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Journal of Food Engineering 86 (2008) 215224 www.elsevier.com/locate/jfoodeng

Optimization of edible whey protein lms containing preservatives for water vapor permeability, water solubility and sensory characteristics
M. Ozdemir a,*, John D. Floros b
a

Department of Chemical Engineering, Section of Food Technology, Gebze Institute of Technology, P.O. Box 141, 41400 Gebze, Kocaeli, Turkey b Department of Food Science, Pennsylvania State University, 111 Borland Laboratory, University Park, PA 16802, USA Received 8 August 2007; received in revised form 20 September 2007; accepted 22 September 2007 Available online 1 October 2007

Abstract The eect of protein, sorbitol, beeswax and potassium sorbate concentrations in whey protein lms on their water vapor permeability, water solubility and organoleptic properties was investigated using mixture response surface methods. All factors including protein, sorbitol, beeswax and potassium sorbate inuenced water vapor permeability and water solubility of the lms. Beeswax was the most important factor inuencing the stickiness and appearance of the lms. Amount of protein (5065%, w/w) had no eect on stickiness and appearance, while the amount of sorbitol (3550%, w/w) in the lms had no inuence on appearance. Mixture proportions of protein = 0.53, sorbitol = 0.38, beeswax = 0.08 and potassium sorbate = 0.01 would yield an edible lm with minimum stickiness, water vapor permeability 6 9 g mm m2 h1 kPa1, water solubility P 39% and appearance score P 80. 2007 Elsevier Ltd. All rights reserved.
Keywords: Edible lms; Whey protein; Potassium sorbate; Response surface; Water vapor permeability; Solubility; Sensory

1. Introduction In recent years, a great deal of research has been dedicated to develop active packages through the use of edible lms and coatings. Edible lms and coatings are usually used to control moisture transfer, limit gas transport, retard oil and fat migration, prevent solute or avor absorption, carry food additives such as antimicrobial agents and antioxidants, and improve structural integrity of foods. The properties of edible lms and coatings composed of hydrocolloids, lipids and proteins have been comprehensively reviewed (Baldwin, Nisperos-Carriedo, & Baker, 1995; Debeaufort, Quezada-Gallo, & Voilley, 1998; Krochta & De Mulder-Johnston, 1997; Miller & Krochta, 1997). Functional properties and potential applications of edible lms and coatings made of milk proteins
*

Corresponding author. Tel.: +90 262 605 3290; fax: +90 262 653 8490. E-mail address: ozdemirm@gyte.edu.tr (M. Ozdemir).

are well known (Chen, 1995; McHugh & Krochta, 1994a; Rhim & Ng, 2007). Whey proteins have exceptional nutritional value and functional properties (Human, 1996; Kinsella, 1984). In addition, liquid whey is produced in large quantities, and its annual production increases continuously (Banerjee & Chen, 1995). The formation of edible lms and coatings from whey proteins can increase the utilization of whey, improve the nutritional value of foods and prolong shelf life. Ozdemir and Floros (2001) produced antimicrobial lms from commercial whey protein isolate and investigated the release mechanism of potassium sorbate from the lms. Ozdemir and Floros (2003) also studied the eect of lm composition on potassium sorbate diusion in whey protein lms using mixture response surface methodology. Increasing the relative amounts of protein and beeswax in the lms decreased potassium sorbate diusivity, while increasing the relative amounts of plasticizer and initial

0260-8774/$ - see front matter 2007 Elsevier Ltd. All rights reserved. doi:10.1016/j.jfoodeng.2007.09.028

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potassium sorbate in the lms increased the diusion of potassium sorbate. McHugh, Aujard, and Krochta (1994) determined the eects of plasticizers, pH and relative humidity on their water vapor permeability. Plasticizers increased water vapor permeability, and this eect was more pronounced as the plasticizer concentration in whey protein lms increased. At constant temperature and relative humidity, high water vapor permeability values were obtained at low pH (<6.0) values. In addition, water vapor permeability values of lms rose exponentially with the relative humidity. Films exhibited moderate barrier properties to water vapor, but they were very good barriers to oxygen (McHugh & Krochta, 1994b). McHugh and Krochta (1994c) developed whey protein lipid emulsion lms by incorporating some lipids into the lm matrix to reduce water vapor permeability. Water vapor barrier properties of lms were signicantly improved by increasing the amount of lipid in the lm. Whey protein emulsion lms containing beeswax had the lowest water vapor permeability among other lipid containing lms. In a similar study carried out by Shellhammer and Krochta (1997), whey proteinbeeswax emulsion lms were found to exhibit signicantly lower water vapor permeabilities than whey protein emulsion lms containing candelilla and carnauba waxes. Control of surface moisture content can signicantly reduce the growth of microorganisms and the rate of deteriorative reactions, thereby increasing the storage stability of foods (Kester & Fennema, 1986). In this respect, the use of edible lms and coatings to suppress microbial growth and limit the rate of deteriorative reactions is of signicant safety and economic value. Edible lms and coatings with good water and/or oxygen barrier properties are usually not adequate by themselves to retard microbial growth. Therefore, the incorporation of antimicrobial agents into edible coating formulations is needed to obtain stronger inhibitory eect against microbial growth. Potassium sorbate and sorbic acid have been known and utilized for years as antimicrobial substances to preserve foods and retard the growth of molds, yeast and bacteria during storage, transportation and handling. Potassium sorbate is eective in most foods at a concentration range of 0.050.3% by weight (Robach, 1979a, 1979b; Robach & Sofos, 1982; Sofos, Busta, & Allen, 1980). Furthermore, both potassium sorbate and sorbic acid are considered as GRAS (Generally Recognized as Safe) substances by the U.S. Food and Drug Administration (FDA) and used commercially as antimicrobial agents. Ozdemir and Floros (2008) developed antimicrobial lms containing potassium sorbate and found that mixture proportions of protein = 0.58, sorbitol = 0.38, beeswax = 0 and potassium sorbate = 0.04 would be recommended to obtain mechanically good and transparent lms. Resistance of edible lms to water is a desired property for lms used as protective layers on intermediate and high moisture foods (Gontard, Guilbert, & Cuq, 1992). Furthermore, edible lms containing antimicrobial agents are also

desired to have low water solubility to avoid the complete dissolution of the lm. The evaluation of sensory properties of edible lms is of important value showing their level of acceptance by consumers. Milk protein-based lms are generally avorless and tasteless (Chen, 1995). However, the incorporation of lipid materials into edible lm formulations to improve their water vapor barrier properties and the addition of additives, such as antimicrobial agents, into lms could adversely aect the sensory characteristics of lms. The objectives of this research were to: (a) investigate the eect of dierent constituents (amounts of whey protein, plasticizer, lipid and antimicrobial agent) of activeedible whey protein lms on their water vapor barrier properties, water solubility and sensory characteristics; and (b) determine the optimal combination of these constituents that provide good water vapor permeability, water resistance and sensory properties using mixture response surface methodology (RSM). 2. Materials and methods 2.1. Experimental design The D-optimal mixture design (Cornell, 1990; Myers & Montgomery, 1995) was used to nd optimum combination of constituents that provide good water vapor permeability, water solubility and sensory characteristics for whey protein lms containing potassium sorbate. Mixture experiment is a special type of response surface experiments in which factors are the proportions of components in a mixture and their levels are not independent of each other. Therefore, the selection of design points was based on the basic constraint for the mixture experimental design:
n X i1

xi x1 x2 xn 1:0

where xi is the proportion of ith component in the mixture and n is the number of components in the mixture. Mixture factors were identied, based on previous experimental studies on whey protein lms (Banerjee & Chen, 1995; Ozdemir, 1999; Shellhammer & Krochta, 1997). The mixture factors were the proportion of whey protein isolate (x1), the proportion of sorbitol (x2), the proportion of beeswax (x3) and the proportion of potassium sorbate (x4) in the mixture. Measured responses were water vapor permeability (WVP), water solubility (WS) and sensory properties of the lms including stickiness (ST) and appearance (AP). The minimum and maximum levels of each mixture component were determined based on preliminary studies (Ozdemir, 1999; Ozdemir & Floros, 2001, 2003). The exact mathematical representation of the functions describing the relationship between mixture factors and responses is either unknown or extremely complex.

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However, either rst or second order models can be used to approximate the true functions: Y n bn0 Y n bn0
4 X i1 4 X i1

bni xi bni xi
4 X i1

2 bnii x2 i
3 4 X X i1 ji1

bnij xi xj

where Yn is the expected response, bn0 is the value of the response at the standard reference mixture according to Cox reference model (Cox, 1971), and bni, bnii and bnij are constant coecients. 2.2. Statistical analysis Cox reference model was tted by multiple regression analysis using Modde software (version 4.0, Umetri AB, Umea, Sweden) to determine the coecients (bs). Model diagnostics were done by generating normal probability plot, and the plots of residual versus predicted values and observed versus predicted values. Trace and contour plots for responses were created with Design-Expert software (version 5.0, Stat-Ease Inc., Minneapolis, MN). The trace plot provides silhouette views of the response surface. In other words, the trace plot shows the eects of changing each component from the reference blend to the vertex. Contour plots are used to see graphically how the responses change with respect to the factors and they show graphical representation of the response surface. 2.3. Materials BiPRO whey protein isolate (WPI) was supplied by Davisco Foods International (Le Sueur, MN). WPI was lactose free with 99.5% protein. Sorbitol and potassium sorbate were obtained from Sigma Chemical (St. Louis, MO). Beeswax (bleached and white) was purchased from Aldrich Chemical Company (Milwaukee, WI). 2.4. Film formation and lm casting Films were formed with the method described by Ozdemir and Floros (2001, 2003). Aqueous solutions of 10% (w/w) WPI were prepared by stirring dry WPI powder in distilled water until the WPI was completely dissolved. Solutions were placed in a water bath (model MW-1120 A-1, Blue M Electric, Blue Island, IL) at 90 C and kept there for 30 min to denature the protein. Solutions were then cooled to room temperature, and various amounts of sorbitol (3550%, w/w) were added to plasticize the lms. For the manufacturing of lipid-containing lms, WPI/ sorbitol solutions were reheated to 85 C, and appropriate amounts of beeswax (015%, w/w) were melted in these hot WPI/sorbitol solutions. The mixture was thoroughly homogenized in a homogenizer (type PT10/35, Brinkmann

Instruments Company, Westbury, NY) equipped with a probe (model PTA 35/2, Brinkmann Instruments Company, Westbury, NY) at 15,000 rpm for 2.5 min. After homogenization, solutions were rapidly cooled to room temperature by placing them in an ice bath to prevent further denaturation of protein (Shellhammer & Krochta, 1997). Various amounts (010%, w/w) of potassium sorbate were nally added and mixed to form antimicrobial lms. All solutions were degassed using a vacuum pump (General Electric, Fort Wayne, IN) at room temperature to remove air bubbles in solutions. Films were casted by pipetting 10 mL of solution on rimmed and smooth low density polyethylene (LDPE) casting plates (Texaco Inc., White Plains, NY) with inner diameters of 10.5 cm. Solutions were spread evenly with a bent L-shaped glass rod, and they were allowed to dry for 24 h at 50 5% relative humidity (RH) and 25 2 C in a controlled temperature and humidity room. After 24 h, dried lms were peeled intact from the casting plates. 2.5. Film conditioning All lms were conditioned in a controlled temperature and humidity storage room at 50 5% RH and 25 2 C for 48 h prior to tests. 2.6. Water vapor permeability Water vapor permeability (WVP) of the lms was measured gravimetrically according to the WVP correction method as described by McHugh, Avena-Bustillos, and Krochta (1993), a modication of ASTM E96 (ASTM, 1997). This modication is needed since ASTM E96 method is designed to determine WVP of synthetic hydrophobic lms. A plastic desiccator cabinet equipped with a fan capable of operating at variable speeds (Tuthill Pump Corp., Concord, CA) was placed into a controlled temperature and humidity room at 50 2% RH and 25 0.5 C. A digital hygrometer and thermometer (Fisher Scientic, Inc., Fair Lawn, NJ) was also placed inside the cabinet to monitor the relative humidity and temperature more precisely. Air speeds inside the cabinet were measured using an anemometer (Davis Instruments, Hayward, CA). The fan speed was set to 182 m min1 to prevent stagnant air layer formation outside test cups and to achieve uniform relative humidity throughout the cabinet. Circular cups with internal diameter of 10.5 cm and depth of 2.1 cm were used to test lm WVP. Cup walls are suciently thick (0.55 0.03 mm) to prevent water vapor transfer through the walls. Three replicates of each lm were tested. Films free of any defects such as pinholes, air bubbles and cracks were used for WVP determinations. The thicknesses of three replicates of each lm formulation were measured with an electronic digital micrometer (MaxCal Inc., Japan) at six random locations on the lm to the

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nearest 0.001 mm. Average lm thickness was 0.181 0.028 mm. Distilled water (20 mL) was placed in the bottoms of the test cups. Test lms cut circular with a razor knife were sealed on the cups using a waterproof adhesive. The lipid side of the lms faced the high relative humidity environment (toward the inner of the test cup) and their protein rich sides faced the room atmosphere (McHugh & Krochta, 1994c). When a thin lm is coated on a food surface, it is desired that the lipid side of the lm has to be in contact with high relative humidity environment so that the coating goes to a minimal interaction with water and thus, the food is protected better from the negative eects of water. A thin coating of high vacuum silicone sealant (Beckman Instruments, Inc., Palo Alto, CA) was applied around the cup circumference to prevent water vapor transfer through the sealant area. Average stagnant air gaps between the water level in the cup and the inner lm surface was measured both before and after each experiment (McHugh & Krochta, 1994c). The mean air gap between the water level in the cup and the inner lm surface after the test was 10 2 mm. After the test began, steady state was attained in almost 2 h. Cups were then periodically weighed for 24 h. The weight loss of the test cups was measured to the nearest 0.0001 g. Water vapor transmission rates (WVTR) of lms at steady state were determined from the slope of weight loss versus time plots using Eq. (4). Linear regression coecients (R2) were greater than 0.99 for all plots. The area of lm exposed in the test cup was 86.5 cm2. WVTR Slope Film area 4

For each experiment, relative humidity values under the lm surface were corrected by using the procedure described by McHugh et al. (1993). 2.7. Film solubility (total soluble matter) A method modied from Stuchell and Krochta (1994) was used to determine the solubility of lms in water. Films were cut into 20 mm 20 mm pieces and dried at 70 C and 96 kPa in a vacuum oven for 24 h. After drying, lms were weighed to the nearest 0.0001 g for the determination of the initial dry weights of lms. Films were individually placed into 20 mL of distilled water in 50 mL screw-top centrifuge tubes. The tubes were capped and placed in a shaking water bath at 25 0.1 C for 24 h. Film pieces were then taken out and dried at 70 C and 96 kPa in a vacuum oven for 24 h to determine the nal dry weights of lms. Three replicates of each lm were done. Percent total soluble matter was calculated from the initial and nal dry weights of lms and reported on dry weight basis. 2.8. Sensory analysis A sensory panel consisting of 12 members was carried out. All panel members were trained by the open discussion method prior to sensory tests. Stickiness and appearance of the lm samples were selected as sensory attributes to be included in the sensory testing. Stickiness and appearance are two critical sensory attributes providing crucial information on the applicability of edible lms and coatings on food surfaces as protective layers. Descriptive analysis (Stone & Sidel, 1993) was used to evaluate selected sensory attributes of the lms. Sensory scores were recorded based on line marking scales where each panelist placed a mark on a 150 mm line to indicate the intensity of each attribute. The endpoints were labeled as not sticky/extremely sticky and dislike extremely/like extremely for stickiness and appearance, respectively. The numerical score for each attribute was determined by measuring the distance in mm with a ruler from the left-hand side of the 150 mm line scale to the point that each panelist placed a mark that best described the attribute. All samples were randomly selected and presented to judges. Panelists were requested to chew the lms in their mouths and evaluate if the lms are sticky. Cups with water were provided for panelists to use during the test to minimize the residual stickiness eect of the lms in the mouth. Appearance was evaluated by visual observation. Sensory analysis was conducted under controlled laboratory conditions in individually partitioned booths. 3. Results and discussion 3.1. Model development Experimental points in terms of actual values and experimental data are shown in Table 1. The comparison of the

The corrected water vapor partial pressure at the lm inner surface (p2) for hydrophilic lms can be calculated from the following equation (McHugh et al., 1993): p2 pT pT p1 expWVTR R T Dz=pT D 5

where pT is the total pressure (kPa), p1 is the water vapor partial pressure at the solution surface (kPa), R is the ideal gas law constant (8.314 kJ kg mol1 K1), T is the absolute temperature (K), D is the diusivity of water through air at 25 C (m2 h1) and Dz is the mean stagnant air gap (m) inside the cup. Permeance is equal to the WVTR divided by the water vapor partial pressure gradient across the lm, as given by Eq. (6): Permeance WVTR p 2 p 3 6

where p3 is the water vapor partial pressure at the lm outer surface (kPa). The corrected water vapor permeability is calculated by multiplying the permeance corrected by Eq. (6) and average lm thickness, as shown in Eq. (7): Permeability Permeance Thickness 7

M. Ozdemir, J.D. Floros / Journal of Food Engineering 86 (2008) 215224 Table 1 Experimental design and experimental data Sample numbera 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20
a

219

Whey protein isolate (%, w/w) 50 65 50 50 55 50 55 60 50 58.3 50 50 60 55 50 50 53.3 53.3 53.3 53.3

Sorbitol (%, w/w) 50 35 35 40 35 35 45 40 43.3 35 45 40 35 35 35 35 38.4 38.4 38.4 38.4

Beeswax (%, w/w) 0 0 15 0 0 5 0 0 0 0 5 10 5 10 11.7 8.3 3.3 3.3 3.3 3.3

Potassium sorbate (%, w/w) 0 0 0 10 10 10 0 0 6.7 6.7 0 0 0 0 3.3 6.7 5 5 5 5

Water vapor permeability (g mm m2 h1 kPa1) 9.64 8.53 5.38 10.17 9.49 9.97 9.00 8.71 10.91 10.31 8.51 7.11 7.63 6.52 8.42 9.72 10.46 10.51 10.02 10.35

Water solubility (% dry matter) 48.24 37.72 36.98 50.85 45.18 45.34 43.90 42.67 50.77 42.32 43.47 41.85 37.33 37.56 39.56 41.56 44.38 43.20 44.75 44.63

Stickiness score 136 90 29 148 120 21 138 97 142 89 28 9 5 3 7 16 12 16 24 28

Appearance score 137 146 68 118 131 44 129 133 120 127 88 75 104 77 32 50 118 111 130 125

Experimental runs were performed in random order.

linear and quadratic models developed for WS and AP (Table 2) revealed that only linear models were signicant (P 6 0.01). These models did not show any signicant lack

of t (P 6 0.05). Although the linear models developed for WVP and ST were signicant at P 6 0.01, they exhibited signicant lack of t (P 6 0.05). The quadratic models

Table 2 Analysis of variance and regression coecients of the rst and second order models for four responses Source Sum of squares Linear models df Model Residual Lack of t Pure error Corrected total R2 Regression coecienta bn0 bn1 bn2 bn3 bn4 bn11 bn22 bn33 bn44 bn12 bn13 bn14 bn23 bn24 bn34
a

Quadratic models AP (n = 4) 17367*** 4832 4626 206 22199 0.78 105.1*** 77.2 18.3 649.9*** 249.7** df 9 10 7 3 19 WVP (g mm m2 h1 kPa1) (n = 1) 41.34*** 0.25 0.11 0.14 41.59 0.99 10.3*** 4.7*** 5.3*** 9.4*** 15.9*** 7.0 9.6 65.4** 433.0*** 20.5 25.1 25.2 18.8 59.5** 210.9*** ST (n = 3) 53947*** 2689 2529 160 56636 0.95 30.7 141.7 293.7** 1533.7*** 281.4* 228.0 287.4 13704.7*** 7177.8* 364.9 64.3 2108.6 2417.4 1098.2 425.1

WS (% dry matter) (n = 2) 295.50*** 14.29 12.77 1.52 309.79 0.95 44.1*** 32.1*** 41.5*** 38.1*** 50.4***

3 16 13 3 19

These are coecients of Eq. (2) for WS and AP, and Eq. (3) for WVP and ST, respectively. bn0 is the value of the response at the standard reference mixture and the numerals 1, 2, 3 and 4 refer to whey protein isolate, sorbitol, beeswax and potassium sorbate, respectively. * Signicant at 10% level. ** Signicant at 5% level. *** Signicant at 1% level.

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for WVP and ST, on the other hand, were signicant (P 6 0.01) without any signicant lack of t (P 6 0.05). Based on these results, linear models were chosen for WS and AP, while quadratic models were selected for WVP and ST. The analysis of variance (ANOVA) and the regression coecients (bs) for the models developed for four responses are shown in Table 2. The analysis of variance for four response variables showed that all models developed were statistically signicant (P 6 0.01) with no signicant lack of t, which suggested that they adequately described the true functions. 3.2. Water vapor permeability A trace plot using Coxs direction (Cox, 1971) was generated based on the model chosen for WVP to investigate the eect of each mixture component on WVP (Fig. 1). The trace plot shows how the response changes when changing the concentration of each mixture component while keeping all others in a constant ratio. This is necessary because when the amount of one mixture component increases, the amounts of all others decrease, but their ratio to one another remains constant. The center of the trace plot, showing the reference mixture, was the centroid of the design. The composition of the reference mixture in terms of actual units was protein = 0.533, sorbitol = 0.384, beeswax = 0.033 and potassium sorbate = 0.050. The trace plot for WVP (Fig. 1) revealed that WVP of the lms were inuenced by all the mixture components. The addition of protein and beeswax decreased WVP,

while the increase in sorbitol increased WVP. Potassium sorbate was the most eective factor that adversely aected WVP. The curvilinear eect of potassium sorbate on WVP implied that quadratic terms for potassium sorbate and the interactions between potassium sorbate and some mixture components could be signicant. As shown in Table 2, not only was the second order term for potassium sorbate signicant, but also the interaction terms between sorbitol and potassium sorbate, and beeswax and potassium sorbate were signicant. 3.3. Water solubility Resistance of edible-antimicrobial lms to water is desirable if the lm is to be used for the preservation of intermediate or high moisture foods. An antimicrobial lm with poor water resistance will dissolve quickly, causing the lm to lose its antimicrobial agent. This will increase the diusion of the preservative from the surface to the bulk of the food, resulting in low preservative concentrations at the food surface. Whey protein lms with potassium sorbate did not dissolve or break apart after they were immersed into water and dried for 24 h. This was an indicative of highly stable protein-polymer network. The inuence of protein, sorbitol, beeswax and potassium sorbate on WS is shown in Fig. 2. Trace plots of each component on WS showed that all mixture components aected WS. Protein and beeswax had negative impacts on WS, while sorbitol and potassium sorbate positively aected WS. This means that the

Fig. 1. Trace plot showing the eects of each mixture component on water vapor permeability (A = protein, B = sorbitol, C = beeswax and D = potassium sorbate).

Fig. 2. Trace plot showing the eects of each mixture component on water solubility (A = protein, B = sorbitol, C = beeswax and D = potassium sorbate).

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addition of more protein and beeswax to the lm formulation produce lms with high water resistance. On the other hand, increasing the amounts of sorbitol and potassium sorbate in the lms reduces the resistance of lms to water. 3.4. Sensory Fig. 3 showed that ST was strongly inuenced by beeswax. However, potassium sorbate and sorbitol also aected ST, but in a lesser extent than beeswax. The addition of potassium sorbate below the reference mixture reduced ST, but further addition increased ST beyond the reference mixture. Increasing the amount of sorbitol in the lms increased ST. Protein seemed to be not an important factor inuencing ST. The presence of curvature in beeswax and potassium sorbate implied that ST could be aected by the second order terms of these components. This behavior was conrmed by the existence of signicant interactions among quadratic terms for beeswax and potassium sorbate in the model developed for ST as shown in Table 2. The eect of protein, sorbitol, beeswax and potassium sorbate on the AP of the lms is shown in Fig. 4. The slope of the trace plot for beeswax was steeper than any of the components in the mixture, indicating that beeswax was the primary factor inuencing AP. Potassium sorbate negatively aected AP similar to that of beeswax, but in a lesser extent. Protein and sorbitol seemed not to have any eects on AP.

Fig. 4. Trace plot showing the eects of each mixture component on appearance (A = protein, B = sorbitol, C = beeswax and D = potassium sorbate).

3.5. Locating the optimum Predictive models were used to graphically represent the system. Contour plots were generated, by plotting each response to three mixture factors, while the fourth factor was kept constant. Numerous contour plots were created for each response to better understand the responses of the system within the experimental region. For each response, a mixture contour plot was obtained at a specic value of protein concentration because ST and AP were not aected by the amount of protein in the lms. Subsequent mixture contour plots were generated at other values of protein concentrations. The contour plots of four responses obtained for each specic value of protein were then superimposed to nd an optimum region with limiting response values (constraints). These constraints were as follows: (a) the stickiness must be minimum based on the sensory scores; (b) the appearance score of at least 80 must be achieved as determined by the panelists; (c) the water vapor permeability must be as low as possible, the value of WVP = 9 g mm m2 h1 kPa1 was chosen as a maximum reference; and (d) the water solubility of at least 39% was chosen based on the solubility values reported by Gontard et al. (1992) and Jangchud and Chinnan (1999) for wheat gluten and peanut protein lms, respectively. Fig. 5 shows the computer generated mixture contour plots for the water vapor permeability, water solubility, stickiness and appearance of lms when the proportion of protein was at its reference value (protein = 0.533).

Fig. 3. Trace plot showing the eects of each mixture component on stickiness (A = protein, B = sorbitol, C = beeswax and D = potassium sorbate).

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Fig. 5. Computer generated mixture contour plots for (a) water vapor permeability (g mm m2 h1 kPa1); (b) water solubility (% dry matter); (c) stickiness and (d) appearance at 53.3% protein concentration.

The shaded area in Fig. 6 shows the optimum region for all four responses based on the constraints given above. The predicted optimum mixture proportions of protein = 0.53, sorbitol = 0.38, beeswax = 0.08 and potassium sorbate = 0.01 would yield an edible lm with minimum stick-

iness, WVP 6 9 g mm m2 h1 kPa1, water solubility P 39% and appearance score P 80. The formation of edible lms and coatings composed of whey is not only important in nding new uses for whey proteins, but also in improving the microbial stability of

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WVP 6 9 g mm m2 h1 kPa1, water solubility P 39% and appearance score P 80. The addition of protein reduced WVP and WS, but it had no eect on ST and AP. Sorbitol had a positive impact on WVP, WS and ST, and it did not aect AP. All four responses were negatively inuenced by beeswax. Potassium sorbate signicantly increased WVP, WS and ST, but it had a negative impact on AP. The lms could be used particularly for bakery products, cookies, individually wrapped cheeses, nuts, biscuits, chocolate surfaces and as casing for sausages and salamis. Edible lms and coatings carrying antimicrobial agents are of particular interest to both food processors and food scientists because they are formed from natural materials. Edible lms with preservatives not only help provide improved food safety and stability like antimicrobial plastic lms, but also overcome the migration of package components from plastic packaging materials. References
Fig. 6. Optimum region obtained by superimposing contour plots of all four responses generated at 53.3% protein concentration. Shaded area represents optimum region with minimum stickiness, WVP 6 9 g mm m2 h1 kPa1, WS P 39% and AP P 80. Dashed lines show the change in potassium sorbate diusivity (1011 m2 s1). ASTM (1997). Designation E 96-95: Standard test method for water vapor transmission of materials. In Annual book of ASTM standards (pp. 752759). Philadelphia, Pennsylvania: American Society for Testing and Materials. Baldwin, E. A., Nisperos-Carriedo, M. O., & Baker, R. A. (1995). Use of edible coatings to preserve quality of lightly (and slightly) processed products. Critical Reviews in Food Science and Nutrition, 35, 509524. Banerjee, R., & Chen, H. (1995). Functional properties of edible lms using whey protein concentrate. Journal of Dairy Science, 78, 16731683. Chen, H. (1995). Functional properties and applications of edible lms made of milk proteins. Journal of Dairy Science, 78, 25632583. Cornell, J. A. (1990). Experiments with mixtures: designs, models, and analysis of mixture data (2nd ed.). New York: Wiley. Cox, D. R. (1971). A note on polynomial response functions for mixtures. Biometrika, 58, 155159. Debeaufort, F., Quezada-Gallo, J.-A., & Voilley, A. (1998). Edible lms and coatings: tomorrows packagings: a review. Critical Reviews in Food Science and Nutrition, 38, 299313. Gontard, N., Guilbert, S., & Cuq, J.-L. (1992). Edible wheat gluten lms: Inuence of the main process variables on lm properties using response surface methodology. Journal of Food Science, 57, 190195, 199. Human, L. M. (1996). Processing whey protein for use as a food ingredient. Food Technology, 50(2), 4952. Jangchud, A., & Chinnan, M. S. (1999). Peanut protein lm as aected by drying temperature and pH of lm forming solution. Journal of Food Science, 64, 153157. Kester, J. J., & Fennema, O. R. (1986). Edible lms and coatings: a review. Food Technology, 40(12), 4759. Kinsella, J. E. (1984). Milk proteins: physicochemical and functional properties. Critical Reviews in Food Science and Nutrition, 21, 197262. Krochta, J. M., & De Mulder-Johnston, C. (1997). Edible and biodegradable lms: Challenges and opportunities. Food Technology, 51(2), 6174. McHugh, T. H., Avena-Bustillos, R., & Krochta, J. M. (1993). Hydrophilic edible lms: Modied procedure for water vapor permeability and explanation of thickness eects. Journal of Food Science, 58, 899903. McHugh, T. H., Aujard, J.-F., & Krochta, J. M. (1994). Plasticized whey protein edible lms: water vapor permeability properties. Journal of Food Science, 59, 416419, 423. McHugh, T. H., & Krochta, J. M. (1994a). Milk-protein-based edible lms and coatings. Food Technology, 48(1), 97103.

foods particularly in presence of preservatives in lm formulations. Ozdemir (1999) showed that whey protein lms containing potassium sorbate eectively suppressed the growth of yeasts and molds by extending the lag period before the growth became apparent. Potassium sorbate incorporated whey protein lms help provide improved food safety and stability, thereby increasing shelf life. These lms are required to have preservative diusion coefcients smaller than those in food products to slow down the diusion of preservative from the food surface to the bulk of the food. Film composition is of primary importance aecting the diusion of preservatives in edible lm formulations (Ozdemir & Floros, 2003). Ozdemir and Floros (2001) determined potassium sorbate diusion coefcients in whey protein lms when the lms are in contact with an intermediate moisture model food system with a water activity of 0.80. Potassium sorbate diusion coecients in the lms ranged from 5.38 to 9.76 1011 m2 s1. The change in the diusivity of potassium sorbate in the optimum region is shown by the dashed lines (Fig. 6). 4. Conclusions Mixture response surface methodology and graphical optimization methods were eective in better understanding the eect of lm formulation on water vapor permeability, water solubility and organoleptic quality of whey protein lms containing potassium sorbate. The predicted optimum mixture proportions of protein = 0.53, sorbitol = 0.38, beeswax = 0.08 and potassium sorbate = 0.01 would yield an edible lm with minimum stickiness,

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M. Ozdemir, J.D. Floros / Journal of Food Engineering 86 (2008) 215224 Rhim, J.-W., & Ng, P. K. W. (2007). Natural biopolymer-based nanocomposite lms for packaging applications. Critical Reviews in Food Science and Nutrition, 47, 411433. Robach, M. C. (1979a). Inuence of potassium sorbate on growth of Pseudomonas putrefaciens. Journal of Food Protection, 42, 312313. Robach, M. C. (1979b). Extension of shelf-life of fresh, whole broilers, using a potassium sorbate dip. Journal of Food Protection, 42, 855857. Robach, M. C., & Sofos, J. N. (1982). Use of sorbates in meat products, fresh poultry and poultry products. A review. Journal of Food Protection, 45, 374383. Shellhammer, T. H., & Krochta, J. M. (1997). Whey protein emulsion lm performance as aected by lipid type and amount. Journal of Food Science, 62, 390394. Sofos, J. N., Busta, F. F., & Allen, C. E. (1980). Inuence of pH on Clostridium botulinum control by sodium nitrite and sorbic acid in chicken emulsions. Journal of Food Science, 45, 712. Stat-Ease, Inc. (1996). Design-Expert software, version 5.0. Minneapolis, MN: Stat-Ease, Inc.. Stone, H., & Sidel, J. L. (1993). Sensory evaluation practices (2nd ed.). San Diego, California: Academic. Stuchell, Y. M., & Krochta, J. M. (1994). Enzymatic treatments and thermal eects on edible soy protein lms. Journal of Food Science, 59, 13321337. Umetri AB (1997). Modde software, version 4.0. Umea, Sweden: Umetri, AB.

McHugh, T. H., & Krochta, J. M. (1994b). Sorbitol- vs glycerolplasticized whey protein edible lms: integrated oxygen permeability and tensile property evaluation. Journal of Agricultural and Food Chemistry, 42, 841845. McHugh, T. H., & Krochta, J. M. (1994c). Water vapor permeability properties of edible whey proteinlipid emulsion lms. Journal of the American Oil Chemists Society, 71, 307312. Miller, K. S., & Krochta, J. M. (1997). Oxygen and aroma barrier properties of edible lms: a review. Trends in Food Science and Technology, 8, 228237. Myers, R. H., & Montgomery, D. C. (1995). Response surface methodology: process and product optimization using designed experiments. New York: Wiley. Ozdemir, M. (1999). Antimicrobial releasing edible whey protein lms and coatings. Ph.D. dissertation, Purdue University, West Lafayette, Indiana, USA. Ozdemir, M., & Floros, J. D. (2001). Analysis and modeling of potassium sorbate diusion through edible whey protein lms. Journal of Food Engineering, 47, 149155. Ozdemir, M., & Floros, J. D. (2003). Film composition eects on diusion of potassium sorbate through whey protein lms. Journal of Food Science, 68, 511516. Ozdemir, M., & Floros, J. D. (2008). Optimization of edible whey protein lms containing preservatives for mechanical and optical properties. Journal of Food Engineering, 84, 116123.