Sorbitan tristearate layers at the air/water interface studied by shear and dilatational interfacial rheology

The shear and dilatational rheology of condensed interfacial layers of the water-insoluble surfactant sorbitan tristearate at the air/water interface is investigated. A new interfacial shear rheometer allows measurements in both stress- and strain-controlled modes, providing comprehensive interfacial rheological information such as the interfacial dynamic shear moduli, the creep response to a stress pulse, the stress relaxation response to a strain step, or steady shear curves. Our experiments show that the interfacial films are both viscoelastic and brittle in nature and subject to fracture at small deformations, as was supported by in-situ Brewster angle microscopy performed during the rheological experiments. Although any large-deformation test is destructive to the sample, it is still possible to study the linear viscoelastic regime if the deformations involved are controlled carefully. Complementary results for the dilatational rheology in area step compression/expansion experiments are reported. The dilatational behavior is predominantly elastic throughout the frequency spectrum measured, whereas the layers exhibit generalized Maxwell behavior in shear mode within a deformation frequency regime as narrow as two decades, indicating the presence of additional relaxation mechanisms in shear as opposed to expansion/compression. If the transient rheological response from stress relaxation experiments is considered, then the data can be described well with a stretched exponential model both in the shear and dilatational deformations.     

Interfacial dilatational elasticity and viscosity of beta-lactoglobulin at air-water interface using pulsating bubble tensiometry

The ability of proteins to provide stability in foams is greatly influenced by their interfacial dilatational rheological properties. Surface tension response of a pulsatingbubble with an adsorbed layer of beta-lactoglobulin was measured for different frequencies and protein concentrations using a pulsating bubble tensiometer. A methodology, accounting for adsorption/desorption as well as variation of surface concentration due to expansion/contraction, was developed for the evaluation of surface dilatational elasticity and viscosity at different frequencies from these measurements. The adsorption rate constants were inferred from the surface pressure dynamics of protein adsorption using a Langmuir minitrough. The desorption rates were shown to be negligible for beta-lactoglobulin from the surface pressure response of a spread monolayer when subjected to compression in a Langmuir minitrough. The proposed model was employed to infer the interfacial dilatational viscosity and elasticity of an adsorbed beta-lactoglobulin layer at the air-water interface from experimental pulsating bubble data for protein concentrations in the range of 0.01-0.5 wt % at pH 7. As expected, the interfacial dilatational rheological properties were found to be higher at higher protein concentrations, this effect being less pronounced for dilatational elasticity. Heating at 80 degrees C for 30 min was found to result in higher interfacial dilatational viscosity and lower interfacial dilatational elasticity though this difference was within experimental error. The traditional approach for the inference of interfacial dilatational rheological properties is found to overpredict the interfacial dilatational elasticity whereas the viscosity values do not differ significantly from those obtained using the current analysis.     

Cross linking and rheological characterization of adsorbed protein layers at the oil-water interface

The dilatational rheological properties of cross-linked protein layers adsorbed at the oil-water interface were investigated with help of a modified drop tensiometer allowing successive replacements of the external phase. This setup enables one to perform cross-linking reactions at the interface only, that is, without any contact between the cross-linking agent and protein molecules in solution, under continuous monitoring of the interfacial tension. The mechanical properties of the resulting interface were investigated with dilatational large strain experiments. Measured rheological properties were related to the expected stability of an emulsion against disproportionation by considering the ratio of the interfacial elasticity to the interfacial tension. In an attempt to increase this ratio to improve the resistance against disproportionation, experiments were performed with densified protein layers obtained via reduction of the droplet area prior to cross linking. To highlight the influence of the protein morphology on the dilatational rheological properties of the cross-linked adsorbed layers, experiments were performed with random coil (beta-casein) as well as globular (beta-lactoglobulin) proteins. Glutaraldehyde was used as a cross-linking agent. Experiments were performed at 55 degrees C and pH 7.0 in 20 mM imidazole buffer for later comparison with enzymatically cross-linked adsorbed protein layers. The present work demonstrated substantial qualitative and quantitative differences in the interfacial rheological properties of cross-linked random coil and globular proteins.     

Dilatational rheology of beta-casein adsorbed layers at liquid-fluid interfaces

The rheological behavior of beta-casein adsorption layers formed at the air-water and tetradecane-water interfaces is studied in detail by means of pendant drop tensiometry. First, its adsorption behavior is briefly summarized at both interfaces, experimentally and also theoretically. Subsequently, the experimental dilatational results obtained for a wide range of frequencies are presented for both interfaces. An interesting dependence with the oscillation frequency is observed via the comparative analysis of the interfacial elasticity (storage part) and the interfacial viscosity (loss part) for the two interfaces. The analysis of the interfacial elasticities provides information on the conformational transitions undergone by the protein upon adsorption at both interfaces. The air-water interface shows a complex behavior in which two maxima merge into one as the frequency increases, whereas only a single maximum is found at the tetradecane interface within the range of frequencies studied. This is interpreted in terms of a decisive interaction between the oil and the protein molecules. Furthermore, the analysis of the interfacial viscosities provides information on the relaxation processes occurring at both interfaces. Similarly, substantial differences arise between the gaseous and liquid interfaces and various possible relaxation mechanisms are discussed. Finally, the experimental elasticities obtained for frequencies higher than 0.1 Hz are further analyzed on the basis of a thermodynamic model. Accordingly, the nature of the conformational transition given by the maximum at these frequencies is discussed in terms of different theoretical considerations. The formation of a protein bilayer at the interface or the limited compressibility of the protein in the adsorbed state are regarded as possible explanations of the maximum.     

Surface shear rheology of saponin adsorption layers

Saponins are a wide class of natural surfactants, with molecules containing a rigid hydrophobic group (triterpenoid or steroid), connected via glycoside bonds to hydrophilic oligosaccharide chains. These surfactants are very good foam stabiliziers and emulsifiers, and show a range of nontrivial biological activities. The molecular mechanisms behind these unusual properties are unknown, and, therefore, the saponins have attracted significant research interest in recent years. In our previous study (Stanimirova et al. Langmuir2011, 27, 12486-12498), we showed that the triterpenoid saponins extracted from Quillaja saponaria plant (Quillaja saponins) formed adsorption layers with unusually high surface dilatational elasticity, 280 ± 30 mN/m. In this Article, we study the shear rheological properties of the adsorption layers of Quillaja saponins. In addition, we study the surface shear rheological properties of Yucca saponins, which are of steroid type. The experimental results show that the adsorption layers of Yucca saponins exhibit purely viscous rheological response, even at the lowest shear stress applied, whereas the adsorption layers of Quillaja saponins behave like a viscoelastic two-dimensional body. For Quillaja saponins, a single master curve describes the data for the viscoelastic creep compliance versus deformation time, up to a certain critical value of the applied shear stress. Above this value, the layer compliance increases, and the adsorption layers eventually transform into viscous ones. The experimental creep-recovery curves for the viscoelastic layers are fitted very well by compound Voigt rheological model. The obtained results are discussed from the viewpoint of the layer structure and the possible molecular mechanisms, governing the rheological response of the saponin adsorption layers.     

Structure and mechanical properties of a polyglycerol ester at the air-water surface

We studied the structure and mechanical properties of surface films resulting from the adsorption of a dispersed L beta phase at the air-water interface. This L beta phase corresponds to multilamellar vesicles and is formed by a commercial polyglycerol fatty acid ester (PGE) in aqueous solution at temperatures below the main chain-melting temperature (Tm=58 degrees C). We measured the adsorption kinetics using the pendant drop technique and mechanical properties of PGE films using oscillatory surface shear and dilatational rheometric methods. Though the adsorption kinetics are very slow, we show that the L beta phase of PGE is surface-active and forms viscoelastic films at the air-water surface after sufficiently long adsorption times. The rheological response functions to shear and dilatational deformation are reminiscent of those of temporary networks, indicating an intermolecular connectivity at the surface. This temporary network is probably created by hydrophobic interactions of alkyl chains. We obtained more detailed information about the properties of this network by comparing the rheological signature of an adsorbed PGE film (unknown structure) with a solvent-spread monolayer (known structure). We characterized the structural features of spread PGE films by recording the Langmuir isotherm and Brewster angle micrographs (BAM).We show that the rheological responses of the adsorbed film and the solvent-spread monolayer are very close to each other, indicating a structural similarity. From this study, we conclude that a dispersed L beta phase of PGE is able to adsorb at the air-water surface at T相似文献   

Correlation between mechanical behavior of protein films at the air/water interface and intrinsic stability of protein molecules

The relation between mechanical film properties of various adsorbed protein layers at the air/water interface and intrinsic stability of the corresponding proteins is discussed. Mechanical film properties were determined by surface deformation in shear and dilation. In shear, fracture stress, sigma(f), and fracture strain, gamma(f), were determined, as well as the relaxation behavior after macroscopic fracture. The dilatational measurements were performed in a Langmuir trough equipped with an infra-red reflection absorption spectroscopy (IRRAS) accessory. During compression and relaxation of the surface, the surface pressure, Pi, and adsorbed amount, Gamma (determined from the IRRAS spectra), were determined simultaneously. In addition, IRRAS spectra revealed information on conformational changes in terms of secondary structure. Possible correlations between macroscopic film properties and intrinsic stability of the proteins were determined and discussed in terms of molecular dimensions of single proteins and interfacial protein films. Molecular properties involved the area per protein molecule at Pi approximately 0 mN/m (A(0)), A(0)/M (M = molecular weight) and the maximum slope of the Pi-Gamma curves (dPi/dGamma). The differences observed in mechanical properties and relaxation behavior indicate that the behavior of a protein film subjected to large deformation may vary widely from predominantly viscous (yielding) to more elastic (fracture). This transition is also observed in gradual changes in A(0)/M. It appeared that in general protein layers with high A(0)/M have a high gamma(f) and behave more fluidlike, whereas solidlike behavior is characterized by low A(0)/M and low gamma(f). Additionally, proteins with a low A(0)/M value have a low adaptability in changing their conformation upon adsorption at the air/water interface. Both results support the conclusion that the hardness (internal cohesion) of protein molecules determines predominantly the mechanical behavior of adsorbed protein layers.     

Surface and interfacial properties of water-soluble wheat proteins

Surface and interfacial properties of water-soluble wheat proteins were investigated and compared with six reference proteins (bovine serum albumin, ovalbumin, β-lactoglobulin, trypsin, cytochrom C and β-amylase). Albumins extracted from wheat flour were separated by the free solution isoelectric focusing. The surface activity at the air/water, dodecane/water interfaces and dilatational rheological behaviour of the adsorbed layers was determined by pendant drop technique. Considerably high surface activity of wheat proteins was found at both interfaces exceeding the corresponding values of most of the reference proteins. Exceptionally, low dilatational moduli (typically  < 10 mN/m) were obtained for wheat fractions in the continuous and the stepwise compression experiments with no age effect (1–20 min) and almost no relaxation. Surface/interfacial activity and rheological properties observed imply that water-soluble wheat proteins are generally characterized by strong hydrophobicity and more flexible molecular structure than the reference proteins.     

Protein adsorption at liquid/liquid interface with low interfacial tension

Adsorption of proteins at the interface of two-liquid systems composed of aqueous ammonium sulfate solution and tert-butanol by phase separation was investigated by drop shape tensiometry. The change of interfacial tension with time and protein concentration as well as upon compression of the adsorbed layer were compared for bovine serum albumin, ovalbumin, β-lactoglobulin, lysozyme, trypsin and horse radish peroxidase. A correlation between the dilatational moduli of various protein films and the partitioning of proteins in the two-liquid system was found, which provides evidence to the role of emulsion stability in protein separation by three-phase partitioning.     

Mechanical properties of protein adsorption layers at the air/water and oil/water interface: A comparison in light of the thermodynamical stability of proteins

Over the last decades numerous studies on the interfacial rheological response of protein adsorption layers have been published. The comparison of these studies and the retrieval of a common parameter to compare protein interfacial activity are hampered by the fact that different boundary conditions (e.g. physico-chemical, instrumental, interfacial) were used. In the present work we review previous studies and attempt a unifying approach for the comparison between bulk protein properties and their adsorption films. Among many common food grade proteins we chose bovine serum albumin, β-lactoglobulin and lysozyme for their difference in thermodynamic stability and studied their adsorption at the air/water and limonene/water interface. In order to achieve this we have i) systematically analyzed protein adsorption kinetics in terms of surface pressure rise using a drop profile analysis tensiometer and ii) we addressed the interfacial layer properties under shear stress using an interfacial shear rheometer under the same experimental conditions. We could show that thermodynamically less stable proteins adsorb generally faster and yield films with higher shear rheological properties at air/water interface. The same proteins showed an analog behavior when adsorbing at the limonene/water interface but at slower rates.     

Mechanical properties of interfacial films formed by lysozyme self-assembly at the air-water interface

We present the first characterization of the mechanical properties of lysozyme films formed by self-assembly at the air-water interface using the Cambridge interfacial tensiometer (CIT), an apparatus capable of subjecting protein films to a much higher level of extensional strain than traditional dilatational techniques. CIT analysis, which is insensitive to surface pressure, provides a direct measure of the extensional stress-strain behavior of an interfacial film without the need to assume a mechanical model (e.g., viscoelastic), and without requiring difficult-to-test assumptions regarding low-strain material linearity. This testing method has revealed that the bulk solution pH from which assembly of an interfacial lysozyme film occurs influences the mechanical properties of the film more significantly than is suggested by the observed differences in elastic moduli or surface pressure. We have also identified a previously undescribed pH dependency in the effect of solution ionic strength on the mechanical strength of the lysozyme films formed at the air-water interface. Increasing solution ionic strength was found to increase lysozyme film strength when assembly occurred at pH 7, but it caused a decrease in film strength at pH 11, close to the pI of lysozyme. This result is discussed in terms of the significant contribution made to protein film strength by both electrostatic interactions and the hydrophobic effect. Washout experiments to remove protein from the bulk phase have shown that a small percentage of the interfacially adsorbed lysozyme molecules are reversibly adsorbed. Finally, the washout tests have probed the role played by additional adsorption to the fresh interface formed by the application of a large strain to the lysozyme film and have suggested the movement of reversibly bound lysozyme molecules from a subinterfacial layer to the interface.     

Dynamic properties of soy globulin adsorbed films at the air-water interface

In this paper we present surface dilatational properties of soy globulins (beta-conglycinin, glycinin, and reduced glycinin with 10 mM of dithiothreitol (DTT)) adsorbed onto the air-water interface, as a function of adsorption time. The experiments were performed at constant temperature (20 degrees C), pH (8.0), and ionic strength (0.05 M). The surface rheological parameters were measured as a function of protein concentration (ranging from 1 to 1x10(-3)% wt/wt). We found that the surface dilatational modulus, E, increases, and the phase angle, phi, decreases with time, theta, which may be associated with protein adsorption. These phenomena have been related to protein adsorption, unfolding, and/or protein-protein interactions (at long-term adsorption) as a function of protein concentration in solution. From a rheological point of view, the surface viscoelastic characteristics of soy globulin films adsorbed at the air-water interface are practically elastic. The main conclusion is that the dilatational properties of the adsorbed films depend on the molecular structure of the protein.     

Nonlinear rheology of complex fluid–fluid interfaces

Fluid–fluid interfaces stabilized by proteins, protein aggregates, polymers, or colloidal particles, tend to have a complex microstructure. Their response to an applied deformation is often highly nonlinear, even at small deformation (rates). The nonlinearity of the response is a result of changes in the interfacial microstructure. Most of the studies on interfacial rheology of complex interfaces currently available in the scientific literature, focus on the linear response regime. Since multiphase systems such as emulsions or foam are routinely exposed to large and fast deformations, characterization of the nonlinear response of complex interfaces is highly relevant. In this paper we review the recent work on nonlinear rheology of complex interfaces, both in shear and dilatational deformations. We also discuss several methods currently available for analyzing nonlinear interfacial rheology data, and recent progress in modeling nonlinear interfacial rheology, using nonequilibrium thermodynamic frameworks.     

Comparisons of the foaming and interfacial properties of whey protein isolate and egg white proteins

Whipped foams (10%, w/v protein, pH 7.0) were prepared from commercially available samples of whey protein isolate (WPI) and egg white protein (EWP), and subsequently compared based on yield stress (τ₀), overrun and drainage stability. Adsorption rates and interfacial rheological measurements at a model air/water interface were quantified via pendant drop tensiometry to better understand foaming differences among the ingredients. The highest τ₀ and resistance to drainage were observed for standard EWP, followed by EWP with added 0.1% (w/w) sodium lauryl sulfate, and then WPI. Addition of 25% (w/w) sucrose increased τ₀ and drainage resistance of the EWP-based ingredients, whereas it decreased τ₀ of WPI foams and minimally affected their drainage rates. These differing sugar effects were reflected in the interfacial rheological measurements, as sucrose addition increased the dilatational elasticity for both EWP-based ingredients, while decreasing this parameter for WPI. Previously observed relationships between τ₀ and interfacial rheology did not hold across the protein types; however, these measurements did effectively differentiate foaming behaviors within EWP-based ingredients and within WPI. Interfacial data was also collected for purified β-lactoglobulin (β-lg) and ovalbumin, the primary proteins of WPI and EWP, respectively. The addition of 25% (w/w) sucrose increased the dilatational elasticity for adsorbed layers of β-lg, while minimally affecting the interfacial rheology of adsorbed ovalbumin, in contrast to the response of WPI and EWP ingredients. These experiments underscore the importance of utilizing the same materials for interfacial measurements as used for foaming experiments, if one is to properly infer interfacial information/mechanisms and relate this information to bulk foaming measurements. The effects of protein concentration and measurement time on interfacial rheology were also considered as they relate to bulk foam properties. This data should be of practical assistance to those designing aerated food products, as it has not been previously reported that sucrose addition improves the foaming characteristics of EWP-based ingredients while negatively affecting the foaming behavior of WPI, as these types of protein isolates are common to the food industry.     

Interfacial rheology of food emulsifiers and proteins

The scientific literature from 1997 (inclusive) to the present on the interfacial rheology of emulsifiers and proteins of relevance to food has been reviewed. Both shear and dilatational rheology of oil–water and air–water interfaces have been covered and the main factors affecting interfacial rheology have been tabulated. Special attention is paid to: the sensitivity of interfacial rheology to film composition and structure; the growing viewpoint of treating proteins films as a two-dimensional gel state; recent theoretical modelling of interfacial rheological effects; those few publications that attempt to relate interfacial rheology to bulk stability. It is concluded that there have been few major advances in the last 4 or 5 years, but the heterogeneity of such adsorbed films seems to be better recognised, both spatially and rheologically, with the challenge remaining to connect this picture to the stability of the corresponding bulk systems.     

Interfacial shear rheology of protein-surfactant layers

The shear rheology of adsorbed or spread layers at air/liquid and liquid/liquid phase boundaries is relevant in a wide range of technical applications such as mass transfer, monolayers, foaming, emulsification, oil recovery, or high speed coating. Interfacial shear rheological properties can provide important information about interactions and molecular structure in the interfacial layer. A variety of measuring techniques have been proposed in the literature to measure interfacial shear rheological properties and have been applied to pure protein or mixed protein adsorption layers at air/water or oil/water interfaces. Such systems play for example an important role as stabilizers in foams and emulsions. The aim of this contribution is to give a literature overview of interfacial shear rheological studies of pure protein and protein/surfactant mixtures at liquid interfaces measured with different techniques. Techniques which utilize the damping of waves, spectroscopic or AFM techniques and all micro-rheological techniques will not discuss here.     

Effect of thermal treatment on interfacial properties of beta-lactoglobulin

The changes in the secondary conformation and surface hydrophobicity of beta-lactoglobulin subjected to different thermal treatments were characterized at pH values of 7, 5.5 and 4 using circular dichroism (CD) and hydrophobic dye binding. Heating resulted in a decrease in alpha-helix content with a corresponding increase in random coil at all pH values, this change being more pronounced for small heating times. Heating also resulted in an increase in surface hydrophobicity as a result of partial denaturation, this increase being more pronounced at pH 4. Thermal treatment resulted in a shift of the spread monolayer isotherm at air-water interface to smaller area per molecule due to increased flexibility and more loop formation. Thermal treatment led to an increase in interfacial shear elasticity and viscosity of adsorbed beta-lactoglobulin layer at pH 5.5 and 7. Interfacial shear elasticity, shear viscosity, stability of beta-lactoglobulin stabilized emulsion and average coalescence time of a single droplet at a planar oil-water interface with adsorbed protein layer exhibited a maximum for protein subjected to 15 min heat treatment at pH 7. At pH 5.5, the interfacial shear rheological properties and average single drop coalescence time were maximum for 15 min heat treatment whereas emulsion stability was maximum for 5 min heat treatment. At pH 7, thermal treatment was found to enhance foam stability. Analysis of thin film drainage indicated that interfacial shear rheological properties do not influence thin film drainage.     

Interfacial layers from the protein HFBII hydrophobin: dynamic surface tension, dilatational elasticity and relaxation times

The pendant-drop method (with drop-shape analysis) and Langmuir trough are applied to investigate the characteristic relaxation times and elasticity of interfacial layers from the protein HFBII hydrophobin. Such layers undergo a transition from fluid to elastic solid films. The transition is detected as an increase in the error of the fit of the pendant-drop profile by means of the Laplace equation of capillarity. The relaxation of surface tension after interfacial expansion follows an exponential-decay law, which indicates adsorption kinetics under barrier control. The experimental data for the relaxation time suggest that the adsorption rate is determined by the balance of two opposing factors: (i) the barrier to detachment of protein molecules from bulk aggregates and (ii) the attraction of the detached molecules by the adsorption layer due to the hydrophobic surface force. The hydrophobic attraction can explain why a greater surface coverage leads to a faster adsorption. The relaxation of surface tension after interfacial compression follows a different, square-root law. Such behavior can be attributed to surface diffusion of adsorbed protein molecules that are condensing at the periphery of interfacial protein aggregates. The surface dilatational elasticity, E, is determined in experiments on quick expansion or compression of the interfacial protein layers. At lower surface pressures (<11 mN/m) the experiments on expansion, compression and oscillations give close values of E that are increasing with the rise of surface pressure. At higher surface pressures, E exhibits the opposite tendency and the data are scattered. The latter behavior can be explained with a two-dimensional condensation of adsorbed protein molecules at the higher surface pressures. The results could be important for the understanding and control of dynamic processes in foams and emulsions stabilized by hydrophobins, as well as for the modification of solid surfaces by adsorption of such proteins.     

Exploring the interfacial structure of protein adsorbates and the kinetics of protein adsorption: an in situ high-energy X-ray reflectivity study

The high energy X-ray reflectivity technique has been applied to study the interfacial structure of protein adsorbates and protein adsorption kinetics in situ. For this purpose, the adsorption of lysozyme at the hydrophilic silica-water interface has been chosen as a model system. The structure of adsorbed lysozyme layers was probed for various aqueous solution conditions. The effect of solution pH and lysozyme concentration on the interfacial structure was measured. Monolayer formation was observed for all cases except for the highest concentration. The adsorbed protein layers consist of adsorbed lysozyme molecules with side-on or end-on orientation. By means of time-dependent X-ray reflectivity scans, the time-evolution of adsorbed proteins was monitored as well. The results of this study demonstrate the capabilities of in situ X-ray reflectivity experiments on protein adsorbates. The great advantages of this method are the broad wave vector range available and the high time resolution.     

Foaming and interfacial properties of hydrolyzed beta-lactoglobulin

beta-lactoglobulin (beta-lg) was hydrolyzed with three different proteases and subsequently evaluated for its foaming potential. Foam yield stress (tau0) was the primary variable of interest. Two heat treatments designed to inactivate the enzymes, 75 degrees C/30 min and 90 degrees C/15 min, were also investigated for their effects on foam tau0. Adsorption rates and dilatational rheological tests at a model air/water interface aided data interpretation. All unheated hydrolysates improved foam tau0 as compared to unhydrolyzed beta-lg, with those of pepsin and Alcalase 2.4L(R) being superior to trypsin. Heat inactivation negatively impacted foam tau0, although heating at 75 degrees C/30 min better preserved this parameter than heating at 90 degrees C/15 min. All hydrolysates adsorbed more rapidly at the air/water interface than unhydrolyzed beta-lg, as evidenced by their capacity to lower the interfacial tension. A previously observed relationship between interfacial dilatational elasticity (E') and tau0 was generally confirmed for these hydrolysates. Additionally, the three hydrolysates imparting the highest tau0 not only had high values of E' (approximately twice that of unhydrolyzed beta-lg), they also had very low phase angles (essentially zero). This highly elastic interfacial state is presumed to improve foam tau0 indirectly by improving foam stability and directly by imparting resistance to interfacial deformation.