foam whipped from sweetened cream with 2.1 wt percentage WPC also tended to have a slightly but not statistically significant lower elastic modulus (G') than fresh foam whipped from sweetened cream with 1.4 wt percentage WPC. This concentration can be considered as the critical value for gel formation in sweetened creams enriched by whey proteins when sheared. This study indicated the potential of WPC powder for reducing foam drainage from whipped homogenized sweetened cream.     

Rheological properties of whey protein isolate stabilized emulsions with pectin and guar gum

Dynamic oscillatory and steady-shear rheological tests were carried out to evaluate the rheological properties of whey protein isolate (WPI) stabilized emulsions with and without hydrocolloids (pectin and guar gum) at pH 7.0. Viscosity and also consistency index of emulsions increased with hydrocolloid concentration. At γ = 20 s⁻¹, the value of viscosity of the emulsion with 0.5% (w/v) pectin was about fivefold higher than that of the emulsion without pectin. Flow curves were analyzed using power law model through a fitting procedure. Flow behaviour index of all emulsions except for containing 0.5% (w/v) guar gum was approximately in the range of 0.9–1.0, which corresponds to near-Newtonian behaviour. The shear thinning behaviour of emulsions containing 0.5% (w/w) guar gum was confirmed by flow behaviour index, n, of 0.396. Both storage (G′) and loss modulus (G″) increased with an increase in frequency. Emulsions behaved like a liquid with G″ > G′ at lower frequencies; and like an elastic solid with G′ > G″ at higher frequencies. Effect of guar gum was more pronounced on dynamic properties. Phase angle values decreased from 89 to <10° with increasing frequency and indicated the viscoelasticity of WPI-stabilized emulsions with and without pectin/guar gum.     

Rheological properties of reduced-fat and low-fat ice cream containing whey protein isolate and inulin

Instrumental analyses were used to evaluate the rheological properties of regular (10%), reduced-fat (6%) and low-fat (3%) ice cream mixes and frozen ice creams stored at −18 °C. The reduced-fat and low-fat ice creams were prepared using 4% whey protein isolate (WPI) or 4% inulin as the fat replacement ingredient. The composition, colour, apparent viscosity, consistency coefficient, flow behaviour index, hardness and melting characteristics were measured. No effect of WPI or inulin was obtained on the colour values. Compared with regular ice cream, WPI changed rheological properties, resulting in significantly higher apparent viscosities, consistency indices and greater deviations from Newtonian flow. In addition, both hardness and melting resistance significantly increased by using WPI in reduced-fat and low-fat ice creams. Inulin also increased the hardness in comparison to regular ice cream, but the products made with inulin melted significantly faster than the other samples.     

Characterization of a chitosan sample extracted from Brazilian shrimps and its application to obtain insoluble complexes with a commercial whey protein isolate

The rheological behaviour of chitosan solutions in 250 mM acetate buffer was studied at different pHs (25 °C). The intrinsic viscosity decreased from ∼17 dL/g to ∼14 dL/g when the pH increased from 4.7 to 6.0. Concentrated solutions (0.5–3.0% w/w) exhibited a shear-thinning behaviour which increased with increasing chitosan concentration and decreasing pH. A good fitting of the experimental data to the Cross and Carreau flow models was obtained. The elasticity of the solutions decreased with increasing pH and decreasing chitosan concentration, as a consequence of increased chain flexibility.     

The influence of different stabilizers and salt addition on the stability of model emulsions containing olive or sesame oil

Stabilizers are widely used in low-fat emulsion production. However, food industry pays attention to ingredients, such as resistant starch (RS) that also present substantial benefits to human health. Low-fat model emulsions of either olive or sesame oil that also contained xanthan gum (XG), whey protein concentrate (WPC), and undigested (resistant) starch (RS) were produced and stored at 5 °C. Salt was added in selected samples. A multiple light scattering technique was applied for investigating destabilization phenomena. Microscopic observations and droplet size measurements took place. Rheological properties performing a heating–cooling cycle experiment (5–25–5 °C) were measured. Olive oil emulsions presented the greatest stability and the lowest droplet size. RS plays the role of solid particle stabilizer, mainly entrapped in the matrix of the continuous phase. By salt addition stability was significantly improved, whereas droplet size was decreased. Those samples had a more pronounced elastic character and significantly greater viscosity values than their counterparts without salt.     

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

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

Effects of ozone on functional properties of proteins

The present study investigates whether the ozone treatment could be an alternative to improve some functional properties of proteins. Ozone treatment was applied on whey protein isolate and egg white proteins which have been extensively used in food products to improve textural, functional and sensory attributes. Ozone treatment of proteins was performed either in aqueous solutions or as gas ozonation of pure protein powders. Foam formation and foam stability of proteins were enhanced extensively. The solubility of proteins were reduced as influenced from the aqueous and gas ozonation medium. The reduction was more pronounced in egg white proteins. Ozone treatment affected emulsion activity of whey protein isolate negatively and reduced the emulsion stability.     

Complex coacervation of soybean protein isolate and chitosan

The formation of coacervates between soybean protein isolate (SPI) and chitosan was investigated by turbidimetric analysis and coacervate yield determination as a function of pH, temperature, time, ionic strength, total biopolymer concentration (TB(conc)) and protein to polysaccharide ratio (R(SPI/Chitosan)). The interaction between SPI and chitosan yielded a sponge-like coacervate phase and the optimum conditions for their coacervation were pH 6.0-6.5, a temperature of 25 °C, and a R(SPI/Chitosan) ratio of four independently of TB(conc). NaCl inhibited the complexation between the two biopolymers. Fourier transform infrared spectroscopy (FTIR) revealed that the coacervates were formed through the electrostatic interaction between the carboxyl groups of SPI (-COO(-)) and the amine groups of chitosan (-NH(3)(+)), however hydrogen bonding was also involved in the coacervation. Differential scanning calorimetry (DSC) thermograms indicated raised denaturation temperature and network thermal stability of SPI in the coacervates due to SPI-chitosan interactions. Scanning electron microscopy (SEM) micrographs revealed that the coacervates had a porous network structure interspaced by heterogeneously sized vacuoles.     

Modulation of rheological properties by heat-induced aggregation of whey protein solution

Heat-induced protein aggregation at low protein concentrations generally leads to higher viscosities. We here report that aggregated protein can yield weaker gels than those from native protein at the same concentration. Aggregated protein was produced by heating a solution of whey protein isolate (WPI) at 3% and 9% w/w. The higher protein concentration resulted in a larger aggregate size and a higher intrinsic viscosity. The protein fraction in native WPI had the smallest size and the lowest intrinsic viscosity. The same trend was observed for the shear viscosity after concentrating the suspensions containing aggregates to around 15% w/w. Suspensions containing aggregates that were produced from a higher concentration possessed a higher viscosity. After reheating the concentrated suspensions, the suspension from the 9% w/w aggregate system produced the weakest gel, followed by the one from 3% w/w, while the native WPI yielded the strongest gel. Reactivity of the aggregates was also an important factor that influenced the resulting gel properties. We conclude that aggregation of whey protein solution is a feasible route to manipulate the gel strength of concentrated protein systems, without having to alter the concentration of the protein.     

壳聚糖修饰植物甾醇脂质体的制备及稳定性研究

采用乙醇注入法制备植物甾醇脂质体(Phytosterol Liposome,PLs),并以不同浓度壳聚糖进行修饰优化制备工艺;通过粒径、Pd I、电位和稳定性指数,分析评价了壳聚糖修饰植物甾醇脂质体(Chitosan modified Phytosterol Liposome,CS-PLs)在不同环境下的稳定性;并对壳聚糖修饰前后PLs进行了体外胃肠消化环境稳定性实验。结果表明:当壳聚糖浓度为0.3 mg/mL时可获得粒径小、分布均一的CS-PLs;且pH、温度和离子强度及种类均对CS-PLs稳定性有显著影响;PLs经壳聚糖修饰前后,胃消化稳定性均良好,但在模拟肠消化环境中,经壳聚糖修饰后的PLs表现出更好的稳定性。     

Study on the rheological properties of heat-induced whey protein isolate-dextran conjugate gel

Effect of glycosylation on the rheological properties of whey protein isolate (WPI) during the heat-induced gelation process was evaluated. Significant changes in browning intensity, free amino groups content and SDS-PAGE profile showed that the conjugate of WPI and dextran (150 kDa) was successfully prepared using the traditional dry-heating treatment. For the conjugate, during the heating and cooling cycle, the curves of G′ and G″ were considerably shifted to lower values and their shapes varied comparing to the corresponding spectra of initial WPI and WPI + dextran mixture. After holding at 25 °C, G' reached a value of about 2200 Pa, only a tenth of the value that obtained in the initial WPI gel. Moreover, frequency sweep measurements revealed that the stiffness of gel was greatly reduced in the conjugate, although a typical elastic gel was still formed. All data showed that the rheological properties of thermal gelation could be modified upon the covalent attachment of dextran.