Food Research International 40 (2007) 347355 www.elsevier.

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Eects of sucrose and sodium chloride on foaming properties of egg white proteins
Vassilios Raikos, Lydia Campbell, Stephen R. Euston
Received 10 April 2006; accepted 2 October 2006

School of Life Sciences, Heriot-Watt University, Riccarton, Edinburgh EH14 4AS, Scotland, UK

Abstract Egg white proteins are extensively utilised in the food industry as foaming agents. A number of factors, singly or in combination, can aect the foaming characteristics of egg albumen. In this study, egg white protein solutions heated at various temperatures in the presence of variable concentrations of sucrose and NaCl were whipped for dierent periods of time. All factors had a signicant impact on the foaming properties of egg albumen. Increasing NaCl content and whipping time enhanced protein adsorption at the airwater interface. The presence of sucrose delayed foam formation but contributed to the stability of the aerated system. Controlled denaturation of the protein solutions induced by mild heat treatment enhanced the foaming properties of egg white proteins. This data indicates that the foaming properties of egg white proteins can be manipulated by altering the eect of extrinsic factors in order to achieve optimal formulations for food industrial applications. 2006 Elsevier Ltd. All rights reserved.
Keywords: Egg white proteins; Airwater interface; Foam stability; Denaturation; Interfacial lm; Overrun; Sucrose; NaCl

1. Introduction Liquid foams are two phase systems which consist of a discontinuous air phase dispersed in a continuous liquid lamellar phase. Globular proteins such as egg white proteins are extensively used in aerated systems to enhance desirable characteristics and food applications include meringues, nougat, bavarois, whipped cream and chocolate mousse among others. Protein foams are characterised by two factors, namely foaming power and foam stability (Kato, Takahashi, Matsudomi, & Kobayashi, 1983). Foaming power or foamability is related to the level of air phase volume upon the introduction of a gas into the protein solution and is determined by measuring the increase in foam volume. Foam stability is determined by measuring the rate of liquid drainage from foam or the rate of decrease in foam volume with time. Foam stability is

Corresponding author. Tel.: +44 131 451 3640; fax: +44 131 451 3009. E-mail address: S.R.Euston@hw.ac.uk (S.R. Euston).

important for the shelf-life and product appearance of food foams and must be maintained when subjected to a variety of processes such as heating, mixing and cutting (Foegeding, Luck, & Davis, 2006). The physicochemical properties of individual proteins determine to a large extent the characteristics of foam-containing food products. However, real food systems very often contain a mixture of proteins. The properties of egg white foams depend on the individual chemical properties (molecular weight, pI, glycosylation, phosphorylation, sulfhydryl/disulde content) of a wide range of proteins which are allowed to interact during foam formation (Li-Chan & Nakai, 1989). Furthermore, the formulations of food products may include other ingredients such as sugars, salt, small-molecule surfactants. The addition of such molecules may aect the functionality of egg white proteins, which may result in gain or loss of handing properties. In addition, the foaming properties of proteins depend on extrinsic factors such as heating conditions (temperature/time), equipment for foam formation and methods of foam production (e.g. whipping time).

0963-9969/$ - see front matter 2006 Elsevier Ltd. All rights reserved. doi:10.1016/j.foodres.2006.10.008

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In previous work (Campbell, Raikos, & Euston, 2003, 2005; Raikos, Campbell, & Euston, 2007) we have found that combinations of sucrose and NaCl protect egg proteins against heat-induced aggregation and gelation. This allows egg white protein to be pasteurised for shorter times and higher temperatures than the normal 58 C for 1 min. This is advantageous for the egg producer. However, it must be ensured that the product is at least as eective in terms of its functional properties compared to the normally pasteurised product. In this study, we set out to investigate whether processing regimes designed to speed up the pasteurisation process for egg have any adverse eect on the foaming properties of egg. The combined eect of sucrose and NaCl and varied heating conditions on the foaming properties of egg albumin was studied. The foam forming ability of pre-heated egg white proteins, as aected by the presence of sucrose and NaCl, at the airwater interface was determined by overrun measurements. The impact of heat-processing conditions and sucrose and NaCl content on foam stability was also investigated. 2. Materials and methods 2.1. Materials Fresh eggs were purchased from Safeways supermarket, UK. Eggs were broken manually and the yolk was separated from the albumen. Sodium chloride and sucrose were obtained from Sigma Aldrich Co. 2.2. Preparation of protein samples Protein samples included the control (egg suspensions with no added sucrose or NaCl) and egg solutions containing dierent sucrose and/or NaCl concentrations (sucrose/ NaCl = 12/0, 0/12, 6/6, w/w). The specic sucrose and NaCl concentrations were chosen to investigate the synergistic eect of both ingredients on egg protein functionality as well as to monitor any changes occurring when one of them (e.g. sucrose) is replaced with an equal amount of the other (e.g. NaCl). 2.3. Heat treatment of egg albumin Suspensions of egg white were subjected to heat treatment (whilst stirred) in a CM4 mashing water bath (Canongate Technology Ltd.). The control samples were heated at 58 C for 2 min (normal egg pasteurisation conditions), whereas the samples containing sucrose and NaCl were heated at dierent temperatures. The heating temperatures were varied to investigate the eect of heat-induced denaturation of egg albumen on protein functionality. Egg white proteins are relatively heat labile, hence the low pasteurisation temperatures used. Previous experience has shown us that in the presence of sucrose and NaCl the pasteurisation temperature can only be increased by a few C above the normal pasteurisation temperature (58 C)

although this was sucient to give a signicant reduction in heating time. Thus, we have limited our experimental heating temperatures to the range 6064 C. Egg suspensions were placed in the water bath during the whole warming up period to ensure they were treated at the required temperature for the given period of time. The samples were placed on ice immediately after heating to prevent any further aggregation of the egg proteins. 2.4. Foaming properties Foaming properties of egg albumen were determined according to the method of Phillips, Haque, and Kinsella (1987). Foams were formed by whipping the pre-heated protein solutions in a household type mixer at ambient temperature. Overrun and stability measurements were made at certain intervals for a total of 15 min whipping using 75 ml dispersions of egg white protein. The whipping times (10 min, 13 min and 15 min) were chosen to ensure that all protein dispersions had adequate whipping time to be incorporated into foam. The protein dispersions were whipped in a double beater Breville SHM1 380watt mixer (COMET, UK). The mixer was calibrated by measuring the beater rotational speed with a stroboscope. The rotational speed of the mixer bowl was set at the high setting and the rotational speed of the beaters was set 5 (maximum). The rotational speed of the beaters was estimated to correspond to 18 ashes/s of the stroboscope and thus, the rotational speed was calculated to be 1080 rpm. 2.4.1. Foaming ability The pre-heated protein dispersions (75 ml) were poured into the bowl (2 l) and whipped for 15 min at ambient temperature. During foam formation, the mixer was stopped momentarily after 10 min and 13 min of whipping and the mixer head was carefully lifted to minimise destruction of the foam structure. Samples of foam were gently scooped out with a plastic spatula and used to quickly ll a pre-weighed weighing boat (100 ml) using small scoops and avoiding the formation of entrapped air pockets. The excess foam was removed from the weighing boat using a metal spatula to level the top of the foam even with the top of the weighing boat to obtain a constant volume of sample for each measurement. The weight of the boat with the foam was recorded and all foam was carefully returned to the bowl to resume whipping. This phase of the procedure was limited to 2 min. All measurements were replicated three times. The % overrun was calculated by the following equation: Overrun W d W f =W f 100 1:1 where Wd is the weight (g) of the unwhipped protein dispersion; Wf is the weight (g) of the whipped protein foam. Fig. 1 is a plot of overrun vs time for three dierent temperatures at a sugar/salt concentration of 0/12. This shows that the maximum in overrun occurs at 13 min.

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1600 % Overrun

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800

0 10 13 Whipping time (min) 15

Fig. 1. Overun vs whipping time for egg white samples with added sugar/ salt in a 0/12 (w/w) ratio at three temperatures. r, 60 C; j, 63 C; m, 64 C.

chosen for microscopic observation. The foam structure of the control (plain egg white, heated at 58 C, 2 min) was also observed for the same whipping times. Traces of the dye were added to the pre-heated protein samples prior to whipping in the Breville SHM1 mixer to ensure an even dispersion throughout the foam structures. Foam samples were placed on a circular plastic slide (observation area 30 mm) which is designed to t in the base plate (81 55.5 5.5 mm) of a POC Chamber System (aluminium, black anodised) to avoid any destruction of the foam structure. Time dependent images were taken at 30 min intervals for an experimental period of 60 min. 2.6. Quantitative determination of sulfhydryl (SH) groups Suspensions of egg white were subjected to heat treatment in a CM4 mashing water bath (Canongate Technology ltd.) The egg samples were heated in the presence of glucose and/or sodium chloride. The concentrations of NaCl and glucose were varied. Egg suspensions were placed in the water bath during the whole warming up period to ensure they were treated at the required temperature for the given period of time. Egg white samples were heated at 58 C, 60 C, 62 C and 64 C for 2 min. Control (non-heated25 C) samples were also prepared for egg white. The samples were placed on ice immediately after heating to prevent any further aggregation of the egg proteins. The total free SH concentration of the egg samples was determined using the standard colorimetricEllmans test (Ellman, 1959). The aim was to quantify the free SH groups as aected by heat treatment and thus, the samples were not exposed to denaturants. Fifty microliter of DTNB reagent (50 mM sodium acetate, 2 mM DTNB in dH2O) were mixed with 100 ll Tris solution (1 M Tris, pH 8) and 500 ll of egg sample and the volume was made up to 1 ml with distilled H2O (350 ll). The solution was mixed carefully and was allowed to stand at room temperature (25 C) for 5 min. The sample solution was introduced into a 1 ml cuvette and absorbance was recorded at 412 nm using a UVVIS spectrophotometer (Novaspec II, Amersham Pharmacia Biotech.). Measurements were carried out in triplicate. A blank sample containing distilled H2O instead of egg sample was used to record background readings each time. The concentration of the SH group (lM) is calculated from A412 readings using Beers law (Eq. (1.2)):    DA412 vlL C lMSH U 1:2 vslL e412 where DA412 is the absorbance change corrected for the reagent blank; vlL is the total volume sample in the measurement cuvette; vslL is the original volume of the food sample; U is the fraction of the assay volume poured into the cuvette (U = 1); e412 is the extinction coecient of the reagent (13,600 M1 cm1).

2.4.2. Foam stability Foam stability was measured by monitoring drainage at ambient temperature. To facilitate continuous measurement of drainage from the foams, the stainless steel bowl was modied by drilling a 0.6 cm hole in the bottom of the bowl 5.0 cm from the centre. The hole was sealed during the whipping process by placing a tape over the hole on the outside of the bowl. The pre-heated protein dispersion (75 ml) was poured into the bowl. The bowl, beaters and protein dispersion were weighed and whipping started at the calibrated setting for a predetermined time e.g. 10 min, 13 min or 15 min. A timer was started immediately after the whipping process was completed to estimate the drainage period. The bowl, beaters and protein foam were quickly weighed to quantify the moisture loss during whipping and obtain an accurate weight of liquid in the foam. The tape was removed, the hole was cleared with a glass rod and the bowl was placed at a 30 angle above a preweighed weighing boat (to ensure that the hole was at the lowest point), standing on the balance pan of a Sartorius 1207 MP2 electronic scale. The liquid was collected in the weighing boat on the balance pan and the increase in weight was recorded over time. Foam stability was determined by measuring the time required to attain 50% drainage (Halling, 1981). All measurements were replicated three times. 2.5. Confocal laser scanning microscopy Microscopic images of the egg albumen foams were obtained using a Leica DM-IRE2 confocal laser scanning microscope (Leica Microsystems, Heidelberg, Germany) equipped with an Ar/HeNe laser and 10 X objective lens (NPLAN 10 0.25 DRY). The uorescence dye was excited at 50% of maximum adsorption at 543 nm and the detection bandwidth was set from 488 nm to 543 nm. Images were recorded at a resolution of 512 512 pixels and analysed by the manufacturers software (Leica Software Development Kit, DM SDK, version 4.2.1). The labelling dye was Fluorescein (isothiocyanate isomer I) and it was purchased from Sigma Aldrich Co. The egg white samples heated at 64 C (2 min) and whipped for 10 min and 13 min were randomly

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3. Results and discussion Figs. 24 present results for the eect of NaCl on foaming ability. The addition of NaCl enhances foaming ability and the samples exhibit signicantly higher overrun compared to the control and the samples containing sucrose. Conversely, the % overrun decreases with increasing sucrose concentration. This is possibly attributed to an
1400 1200 (%) Overrun 1000 Sugar/Salt=12/0 800 600 400 200 0 10 13 Time (min) 15 Sugar/Salt = 0/12 Sugar/Salt = 6/6 Control

Fig. 2. Eect of sugar and salt content and whipping time on % overrun of egg white aerated systems heated at 60 C (2 min). Data are the mean of three replicates.

1400 1200 (%) Overrun 1000 800 600 400 200 0 10 13 Time (min) 15 Sugar/Salt=12/0 Sugar/Salt=0/12 Sugar/Salt=6/6 Control

Fig. 3. Eect of sugar and salt content and whipping time on % overrun of egg white aerated systems heated at 63 C (2 min). Data are the mean of three replicates.

1400 1200 (%) Overrun 1000 800 600 400 200 0 10 13 Time (min) 15 Sugar/Salt=12/0 Sugar/Salt=0/12 Sugar/Salt=6/6 Control

Fig. 4. Eect of sugar and salt content and whipping time on % overrun of egg white aerated systems heated at 64 C (2 min). Data are the mean of three replicates.

increase in the medium viscosity, which allows less air to be incorporated. The foaming ability is enhanced as whipping time increases, apart from the sample with the highest NaCl concentration. In this case, the maximum overrun is obtained after 13 min of whipping and overbeating occurs after this stage. The same phenomenon is observed for the sample containing sucrose/NaCl in a 6/6 (w/w) ratio when heated at 64 C (Fig. 4). This improved functionality may be attributed to the input of higher amounts of mechanical energy with increasing whipping time, which in turn enables more protein to adsorb and aggregate on the interfacial lamellae resulting in decreased surface tension. In some cases, a maximum in the overrun is observed after several minutes of whipping and further input of mechanical energy results in overrun decline. For instance, the samples containing sucrose/NaCl in a 0/12 weight ratio reach a maximum overrun after 13 min of whipping and longer whipping times lead to lower overrun values. The same phenomenon (maximum overrun after 10 min of whipping) is observed for the sample heated at 64 C (2 min) in the presence of sucrose and NaCl in a 6/6 weight ratio. The mechanism responsible for the overrun decline after a specied time of whipping, also known as overbeating, has been known for a long time (Halling, 1981; Kinsella, 1981). According to the theory, overbeating due to prolonged whipping periods leads to excessive coagulation of globular proteins at the airwater interface, which is usually associated with the formation of insoluble aggregates that exhibit little water-holding ability. The impaired water-holding ability of proteins at the interface is in turn responsible for the foam collapse, which is reected by the overrun decline. To explain the eects of the cosolutes on foaming, we can speculate that NaCl counter ions may screen the charged protein molecules, reduce the electrostatic repulsion between adsorbed and non-adsorbed protein molecules and thus facilitate adsorption at the airwater interface. Davis, Foegeding, and Hansen (2004) reported similar ndings with respect to the eect of NaCl on the adsorption of whey protein isolate at the airwater interface. On the contrary, sucrose delays foam formation and decreases the foaming power of egg white samples, especially in the rst part of the beating period (Lomakina & kova , 2006). It has been documented that the presence M of sugar can aect the thermodynamic and functional properties of food proteins, especially the adsorption and aggregation behaviour (Dickinson & Matia-Merino, 2002). With increasing sucrose concentration the amount of air incorporated decreases and the overrun values of the samples containing sucrose/NaCl in a 12/0 weight ratio are lower compared to the control. According to Lau and Dickinson (2005), the addition of sugar results in increased continuous phase viscosity, which in turn is disadvantageous for air incorporation and the rapid diusion and unfolding of the protein in the vicinity of the interface. Moreover, Antipova, Semenova, and Belyakova (1999) stated that the adsorption of ovalbumin decreases in the pres-

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ence of sucrose possibly because ovalbumin forms hydrogen bonds with the sugar molecules, which results in increased hydrophilicity and decreased surface activity. Thereby the ovalbumin molecules which participate in hydrogen bond formation with sucrose preferentially remain in bulk rather than adsorb to the interface. We should note that egg white is not a pure ovalbumin suspension; rather it is a complex mixture of many proteins, the most abundant of which is ovalbumin. The functional properties of most of these proteins have not been studied in the presence of sugar and salt. The eects of sugars on protein functional properties is, however, a general property which we would expect to apply to all of the component proteins in the egg white, and thus a comparison with ovalbumin is pertinent. The eect of heat treatment in the presence of sucrose and NaCl was, in most of the cases, benecial for the foaming capacity of the egg white samples. This eect is more evident for the samples that contain high sucrose content. The % overrun of the samples containing sucrose/NaCl in a 12/0 weight ratio clearly improves when heated at 64 C. The negative eect imposed by the high sucrose concentration is less severe when the samples are subjected to heat treatment at elevated temperatures. Hagolle, Launay, and Relkin (1998) stated that preheating treatment increased the ability of ovalbumin to adsorb to the air water interface and thereby decrease the surface tension, provided that the formation of large aggregates was prevented. This enhanced foaming ability induced by heat treatment was attributed to higher surface hydrophobicity and increased chain exibility (Kato, Komatsu, Fujimoto, & Kobayashi, 1985). Furthermore, Relkin, Hagolle, Dalgleish, and Launay (1999) suggested that mild heat treatments of ovalbumin solutions may result in molecular species with partially-denatured structures which demonstrate enhanced foaming properties. The extent of denaturation of the native structures of protein molecules under controlled heating conditions could explain the enhanced adsorption capacity of egg white proteins at 64 C. It has been established that globular proteins, under certain conditions, may exist in stable conformations which are intermediate between the native state and the highly disordered state (Kuwajima, Hiraoka, Ikeguchi, & Sugai, 1985). This congurational state, also known as the molten globule state, exhibits a native-like secondary structure and a disordered tertiary structure (Ptitsyn, 1992). Globular proteins when adsorbed at the airwater interface unfold and undergo various structural rearrangements in order to decrease the surface tension. Thus, it can be speculated that the congurational state of adsorbed protein molecules resembles the molten globule state. Nevertheless, the available time for the compact globular proteins to unfold properly at the interface is rather limited because they are rapidly surrounded by other protein molecules, producing a close-packed structure in which there is little scope for further congurational adjustments (Matsumura, Mitsui, Dickinson, & Mori, 1994). Therefore, one may speculate

that the ability of proteins to unfold rapidly at the interface and obtain a molecular conformation which is thermodynamically favourable can be enhanced if they exist in the molten globule state prior to adsorption. The latter may be achieved by various means including heating and it has been stated that the optimum conditions for converting ovalbumin in the monomeric molten globule state is by heating at neutral pH (Matsumura et al., 1994). Although the results obtained on single-protein systems are not easily extrapolable to complex systems (Lechevalier et al., 2005), we could hypothesise that the enhanced adsorption capacity of the egg white proteins following heat treatment observed in this study may be attributed to the conversion of the protein structures from the native state into the molten globule state. Additional evidence for this hypothesis may come from the fact that, as shown in Fig. 5, the concentration of free sulfhydryl groups increases when egg white samples containing various concentrations of glucose and NaCl are heated at temperatures above 60 C (2 min). The concentration of SH groups is not aected to any great extent by the heat treatment when the samples are thermally processed at temperatures as high as 60 C (Fig. 5). Nevertheless, at 62 C and above the reactive SH group concentration increases rapidly. At 62 C, the denaturation of egg white proteins is initiated in the presence of sugar and salt, which results in the exposure of SH groups that remained hidden in the interior core of the protein molecule. Again, the samples containing NaCl appear to be more resistant to heat denaturation compared to the samples heated in the presence of sugar. The increased concentration of free SH groups following heat treatment indicates protein conformational changes associated with transformation from the native state to the denatured state. SH groups which were previously buried in the interior of the protein molecules are now exposed and free to interact with other neighbouring molecules through free thiol/disulde interchange reactions. According to Doi, Kitabatake, Hatta, and Koseki (1989) the signicance of disulde bond formation between denatured ovalbumin molecules at the airwater interface with respect to the foaming properties is limited. However, the

0.0002 0.00018 0.00016 0.00014 0.00012 0.0001 0.00008 0.00006 0.00004 0.00002 0 25 58 60 62 64 Temperature (C)

-SH conc. ( M)

10% glucose 7.5% glucose + 2.5% salt 5% glucose + 5% salt

Fig. 5. Eect of temperature and sugar and salt content on the concentration of free SH groups of egg white. Data are the mean of three replicates.

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increased hydrophobicity and surface activity as indicated by the increase in the SH group concentration following heat treatment at temperatures above 60 C may be associated with molecular conformations that enable the ease of protein unfolding at the interface, thereby exhibiting enhanced adsorption capacity and eciently decreasing the surface tension. Figs. 6 and 7 illustrate the eect of sucrose and NaCl content, whipping time and heating temperature on the stability of the aerated systems. When the samples are subjected to whipping for 10 min, the sample containing sucrose/NaCl in a 0/12 weight ratio demonstrates clearly the highest stability compared to all the other samples. Although there is no evidence of direct correlation between foaming ability and stability, it may be hypothesised that the remarkable dierence in foam stability exhibited between the sample with high NaCl content and the rest of the samples derives from their relative ability to rapidly adsorb at the airwater interface. As described above the samples containing high NaCl concentration are able to rapidly adsorb at the airwater interface and lower the surface tension even when the whipping period is limited to 10 min. Conversely, the samples containing sucrose and the control exhibit a lag phase with respect to protein diffusion and adsorption, which possibly means that after a period of whipping time of 10 min, the protein concentration at the interface is rather limited leading to low levels
100 90 80 70 60 50 40 30 20 10 0 10 13 Whipping time (min) 15

Sugar/Salt=12/0 Sugar/Salt=0/12 Sugar/Salt=6/6 Control

Fig. 6. Eect of sugar and salt content and whipping time on half-life of egg white aerated systems heated at 60 C (2 min). Data are the mean of three replicates.

160 140 50% Drainage (min) 120 100 80 60 40 20 0 10 13 Whipping time (min) 15 Sugar/Salt=12/0 Sugar/Salt=0/12 Sugar/Salt=6/6 Control

Fig. 7. Eect of sugar and salt content and whipping time on half-life of egg white aerated systems heated at 64 C (2 min). Data are the mean of three replicates.

of stability compared to the samples with high NaCl content. This dierence in the foam stability between the sample containing high NaCl concentration and the rest of the samples minimises under experimental conditions (e.g. increasing heating temperature) which enhance protein adsorption and result in higher overrun values. Furthermore, when the samples are subjected to whipping for longer periods of time (13 min and 15 min), the situation with respect to foam stability changes. The samples containing high sucrose content are far more stable compared to all other samples. This increased foam stability demonstrated by the samples with high levels of sucrose may be attributed to the eect of sugars on the viscosity of the bulk phase. Sugars contribute to foam stability by increasing the viscosity of lamella water and thereby retarding drainage (Lau & Dickinson, 2005). Increasing the heating temperature of the egg white dispersions increases the stability of the aerated systems. The only exception was the sample containing high NaCl levels when whipped for 10 min. It has been stated that mild heat treatments of protein solutions, under certain conditions (pH, ionic strength) are correlated with enhanced foaming properties. This may be attributed to the unfolding of protein structure, which may lead to the formation of soluble aggregates through inter-molecular linking. According to Vardhanabhuti and Foegeding (1999), whey protein polymers resulting from mild heat treatments under controlled conditions have higher intrinsic viscosity than the native globular protein. Furthermore, Davis and Foegeding (2004) stated that heat-induced whey protein polymers in solution decreased the air phase volume but made the foams more stable compared to native whey protein suspensions. The increase in stability was directly associated with an increase in bulk viscosity, thereby retarding the rate of drainage. These results are in agreement with the ndings in the present study. As suggested by the determination of the free SH concentration of the protein molecules, heating at 64 C (2 min) in the presence of sugar and NaCl induces the unfolding to some extent of the protein structure. These denatured protein molecules may interact inter-molecularly to form soluble proteinprotein aggregates, which remain in bulk increasing the solution viscosity and contributing to the stability of the foam microstructure. Finally, longer whipping times have a negative impact on foam stability for the samples containing NaCl whereas the samples containing sucrose and the control appear to be fairly unaected. It has been stated that prolonged whipping times are associated with liquid lm thinning, mechanical deformation and bubble-wall rupture (Gassmann, Kroll, & Cifuentes, 1987). All these mechanisms, singly or in combination, may contribute to foam collapse and cause instability. The microstructures of the newly-formed foams as revealed by confocal microscopy (Figs. 811) complement the overrun and foam stability measurements and provide further experimental evidence on the eect of sucrose and

50% Drainage (min)

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Fig. 8. Confocal microscopy of aerated egg white systems containing sugar/salt in a 12/0 ratio (w/w) during heating at 64 C (2 min) and whipped for 10 min and 13 min. Evolution of microstructure was monitored by scanning at 30 min intervals. The scale bar is 300 lm.

Fig. 9. Confocal microscopy of aerated egg white systems containing sugar/salt in a 0/12 ratio (w/w) during heating at 64 C (2 min) and whipped for 10 min and 13 min. Evolution of microstructure was monitored by scanning at 30 min intervals. The scale bar is 300 lm.

NaCl on the adsorption capacity of egg white proteins. The evolution of the microstructure of control samples (plain egg white, heated at 58 C, 2 min) and samples heated for 2 min at 64 C and whipped for 10 min and 13 min are illustrated in Figs. 811. The sample containing sucrose/ NaCl in a 12/0 ratio (w/w) and the control exhibit reduced foaming capacity, especially when the whipping time was limited to 10 min. This conclusion can be drawn from the number and size of air bubbles immediately after foam formation (0 min). When the whipping time increases to

13 min, the adsorption of egg white proteins at the air water interface improves, resulting in enhanced foaming power. These ndings are consistent with % overruns of the specic samples under the given conditions (Fig. 4). On the other hand, the microstructures of the aerated samples containing NaCl during the heating process suggest enhanced adsorption at the airwater interface after a whipping period of 10 min. This is revealed by the density and the size of the air bubbles of the microscopic pictures taken immediately after foam formation (Figs. 9 and 10).

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Fig. 10. Confocal microscopy of aerated egg white systems containing sugar/salt in a 6/6 ratio (w/w) during heating at 64 C (2 min) and whipped for 10 min and 13 min. Evolution of microstructure was monitored by scanning at 30 min intervals. The scale bar is 300 lm.

Fig. 11. Confocal microscopy of aerated plain egg white (control) systems heated at 58 C (2 min) and whipped for 10 min and 13 min. Evolution of microstructure was monitored by scanning at 30 min intervals. The scale bar is 300 lm.

With respect to foam stability, the microscopic observation of the aerated systems indicates that the samples containing sucrose show a lower rate of increase of the bubble size during the standing period of 60 min compared to the samples containing only NaCl and the control. 4. Conclusions All the factors involved in this study (amount of sucrose and NaCl present during the heating process, heating tem-

perature, whipping time) have a signicant impact on the foaming characteristics of egg white protein dispersions. Increasing NaCl concentration, heating temperature and whipping time enhances foam formation, whereas increasing the amount of sucrose confers foam stability for prolonged periods of whipping. The degree of heat-induced denaturation of protein molecules with respect to their molecular conguration in bulk may be crucial for protein adsorption and rearrangement at the airwater interface. The increase in the viscosity of the medium due to the pres-

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ence of sucrose and the controlled formation of protein polymers induced by mild heat treatments, are correlated with enhanced stability. Studies on the heat-induced changes of the secondary structure of egg white proteins would be benecial to elucidate the structurefunction relationship regarding the foaming properties of egg albumen. Further investigations on the interfacial lm composition are required to verify the ndings of the present study. Acknowledgement The authors would like to thank Dr. Hershell Ball, MICHAEL FOODS INC, Minnesota, USA for funding this work and his permission to publish is gratefully acknowledged. References
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