Journal of Food Engineering 113 (2012) 136142

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Journal of Food Engineering

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Effect of homogenization conditions on properties of gelatinolive oil

composite lms
Wen Ma a, Chuan-He Tang a, Shou-Wei Yin a,b,, Xiao-Quan Yang a, Jun-Ru Qi a, Ning Xia c
a

Department of Food Science and Technology, South China University of Technology, Guangzhou 510640, PR China
State Key Laboratory of Pulp and Paper Engineering, South China University of Technology, Guangzhou 510640, PR China
c
Department of Light Industry and Food Engineering, Guangxi University, Nanning 530004, PR China
b

a r t i c l e

i n f o

Article history:
Received 13 February 2012
Received in revised form 6 May 2012
Accepted 7 May 2012
Available online 15 May 2012
Keywords:
Microuidization
Olive oil
Particle size
Microstructure
Physical properties
Films

a b s t r a c t
Gelatinolive oil composite lms were prepared through emulsication to improve water barrier ability
of gelatin-based lms. The effects of homogenization conditions of lm-forming dispersions (FFD) on
lipid droplets distributions in the FFD and lms were evaluated and compared. Some selected physical
properties, e.g., water vapor permeability (WVP), microstructure of the lms were also evaluated. The
rotorstator homogenizer provided a lower energy input and so the largest particles were observed in
the related FFD and lms. These lms exhibited excellent water barrier ability, but poor mechanical resistance, extensibility and transparency. The microuidizer provided the FFD with lower and narrower particle size distributions, promoting mechanical resistance, extensibility and transparency of the lms. The
physical properties of the emulsied lms were dependent on the special microuidization pressure or
cycle used, e.g., the WVP of the lms decreased upon increasing microuidization pressure or cycle.
The present results indicated that the microuidizer can be used to modulate lipid droplets in the FFD,
thus lms properties.
2012 Elsevier Ltd. All rights reserved.

1. Introduction
The large majority of the polymers presently utilized are of synthetic origin, their biocompatibility and biodegradability are extremely limited with respect to the biopolymers such as protein,
polysaccharide and lipid. Biopolymer-based lms have shown
promising potential as a substitute of petroleum-based plastic
package lms to reduce environmental impact. The characteristics
of biopolymer-based lms are determined mainly by chemical nature of the biopolymer. Proteins or polysaccharide-based lms have
good mechanical behavior, but poor moisture barrier properties
due to their hydrophilic nature (Kristo et al., 2007). On the other
hand, although lipids yield lms with good water vapor barrier
properties, they are brittle (Perez-Gago and Krochta, 2001). Among
the biopolymer-based lms, protein-based lms are the most
attractive due to their nutritional value as well as impressive gas
barrier properties (Ou et al., 2004). Gelatin, a natural polymer from
animal by-products through acid or alkaline hydrolysis, was possessed of great potential to form edible/biodegradable lms due
to its abundance, biodegradability, low cost (Bigi et al., 2002).
However, the gelatin lms prepared from bovine and/or porcine
Corresponding author at: Department of Food Science and Technology, South
China University of Technology, Guangzhou 510640, PR China. Tel.: +86 20
87114262; fax: +86 20 87114263.
E-mail address: feysw@scut.edu.cn (S.-W. Yin).
0260-8774/$ - see front matter 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.jfoodeng.2012.05.007

skins presented poor water vapor barrier ability (Sobral et al.,

2001).
In order to obtain protein-lipid composite lms with improved
water barrier ability, a lipid can be incorporated into the protein
matrix by laminate techniques to form bi-layer lms, or dispersing
a lipid in the protein aqueous solutions to obtain an emulsied
lm. Bi-layer lms tend to delaminate over time, develop pinholes
or cracks and exhibit non-uniform surface (Quezada Gallo et al.,
2000). As concerns emulsied lms, they require only one lmogenic emulsion casting and one drying process, those lms exhibit
good mechanical and adhesive properties. However, they are less
efcient water vapor barriers due to the fact that water vapor
can migrate through the continuous hydrophilic matrix (Quezada
Gallo et al., 2000). Thus, small particles and a high emulsion stability during the lm drying give rise to a homogeneous distribution
of the lipid particles in the lm, which in turn contributes to a
more efcient control of water transfer (Debeaufort et al., 1993).
Various homogenization techniques, e.g., the rotorstator
homogenization and microuidization techniques, may be used
to modulate lipid droplets in the lmogenic dispersions. Rotor
stator systems are often used in the food industry to homogenize
different media and high viscosity immiscible liquids, reaching
particle sizes in the range of 1 lm (Urban et al., 2006). High pressure homogenizers with process pressures of up to 100 MPa are
commonly used to reduce the size of lipid particles, giving rise to
particles much smaller than 1 lm. Nowadays, novel homogenizers

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W. Ma et al. / Journal of Food Engineering 113 (2012) 136142

such as microuidizers can produce pressures of 200 MPa or higher, providing dispersions or emulsions with narrower particle distributions due to the high shear stresses developed in the
microchannels of the interaction chamber (Sherwin et al., 1998;
Strawbridge et al., 1995). The presence of a large number of uniformly dispersed spherical particles in lms may provoke an increase of the tortuosity factor for mass transfer in the continuous
matrix. However, there are few studies about the effects of various
homogenization techniques, especially the microuidizer on the
properties of lm-forming dispersions as well as the related lms.
Of vegetable oils, olive has received the most attention in meat
products reformulation, chiey as a source of MUFAs (mono unsaturated fatty acids). Olive oil has a high biological value due to the
presence of a favorable mix of predominantly MUFAs and natural
antioxidants, and olive oil intake is associated with various positive
health benets in human health (Lpez-Miranda et al., 2006). Thus,
the objective of this work is to develop gelatin/olive oil emulsied
lms through emulsication technique to improve water barrier
ability of gelatin-based lms. The effects of homogenization conditions of the FFD on lipid droplets size in lmogenic dispersions and
emulsied lms were evaluated and compared. Some selected
physical properties, e.g., water vapor permeability (WVP), microstructure of the lms were also evaluated. In addition, the possible
relationship between some selected physical properties and microstructure (lipid droplets size) of emulsied lms will also be
discussed.

Table 1. The temperature of the FFD increased to 5060 C after

microuidization treatment, in a pressure or cycle dependent manner. The control samples were prepared using the same mentioned-above procedure, except without addition of olive oil.

2. Materials and methods

Following degassing under vacuum, the lm-forming solution

was cast onto rimmed, leveled glass plates coated with polyethylene lms (Clorox China Co. Ltd., Guangzhou, China). The lm thickness was controlled by casting the same solids (2.8 g) on each plate
(18  20 cm). The castings were air-dried in a thermostatic and
humidistatic chamber [25 1 C, 50 5% relative humidity (RH)]
for 48 h, then the lms were peeled off the plates, and various
specimens for physical property testing were cut. Specimens of
2.5  8 cm rectangular strips were for tensile testing, and
2  2 cm squares for moisture content (MC) and total soluble mass
(TSM) measurements.

2.1. Materials
Bovine hide gelatin type B (Bloom 150), Glycerol (98% reagent
grade) was purchased from Sigma Chemical Co. (St. Louis, MO,
USA). Original olive oil was purchased from a local supermarket
(Guangzhou, China). All other chemicals used were of analytical
or better grade.
2.2. Preparation of lm-forming emulsions
The lm-forming dispersions (FFD) contained 5% (w/w) gelatin
and glycerol was used as a plasticizer by using a glycerol-to-protein ratio of 0.4:1 (w/w). The lipid-to-protein ratio in the emulsied lms was 0.2. So, the mass fraction of solid components in
the lms was: 62.5% gelatin, 25% glycerol, and 12.5% lipid. The
pH of the solution was appropriately adjusted to 8 with 0.1 M
NaOH. The FFD was heated (60 C) to favor the oil dispersion in
the system. The emulsication process was performed by using a
rotorstator homogenizer (Ultraturax T25, Janke & Kunkel, Germany) in a rst step (at about 40 C). Some FFD were homogenized
in a second step using a M-110EH microuidizer (Microuidics
International Corp., Newton, Massachusetts, USA) as described in

2.3. Particle size distribution of the lm-forming dispersions

The particle size distribution of the FFD was measured by using
a laser diffractometer (MasterSizer 2000, Malvern Instruments,
UK). The samples were diluted in de-ionized water at 2000 rpm
until an obscuration rate of 10% was obtained. The Mie theory
was applied by considering the following optical properties for olive oil droplets: a refractive index of 1.46. Each FFD was measured
in triplicate. Weight mean diameter (D4,3) and volume-surface
mean diameter (D3,2) were obtained according to eqs. (1) and (2).

P
4
ni di
D4;3 P
3
ni di

P
3
ni di
D3;2 P
2
ni di

ni is the number of droplets of a determined size range and di is the

droplet diameter.
2.4. Preparation of lms

2.5. Moisture content (MC) and total soluble mass (TSM)

MC of lms was determined by oven-drying at 105 2 C for
24 h, and expressed as the percentage of initial lm weight lost
during drying. TSM of each lm was determined as the percentage
of lm dry matter solubilized after 24 h immersion in de-ionized
water. Three randomly selected 2  2 cm samples from each type
of lm were used to determine the TSM. The lm specimens were
placed in a 50 mL cuvette containing 30 mL of de-ionized water.
The cuvettes were covered with polyethylene lms (Clorox China
Co. Ltd., Guangzhou, China) and incubated in a shaking water bath
at 25 C for 24 h with gentle vibrating. Undissolved dry matter was

Table 1
Volume-surface mean diameter (D3,2) and weight mean diameter (D4,3) of gelatin/olive oil lm-forming dispersion (FFD) as affected by homogenization conditions.
Method

First stepA

Second stepB

D3,2C

D4,3C

A
B
C
D
E
F

4 min
4 min
4 min
4 min
4 min

1
3
1
3

3.090 0.005a
0.570 0.007c
0.396 0.007e
0.433 0.003d
1.621 0.003b

13.99 0.044a
0.871 0.002d
0.931 0.001c
0.667 0.003e
3.680 0.023b

10000 rpm
10000 rpm
10000 rpm
10000 rpm
10000 rpm

Cycle at 69 MPa
Cycles at 69 MPa
Cycle at 138 MPa
Cycles at 138 MPa

Superscript letters (a-e) indicate signicant (P < 0.05) difference within the same column.
A
The FFD was homogenized by rotorstator homogenizer.
B
The FFD was homogenized by a M-110EH microuidizer.
C
Values were expressed as means and standard deviations of triplicate measurements.

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W. Ma et al. / Journal of Food Engineering 113 (2012) 136142

determined by removing the lm pieces from the beakers, gently

rinsing them with de-ionized water, and then oven-drying the
rinsed lms (105 C, 24 h).
2.6. Tensile strength (TS) and elongation at break (EAB)

2.11. Modulated differential scanning calorimetry (MDSC)

TS and EAB were measured using a TA-XTplus texture analyzer

(Stable Micro Systems, London, UK). Samples were preconditioned
at 25 C and 50 3% RH in a desiccator containing magnesium nitrate saturated solution [Mg(NO3)26H2O] for at least 2 days prior
to analysis. Initial gap separation and cross-head speed were set
at 50 mm and 1 mm/s, respectively. TS was calculated by dividing
the maximum load at break by initial specimen cross-sectional
area (ASTM, 2001). EAB was calculated by dividing the extension
at break of the specimen by the initial gage length of the specimen
(50 mm) and multiplying by 100 (ASTM, 2001). Each trial was replicated at least eight times, and the averages were taken as the
data.
2.7. Water vapor permeability (WVP)
A modication of the ASTM E96-95 gravimetric method (1995)
was employed to measure WVP of exible lms (McHugh et al.,
1993). Films samples were chosen for WVP testing based on a lack
of physical defects such as cracks, bubbles or pinholes. Distilled
water (15 mL) was placed in each test cup to expose the lm to
high relative humidity on one side through a circular opening of
4.5 cm diameter. Once the lms were secured, the test cups containing lms were inserted into the pre-equilibrated 0% RH desiccator cabinets and a fan was placed over the cups to ensure
maintenance of moisture conditions of the lms surface. Within
two hours, steady state had been achieved, the cups were weighed
periodically using an analytical balance (0.0001 g), ve weights
were then taken for each cup at >3h intervals.WVP was calculated
according to the WVP method of McHugh et al. (1993).
2.8. Light transmission and opaque
The barrier properties of lms against ultraviolet (UV) and visible light were measured according to the method described by
Limpisophon et al. (2009), with some modications. Film opaque
was calculated by the following equation:

Film opaque A=mm logT=x

slips, and examined with a 100 magnication lens and an argon/

krypton laser having an excitation line of 514 nm and a Helium
Neon laser (HeNe) with excitation at 633 nm.

where A = absorbance at each wavelength; T = transmittance (%) at

each wavelength; x = lm thickness (mm).
2.9. Scanning electron microscopy
The surface of the lms was analyzed using a TM-3000 scanning
electron microscope (Hitachi, Japan) in the charge-up reduction
mode at an accelerating voltage of 15 kV. The pieces of the lms
were attached to the SEM aluminum sample plate using a double-sided tape.
2.10. Confocal laser scanning microscopy (CLSM)
CLSM observations of the emulsions were performed on a Leica
TCS SP5 Confocal Laser Scanning head mounted on a Leica DMRE-7
(SDK) upright microscope (Leica Microsystems Inc., Heidelberg,
Germany) equipped with a 20HC PL APO/0.70NA oil immersion
objective lens. The various emulsion samples were stained with
an appropriate amount of 1.0% (w/v) Nile Blue A (uorescent
dye) in distilled water. The stained emulsions were placed on concave confocal microscope slides (Sail; Sailing Medical-Lab Industries Co. Ltd., Suzhou, China), covered with glycerol-coated cover

The glass transition temperature (Tg) and helixcoil transition

temperature (melting point Tm) of the lms were determined by
MDSC experiments, using a TA Q100-DSC thermal analyser (TA
Instruments, New Castle, DE, USA), according to the method of Bell
and Touma (1996), with slight modication. The enthalpy (DH) of
the helixcoil transition was also calculated. Indium was used to
calibrate the equipment. Samples were preconditioned at 25 C
and 50 3% RH in a desiccator containing magnesium nitrate saturated solution for at least 2 days prior to analysis. The lms samples (78 mg, with 0.1 mg accuracy) were hermetically sealed in
solid pans, and an empty pan was taken as the reference. The scan
rate was 5 C/min over an appropriate temperature range: equilibrate at 80 C, modulate 1.0 C every 60 s, isothermal during
5 min, heating at 5 C/min to 150 C. The peak temperature of the
endotherm on total heat ow proles was taken as the Tm. The Tg
was characterized on the reversing heat ow signals. The Tg, Tm
and DH were computed from the thermograms by the universal
analyser 2000, version 4.1D (TA Instrument-Waters LLC, New Castle, DE, USA). All experiments were conducted in triplicate.
2.12. Film thickness determination
Film thickness was measured with a digital micrometer (TAIHAI
apparatus Co. Ltd., Shanghai, China) to the nearest 0.001 cm. Measurements were taken along the length of the specimen ve times,
and the mean values were used to calculate lm tensile strength.
2.13. Statistics
An analysis of variance (ANOVA) of the data was performed
using the SPSS 13.0 statistical analysis system, and a least signicant difference (LSD) with a condence interval of 95 or 99% was
used to compare the means.
3. Results and discussion
3.1. Characterization of lms-forming solution
The size of the lipid particles in the lm-forming dispersions
(FFD) affects important characteristics of the lms, such as their
mechanical and barrier properties. The particle size distributions
of the FFD are plotted in Fig. 1, and the related D3,2 and D4,3 values
of the FFD are also listed in Table 1. The FFD coded as A in Table 1
has no oil in the lm formulation. The FFD homogenized by the RS
homogenizer showed a bimodal particle size distribution, and two
maxima occurred close to 1 and 7 lm in volume distributions. This
distribution changed drastically when the FFD were submitted to
microuidization treatment, depending on the microuidization
pressure or cycles. The FFD homogenized by method C or E
exhibited nearly monomodal particle size distributions. Unexpectedly, the FFD obtained by method F recovered a bimodal particle
size distribution (Fig. 1). Those results the moderate microuidization treatment (e.g., pressure or cycle) can provide the FFD with
narrower particle size distributions, while high process pressure
or cycles made make dispersed lipid particles more susceptible
to occulation phenomena. Similar results were reported by Jafari
et al. (2008), who reviewed the re-coalescence of emulsion droplets during high-energy emulsication and pointed out that if the

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Relative volume fraction (% v/v)

W. Ma et al. / Journal of Food Engineering 113 (2012) 136142

B
C
D
E
F

10
8
6
4
2
0
0.01

0.1

10

100

1000

Particle size( m)
Fig. 1. Particle size distribution of lm-forming dispersion containing olive oil as a
function of homogenization conditions.

process pressures are too high, they tend to make dispersed particles more susceptible to occulation phenomena.
The D3,2 and D4,3 of the FFD was a function of the energy input.
The D3,2 and D4,3 of the FFM by the RS homogenizer were 3.090 lm
and 13.99 lm, respectively. The lipid droplets parameters (D3,2 and
D4,3) of the FFD showed a markedly reduced trend when the microuidizer was used in a second step (Table 1). In fact, among the
process techniques used, the rotorstator homogenizer provides
a lower energy input and so produced the largest particles (method
B). Both microuidization pressure and cycle times affected lipid
droplets distribution in the FFD. When the homogenization pressures increase from 69 to 138 Mpa, the D3,2 and D4,3 of the FFD
exhibited a distinct trend for different microuidization cylces. In
1 cycle case, the D3,2 and D4,3 of the FFM decreased (methods C
and E). In 3 cycles case, the D3,2 and D4,3 of FFD, on the contrary,
increased signicantly (methods D and F). Moreover, the effects of the microuidization cycles on the lipid droplets distribution were also dependent on microuidization pressures. Upon
increasing cycles, the D3,2 and D4,3 decreased at the lower pressure
(69 MPa), whereas increased at the higher pressure (138 MPa). The
mean diameter data further supported that high process pressure
or cycles made dispersed lipid particles more susceptible to occulation phenomena due to the incomplete coating of the generate
interfacial surface with the stabilizing polymer (gelatin), giving rise
to bridging occulation.
3.2. Characterization of the lms
3.2.1. MC and TSM
Packaging lms should maintain moisture levels within the
packaged product. Therefore, the knowledge of water solubility
and moisture content of the lm is very important for food packaging applications. MC and TSM of gelatinolive oil emulsied lms
as a function of homogenization conditions are shown in Table 2.

The MC values signicantly decreased due to the inclusion of olive

oil. This is due to the hydrophobic nature of olive oil. Similar results were observed for pistachio globulin protein and fatty acids
emulsied lms (Zahedi et al., 2010). Partial proteinwater interactions replaced with proteinolive oil interactions may account
for MC reduction occurred in emulsied lms. The method B
exhibited the lowest MC in all preparations (Table 2). This phenomenon may be attributed to the formation of oil layer on the
surface of emulsied lms.
Solubility of a lm in water is an important property of edible
lms, and water insolubility or resistance is usually required for
a potentially commercial lm. Water solubility is an indication of
a lms hydrophilicity. After 24 h incubation in de-ionized water,
the gelatinolive oil emulsied lms were found to still maintain
their integrity, while the control (gelatin) lm changed its shape.
The TSM of emulsied lms decreased with increasing pressures
or cycle times (Table 2). Perez-Mateos et al. (2009) also reported
that the water solution (WS) of cod gelatin lms decreased due
to the incorporation of sunower oil. In contrast, Gontard et al.
(1994) found that the addition of lipids to a gluten lm caused
an increase in its WS. They suggested that the breakage of strong
intermolecular bonds in the protein network and the formation
of brittle bonds with lipids resulted in structural instability and
made the emulsied lm be more susceptible to dissolution. The
present results supported that the inclusion of olive oil in gelatin
lms had little effect on their structural integrity incorporation.
3.2.2. Scanning electron microscopy (SEM) and confocal scanning laser
microscopy (CSLM)
The lm surface microstructure, analyzed through SEM, is
shown in Fig. 2A. The control (gelatin) lms had dense, homogeneous and smooth structure without grainy and porous structure.
The emulsied lms had the irregular surface with the distribution
of oil droplets, especially the lms prepared by the RS homogenizer
(Fig. 2A). When the microuidizer was used in a second step, the
irregularity (e.g., caves, holes) of the emulsied lms decreased
markedly. The application of high pressure homogenization promoted relevant changes in the lm microstructure even at the lower pressure (69 MPa, 1 cycle), since differences between both
phases became less marked after the application of the microuidization. Lipid particles become embedded in the network and
the smaller particles made them more closely connected with
the polymer. Interestingly, the surface microstructure of the emulsied lm is a function of homogenization conditions. The irregularity (e.g., caves, holes) of the lms showed a descending trend as
the average lipid droplets diameter (D3,2, D4,3) in the FFD decreased
upon various homogenization treatments (Fig. 1A, Table 1), indicating that the lower lipid droplets in the FFD, the less irregularity
of the lms surface.
The combination of the principles of measuring using uorescence and electronic microscopy allows visualizing the morphological structure of the lms, including the distribution of the
constituents in the lmogenic matrix. Fig. 2B shows the CLSM

Table 2
Tensile properties, water vapor permeability and surface hydrophobicity of gelatin lms containing olive oil as a function of the homogenization method.
Method
A
B
C
D
E
F

MC(%)

TSM(%)
a

19.3 0.8
11.5 0.2d
12.2 0.1c
12.9 0.4c
12.4 0.2c
13.64 0.12b

44.7 6.1
39.3 2.9b
37.5 2.9b
33.2 0.6c
33.4 0.9c
31.8 0.2d

WVP  1010gs1m1Pa1

TS (MPa)

EAB (%)

5.61 0.07a
4.22 0.0293c
4.67 0.16b
4.19 0.04c
3.96 0.23cd
3.74 0.11d

17.1 2.6b
12.0 1.7c
24.2 2.5a
21.3 2.8a
18.2 1.4b
23.2 3.0a

39.5 7.9c
50.5 8.1c
50.5 4.7c
106.5 8.3a
58.4 9.2b
106.7 4.7a

Values were expressed as means and standard deviations of triplicate measurements.

Superscript letters (a-d) indicate signicant (P < 0.05) difference within the same column.

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W. Ma et al. / Journal of Food Engineering 113 (2012) 136142

Fig. 2. Microstructure of gelatinolive oil emulsied lms as a function of homogenization conditions. Panel A: SEM observations of surface of the emulsied lms. Panel B:
CLSM images of lms. Letters af within the gures represented the homogenization methods as shown in Table 1.

images of gelatin or emulsifying lms. The distribution of olive oil

(green) in the lm matrix was dependent on the homogenization
techniques used. CLSM showed a marked decreasing in lipid droplets size in lms matrix when the microuidizer was used in a second step as compared with the lms prepared by the RS
homogenizer, and the olive oil was non-homogenously distributed
as discrete droplets within the continuous gelatin phase (red).
Interestingly, the lipid droplets size of emulsied lms presented a similar trend as they did in the FFD upon various microuidization treatments. e.g., when the homogenization pressures
increase from 69 to 138 Mpa, the lipid droplets decreased for 1 cycle case, whereas the lipid droplets increased for 3 cycles case
(Fig. 2B). This result was also supported by SEM micrographs
(Fig. 1A). CLSM data further indicated that high process pressure
or cycles made dispersed particles more susceptible to occulation
phenomena during the drying process of lms.

3.2.3. Tensile properties

Table 2 shows tensile parameters (TS: tensile strength, EAB:
elongation at break) of gelatin lms with or without olive oil.
The TS and EAB of lms was dependent on the special homogenization conditions. The TS of lms produced the RS homogenizer
treatments decreased as compared gelatin alone lms. The reduced
continuity and cohesion of the protein network due to the large lipid globules within lms matrix might be the reason of this behavior. When the microuidizer was used in a second step, the TS of
the lms increased (Table 2). Since olive oil constituted a nonmiscible emulsied phase in the FFD, the protein chains segregated
to form a protein-rich phase. The microuidization treatment favored proteinprotein interactions to a great extent and, as a consequence, higher TS were observed. These observations are
consistent with the results of proteincinnamon oil and gelatin
corn oil lms systems (Atars et al., 2010; Wang et al., 2009).

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W. Ma et al. / Journal of Food Engineering 113 (2012) 136142

Table 3
Light transmission (%) and transparency (A600/mm) values for the control lms and gelatin olive oil lms as a function of homogenization conditions.
Sample
A
B
C
D
E
F
LDPEA
A

200

280
a

4.33 0.11
3.10 0.09c
4.30 0.11a
3.24 0.23c
3.23 0.07c
3.65 0.12b
0.45

46.43 1.54
5.80 0.90b
2.35 0.28c
2.52 0.06c
2.56 0.06c
3.04 0.13c
27.65

400
a

500
a

87.46 0.68
15.11 0.88b
8.40 0.42e
11.14 0.83d
12.52 0.42c
15.36 0.42b
39.98

600
a

90.26 0.57
27.68 2.89c
19.72 0.54e
28.50 0.85c
30.90 0.96b
25.59 0.65d
45.53

800
a

91.03 0.39
22.38 0.89f
28.73 0.31e
34.51 1.01c
43.30 1.21b
32.52 0.89d
49.81

A600/mm
a

91.48 0.73
28.22 1.70e
40.99 0.96d
48.34 1.00c
59.82 0.37b
41.80 0.72d
56.33

0.53 0.03f
9.85 0.23a
6.37 0.06b
5.57 0.16d
4.54 0.16e
5.95 0.15c
4.26

LDPE, low-density polyethylene (data from Guerrero et al., 2011).

Similar results were also found by Fabra et al. (2009), when adding
different lipids to casein lms.
The EAB increased as the microuidization pressure or cycle increased, producing more extensible lms (Table 2). This effect may
be attributed to the promotion of proteinlipid interactions when
homogenization conditions were more intense, and some small lipid particles became embedded in the protein network, which
seemed to have some plasticizing effect on the lms. Similar results were observed by Atars et al. (2010), cinnamon oil had some
plasticizing effect on the SPI lms, making the lms more extensible as the oil content increases.
3.2.4. Water vapor permeability
The WVP of the lms are shown in Table 2. The WVP signicantly decreased due to the inclusion of olive oil, regardless of
homogenization conditions. Unexpectedly, the lms prepared by
method B exhibited good water resistance (Table 2). In fact, the
RS homogenizer provides a lower energy input and so produced
the largest particles in the related FFD (Table 1) and lipid droplets
with the diameters about 930 lm appeared in the CLSM images of
the lms (Fig. 2B). This behavior may be associated with the formation of analogous proteinlipid bi-layer structure, evidenced by
SEM data (Fig. 2). In fact, hydrocolloid-lipid bilayer lms showed
better barrier properties (Fabra et al., 2011).
When the microuidizer was used in a second step, the WVP of
the lms decreased from 5.61  1010 gm1 s1 Pa1 (control) to
(3.744.67)  1010 gm1 s1 Pa1 (emulsied lms). Furthermore,
the WVP of the lms decreased upon increasing microuidization
pressure or cycle, and the lms obtained by method F exhibited
the best water resistance (Table 2). However, high process pressure
or cycles made dispersed particles more susceptible to occulation
phenomena, with subsequent formation of the agglomerates of lipid droplets in the FFD and the obtained lms (method F) (Fig. 1
and Fig. 2B). The reason for this abnormal response of WVP to the
lipid droplets was unknown. However, this may be associated with
the fact that the intensive microuidization treatment may enhance the interactions between oil phase and hydrophobic sections
of protein molecule due to the high shear stresses developed in the
microchannels of the interaction chamber. This interaction may
weaken the hydrophilic properties of the continuous protein matrix, with subsequent decrease in the WVP.
3.2.5. Light absorption
Transparency of lms designed for packaging is a relevant property since it has a direct impact on the appearance of the coated
product. Table 3 shows light transmission values at wavelengths
from 200 to 800 nm and transparency for lms, in comparison to
a commonly used synthetic lms (LDPE). Light transmission of
lms decreased at almost all wavelengths due to the inclusion of
olive oil, indicating that the lms are less transparent. Concomitantly, the addition of olive oil produced a sharp increase in the
lm opaque related to the control lm (Table 3). However, lm
opaque decreased from 9.85 to 4.546.37 when the microuidizer

was used in a second homogenization step (Table 3). Transparency

of lms is an auxiliary criterion to judge the compatibility of the
components. If compatibility between lm constituents is not
good, then the light transmittance is low due to light dispersion
or reection at the two-phase interface (Rhim et al., 2007). Since
olive oil constituted a non-miscible emulsied phase in the emulsied lms (Fig. 2B), differences between both phases became less
marked after the application of the microuidization, with
subsequent decrease in lm opaque. Interestingly, lm opaque
presented a decreasing trend when the lipid droplets in the FFM
and lms decreased. Moreover, the opaque values for the lms
prepared by microuidization techniques were similar or slight
higher than the values of LDPE, a commercial lm used for packaging purposes (Table 2).
It is also noteworthy that the emulsied lms showed excellent
barrier ability to UV light at 280 nm in comparison with the gelatin
or LDPE lms (Table 2). Some plant protein lms (e.g., SPI lms)
showed excellent barrier properties to UV light in the range of
200280 nm due to the presence of aromatic amino acids (e.g.,
tyrosine, phenylalanine and tryptophan) (Li et al., 2004). In contrast, gelatin lms showed poor barrier ability to UV light at
280 nm, this may be attributed to the scarcity of aromatic amino
acids in gelatin. Olive oil intake has been associated with various
positive health benets in human health due to the presence of
some antioxidants (Lpez-Miranda et al., 2006) providing with
strong UV absorption. Those results suggested the potential preventive effect of gelatinolive oil lms on the retardation of product oxidation induced by UV light.
3.2.6. Modulated differential scanning calorimetry
Fig. 3 shows the typical MDSC thermogram of gelatin lms with
or without olive oil. The total heat signal of gelatin lm was characterized by the presence of an endothermic peak (Tm) at about
65.8 C, attributed to the melting of triple-helix crystalline structure of gelatin. The thermogram for the lms with olive oil showed
an additional endothermic peak at 8 C, which can be attributed
to the melting peak of olive oil. This is consistent with its nonhomogenous incorporation of olive oil into the protein matrix, as
evidenced by SEM micrographs and CLSM images (Fig. 2). Concomitantly, the inclusion of olive oil in lms matrix resulted in the increase in the Tm of the helixcoil transition of gelatin (Fig. 3A).
Reverse heat ow signals of MDSC were used to characterize the
glass transition of the lms with or without olive oil. The glass
transition temperature (Tg) of gelatin lms was about 61 C and
this transition temperature tended to increase due to the incorporation of olive oil (Fig. 3B). Bell and Touma (1996) determined the
Tg of gelatin by separating it from other irreversible thermal
changes using MDSC technique, and reported that its Tg was about
60 C. Since olive oil constituted a non-miscible emulsied phase
in the FFD. Thus, the protein chains may segregate to form a protein-rich phase, which may favor proteinprotein interactions to
a great extent. As a consequence, the emulsied lms possessed
higher Tm and Tg in comparison with those of the lms without oil.

142

W. Ma et al. / Journal of Food Engineering 113 (2012) 136142

Total heat flow (w/g)

G-film
RS-film
M-film

-1
-2
-3
-4
-50

-25

25

50

75

100

Reversable heat flow(w/g)

Temperature (C)
-0.4

G-film
RS- film
M-film

-0.8

-1.2

-1.6

-2.0
0

25

50

75

100

Temperature (C)
Fig. 3. DSC proles of gelatin and gelatinolive oil lms. Panel A: total heat ow.
Panel B: reverse heat ow. G-lm: gelatin lm. RS-lm: the lm prepared from the
FFD by method B using a RS homogenizer. D-lm: the lm prepared from the FFD
by method D using a rotorstator homogenizer and a M-110EH microuidizer.

4. Conclusion
Homogenization techniques including the RS homogenizer and
microuidizer were used to prepare lmogenic dispersions. The RS
homogenizer provided a lower energy input and so produced the
largest particles, olive oil molecules tended probably to form
bi-layer on air side of lms. These lms exhibited excellent water
barrier ability, but poor mechanical resistance, extensibility and
transparency. The microuidizer can provide lmogenic dispersions with narrower particle size distributions, promoting
mechanical resistance, extensibility and transparency of the lms.
Concomitantly, the WVP decreased upon increasing pressure or
cycle. In addition, the obtained gelatinolive oil lms exhibited
excellent barrier ability to UV light. The transparency values and
surface irregularity of the lms presented a reduced trend when
lipid droplets in the FFD and lms decreased. Thus, the present
results indicated that the microuidizer appeared to be a suitable
approach to modulate lipid droplets distribution of lmogenic
mitrix, thus the lms properites, and to develop edible lms
adapted to a specic target application.
Acknowledgements
The authors thank for the nancial support from the Natural
Science Fund of Guangdong Province, China (serial number:10451064101005149), the Fundamental Research Funds for
the Central Universities (SCUT, 2012ZZ0082) and the Open Project
Program of State Key Laboratory of Pulp and Paper Engineering,
South China University of Technology (No.201023).

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