Food Hydrocolloids 23 (2009) 10621064

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Food Hydrocolloids
journal homepage: www.elsevier.com/locate/foodhyd

Short Communication

The inuence of hydrophobic substances on water vapor permeability of sh

gelatin lms modied with transglutaminase or 1-ethyl-3-
(3-dimethylaminopropyl) carbodiimide (EDC)
Katarzyna Sztuka, Ilona Ko1odziejska*
Department of Food Chemistry, Technology and Biotechnology, Chemical Faculty Gdansk University of Technology, G. Narutowicza 11/12, 80-952 Gdansk, Poland

a r t i c l e i n f o a b s t r a c t

Article history: The effect of different hydrophobic substances on water vapor permeability (WVP) of unmodied sh
Received 15 May 2008 gelatin lms and cross-linked with transglutaminase (TGase) or with 1-ethyl-3-(3-dimethylamino-
Accepted 4 August 2008 propyl) carbodiimide (EDC) was determined. Both unmodied and cross-linked lms were characterized
by very poor water barrier properties. Amaranth oil, rapeseed oil, lanolin, beeswax and ozococerite at
Keywords: concentration of 10% decreased WVP of unmodied gelatin lms by 42, 15, 37, 53 and 36%, respectively.
Fish gelatin
Increasing concentration of these substances up to 60% caused further improving of the water barrier
Hydrophobic substances
properties. Addition of lecithin into lm-forming emulsions prevented separation of lipid layer on the
Transglutaminase (TGase)
1-ethyl-3-(3-dimethylaminopropyl) lm surface. Among unmodied lms with lecithin and 60% of lipids the highest decrease of WVP was
carbodiimide (EDC) found in case of amaranth oil and beeswax, by 73 and 87%, respectively, in comparison to only-gelatin
Water vapor permeability (WVP) lms. WVP of chemically modied lms in the presence of 60% of beeswax with addition of lecithin was
decreased by about 65%. Enzymatically modied lms with beeswax were very brittle and broke during
analyzes, similarly as those with amaranth oil and lecithin. WVP of enzymatically modied lms with
lecithin and 60% of rapeseed oil and lanolin was respectively, about 60 and 47% lower than that of lms
without hydrophobic substances.
2008 Elsevier Ltd. All rights reserved.

1. Introduction However, gelatin lms, similarly as made of other proteins, have

During the last two decades the interest in edible and biode-
gradable packaging materials has increased due to their environ-
mental benets. Such edible materials can be made from natural and
easily accessible polymers like proteins and polysaccharides, very
often obtained from by-products of food industry. Among others
mammal and sh gelatins are used for this purpose (Bertan, Tanada-
Palmu, Siani, & Grosso, 2005; Bigi, Panzavolta, & Rubini, 2004; de
Carvalho et al., 2008; Jongjareonrak, Benjakul, Visessanguan,
Prodpran, & Tanaka, 2006; Jongjareonrak, Benjakul, Visessanguan, &
Tanaka, 2006; Ko1odziejska & Piotrowska, 2007; Perez-Mateos,
Montero, & Gomez-Guillen, 2007; Piotrowska, Sztuka, Ko1odziejska,
& Dobrosielska, in press; Sobral, Menegalli, Hubinger, & Roques,
2001; Vanin, Sobral, Menegalli, Carvalho, & Habitante, 2005).
Films based on natural polymers are generally a good barrier to
oxygen and carbon dioxide as well as to many organic vapors. The
development of edible lms and coatings with selective gas
permeability is potentially of great interest for controlling the
exchange of carbon dioxide and oxygen.
* Corresponding author. Tel.: 48 58 347 2656; fax: 48 58 347 2694.
E-mail address: i.kolodziejska@chem.pg.gda.pl (I. Ko1odziejska).
higher water vapor permeability (WVP) than those from synthetic
materials (Bertan et al., 2005; Bigi et al., 2004; de Carvalho et al.,
2008; Jongjareonrak, Benjakul, Visessanguan, Prodpran, et al.,
2006; Perez-Mateos et al., 2007; Piotrowska et al., in press; Sobral
et al., 2001; Vanin et al., 2005). In order to limit the WVP hydro-
phobic substances, such as fatty acids, lipid alcohols, vegetable oils,
milk fat or waxes have to be incorporated into the polymer network
(Ayranci & Tunc, 1997; Gontard, Duchez, Cuq, & Guilbert, 1994;
Perez-Gago & Krochta, 1999; Sapru & Labuza, 1994; Shellhammer &
Krochta, 1997; Sherwin, Smith, & Fulcher, 1998; Tanaka, Ishizaki,
Suzuki, & Takai, 2001). The WVP of lms containing lipids depends
on the concentration, polarity, character of the chemical groups,
length of the aliphatic chain and degree of fatty acid unsaturation of
the lipids and quality of dispersion of the lipids in the lm matrix
(Ayranci & Tunc, 1997; Hagenmaier & Baker, 1997; Kester &
Fennema, 1989; Koelsch & Labuza, 1992; Sapru & Labuza, 1994;
Shellhammer & Krochta, 1997; Tanaka et al., 2001).
The objective of this work was to reduce WVP of sh gelatin
lms by incorporating different hydrophobic substances, such as
amaranth and rapeseed oil, lanolin, beeswax and ozocerite into
their structure. The effect was determined for unmodied lms and
those cross-linked with transglutaminase (TGase) or 1-ethyl-3-
(3-dimethylaminopropyl) carbodiimide (EDC).

0268-005X/$ see front matter 2008 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodhyd.2008.08.002
 K. Sztuka, I. Koodziejska / Food Hydrocolloids 23 (2009) 10621064 1063

Our previous study showed that enzymatic or chemical cross- Table 1

linking of sh gelatin effectively limits the solubility of lms, but Effect of hydrophobic substances on WVP of unmodied sh gelatin lms

did not improve their water barrier properties (Piotrowska et al., Lipid WVP [g mm/h kPa m2]a
in press). There is also no data in literature about effect of hydro- concentration
Amaranth Rapeseed Lanolin Beeswax Ozocerite
phobic substances on lms cross-linked by TGase or EDC. [%]
oil oil
0 3.60a/u 3.60a/u 3.60a/u 3.60a/u 3.60a/u
10 2.09b/u 3.05b/w 2.27b/x 1.68b/y 2.30b/x
2. Materials and Methods 60 1.18c/uw 2.85b/x 1.31c/u 1.09c/w 1.74c/y
10b 1.96b/uw 2.05c/u 2.59b/x 1.87b/w 2.59b/y
2.1. Materials 60b 0.97c/u 1.90c/w 2.28b/x 0.46d/y 1.52c/z
a
The values for a particular columns and lines followed by different letters (ad)
Fish gelatin was obtained from the Baltic cod skins as described and (uz), respectively, differ signicantly (P < 0.05).
b
by Ko1odziejska, Kaczorowski, Piotrowska, and Sadowska (2004). Films to that additionally lecithin was introduced (10% of hydrophobic substance
EDC was purchased from Sigma Chemical Co., Germany. For the mass).

enzymatic modications the preparation of commercial TGase

containing 1% of enzyme and 99% of maltodextrin (Ajinomoto Cos
Transglutaminase Activa-WM, Japan) was used. Rened rapeseed
oil was bought from a local shop, beeswax, ozocerite and synthetic
lanolin were purchased from Kahl Wax Renery, Poland, and leci-
thin was from BDH, Poland. Amaranth oil was kindly provided by
Professor Andrzej Sto1yhwo from Warsaw University of Life
Science, Poland.
2.5. Statistical Analysis
Results in tables are averages from three replications. Data were
evaluated by analysis of variance (one-way procedure) using the
program Statgraphics, Statistical Graphic Corporation and the
differences between the means determined by Tukey multiple test
(P < 0.05).

2.2. Enzymatic and Chemical Modications 3. Results and Discussion

Fish gelatin was dissolved in water at room temperature to
achieve the nal concentration of 5% (w/w).
Lipids at their melting temperature, declared by the producer
(amaranth and rape seed oil at room temperature; lanolin at
3237; bees wax 6265; ozocerite 69), and concentrations of 10
and 60% (% of the gelatin mass, w/w) were homogenised with
gelatin solution for 4 min at 26 000 rpm. In some experiments
homogenisation was conducted additionally in the presence of
lecithin (10% of the lipids mass, w/w).
To the lm-forming solutions, TGase or EDC was added to the
nal concentration of 0.2 mg/mL and 30 mM, respectively. As we
have shown earlier, these concentrations were the most efcient in
decreasing the gelatin lms solubility (Piotrowska et al., in press).
The lms were formed just after adding of TGase or EDC as
described below. The enzymatic and chemical reactions were
running during that process.
2.3. Film Formation
In all experiments, 20 g of solutions were cast on a square area
9.5  13.5 cm of a polyester surface and spread manually to the
outside borders. The lms were dried at room temperature for 24
48 h at 3545% relative humidity (RH). The thickness of the lms
was measured at ve random locations with a handheld microm-
eter. The average thickness of the lms was 0.155  0.005 mm.
Unmodied sh gelatin lms were characterized by very poor
water barrier properties (Table 1). It was shown previously that
WVP for lm of thickens 0.12 mm was about 2.64 g mm/h kPa m2
(Piotrowska et al., in press). In this work all lms were 0.16 mm
thick therefore WVP of unmodied gelatin lm was higher and
amounted 3.6 g mm/h kPa m2. Much lower WVP of gelatin lms,
even in spite of the presence of plasticizer, was obtained by de
Carvalho et al. (2008), Bertan et al. (2005). The discrepancy in these
data may be caused by over two times higher thickness of the lms
in our experiments, as well as by different conditions used during
preparation of the lms.
Cross-linking gelatin with TGase did not improve considerably
water barrier properties of lms (Tables 1 and 2). Differently, modi-
cation with TGase of whey proteins isolate, soybean globulin 11S or
mixture of these components increased the WVP of lms by about
60% (Yildirim & Hettiarachchy, 1998). On the other hand, de Carvalho
and Grosso (2004) found that WVP of TGase cross-linked bovine
gelatin lms was about 40% lower than that of unmodied lms.
Cross-linking of sh gelatin lms with EDC decreased WVP from
3.60 to 3.04 g mm/h kPa m2 (Table 3).
Improving of water barrier properties can be achieved by an
addition of hydrophobic substances to the lm structure. All tested
hydrophobic substances at concentration of 10% decreased WVP of
unmodied gelatin lms, particularly amaranth oil and beeswax, by
42 and 53%, respectively (Table 1). Increasing the concentration of

Table 2
2.4. Water Vapor Permeability Effect of hydrophobic substances on WVP of lms cross-linked with TGase at
concentration of 0.2 mg/mL

WVP was determined according to the ASTM method E 96-95 Lipid WVP [g mm/h kPa m2]a
(ASTM, 1995). The lms were conditioned for 24 h at 25  C and 50% concentration
Amaranth Rapeseed Lanolin Beeswax Ozocerite
[%]
RH before determination. RH was measured using hygrometer LA8 oil oil
02/96 (Rotronic). Film samples were mounted on cups lled with 0 3.30a/x 3.30a/x 3.30a/x 3.30a/x 3.30a/x
water. The cups were placed at 25  C and 50% RH in a desiccator. 10 2.80b/x 3.40a/y 2.88b/x eb ih
The weight of the cups was measured at 1 h intervals for 9 h. The 60 2.21c/x 2.06b/x 2.29c/x eb ih
10b eb 2.00b/x 2.15c/x eb 2.61b/y
simple linear regression was used to estimate the slope of weight
60b eb 1.37c/x 1.75d/y eb 2.04c/z
loss vs. time plot.
WVP was calculated from: WVP (WVTR L)/Dp; where WVTR Abbreviations: eb, excessively brittle; ih, inhomogeneous.
a
The values for a particular columns and lines followed by different letters (ad)
(Water vapor transmission rate) Slope/Film area [g/m2  h], L and (xz), respectively, differ signicantly (P < 0.05).
lm thickness [mm], Dp partial water vapor pressure difference b
Films to that additionally lecithin was introduced (10% of hydrophobic substance
[kPa] between the two sides of the lm. mass).
1064 K. Sztuka, I. Koodziejska / Food Hydrocolloids 23 (2009) 10621064
Table 3
Effect of hydrophobic substances on WVP of lms cross-linked with EDC at
concentration of 30 mM
Lipid WVP [g mm/h kPa m2]a
concentration [%]
Amaranth oil Rapeseed oil Lanolin Beeswax
0 3.04a/w 3.04a/w 3.04a/w 3.04a/w
10 2.58b/w 2.44b/w 2.57b/w 2.57b/w
60 1.46c/w 2.80a/x 2.15c/y 1.98c/y
10b 2.44bd/w 2.01c/x 2.33b/w 2.79a/y
60b 2.27d/w 1.96c/x 2.35b/w 0.90d/y
a 470473.
The values for a particular columns and lines followed by different letters (ad)
and (wz), respectively, differ signicantly (P < 0.05).
b
Films to that additionally lecithin was introduced (10% of hydrophobic substance
mass).
these substances up to 60% caused further reduction of WVP, by
about 1725% (Table 1). Yang and Paulson (2000) found that WVP of
gellan emulsion lms containing 20% of beeswax was about two
times lower than that for unmodied lms.
The presence of lecithin in lm-forming emulsions prevented
separation of lipid layer on the lm surface. Addition of emulsier
had no inuence on WVP of unmodied lms with 10% of lanolin or
beeswax (Table 1). However, WVP of lms containing 10% of
rapeseed oil and lecithin was lower than those without lecithin. The
highest decrease of WVP was found in case of lms with lecithin
and beeswax in concentration of 60%, by 87% (Table 1). According to
Krochta and De Mulder-Johnston (1997) lms with such value of
WVP show moderate water barrier properties.
Among enzymatically modied lms with lecithin and 60% of
lipids, those with rapeseed oil showed the lowest WVP, simulta-
neously about 30% lower than lms without cross-linking. Simi-
larly, WVP of lms modied with TGase containing lecithin and 60%
of lanolin was considerably lower than that of unmodied lms
(Tables 1 and 2). However, WVP of these lms was still too high to
serve as materials with moderate water barrier properties.
Enzymatically modied lms with beeswax were brittle and
broke during analyzes, similarly as those with amaranth oil and
lecithin.
Incorporation of 10% of the tested hydrophobic substances, with
or without of lecithin, into lms cross-linked with EDC decreased
WVP by about 20%, except of lms containing ozocerite (Table 3).
All variants in that case showed similar WVP as gelatin lms
without this substance (Table 3). Increasing amaranthus oil and
beeswax concentration up to 60% decreased WVP from 3.04 to 1.46
and 1.98, respectively. The highest decrease of WVP of chemically
modied lms was found in the presence of 60% of beeswax with
addition of lecithin by about 70% (Table 3). However, this WVP is
twice higher than that of uncross-linked lms of the same ingre-
dient composition.
4. Conclusions
Unmodied, enzymatically and chemically cross-linked sh
gelatin lms were characterized by very poor WVP. Among selected
hydrophobic substances beeswax was the most effective in reducing
of WVP. Films containing 60% of beeswax and lecithin show moderate
water barrier properties. Similarly, addition of these substances
caused the highest reduction of WVP of lms cross-linked with EDC.
In case of enzymatically modied lms the lowest WVP was observed
for lms with 60% of rapeseed oil and lecithin. The presence of
beeswax in TGase cross-linked lms increased their fragility.
Acknowledgements
This research project was nanced by the national research
budget provided for the years 20072009.
The authors also wish to express their gratitude to Professor
Zdzis1aw E. Sikorski for the nal correction of this manuscript.
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