Food Chemistry 122 (2010) 10951101

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Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Characterisation of gelatinfatty acid emulsion lms based on blue shark

(Prionace glauca) skin gelatin
Kanokrat Limpisophon a, Munehiko Tanaka b, Kazufumi Osako a,*
a
b

Department of Food Science and Technology, Tokyo University of Marine Science and Technology, 5-7 Konan 4, Minato-ku, Tokyo 108-8477, Japan
Department of Home Economics, Kokugakuin Tochigi Junior College, 608 Hirai, Tochigi 328-8588, Japan

a r t i c l e

i n f o

Article history:
Received 9 July 2009
Received in revised form 17 January 2010
Accepted 23 March 2010

Keywords:
Gelatin
Emulsion lm
Shark skin
Stearic acid
Oleic acid

a b s t r a c t
Incorporation of fatty acids (stearic and oleic) into edible lms based on blue shark (Prionace glauca) skin
gelatin was investigated to modify properties such as water vapour barrier and exibility due to their
hydrophobicity and plasticizing effect, respectively. Addition of stearic acid from 0% to 100% of protein
concentration in the lm-forming solution considerably decreased water vapour permeability of gelatinfatty acid emulsion lms compared to addition of oleic acid at the same fatty acid concentration.
Increasing concentrations of both fatty acids decreased tensile strength, but increased elongation at break
due to their plasticizing effect. At the same concentration, oleic acid gave a greater plasticizing effect than
did stearic acid. On the other hand, transparency of the gelatinstearic acid emulsion lm was lower than
that of the gelatinoleic acid emulsion lm. Faster stirring speed of homogenisation improved properties
of only the gelatinstearic acid emulsion lm.
2010 Elsevier Ltd. All rights reserved.

1. Introduction
Edible lm, a thin layer of biomaterial, has attracted the packaging industry, since it can provide a barrier to moisture, oxygen and
solute movement of foods (Guilbert, 1986). Amongst protein-based
edible lms, gelatin lm has received attention for edible and pharmaceutical products, such as capsules, meats and sausage casing
components, since gelatin is unique amongst hydrocolloids in
forming thermo-reversible gels with a melting point close to body
temperature (Achet & He, 1995). Gelatin lm properties depend on
several factors, including the gelatin source and the gelatin lmmaking process. Non-bovine gelatin sources, especially marine
species, have currently received attention in response to the bovine
spongiform encephalopathy (BSE) crisis. In our previous study, gelatin lm from blue shark (Prionance glauca) skin was characterized
in order to enhance the utilisation of by-products from shark processing (Limpisophon, Tanaka, Weng, Abe, & Osako, 2009). Gelatin
lm from shark skin has a lower water vapour permeability (WVP)
and higher opacity at 280 nm (UV wavelength) than have gelatin
lms from other sh. This gelatin lm can thus be applied to pharmaceutical products or foods rich in fat due to its excellent barrier
properties against water vapour and UV. However, WVP of shark
skin gelatin lm was higher than that of other edible lms.
Materials, such as fatty acids, lipids and waxes, are commonly
used as edible coatings to reduce WVP due to their nonpolar or
* Corresponding author. Tel./fax: +81 3 5463 0620.
E-mail address: osako@kaiyodai.ac.jp (K. Osako).
0308-8146/$ - see front matter 2010 Elsevier Ltd. All rights reserved.
doi:10.1016/j.foodchem.2010.03.090

hydrophobic characteristics. Moreover, they bring about visual appeal (gloss) to products (Talens & Krochta, 2005). There are many
reports of lipid incorporation in protein-based lms to retard water
migration (Anker, Berntsen, Hermansson, & Stading, 2002; Bertan,
Tanada-Palmu, Siani, & Grosso, 2005; Perez-Gago & Krochta, 2001).
Proteinlipid lms can be prepared by two methods: lamination
and emulsication. During casting of lipid onto protein lm for
the lamination, temperatures of lm and lipid can be easily and
separately controlled. On the other hand, the temperature of the
emulsion must be above the melting point of the lipid but below
the melting temperature of gelation and solvent volatilisation temperature for the structural network of emulsion lms. The main
disadvantage of laminate lms is that the preparation technique
requires four stages; protein lm casting, protein lm drying, lipid
casting onto protein lm and nal lipid lamination. This is the reason why laminate lms are less popular in the food industry, despite their good barrier property against water vapour.
On the other hand, the preparation of emulsion lms requires
only one emulsion solution casting and one drying step. However,
the water vapour barrier property of nished lms is rather poor,
since water molecules still permeate through the non-lipid phase
(Anker et al., 2002). In the case of emulsion lms, the water barrier
efciency increases with a decrease in lipid globule size and with
distribution of homogeneous hydrophobic substances (Debeaufort
& Voilley, 1995; Perez-Gago & Krochta, 2001). Stirring speed during the emulsication process can thus be considered as an important factor for obtaining homogeneous lms and it has a great
inuence on their performance (Garcia, Martino, & Zaritzky,

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K. Limpisophon et al. / Food Chemistry 122 (2010) 10951101

2000). Moreover, many researchers have reported that an increase

in the carbon number of fatty acids decreases the WVP property of
the lipidprotein edible lms (Bravin, Peressini, & Sensidoni, 2004;
Bertan et al., 2005; Yang & Paulson, 2000). Stearic acid is one of the
most common fatty acids used in emulsion lms, since it overcomes the poor moisture resistance of the edible lms (Colla, Sobral, & Menegalli, 2006; Karnnet, Potiyaraj, & Pimpan, 2005;
Morillon, Debeaufort, Blond, Capelle, & Voilley, 2002). However,
there has been no research on gelatin lms about the effect of fatty
acids with the same carbon number on water barrier efciency, e.g.
stearic acid (C 18:0, m.p. = 69 C) and oleic acid (C 18:1, m.p. = 13
14 C). Therefore, this study was carried out to elucidate properties
of emulsion lms based on shark skin gelatin by the incorporation
of fatty acids (stearic and oleic acids) from the standpoint of their
concentration and stirring speed of homogenisation.

2. Materials and methods

2.1. Materials
Sucrose stearate (Ryoto Sugar Ester S970), provided by the Mitsubishi Chemical Foods, Co. Ltd. (Tokyo, Japan), was used as an
emulsier. Stearic acid and oleic acid were purchased from Wako
Pure Chemical Industries, Ltd. (Tokyo, Japan). Glycerol, as a plasticizer, was purchased from Kokusan Chemical Works, Ltd. (Tokyo,
Japan).

2.2. .Extraction of gelatin from shark skin

Frozen blue shark (P. glauca) skin was obtained from Kyodo Suisan Co. Ltd. in Kesennuma, Miyagi Prefecture, Japan. Thawed shark
skin was cut, minced, and stored at 30 C. Extraction of gelatin
from shark skin followed the procedure of Limpisophon et al.
(2009).

2.3. Preparation of gelatin lms

The studied levels of fatty acids (stearic and oleic acids) in lmforming solution (FFS) were 25%, 50%, and 100% (w/w) of the protein. Gelatin was dissolved at 60 C for 30 min to obtain the FFS
with 2% (w/v) protein concentration, and then mixed with glycerol
(50% w/w of the protein). Sucrose stearate and stearic acid were
homogenised at 24,000 rpm with a ratio of 1:3 (w/w) for 3 min,
using a homogenizer (Heidolph DIAX 600, Heidolph Co., Kelheim,
Germany) at 85 C, since this temperature is higher than melting
point of stearic acid (m.p. = 69 C). The homogenate was called
mixing solution in this study. In the case of oleic acid, the mixing
solution was prepared at room temperature, since oleic acid is in
the liquid state at room temperature. Gelatin solution and mixing
solution were homogenised together at room temperature, and
24,000 rpm for 3 min using the homogenizer. The air bubbles in
the FFS were removed by a hybrid mixer (HM-500; Keyence Co.,
Tokyo, Japan). De-aerated FFS (4 g) was cast onto a rimmed silicone
resin plate (50  50 mm) and dried in a ventilated oven (EYELA,
KCL-2000A, Tokyo Rikakikai Co., Ltd., Japan) at 25 0.5 C and
50 5% relative humidity (RH) for 24 h. All experimental treatments were compared to 2% (w/v) gelatin lm with glycerol (50%
w/w of the protein) as a control.
To study the effect of stirring speed on gelatinfatty acid emulsion lm, fatty acid (100% w/w of the protein) was used. Stirring
speeds at two homogenisation steps were 8000, 13,500 or
24,000 rpm.

2.4. Laser diffraction size analysis

In the study, the effect of stirring speed on gelatinfatty acid
emulsion lms, and size of oil droplets in the FFS was determined.
FFS, prior to casting, was warmed to 60 C for 15 min and then diluted with distiled water at a ratio of 1:20 (v/v). Size of oil droplets
in diluted FFS was determined using a laser diffraction particle size
analyser (Violet Laser SALD-300 V, Shimadzu Co., Kyoto, Japan).
2.5. Characterisation of gelatin lms
2.5.1. Film thickness
Film thickness was measured as described in the previous study
of Limpisophon et al. (2009). After measurement of lm thickness,
lms were conditioned at 25 0.5 C and 50 2%RH for 48 h.
2.5.2. Water vapour permeability (WVP)
WVP of lms was measured as described by Limpisophon et al.
(2009). WVP of gelatin lm was calculated by the following equation (McHugh, Avena-Bustillos, & Krochta, 1993):

WVPg m1 s1 Pa1 w  x  A1  t 1  P2  P1 1

where w = weight gain (g) of the cup; x = lm thickness (m);
A = area of exposed lm (m2); t = time of gain (s); (P2  P1) = vapour
pressure differential (Pa) across the lm.
2.5.3. Mechanical properties
Tensile strength (TS) and elongation at break (EAB) were also
determined by using the method of Limpisophon et al. (2009).TS
(MPa) was calculated by the following equation:

TSMPa Fmax=A
where F max = maximum load (N) needed to pull the sample apart;
A = cross-sectional area (m2) of the samples; EAB (%) was calculated
by following equation:

EAB% E=30  100

where E = lm elongation (mm) at the moment of rupture; 30 = initial grip length (mm) of samples.
2.5.4. Transparency
The barrier properties of gelatin lms against ultraviolet (UV)
and visible light were measured according to the method described
by Limpisophon et al. (2009). The transparency of the lms was
calculated by the following equation:

Transparency A=mm  log T=x

where A = absorbance at each wavelength; T = transmittance (%) at
each wavelength; x = lm thickness (mm).According to the equation, high transparency indicates opaque.
2.5.5. Scanning electron microscopy (SEM)
The upper surface and lower surface, which faced air and silicone plate during dehydration, respectively, were analysed by
SEM according to the method described by Colla et al. (2006). Films
were maintained in a desiccator with silica gel for 7 days to dehydrate and then stuck onto a cylindrical aluminium stub by a double-sided tape. The stub with the lm was coated with PtPd in
an ion sputter coater (E-1030, Hitachi Co., Tokyo, Japan) for 100 s
and then placed into a SEM (S-4000, Hitachi Co., Tokyo, Japan) to
observe the morphology of the surfaces.

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K. Limpisophon et al. / Food Chemistry 122 (2010) 10951101

2.6. Statistical analysis

Statistical analysis on a completely randomized experimental
design was performed using the General Linear Models procedure
in SPSS computer programme (SPSS statistical software, Arizona,
USA). One-way analyses of variance (ANOVA) were carried out
and mean comparisons were run by Duncans multiple range tests
(Stell & Torrie, 1980).
3. Results and discussion
3.1. Effect of fatty acid concentration on gelatinfatty acid emulsion
lm
3.1.1. Thickness of lm
Film thickness of the gelatin-stearic acid emulsion lm increased from 27.00 to 71.85 lm with increasing stearic acid concentration (p < 0.05), as shown in Table 1. The results of the lm
with oleic acid were similar to those of the lm with stearic acid,
as is also given in Table 1. However, the thickness of lm with oleic
acid was smaller (27.0054.83 lm) than that of lm with stearic
acid at the same fatty acid concentration. This might be due to
the difference of specic gravity between stearic acid and oleic acid
at 25 C (conditioning temperature).
3.1.2. WVP
WVP of the gelatinstearic acid emulsion lm (Table 1) decreased markedly from 1.30 to 0.70  1010 g m1 Pa1 s1
(p < 0.05) upon increasing stearic acid concentration from 0 to
100% of protein concentration. WVP of the gelatin-oleic acid emulsion lm also decreased from 1.30 to 0.91  1010 g m1 Pa1 s1
(p < 0.05) upon increasing oleic acid concentration from 0% to
100% of protein concentration, as shown in Table 1. Incorporation
of 100% fatty acid into the emulsion lm successfully reduced
WVP, about 46% for stearic acid and about 30% for oleic acid. From
these results, addition of stearic acid to gelatin lm could improve
the water barrier property better than could oleic acid addition at
the same fatty acid concentration. Although oleic acid and stearic
acid have the same carbon number in the molecule, oleic acid is
in a liquid state in the emulsied lm and has a lower melting
point, indicating a higher water solubility coefcient of the lm.
Bertan et al. (2005) also observed that the WVP of gelatin lm with
stearic acid (melting point = 69 C) was lower than that of gelatin
lm with palmitic acid (melting point = 6364 C). The potential
of lipid melting point as an indicator for WVP of the lm was also

studied by McHugh and Krochta (1994). WVP of the fatty acid

emulsion lm seemed to decrease as melting point of the added
fatty acid increased.
3.1.3. Mechanical properties
Mechanical properties of the gelatinfatty acid emulsion lms,
with varied fatty acid concentrations, are presented in Table 1.
Addition of fatty acid, from 0% to 100% of the protein concentration, signicantly reduced TS of the lm from 23.26 to 13.62 MPa
for the gelatinstearic acid emulsion lm and from 23.26 to
9.46 MPa for the gelatinoleic acid emulsion lm (p < 0.05). By
contrast, addition of stearic acid improved EAB of the lm from
80.78% to 108.41% (p < 0.05). Addition of oleic acid markedly increased EAB from 80.78% to 257.17% (p < 0.05). Lipids lack cohesive
structural integrity (Gontard, Marchesseau, Cuq, & Guilbert, 1995).
Incorporating lipid in protein-based or polysaccharide-based lms
may interfere with polymer chain-to-chain interactions and provide exible domains within the lm. The result can be a plasticizing effect, including reduction of lm strength and increase of lm
exibility (Talens & Krochta, 2005). From these results, it can be
concluded that the addition of oleic acid to the gelatin-based lm
had a greater plasticizing effect than had addition of stearic acid.
This phenomenon may be because of the different physical states
of the fatty acids at room temperature. Morillon et al. (2002) also
reported that oleic acid is liquid and can ow during lm deformation, whilst beeswax is solid and so less deformable during the lm
stretching.
3.1.4. Transparency
Transparency of gelatinstearic acid emulsion lms decreased
at all wavelengths with increasing stearic acid concentration, as
shown in Table 2, indicating that the emulsion lms are more
transparent. Transparency of gelatinoleic acid emulsion lm (Table 2) also decreased, at almost all wavelengths, with increasing
oleic acid concentration; however, transparency values of the lms
were similar at 280 nm and 350 nm. Transparency, at 280 nm (UV
light), of the gelatinfatty acid emulsion lm at each fatty acid concentration was considerably higher than that in the visible wavelength range (400780 nm). This was because the highmolecular-weight proteins of gelatin lm from shark skin can absorb UV light, as shown by Limpisophon et al. (2009). Gelatin lm
with stearic acid gave a lower transparency than did gelatin lm
with oleic acid at the same fatty acid concentration. This may be
because the stearic acid at the upper surface of the emulsion lm
had a smaller particle size than had oleic acid, as shown by SEM

Table 1
Effect of fatty acid concentration on thickness, water vapour permeability (WVP), tensile strength (TS), and elongation at break (EAB) of gelatinfatty acid emulsion lms.a,b
WVPc (1010 g m1 Pa1 s1)

TSd (MPa)

EABe (%)

Stearic acid concentration (%)

27.00 2.10a
Controlf
25
42.22 3.30b
50
52.01 5.17c
100
71.85 7.22d

1.30 0.12c
0.95 0.07b
0.96 0.04b
0.70 0.06a

23.26 2.03c
17.21 0.76b
16.74 0.34b
13.62 0.32a

80.78 4.54a
99.31 5.96b
110. 32 6.90c
108.41 9.77c

Oleic acid concentration (%)

27.00 2.10a
Controlf
25
38.24 4.46b
50
41.45 2.16b
100
54.83 4.71c

1.30 0.12c
1.17 0.14bc
1.01 0.03ab
0.91 0.06a

23.26 2.03c
13.23 1.58b
14.16 0.87b
9.46 0.36a

Thickness (lm)

80.78 4.54a
178.56 12.38b
222.12 11.04c
257.17 7.13c

Data are expressed as means standard deviation.

Different superscripts in the same column and the same section indicate statistical differences (p < 0.05).
c
WVP (g m1 s1 Pa1) = w  x  A1  t1  (P2  P1)1 (w, weight gain (g) of the cup; x, lm thickness (m); A, area of exposed lm (m2); t, time of gain (s); (P2  P1),
vapour pressure differential (Pa) across the lm).
d
TS (MPa) = F max/A (F max, maximum load (N) needed to pull the sample apart; A, cross-sectional area (m2) of the samples).
e
EAB (%) = (E/ 30)  100 (E, lm elongation (mm) at the moment of rupture; 30, initial grip length (mm) of samples).
f
Control is 2% gelatin lm with 50% glycerol.
b

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Table 2
Effect of fatty acid concentration on transparencya of gelatinfatty acid emulsion lms.
Transparency (A/mm) at wavelength (nm)
200c
b

Control
Stearic acid concentration (%)
25
50
100
Oleic acid concentration (%)
25
50
100
a
b
c

280

350

400

500

600

800

28.69

10.09

6.83

5.69

5.24

4.65

18.85
14.80
10.23

7.09
5.49
3.84

5.67
4.32
2.91

4.75
3.47
2.31

4.24
2.95
1.99

3.70
2.40
1.72

25.07
25.11
24.92

9.22
9.50
8.57

7.12
7.06
5.96

5.56
5.30
4.09

4.68
4.42
3.15

3.62
3.53
2.26

Transparency (A/mm) = log T/x (A, absorbance at each wavelength; T, transmittance (%) at each wavelength; x, lm thickness (mm)).
Control is 2% gelatin lm with 50% glycerol.
Value could not be determined.

micrographs in Fig. 1. Osipow and Rosenblatt (1967) also reported

that a micro emulsion, composed of smaller particles, is more
transparent than a macro emulsion.
3.1.5. SEM
SEM revealed the presence of crystalline lipid particles at various stearic acid concentrations within the protein matrix of the
emulsion lm (Fig. 1). There were discrete crystalline particles of
stearic acid on the upper surface of gelatinstearic emulsion lms.
Lower surfaces of all emulsion lms also revealed small crystalline
particles. These physical surfaces may support lower WVP of gelatinstearic acid emulsion lms compared to those without fatty
acid (control), as shown in Table 1. This may be because fat droplets hindered the transfer of water molecules inside the lm; therefore, a homogeneous distribution of oil droplets would give the
lm a lower WVP (Karbowiak, Debeaufort, & Voilley, 2007).

Control

Upper surface

Upper surfaces of all gelatin lms formulated with different levels of oleic acid showed even distributions of oil droplets (Fig. 1).
Lower surfaces of all lms did not have any oil droplets, indicating
that oleic acid was dispersed only on the top surface. This phenomenon would occur during FFS dehydration. Films with oleic acid did
not have crystalline particles like lms with stearic acid, since oleic
acid is in a liquid state at room temperature (m.p. = 1314 C).
This study revealed that gelatin emulsion lms with 100% fatty
acid (stearic acid or oleic acid) effectively gave the lowest WVP.
3.2. Effect of stirring speed on gelatinfatty acid emulsion lm
3.2.1. General
The gelatin emulsion lms with 100% stearic acid or 100% oleic
acid were used to study the effect of stirring speed on gelatinfatty
acid emulsion lm.

Lower surface

Fatty acid
concentration
(%)

Gelatin-stearic acid emulsion film

Upper surface

Lower surface

Gelatin-oleic acid emulsion film

Upper surface

Lower surface

25

50

100

Fig. 1. SEM micrographs of gelatin lm surfaces formulated with different fatty acid concentrations.

K. Limpisophon et al. / Food Chemistry 122 (2010) 10951101

3.2.2. Particle size of fatty acid in gelatinfatty acid emulsion FFS

The effect of stirring speed on distribution of stearic acid particles in FFS is depicted in Fig. 2. Increasing stirring speed of homogenisation from 8000 rpm to 24000 rpm decreased size of fat
particles in FFS from 36.84 to 0.54 lm, indicating that lms formed
by homogenisation at faster stirring speeds gave smaller fat particles. Distribution of oleic acid particles in FFS with oleic acid was
also affected by stirring speed of homogenisation, as shown in
Fig. 2. Size of oleic acid particles decreased from 34.84 to
0.28 lm with increasing stirring speed.
3.2.3. Thickness of lm
Thickness of gelatinstearic acid and gelatin-oleic acid emulsion lms, prepared by different stirring speeds, did not show statistical difference (p > 0.05), as presented in Table 3. From these
results, it could be concluded that stirring speed did not affect
thickness of gelatin lmfatty acid emulsion lm.
3.2.4. Water vapour permeability (WVP)
WVP of gelatin lm with stearic acid signicantly decreased
upon increasing stirring speed from 8000 to 24,000 rpm, as shown
in Table 3 (p < 0.05). This suggests that stirring speed for homogenisation inuences WVP of the gelatin lm with stearic acid, since
proteinlipid interactions at the lipid interface increase with reduced lipid particle size. On the other hand, in the case of gelatin
lm with oleic acid, the stirring speed did not result in a decrease
in WVP, as presented in Table 3 (p > 0.05). This may be caused by
the aggregation of oleic acid particles during the drying process
after homogenisation, as depicted in Fig. 3.
3.2.5. Mechanical properties
The effect of stirring speed on TS of gelatin lm with stearic acid
is presented in Table 3. TS increased from 11.03 to 13.62 MPa upon
increasing stirring speed from 8000 to 24000 rpm (p < 0.05). In
addition, EAB increased from 74.35% to 108.41% with increasing
stirring speed (p < 0.05). Therefore, faster stirring speed could im-

Fig. 2. Effect of stirring speed on distribution of fatty acid particles in FFS (A:
distribution of stearic acid particles in FFS, B: distribution of oleic acid particles in
FFS.).

1099

prove mechanical properties of the gelatin lms with stearic acid.

Colla et al. (2006) also explained that better stearic acid distribution of the emulsion lm-based Amaranthus cruentus our avoided
formation of fatty acid accumulation during the drying process.
The results obtained in this study agree with the study of PerezGago and Krochta (2001) that TS and EAB signicantly increased
with decreasing particle size of beeswax in emulsion lm with
60% beeswax (dry basis), due to an increase in protein immobilization at the beeswax particle interface. On the other hand, a faster
homogenisation process could not improve mechanical properties
of the gelatinoleic acid emulsion lm (Table 3). There were no statistical differences in TS and EAB of the gelatinoleic acid emulsion
lm amongst the different stirring speeds during homogenisation
(p > 0.05). This phenomenon may be caused by accumulation of
oleic acid during the lm drying process. Size of oleic acid droplets
in the emulsion lm was not inuenced by the stirring speed, as
shown by SEM (Fig. 3), although faster homogenisation resulted
in a decrease in oleic acid particles in the FFS, as shown in Fig. 2.
However, higher EAB and lower TS of gelatinoleic acid emulsion
lms were observed in comparison with those of gelatinstearic
acid emulsion lms homogenised at the same stirring speed. This
may be because the liquid state of oleic acid in the lm has a greater plasticizing effect than has the solid state of stearic acid. The
smaller plasticizing effect of stearic acid was also reported by Sakanaka, Sobral, and Menegalli (2001).
3.2.6. Transparency
Table 4 shows that faster stirring speed of homogenisation reduced transparency of gelatinfatty acid emulsion lms, suggesting that these emulsion lms become clearer.
However, the transparency of gelatinoleic acid emulsion lms
at 280 nm did not decrease with increasing stirring speed. Gelatin
stearic acid emulsion lm, homogenised at 8000 rpm, showed a
higher transparency than did gelatin-oleic acid emulsion lm at
each wavelength, since the particle size of stearic acid in the upper
surface of the lm was larger than that of oleic acid (Fig. 3). However, the gelatin-stearic acid emulsion lm homogenised at 13,500
or 24,000 rpm had a lower transparency than had the gelatinoleic
emulsion lm at each wavelength. This may be due to the smaller
particle size of stearic acid at 13,500 or 24,000 rpm compared to
that of oleic acid, as shown by SEM (Fig. 3). Although particle size
of oleic acid in FFS was smaller than that of stearic acid at each stirring speed (Fig. 2), oleic acid particles, which are in a liquid state,
may be aggregated during lm drying.
3.2.7. SEM
Physical surfaces of each lm with stearic acid at different levels
of stirring speed are presented in Fig. 3. Film formed by homogenisation at 8000 rpm had the largest lipid crystalline particles. There
were large lipid particles present in the lower surface of lm
homogenised at 8000 rpm, whereas smaller lipid particles were
observed in those homogenised at 13,500 and 24,000 rpm. These
results signicantly supported the conclusion that WVP was reduced when lm was more homogeneous (Table 3). Smaller oil
droplets were also present on upper surfaces of the gelatinoleic
acid lm homogenised at faster stirring speed; however, homogenisation at different stirring speed did not considerably inuence
particle size compared to gelatinstearic acid lm. For each lm
prepared by different stirring speeds of homogenisation, oil droplets did not appear on the lower surfaces.
Based on these results, it was elucidated that a faster stirring
speed of homogenisation effectively resulted in smaller sizes of
oil droplets in emulsion lms. Film properties with stearic acid,
such as TS, EAB, WVP, transparency and microstructures were also
affected by stirring speed (p < 0.05). Even though microstructures
of lms with oleic acid at different stirring speeds were different,

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Table 3
Effect of stirring speed on thickness, water vapour permeability (WVP), tensile strength (TS), and elongation at break (EAB) of gelatinfatty acid emulsion lms.a,b
Stirring speed (rpm)

Thickness (lm)

WVPc (1010 g m1 Pa1 s1)

TSd (MPa)

EABe (%)

Gelatinstearic acid emulsion lms

8000
13,500
24,000

68.83 3.07a
68.65 2.26a
71.85 7.22a

1.04 0.09c
0.87 0.01b
0.70 0.06a

11.03 0.71a
11.83 0.48b
13.62 0.32c

74.35 7.06a
77.11 4.40a
108.41 9.77b

Gelatinoleic acid emulsion lms

8000
13,500
24,000

57.39 2.09a
56.10 3.33a
54.83 4.71a

1.02 0.06a
1.01 0.04a
0.91 0.06a

10.23 0.85a
9.73 0.66a
9.46 0.36a

259.80 27.75a
261.40 15.84a
257.17 7.13a

Data are expressed as means standard deviation.

Different superscripts in the same column and the same section indicate statistical differences (p < 0.05).
c
WVP (g m1 s1 Pa1) = w  x  A1  t1  (P2  P1)1 (w, weight gain (g) of the cup; x, lm thickness (m); A, area of exposed lm (m2); t, time of gain (s); (P2  P1),
vapour pressure differential (Pa) across the lm).
d
TS (MPa) = F max/A (F max, maximum load (N) needed to pull the sample apart; A, cross-sectional area (m2) of the samples).
e
EAB (%) = (E/30)  100 (E, lm elongation (mm) at the moment of rupture; 30, initial grip length (mm) of samples).
b

Stirring speed
(rpm)

Gelatin-stearic acid emulsion film

Upper surface

Gelatin-oleic acid emulsion film

Lower surface

Upper surface

Lower surface

8,000

13,500

24,000

Fig. 3. SEM micrographs of surfaces of gelatinfatty acid emulsion lms prepared by different stirring speeds.

these did not signicantly inuence WVP, TS or EAB (p > 0.05). This
may be because the particle size of oleic acid in the lm which was
prepared at a faster stirring speed was not considerably smaller
than that of stearic acid. Although faster homogenisation impacted
smaller particle size in FFS of oleic acid (Fig. 2), its particles became
aggregated after homogenisation, as shown in Figs. 2 and 3. The
melting point of oleic acid is around 1314 C, whereas melting

point of stearic acid is 69 C. Consequently, stearic acid is in the solid state whilst oleic acid is liquid in the emulsion lms during
casting and conditioning. It might be concluded that the fatty acid
state in the emulsion lm has a greater effect on the lm characteristics than has homogenisation speed.
From a practical point of view, gelatin lms with stearic acid or
oleic acid would be applied as wrapping of food to reduce moisture

Table 4
Effect of stirring speed on transparencya of gelatinfatty acid emulsion lms.
Stirring speed (rpm)

a
b

Transparency (A/mm) at wavelength (nm)

200b

280

350

400

500

600

800

Gelatinstearic acid emulsion lms

8000
13,500
24,000

26.09
12.31
10.23

36.65
6.10
3.84

14.99
4.84
2.91

13.87
3.93
2.31

13.17
3.37
1.99

12.23
2.67
1.72

Gelatinoleic acid emulsion lms

8000
13,500
24,000

24.17
24.54
24.92

10.93
9.07
8.57

8.39
6.44
5.96

7.14
4.80
4.09

6.49
3.94
3.15

5.68
2.97
2.26

Transparency (A/mm) = log T/x (A, absorbance at each wavelength; T, transmittance (%) at each wavelength; x, lm thickness (mm)).
Value could not be determined.

K. Limpisophon et al. / Food Chemistry 122 (2010) 10951101

loss. In real application, gelatin lms can potentially preserve meat

products, such as frozen meat cuts and sausages, as reported in the
review of Gennadios, Hanna, and Kurth (1997). To compare
mechanical properties of both gelatinfatty acid lms with plastic
lms, TSs were relatively close to low-density polyethylene (LDPE),
having TS 16.5 MPa. EABs were higher than oriented polypropylene
(OPP), having 73%, as shown in the report of Shiku, Hamaguchi, and
Tanaka (2003). With the effective UV barrier property of shark skin
gelatin, the lms could prevent lipid oxidation. Furthermore, the
emulsion lms provide an ameliorating appearance due to their
transparent glossyness.
4. Conclusions
Addition of both fatty acids; stearic acid and oleic acid, to lm
based on blue shark (P. glauca) skin gelatin reduced WVP. Increasing fatty acid concentration signicantly decreased WVP (p < 0.05).
Addition of stearic acid to the gelatin lm reduced WVP more than
did oleic acid at the same fatty acid concentration. The addition of
oleic acid to the gelatin lm had a greater plasticizing effect than
had stearic acid, which reduced TS, but increased EAB. Transparency of gelatin-stearic acid emulsion lm was lower than that of
gelatinoleic acid emulsion lm at the same fatty acid concentration. Increasing stirring speed of homogenisation from 8000 to
24,000 rpm improved properties of gelatinstearic acid emulsion
lm (p < 0.05); however, it did not enhance those of gelatinoleic
acid emulsion lm (p > 0.05).
Acknowledgement
This study was supported by the Japanese Ministry of Education, Culture, Sports, Science and Technology (MEXT).
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