Materials Science and Engineering C 33 (2013) 1819–1841

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Materials Science and Engineering C

journal homepage: www.elsevier.com/locate/msec

Review

Chitosan based edible films and coatings: A review

Maher Z. Elsabee a,⁎, Entsar S. Abdou b, c
a
Chemistry Department, Faculty of Science, Cairo University, Giza 12613, Egypt
b
Food Packaging and Engineering Department, Food Technology Research Institute, Agriculture Research Center, Giza 12613, Egypt
c
Chemistry Department, Faculty of Science and Humanities, Salman bin Abdulaziz University, Hotet Bany Tamim 11941, Saudi Arabia

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

Article history: Chitosan is a biodegradable biocompatible polymer derived from natural renewable resources with numer-
Received 9 August 2012 ous applications in various fields, and one of which is the area of edible films and coatings. Chitosan has
Received in revised form 11 December 2012 antibacterial and antifungal properties which qualify it for food protection, however, its weak mechanical
Accepted 9 January 2013
properties, gas and water vapor permeability limit its uses. This review discusses the application of chitosan
Available online 18 January 2013
and its blends with other natural polymers such as starch and other ingredients for example essential oils,
Keywords:
and clay in the field of edible films for food protection. The mechanical behavior and the gas and water
Chitosan vapor permeability of the films are also discussed. References dealing with the antimicrobial behavior of
Blends these films and their impact on food protection are explored.
Essential oils © 2013 Elsevier B.V. All rights reserved.
Nanoclay
Water vapor and gas permeability
Antibacterial and antifungal properties

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1820
2. Chitin and chitosan . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1820
2.1. Blending . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1820
2.1.1. Blending of chitosan with starch . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1821
2.1.2. Chitosan and gelatin based edible films . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1826
2.1.3. Chitosan with alginate and carageenan . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1827
3. Chitosan/essential oil films . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1827
4. Chitosan and clay . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1830
5. Gas permeation properties of edible coatings . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1831
6. Effect of electric field on film formation . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1833
7. Antibacterial activity of chitosan and chitosan blends . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1833
8. Increasing the shelf life of foods . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1837
9. Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1839
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1839

Abbreviations: DD, degrees of deacetylation; EMC%, Equilibrium moisture content; CH, high molecular weight chitosan; CM, medium molecular weight chitosan; CL, low molecular
weight chitosan; MD, machine direction; TD, transverse direction; TS, tensile strength; CS, chitosan; Ec, elastic modulus; E, Elongation at the break; BCC, bacterial cellulose–chitosan; BC,
bacterial cellulose; FS, Film solubility; MW, molecular weight; WVP, water vapor permeability; M, (1→ 4)-linked-β-D-mannuronate; G, (1 →4)-linked-α-L-guluronate; CEO, cinnamon
essential oil; TVC, total viable counts; BO, Bergamot essential oil; FFD, Film-forming dispersions; TTO, tea tree oil; PCL, poly-caprolactone; PBS, poly(butylene succinate); PLA, poly(lactic
acid); PBTA, poly(butylene terephthalate adipate); PBSA, poly(butylene succinate adipate); WVT, Water vapor transmission rate; MC, methylcellulose; AFM, Atomic force microscopy;
GO, Garlic oil; PS, potassium sorbate; N, nisin; AM, antimicrobial; LPSSD, low-pressure superheated steam drying; PDA, potato dextrose agar; CWC, Chinese water chestnut; ACS,
acid-soluble chitosan; WCS, water-soluble chitosan; PVDC, polyvinyl dichloride; TVC, total viable counts; MAP, modified atmosphere packaging; CMC, carboxymethyl-cellulose.
⁎ Corresponding author. Tel.: +20 26352316, +20 1006680474 (mobile).
E-mail address: mzelsabee@yahoo.com (M.Z. Elsabee).

0928-4931/$ – see front matter © 2013 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.msec.2013.01.010
1820 M.Z. Elsabee, E.S. Abdou / Materials Science and Engineering C 33 (2013) 1819–1841
1. Introduction
Recently, considerable research has been conducted to develop and
apply bio-based polymers made from a variety of agricultural commod-
ities and/or of food waste products [1]. This increased interest was in-
tensified due to concerns about limited natural resources of the fossil
fuel reserve and the environmental impact caused by the use of non-
biodegradable plastic-based packaging materials [2]. Such biopolymers
include starches, cellulose derivatives, chitosan/chitin, gums, proteins
(animal or plant-based) and lipids [3]. These materials offer the possi-
bility of obtaining thin films and coatings to cover fresh or processed
foods to extend their shelf life.
Edible films and coatings offer extra advantages such as edibility,
biocompatibility, esthetic appearance, barrier to gasses properties,
non-toxicity, non-polluting and its low cost [4]. In addition, biofilms
and coatings, by themselves or acting as carriers of foods additives
(i.e.: antioxidants, antimicrobials), have been particularly considered
in food preservation due to their ability to extend the shelf life [5].
Seafood products are more perishable than chicken or red meat as
they contain relatively large quantities of free amino acids and vola-
tile nitrogenous bases compared with other meats [5]. During storage,
fish quality is quickly reduced as chemical and enzymatic reactions
lead to the initial loss of freshness, while microbial spoilage produces
the end of the shelf life. The increasing demand for high quality fresh
seafood has intensified the search for new methods and technologies
for better fish preservation. One of the possibilities, not much
explored, is the application of an edible film or coating, in combina-
tion with other microbial stress factors, on the fresh fish muscle.
Gomez-Estaca et al. [6] reported that a gelatin–chitosan-based edible
film together with refrigeration and high pressure have lowered the
microbial growth of cold-smoked sardine in comparison to uncoated
samples.
Over the past several decades, several biopolymers have received
increased attention for their applications in chemical, biomedical,
and food industries, [7]. For example, chitin suture is resorbable in
human tissues from which chitosan–collagen composites for an artifi-
cial skin are commercially produced [8]. These polymers are not only
biodegradable, but also edible. Another area of growing interest is the
preparation of antimicrobial edible films and coatings [2,9] where
chitosan plays an important role due to its well-documented antimi-
crobial properties [10].
2. Chitin and chitosan
Chitin is an abundant naturally occurring biopolymer and is found in
the exoskeleton of crustaceans, in fungal cell walls and in other biolog-
ical materials. It is mainly poly(β-(1–4)-2-acetamido-D-glucose), which
is structurally identical to cellulose except that a secondary hydroxyl on
the second carbon atom of the hexose repeat unit is replaced by an acet-
amide group. Chitosan is derived from chitin by deacetylation in an
alkaline media [11]. Actually, chitosan is a copolymer consisting of
β-(1–4)-2-acetamido-D-glucose and β-(1–4)-2-amino-D-glucose units
with the latter usually exceeding 60%. Chitosan is described in terms
of degree of deacetylation and average molecular weight and its impor-
tance resides in its antimicrobial properties in conjunction with its
cationicity and film-forming properties.
The potential of chitosan to act as a food preservative of natural origin
has been widely reported on the basis of in vitro trials as well as through
direct application on real complex matrix foods [12–17]. Chitosan is also
an excellent film forming material [18]. Chitosan films have a selective
permeability to gasses (CO2 and O2) and good mechanical properties.
However, the fact that chitosan films are highly permeable to water
vapor limits their use as being an important drawback since an effective
control of moisture transfer is a desirable property for most foods, espe-
cially in moist environments. Therefore, several strategies have been
used to improve the physical properties of biopolymer based films.
Among them, promising results in increasing the hydrophobicity have
been obtained by addition of neutral lipids, fatty acids waxes [19,20]
and clay [21] although often compromising their mechanical and chem-
ical stability and/or of their organoleptic attributes. Moreover, various
chemical and physical means have been demonstrated as good strategies
to improve their mechanical properties, such as the addition of
cross-linking agents, irradiation and ultrasonic treatments [22].
The antifungal and antimicrobial activities of chitosan are believed to
originate from its polycationic nature [23]. The antimicrobial action of
chitosan is hypothesized to be mediated by the electrostatic forces
between the protonated amino group (NH2) in chitosan and the negative
residues at cell surfaces [24]. The number of protonated amino groups
(NH2) present in chitosan increases with increased degrees of
deacetylation (DD) which influences the antimicrobial activity [25]. Liu
et al. (2004) [25] state that the bactericidal activity of chitosan is caused
by the electrostatic interaction between NH3+ groups of chitosan and the
phosphoryl groups of the phospholipid component of the cell mem-
brane. However, it was found that water-soluble chitosan promoted
the growth of Candida albicans even in acidic media whereas water-
insoluble chitosan exhibited inhibitory effect [26]. In addition, a strong
interaction between microbial proteins and chitosan at very acidic pH
values is low and adsorption of chitosan to Escherichia coli cells increased
strongly with increasing pH. This means that the protonated NH3+ is not
the predominant factor in the antibacterial capacity of chitosan. Park et
al. suggested that the antimicrobial activity of chitosan is not proportion-
al to its DD value [27]. As the water soluble chitosan was not efficient as
antibacterial agent it is thus conceivable that chitosan molecules have
the ability to interact with bacterial surface compounds, and is absorbed
on surface of the cells. However, physiological pH in the cell is around
neutral, so water-insoluble chitosan molecules can precipitate, and
stack on the microbial cell surface, thereby forming an impervious
layer around the cell and blocking the channels, which are crucial for liv-
ing cells. Such a layer can be expected to prevent the transport of essen-
tial solutes and may also destabilize the cell wall beyond repair thereby
causing severe leakage of cell constituents and ultimately cell death
[26]. In support of this last idea, it was found that the cell membranes
of Gram-negative and Gram-positive bacteria showed significant mor-
phological changes and shrinking after contact with chitosan treated cot-
ton fabrics [28].
2.1. Blending
The functional properties of chitosan-based films can be improved
by combining them with other hydrocolloids [29–31]. Chitosan/pectin
laminated films have been developed by the interaction of the cationic
groups of chitosan with the anionic groups of pectin. A decrease in
water vapor transmission rates (WVTRs) by combining chitosan with
two thermally gelatinized corn starches has been observed [31].
An alternative way to improve the mechanical and physical proper-
ties of these biofilms is by combining proteins (e.g. milk proteins, soy
protein, collagen and gelatin) with polysaccharides (e.g. starches, algi-
nates, cellulose and chitosan). Chitosan–gelatin blend films have been
shown to be homogeneous due to the good miscibility between both
biopolymers [32–34] leading to improved material properties of the
blend films as compared to those obtained from the pure polymers.
This is explained by the formation of electrostatic interactions between
the ammonium groups of the chitosan and the carboxylate groups of
the gelatin. On the other hand, chitosan/soy protein blended mem-
branes [33], are not completely miscible. The blended membranes
became more brittle with increasing soy protein content, and showed
a rougher surface morphology, this is probably related to phase separa-
tion among blend components. Chitosan/sodium caseinate films have
also been studied; in this case no phase separation was observed due
to the complexation of the two polymers within the blend film matrix
[34]. Some polysaccharide–whey protein films have also been prepared
and characterized [35]. Also, the addition of Pullulan to a whey protein
 M.Z. Elsabee, E.S. Abdou / Materials Science and Engineering C 33 (2013) 1819–1841 1821

film has shown to decrease water vapor and oxygen permeability, al-
though these barrier properties got worse as the amount of polysaccha-
ride increased [36]. Chitosan–whey protein films have been prepared at
pH 6 with different protein concentrations, in the absence or presence
of transglutaminase as a cross-linking agent [37]. The chitosan was
the main film component and its amount was kept constant, the protein
was from a spray dried whey product still rich in lactose and the amount
of whey protein did not exceed the proportion of 1:9 (protein:chitosan)
in the final films. In more recent work [2], films of chitosan–whey pro-
tein blend with a high amount of protein has been prepared in order to
obtain a blend with new functionality out of the interaction of the cat-
ionic polyelectrolyte chitosan with protein. The study was aimed also
to prepare edible film-forming material with anti-microbial properties.

2.1.1. Blending of chitosan with starch

The main differences between starch and chitosan are the glucoside
linkage: α (1, 4) for starch and β (1, 4) for chitosan and, the hydroxyl
group of the second carbon is replaced by the amine group which
appears acetylated in the case of the natural polymer chitin.
Fernandez et al. [38] studied the physical stability and moisture
sorption of aqueous chitosan–amylase starch films plasticized with
polyols. They used high, medium and low molecular weight chitosan
with amylose-rich corn starch as a co-film former in the presence of
glycerol and i-erythritol.
In comparison to regular corn starch which contains approximately
28% amylose, Hylon VII is a corn hybrid containing approximately 70%
amylose. Since amylose is a linear polymer, it can closely align or associate
through hydrogen bonding. This characteristic of amylose is primarily re-
sponsible for the gelling and film-forming ability of starches. Since Hylon
VII contains more than twice as much amylose as regular corn starch it
can form more rigid gels and contribute to the formation of stronger
and tougher films. Chitosan–Hylon VII solutions plasticized with glycerol
or erythritol were prepared in a high-pressure reactor equipped with a
blade mixer.
The steady-state moisture in the starting materials was measured
after 9 days of storage of the samples at different relative humidity.
As seen in Fig. 1A, the moisture increase of low molecular weight chi-
tosan was lower than that of high and medium molecular weight chi-
tosan at a relative humidity of 95%. The moisture increase of Hylon VII
was lower than that of other starting materials from relative humidity
of 52–95%. The storage of the erythritol at a high humidity resulted in
a significant increase in water uptake, causing a liquefaction of the
substance even higher than that of the chitosan. Starch and chitosan
are hydrophilic and retain a considerable amount of water which
depends on the relative humidity. At least in chitosan, there exist
three predominant absorption sites such as the hydroxyl group, the
amino group, and the polymer chain end. The polymer chain end is
supposed to be composed of a hydroxyl or an aldehydic group [39].
Usually, the amine content increases with increasing molecular
weight. In the case of chitosan, the water is bound to the hydroxyl
group more strongly than to the amine group. Therefore, the release
of water molecules could preferentially occur via the amine group.
The crystallinity of various film samples plasticized with erythritol
started to increase after 2 months. The crystallinity of the films stored
at 25 °C/60% RH was higher than those of the respective films stored at
40 °C/75% RH. The diffraction pattern of the 40 °C/75% RH sample after
2 months has a strong amorphous background and only two reflections
of crystalline erythritol at about 24.6 and 28.3 (2θ). Until 3 months, the
diffraction pattern has a strong amorphous background and three reflec-
tions at about 19.6, 20.3, and 37.58 (2θ), while the diffraction patterns of
the 25 °C/60% RH samples after 2 and 3 months showed a slightly amor-
phous background and almost all of reflections of crystalline erythritol.
Bangyekan et al. [40] prepared chitosan-coated starch film by coating
chitosan solution on cassava starch film containing glycerol as a plasticiz-
er. A mixture of cassava starch dispersion (6% w/v in water) and glycerol
used as a plasticizer was heated at the starch gelatinization temperature
Fig. 1. Equilibrium moisture content (EMC%) of high molecular weight chitosan (CH),
medium molecular weight chitosan (CM), low molecular weight chitosan (CL), amy-
lose corn starch (Hylon VII) starting materials (A), and chitosan–Hylon VII films (B)
stored at different relative humidity for 9 days [38].
of 70 °C under stirring until viscous and transparent solution was
observed. After homogeneously mixing for 10 min this solution was
poured into 5 mm thickness acrylic mold with removable edge strips
and allowed to dry freely at room temperature. After air drying, the
edges of the mold were removed and four sides of the film were sealed
with adhesive tape to prevent the underneath of starch film from getting
contact with chitosan coating solution in the next step. Chitosan (varied
from 1 to 4% (w/v)) was dissolved in 2% (v/v) acetic acid solution then
filtered and poured onto the starch film and the coating was carried
out by an automatic film coater with wire bar coating rod. After coating,
the acrylic mold support containing chitosan-coated starch film was
taken out from the automatic film coater and stored at room tempera-
ture, allowing the coated film to dry for at least 72 h. Three surface prop-
erties of the obtained films were studied; gloss, transparency and
hydrophobicity.
Gloss is one of the esthetic factors enhancing general appearance
as well as consumer acceptance. The gloss values of chitosan-coated
starch films including free starch film and free chitosan film are
graphically shown in Fig. 2. The gloss values of free chitosan films
are found in the range of 132.5–145.6 units, indicating a highly glossy
film probably due to a smooth surface. On the other hand, the gloss
values of free starch films are between 52.4 and 60.1 units, reflecting
the likelihood of uneven film surface.
For the coated films, it can be seen that the gloss values increase
with an increase in the chitosan coating solution content. From the
results, only 1 wt.% chitosan coating solution brings about a signifi-
cant increase in gloss values of film compared with those of free
starch film. The optimum gloss value is achieved when applying
1822 M.Z. Elsabee, E.S. Abdou / Materials Science and Engineering C 33 (2013) 1819–1841
Fig. 2. The relationship between gloss and chitosan coating content of coated films
containing 2, 3, 4, 5, and 6 wt.% glycerol [40].
4 wt.% chitosan coating solution indicating a complete coverage by
chitosan layer with an increase in chitosan coating concentration. It
could be concluded that the smoother surface of chitosan film
enhanced the regularity of coated film surface leading to increasing
in the gloss value.
Transparency may be affected by various factors including film
thickness. The percent transmittance values of coated films including
starch and chitosan free films are presented in Fig. 3. The transmit-
tance of chitosan film is slightly higher than that of starch film. The
smoother surface combined with relatively more amorphous struc-
ture of chitosan film (from X-ray evidence) may be responsible for
this transparency change.
The surface hydrophilicity of the chitosan-coated starch film was
evaluated by means of contact angle determination. The contact angles
of coated films including starch and chitosan free films are shown in
Fig. 4.
When considering the particular glycerol content, the free starch
film exhibits the smallest contact angle. It can be clearly seen that
an increase in concentration of chitosan coating solution brought
about a significant increase in contact angle values of the coated
films. These results indicate that the wettability of the coated films
decreased with an increase in the chitosan coating concentration.
This phenomenon was attributed to the higher hydrophobicity of chi-
tosan surface layer, which was attributed to the role of available
Fig. 3. The relationship between % transmittance and chitosan coating content of coat-
ed films containing 2, 3, 4, 5, and 6 wt.% glycerol [40].
Fig. 4. Effect of chitosan coating contents on contact angles of coated films containing
2, 3, 4, 5, and 6 wt.% glycerol [40].
residual hydrophobic acetyl groups present in chitosan chain. This
finding suggests that chitosan film, with this particular degree of
deacetylation, was more hydrophobic than starch film.
Mechanical properties are very important for edible films and
coating to improve mechanical handling of foods [41] or pharmaceu-
tical products [42].
The effect of CS coating contents on the tensile properties of CS-coated
cassava starch films in both machine direction (MD) and transverse direc-
tion (TD) has been investigated [40]. As shown in Fig. 5, it was found that,
at individual glycerol content in the coated film, there is a significant
change of the tensile stress at maximum load and tensile modulus in
both the MD and TD upon increasing the amount of CS coating.
The tensile stress and tensile modulus values in MD were higher
than those values in TD. This is probably because polymer chain of
chitosan aligned along the MD of automatic film applicator during ap-
plying force to wire bar coater. The % elongation at break values in
both directions tended to be lower than that of the uncoated or free
starch films. In addition, % elongation at break in MD was found to
be lower than in TD. Therefore chitosan improves the tensile stress
at maximum load and tensile modulus, of coated starch films, where-
as % elongation at break tends to decrease. In addition, with increas-
ing CS coating concentration, a remarkable decrease in % elongation
at break compared to coated films containing 3, 4, 5, and 6 wt.% glyc-
erol may be attributed to the less plasticizing effect due to the mini-
mized concentration of plasticizer in starch base film, including the
effect of brittleness of chitosan film.
It is observed that there is a little effect on tensile strength of the
coated film containing 5 and 6 wt.% glycerol upon increasing CS
coating concentration. Fig. 6 presents the effect of CS coating contents
on tensile properties of coated film containing 5 wt.% glycerol in both
directions. At 6 wt.% glycerol, although the improved tensile proper-
ties of coated films in MD and TD are attributed to an increase in CS
coating concentration, the greatest tensile strength values obtained
from 4 wt.% CS coating are relatively low, i.e., in MD, the tensile stress
at maximum load and tensile modulus are about 1.2 and 11.7 MPa,
respectively. The % elongation at break in both directions of coated
films containing high glycerol content, especially in TD, tended to in-
crease with increasing the amount of chitosan coating.
Mathew and Abraham [43] modified starch–chitosan blend films
by incorporating ferulic acid, to find possible application as an edible
film or coating.
Ferulic acid ((E)-3-(4-hydroxy-3-methoxy-phenyl)prop-2-enoic
acid) is an abundant phenolic phyto-chemical found in plant cell wall
 M.Z. Elsabee, E.S. Abdou / Materials Science and Engineering C 33 (2013) 1819–1841 1823

Fig. 5. Effect of chitosan coating contents on tensile properties of coated films containing 5 wt.% glycerol. (A) Tensile stress at maximum load and % elongation at break. (B) Tensile modulus [40].

components, like many phenols, it is an antioxidant in the sense that it
is reactive toward free radicals such as reactive oxygen species, it also
may have pro-apoptotic effects in cancer cells, thereby leading to their
destruction and may be effective at preventing cancer induced by expo-
sure to the carcinogenic compounds. The starch–chitosan blend films
were fabricated by means of a casting/solvent evaporation method.
Ferulic acid was oxidized by adding different concentrations of ferulic
acid (25, 50, 75, and 100) to hydrogen peroxide solution (0.1%, v/v)
and kept at room temperature for 1 h under stirring.
Thermal analysis of the prepared films was conducted to investi-
gate the effect of ferulic acid on the stability of the films since this is
a vital property to be considered for their application in food and
pharmaceutical industry as the edible films may be subjected to
heat processes during their preparation, processing or consumption
[43]. The TGA curves of starch–chitosan control film, ferulic acid and
ferulic acid incorporated blend films are shown in Figs. 7 and 8. It
can be seen from these figures that incorporation of up to 100 mg
of ferulic acid did not affect the thermal stability.
The surface of the control blend films and ferulic acid incorporated
starch–chitosan films were found to be relatively smooth and
homogenous.
The tensile strength (TS) values of ferulic acid incorporated starch–
chitosan composite films have also been investigated [43]. Compared
to the control film, the TS values of the blend films increased to a
value of 62.71 MPa with the incorporation of ferulic acid at a concentra-
tion of 75 mg as shown in Fig. 9. However, there was a slight reduction
in strength as the ferulic acid concentration reached 100 mg. Increase in
TS could be due to the formation of a stable network on account of the
cross linkages introduced by ferulic acid. Ferulic acid can enhance the
cross linking between polysaccharides through several mechanisms;
through free radical mediated cross linking, by esterification with the
hydroxyl groups of chitosan and starch or by quinone-mediated reac-
tions [44]. There has been reports on the cross linking property of ferulic
acid in the preparation of edible films from soy protein isolate [44] and
gelatin [45]. The TS increased with the increase in the level of cross-
linking agent until the ratio of the ferulic acid/carbohydrate moiety
became too high. This may be attributed to the redundant hydroxyl
groups which may interact with similar hydroxyl groups and reduce
the attractive force.
The flexibility of the film is indicated by the percentage elongation (E)
value and it was found to be influenced by the ferulic acid content. The
average E values of the films decreased from 29.3% for the control film
to a minimum of 21.6% and 22.9% for the blend films containing 75 and
100 mg of ferulic acid (Fig. 9). The reduction in percentage elongation
with increase in ferulic acid content might be due to the increase in the
number of intermolecular crosslinks and decrease in the intermolecular
distance.
Starch from two sources, tapioca and rice, has been used with chito-
san to make blends with better qualities than from the individual poly-
mers. Tapioca is a significant crop in South America, [46]. Its edible
films exhibit appropriate physical characteristics, since they are odorless,
tasteless, colorless and impermeable to oxygen. However, films show

Fig. 6. Effect of chitosan coating contents on tensile properties of coated film containing 6 wt.%glycerol in MD and TD [40].
1824 M.Z. Elsabee, E.S. Abdou / Materials Science and Engineering C 33 (2013) 1819–1841
Fig. 7. Thermogravimetric curves of (a) blend film; (b–d) ferulic acid incorporated
films (50, 75 and 100 mg); (e) ferulic acid and (f) potato starch [43].
brittleness with inadequate mechanical properties. Chillo et al. [47] have
blended chitosan (CS) with tapioca starch and glycerol as a plasticizing
agent. The apparent viscosity of the film-forming solution, mechanical
and dynamic-mechanical properties, water vapor permeability (WVP)
and color of the blend films have been investigated. The mechanical
measurements and viscoelastic properties of the chitosan/tapioca
starch-based edible films have been measured using a stress-controlled
Dynamic-Mechanical Analyzer equipped with a tension clamp. Mechan-
ical tests under static, transient and dynamic conditions were performed,
i.e. uniaxial tension, stress relaxation and oscillatory stress, respectively.
The influence of CS and glycerol concentrations on the properties of tap-
ioca starch-based edible film was analyzed. The obtained data inferred
that, the elastic modulus (Ec) values were positively affected by the linear
and quadratic terms of CS and glycerol contents, respectively, while neg-
atively influenced by glycerol and CS–glycerol interaction. In addition, the
tensile strength values were influenced by the individual positive term of
the CS and by the negative influence of the CS–glycerol interaction.
Previous studies have shown that the starch–chitosan blend films
to exhibit significantly higher elongation values compared to films
made from starch or chitosan alone [48].
Xu et al. [31] observed a dependence of the elongation at break by
starch to CS ratio with a maximum value corresponding to a ratio of
Fig. 8. Derivative thermogravimetric curves of (a) blend film; (b–d) ferulic acid incor-
porated films (50, 75 and 100 mg); (e) ferulic acid and (f) potato starch [43].
Fig. 9. Effect of ferulic acid concentration on the tensile strength and percentage elon-
gation of the blend films [43].
1.5:1. Probably, in this study, the higher ratio starch to CS was responsible
for a negligible effect of the CS on the elongation at break.
A biodegradable or edible film must withstand the normal stress
encountered during its application and subsequent shipping and han-
dling of the food to maintain its integrity and barrier properties [49].
High tensile strength is generally required, but deformation values
must be adjusted according to the intended application of the films.
That is, whether it is undeformable material to provide structural integ-
rity or reinforce structure of the food. The TS of biodegradable blend
films from rice starch/chitosan with different chitosan ratios is shown
in Fig. 10A. The TS of biodegradable blend films was affected by the chi-
tosan ratios. The results demonstrated that the TS of biodegradable
blend films increased with the addition of chitosan, and the maximum
occurred at the rice starch and chitosan ratio of 1:1 and 0.5:1.0. The in-
creasing TS values of the biodegradable blend films, with the increase of
rice starch and chitosan ratios from 2:1 to 0.5:1, are attributable to a
high formation of intermolecular hydrogen bonding between NH2 of
the chitosan backbone and OH of the rice starch. The amino groups
(NH2) of chitosan were protonated to NH3+ in the acetic acid solution,
whereas the ordered crystalline structures of starch molecules were
destroyed with the gelatinization process, resulting in the OH− groups
being exposed to readily form hydrogen bonds with NH3+ of the
chitosan. However, the TS of biodegradable blend film prepared at the
starch to chitosan ratio of 1:1 and 0.5:1 was not significantly different.
This phenomenon indicated the critical ratios of the greatest miscibility
of the two main film-forming components.
Elongation at the break (E) is an indication of the films flexibility and
stretchability (extensibility), which is determined at the point when the
film breaks under tensile testing. The value of E was affected by the chito-
san ratios (Fig. 10B). The average E values of the biodegradable blend film
behaved inversely to the TS value, decreasing from 12.99% to a minimum
8.06% when the rice starch and chitosan ratio was 0.5:1 (Fig. 10B).
E was reduced in the presence of chitosan, probably due to the in-
creased crystallinity of starch in the blend film.
Liu et al. [50] studied the mechanical properties of starch/chitosan
blending membrane. Elongation-at-break (E), values of chitosan/starch
blend membranes with the different starch masses are shown in Fig. 11.
The E membranes' values were affected by the starch contents. This
phenomenon has also been reported by Xu et al. [31]. With the mass
of starch increasing, the E value of obtained membrane increased in
the initial stage until reaching a maximum after which the curve started
bending downwards. The data in Fig. 11 could be explained as follows:
with the addition of starch, the E value of blend membrane increases
due to the formation of hydrogen bonds between NH3+ of the protonat-
ed chitosan and OH− of the starch. However, when the addition of
starch was too high, the flexibility of obtained membrane was lowered
 M.Z. Elsabee, E.S. Abdou / Materials Science and Engineering C 33 (2013) 1819–1841
Fig. 10. Tensile strength (MPa) (A), and elongation at break (%) (B) as a function of
starch:chitosan ratio [49].
and the E value also decreased for the brittle nature of starch membrane
[51]. Thus, it is comparatively difficult to form homogeneous starch/chi-
tosan membrane with higher content of starch.
Effects of potassium sorbate (KS) and chitosan incorporation on
the tensile strength (TS) and elongation at break (E) of sweet potato
starch film was studied by Shen et al. [52] as seen in Fig. 12. It has
been observed that the E values of the films significantly decreased
when (KS) was added, and the higher the addition of potassium sor-
bate, the lower the E of the films.
Flores et al. [53] also verified the fact that (KS) could decrease the TS
of tapioca-starch edible films. This was attributed to the interaction be-
tween potassium sorbate and the starch molecules resulting in a modi-
fication of the starch network in the films. It is a well-known fact that
the mechanical behavior of starch films is affected by the presence of
crystalline phases. Fama, et al. showed that the control films have
higher degree of crystallinity than those with potassium sorbate (KS),
Fig. 11. Elongation-at-break (E) values of chitosan/starch blend membranes with the
different chitosan masses [50].
1825
Fig. 12. Tensile strength (TS) and elongation at break (E) of sweet potato starch films
as a function of potassium sorbate (K) and chitosan (C) addition. Different lowercase
letters in the same curve indicate significant differences (p b 0.05). Data shown in
mean ± standard deviation (n = 5) [52].
which contributed to the better mechanical behavior of the control
starch film [54]. This was also proved by FT-IR spectra analysis.
The addition of chitosan significantly improved both TS and E of
sweet potato starch films. The TS of films with 5% and 10% chitosan
was lower than those of films with 15% chitosan, but no significant
difference was observed between TS of films with 5% and 10% chito-
san. Films containing 10% and 15% chitosan had higher values of E
than those with 5% chitosan.
Gallstedt and Hedenqvist [55] studied the mechanical and barrier
properties of pulp–fiber–chitosan sheets. They used five methods to
prepare pulp–fiber–chitosan sheets, optical micrographs representing
the produced sheets are shown in Fig. 13.
The results of their work show that, the fracture strain and the frac-
ture stress decreased with increasing chitosan solution content. At chi-
tosan solution contents above 50 wt.%, the shrinkage during the buffer
treatment could be reduced effectively by the presence of pulp fibers.
The buffered sheets also had the highest Young's modulus.
Considering that chitosan is a disintegrating polysaccharide with a
similar structure to that of cellulose Phisalaphong and Jatupaiboon
[56] studied the supplement of chitosan during Acetobacter xylinum cul-
tivation of cellulose. The aim of their study was to develop a new nano-
structure film composed of chitosan and cellulose. Microstructure and
mechanical properties of the developed films are then characterized
to provide indications for the modification of bacterial cellulose–chito-
san (BCC) film. The blank sample of the bacterial cellulose (BC) film is
the sample with 0% of chitosan content. In comparison to that of the
BC film, the tensile strength of BCC films (samples containing chitosan)
increased with an increase of chitosan content Fig. 14.
The tendency of tensile strength of the re-swollen (wet) films rather
corresponded to the dry films but in a lower range. Fig. 15 shows that the
Young's modulus increased with the increase of chitosan supplement
correspondingly to the effect on tensile strength.
Fig. 16 shows that in contrast to the effect on tensile strength, the
percentage of elongation at break decreased with increasing chitosan
concentration.
Rice is the most widely consumed basic food in the world [49]. Each
year over 500 million tons of rice are harvested, providing sustenance
to many countries and people throughout the world. The unique proper-
ties of rice starches are found in its many varieties. Due to different cli-
mates, soil characteristics and cultures, over 240,000 registered varieties
of rice exist in the world. These varieties lead to a wide range of rice
starches with many different characteristics including: different starting
gelatinization temperatures, textures, processing stabilities and viscosi-
ties. Rice starch and its major components, amylose and amylopectin,
are biopolymers, which are attractive raw materials for use as barriers
in packaging materials. They have been used to produce biodegradable
films to partially or entirely replace plastic polymers because of its low
1826 M.Z. Elsabee, E.S. Abdou / Materials Science and Engineering C 33 (2013) 1819–1841
Fig. 13. Optical micrographs representing sheets produced according to (1) Method 1 (10 wt.% chitosan solution), (2) Method 2 with 20 wt.% chitosan solution and pressure
(3) Method 4 with 90% chitosan solution and pressure. The sides of the micrographs correspond to 16.5 cm (A) and (B) and 8.25 cm (C) [55].
cost and renewability, as well as good mechanical properties [31]. Howev-
er, wide application of starch film is limited by its mechanical properties
and efficient barrier against low polarity compounds [57]. This constraint
has led to the development of the improved properties of rice-based films
by modifying its starch properties and/or incorporating other materials.
Starch was blended with different proteins to decrease the water vapor
permeability of the films and to increase their tensile strength [58].
Bourtoom and Chinnan [49] prepared biodegradable films using chito-
san with starch. A series of films was prepared by mixing 100 ml of the
starch solutions (1, 2, 3 and 4 g/100 ml) with 100 ml of the chitosan so-
lution (2 g/100 ml). Sorbitol was added as 40% (w/w) of the total solid
weight in solution. The mixtures were cast onto flat, leveled, non-stick
trays to set. Film solubility (FS) is a very important property where
water resistance is an important property of biodegradable or edible
films for applications as food protection where water activity is high, or
when the film must be in contact with water during processing of the
coated food (e.g. to avoid exudation of fresh or frozen products). Howev-
er, a high solubility may be an advantage for some applications. Film sol-
ubility is advantageous in situations when the films will be consumed
with a product that is heated prior to consumption and may also be an im-
portant factor that determines biodegradability of films when used as
packaging wrap. Biodegradable blend film produced from rice and chito-
san maintain their integrity (i.e., did not dissolve or break apart) even
after 24 h of incubation with gentle motion. This indicates that the rice
starch and chitosan intra and/or intermolecular network remained intact
and only the monomers, small peptides, and non-protein material were
soluble [49].
Fig. 14. The tensile strength of BCC films in re-swollen (wet) form: (a) BCC-MW 30,000
and (b) BCC-MW 80,000 and in dry form (c) BCC-MW 30,000 and (d) BCC-MW 80,000
as a function of chitosan content (% w/v) in culture medium [56].
Color of the packaging is also an important factor in terms of gen-
eral appearance and consumer acceptance [59]. The results of the
measurements performed on the blend film's color were expressed
in accordance with CIELAB system and the rectangular coordinates;
the main difference observed was that biodegradable blend films
with higher content of chitosan had lighter color [49].
2.1.2. Chitosan and gelatin based edible films
Composite edible films and coatings can be formulated to combine
the advantages of each component. Whereas biopolymers, such as pro-
teins and polysaccharides, provide the supporting matrix, lipids provide
a good barrier to water vapor [60]. Since gelatin and chitosan are hydro-
philic biopolymers with good affinity and compatibility, they are
expected to form composite films with good properties [61]. Gelatin/
chitosan blends have been used extensively for the production of scaf-
folds and bi-layers for biomedical applications [62,63]. Rivero et al.
[64] developed composite, bi-layer and laminated biodegradable films
based on gelatin and chitosan, to determine film barrier and mechanical
properties and to characterize their microstructure. The gelatin blends
with chitosan and chitosan gel to form wound healing materials [65].
The blend has a smooth and homogeneous surface as revealed by SEM
and X-ray measurements [66].
The effect of chitosan molecular weight (MW) and degree of
deacetylation (DD) on the physicochemical properties of gelatin-
based films were studied [67]. Determination of the dynamic
Fig. 15. The Young's modulus of BCC films in re-swollen (wet) form: (a) BCC-MW
30,000 and (b) BCC-MW 80,000 and in dry form (c) BCC-MW 30,000 and
(d) BCC-MW 80,000 as a function of chitosan content (% w/v) in culture medium [56].
 M.Z. Elsabee, E.S. Abdou / Materials Science and Engineering C 33 (2013) 1819–1841
Fig. 16. The elongation at break of BCC films in re-swollen (wet) form: (a) BCC-MW
30,000 and (b) BCC-MW 80,000 and in dry form (c) BCC-MW 30,000 and
(d) BCC-MW 80,000 as a function of chitosan content (% w/v) in culture medium [56].
viscoelastic properties (elastic modulus G′ and viscous modulus G″) of
the film-forming solutions revealed that the interactions between gela-
tin and chitosan were stronger in the blends made with chitosan of
higher molecular weights or higher degrees of deacetylation than the
blends made with lower molecular weights or degrees of deacetylation.
Fish gelatin films modified with chitosan of higher molecular weights or
higher degrees of deacetylation had higher tensile strengths.
Films of chitosan and gelatin were prepared by casting their aque-
ous solutions (pH b 4.0) at 60 °C and evaporating at 22 or 60 °C (low-
and high-temperature methods, respectively). The physical (thermal,
mechanical and gas/water permeation) properties of these composite
films, plasticized with water or polyols, were studied [68]. An in-
crease in the total plasticizer content resulted in a considerable de-
crease of elasticity modulus and tensile strength (up to 50% of the
original values when 30% plasticizer was added), whereas the elonga-
tion percentage increased (up to 150% compared to the original
values). The low-temperature preparation method led to the devel-
opment of a higher percentage crystallinity of gelatin which resulted
in a decrease, by one or two orders of magnitude, of CO2 and O2 per-
meability in the chitosan/gelatin blends. An increase in the total plas-
ticizer content (water and polyols) of these blends was found to be
proportional to an increase in their gas permeability. This blend
should be a promising candidate for good edible biodegradable films.
It has been found that gelatin origin plays a role in improving the
physical characteristics of films with chitosan [69]. The data revealed
that the interactions between gelatin and chitosan were stronger in
the blends made with tuna-skin gelatin than in the blends made with
bovine-hide gelatin. As a result, the fish gelatin chitosan films were
more water resistant (w18% water solubility for tuna vs. 30% for bovine)
and more deformable (w68% breaking deformation for tuna vs. 11% for
bovine) than the bovine gelatin chitosan films. The breaking strength of
gelatin chitosan films, whatever the gelatin origin, was higher than that
of plain gelatin films. Bovine gelatin chitosan films showed a significant
lower water vapor permeability (WVP) than the corresponding plain
films, whereas tuna gelatin chitosan ones were only significantly less
permeable than plain chitosan film. Complex gelatin chitosan films be-
haved at room temperature as rubbery semi-crystalline materials. In
spite of gelatin chitosan interactions, all the chitosan-containing films
exhibited antimicrobial activity against Staphylococcus aureus, a rele-
vant food poisoning. Mixing gelatin and chitosan may be a means to im-
prove the physico-chemical performance of gelatin and chitosan plain
films, especially when using fish gelatin, without altering the antimicro-
bial properties.
The effect of glycerol on the mechanical and water barrier properties,
as well as on the water solubility, of fish gelatin–chitosan films was stud-
ied by Kołodziejska and Piotrowska [70]. The final goal of their study was
to design biodegradable material with good mechanical and barrier
1827
properties, suitable for packages of many kinds of food products with dif-
ferent acidities and contents of moisture.
More research should be directed to the exploitation of these
blends with potential applications for food protection.
2.1.3. Chitosan with alginate and carageenan
Alginate is a linear copolymer extracted from brown seaweeds known
as Phaeophyceae and composed of (1→4)-linked-β-D-mannuronate (M)
and (1→4)-linked-α-L-guluronate (G) units. These units are arranged in
G-blocks, M-blocks and alternating sequences of GM-blocks forming the
polymeric structure, where the sequential arrangement depends on dif-
ferent factors such as the specie, age or parts of the seaweeds from
which this material was obtained [71]. It has been well characterized in
both the liquid and gel state, making this biopolymer unique compared
to other gelling polysaccharides. Alginates possess good film-forming
property, producing uniform, transparent, and water soluble films.
Alginate-based films are impervious to oils and fats but, as other hydro-
philic polysaccharides, have high WVP. However, alginate gel coating
can act as a sacrificing agent, where moisture is lost from the coating be-
fore the food significantly dehydrates. The coating can also improve the
adhesion of batter to the surface of fruits and vegetables [72]. Alginate
coatings are good oxygen barriers that can retard lipid oxidation in vari-
ous fruits and vegetables, and have been found to reduce weight loss
and natural micro flora counts in minimally processed carrots [73]. Calci-
um alginate coatings were found to improve the quality of fruits and veg-
etables, such as reducing shrinkage, oxidative rancidity, moisture
migration, oil absorption, and sealing-in volatile flavors, improving ap-
pearance and color, and reducing weight loss of fresh mushrooms in com-
parison with uncoated ones [74].
Carrageenans are anionic linear polysaccharides extracted from red
seaweed (Rhodophyceae), consisting of alternating α-1, 4 and β-1, 3
linked anhydrogalactose residues. There are three major fractions (κ —
kappa, ι — iota and λ — lambda) with varying number and position of
sulfate groups on the galactose dimer. Carrageenan-based coatings
have been applied to fresh fruits and vegetables such as fresh apples
for reducing moisture loss, oxidation, or disintegration of the apples
[72,75]. In combination with anti-browning agents such as ascorbic
acid, carrageenan-based coatings resulted in positive sensory results
and reduction of microbial levels on minimally processed apple slices.
By acting as a sacrificial moisture layer, carrageenan coating was able
to protect moisture loss of grapefruits. In addition, κ-carrageenan films
can effectively carry food-grade antimicrobials such as lysozyme, nisin,
grape fruit seed extract, and EDTA for a wide range of applications as a
food package material [76].
Both the alginates and the carrageenans can interact with chitosan
forming polyelectrolyte complexes [77] which were used to obtain
microcapsules for cell encapsulation and devices for the controlled re-
lease of drugs or other substances. It seems there is good potential to
investigate this interaction to produce edible films from these mate-
rials which could be of great value, however not yet explored.
3. Chitosan/essential oil films
Many spices and herbs and their extracts possess antimicrobial ac-
tivity. The composition, structure as well as functional groups of the
oils play an important role in determining their antimicrobial activity
[78]. Usually compounds with phenolic groups are most effective [79].
Among these, the oils of clove, thyme, cinnamon, rosemary, sage and
vanillin have been found to be most consistently effective against mi-
croorganisms. Besides antibacterial properties [80,81] essential oils,
Eos, or their components have been shown to exhibit antimycotic
[82], antitoxigenic [83] and antiparasitic [84] properties. These charac-
teristics are possibly related to the function of these compounds in
plants [85]. A comprehensive review dealing with the more common
synthetic and natural antimicrobial agents derived from essential oils
incorporated into or coated onto synthetic packaging films for
1828 M.Z. Elsabee, E.S. Abdou / Materials Science and Engineering C 33 (2013) 1819–1841
antimicrobial packaging applications has been published by Kuorwel
et al. [86]. The focus is on the widely studied herb varieties including
basil, oregano, and thyme and their essential oils (EOs).
Because of the effect of direct addition of essential oils to food incor-
poration of essential oils to films may have supplementary applications
in food packaging [87,88]. Combining antimicrobial agents such as plant
essential oils directly into a food packaging is a form of active packaging.
Rosemary, a good source of antioxidant compounds, is widely used in
the food industry to prevent oxidative degradation of foods [89–91]. The
antioxidant activity of rosemary extracts is associated with the presence
of several phenolic diterpenes, which break free radical chain reactions
by hydrogen donation. Oxygen pretreatment and chitosan coating
0.03% rosemary extracts have improved the quality of fresh-cut pears
and extend the shelf-life [92]. The chitosan film was prepared by dispers-
ing chitosan in an aqueous solution of 1% glacial acetic acid (v/v) at 4 °C,
1.5% glycerol (w/v) and 0.2% Tween-80 (v/v) were added to the mixture
and the mixture was homogenized at 1000 rpm for 10 min. The pH
value was adjusted to 5.6 with 1 M NaOH. The solution was strained
through layers of cheesecloth and degassed under vacuum at room tem-
perature 0.03% rosemary extract was added to the final chitosan solution.
The chitosan film solutions were stored in a refrigeration condition
(4 °C) for 1–2 days [92].
In spite of the extensive use of chitosan as edible films for coating, it
still suffers from high water vapor permeation which lowers its protec-
tive action, therefore trials were made to add oils to increase its hydro-
phobicity and improve its water vapor permeation. Chitosan was mixed
with increasing concentration of olive oils to prepare homogeneous
films with decreasing moisture sorption, lower water vapor permeation
through the films and smaller effective diffusion coefficients of the films
as the oil concentration increases [93]. All the tensile properties (Young
modulus, strength and maximum elongation) increased with olive oil
concentration and were explained considering the interactions devel-
oped between lipid and carbohydrate phases in addition to the lubricant
characteristics of the oil [93].
Ojagh et al. [94] studied the effect of adding cinnamon essential oil
(CEO) into chitosan-based films. It was found that CEO increased the
antimicrobial activity, while decreased the moisture content, the sol-
ubility in water, the WVP and elongation at break of the chitosan
Fig. 17. Scanning electronic microscopic images of chitosan control film (A) and (B), and film containing CEO at level of 1.5% (C) and (D) surfaces and cross sections, respectively [95].
films. Incorporating CEO at level of 0.4%, 0.8%, 1.5% and 2% (v/v)
into chitosan films increased the tensile strength values significantly.
The authors claimed that a strong interaction between the polymer
and the CEO produced a cross-linker effect, which decreases the free
volume and the molecular mobility of the polymer. This phenomenon
led to a sheet like structure as seen in (Fig. 17C).
Arrangement of stacking layers of CEO added chitosan sheets
(Fig. 17D) means that in these films a compact structure has formed
leading to a decrease in elongation at break and increasing of the ten-
sile strength. There are also possibilities that such structure enhances
the decrease in moisture content of the films incorporated with CEO.
The active component of CEO is cinnamaldehyde (~ 60%) [95].
Chitosan control films did not show inhibitory zone in bacterial
strains tested. Despite antimicrobial activity of chitosan because of its
innate characteristic, this effect of chitosan occurred without migration
of active agents. Chitosan does not diffuse through the adjacent agar
media in agar diffusion test method; so that only organisms in direct
contact with the active sites of chitosan are inhibited [13].
Cinnamon oil has also been used by the same authors to blend with
chitosan for the preservation of trout fish filet [87]. This treatment
could maintain trout filet shelf life till the end of the storage period
(16 days) without any significant loss of texture, odor, color or overall
acceptability and without significant microbial growth. Variations in
the value of total viable counts (TVC) on the fish surface during the
refrigerated storage are presented in Fig. 18. The initial TVC (log CFU/g)
in trout filet ranged from 3.51 in Ch +C-coated samples to 3.86 in con-
trols. In the meantime, the control samples had a shelf life of only
12 days. Therefore, chitosan coating together with cinnamon oil proved
to be an efficient method of protecting the fish under refrigerated storage
for a longer period f time.
Bergamot essential oil (BO) is citrus oil (from Citrus bergamia),
whose major chemical compounds are limonene (32–45%) and linalool
(around 10.23%) [96–98]. The antimicrobial efficiency of BO, and its
components, linaool and citral, have been found to be effective against
Campylobacter jejuni, E. coli O157, Listeria monocytogenes, Bacillus cereus,
S. aureus, Arcobacter butzleri and Penicillium digitatum [99], among
others, both when oil is applied directly and when in contact with the
oil vapor. Chitosan-based films containing BO (CS–BO) at 0.5, 1.2 and
 M.Z. Elsabee, E.S. Abdou / Materials Science and Engineering C 33 (2013) 1819–1841
Fig. 18. Changes in total viable counts (TVC) of fish samples during refrigerated storage
[87].
3% w/w were prepared to evaluate their physical and antifungal proper-
ties. Film-forming dispersions (FFD) were also characterized in terms of
rheological properties, particle size distribution and ζ-potential. Fur-
thermore, the antifungal effectiveness of CS–BO composite films against
Penicillium italicum was studied. Results showed that incorporation of
BO provoked a decrease in the water vapor permeability, this reduction
being around 50% when using a BO–CS ratio of 3:1. Concerning mechan-
ical and optical properties, CS–BO composite films were less resistant to
break, less deformable and less glossy. The load parameters (TS and EM)
decreased more than 50% and the percentage of elongation at break was
also dramatically reduced from 22% to 5%, as compared with the pure
chitosan films. CS–BO composite films showed a significant inhibitory
effect on the growth of P. italicum, which depended on the BO concen-
tration. Chitosan films with the maximum bergamot oil content (3:1
BO–CS ratio) led to a total inhibition of the fungus growth during the
first 5 days at 20 °C.
Although the antifungal effectiveness of the films decreased through-
out the storage time, a significant reduction of 2 log units as compared
with the control remained possible, after 12 days at 20 °C, using the
highest BO content. The mechanisms by which essential oils bring
about their antimicrobial effect are not clear, Holley and Patel [78] had
suggested that terpenes have the ability to disrupt and penetrate the
lipid structure of the cell membrane, as well as the mitochondrial mem-
brane, leading to the denaturation of proteins and the destruction of cell
membrane, cytoplasmatic leakage, cell lysis and eventually, cell death.
The essential oil of Melaleuca alternifolia, also named as tea tree oil
(TTO), is a complex mixture of terpen hydrocarbons and tertiary alcohols
[100]. The main compounds responsible for the antimicrobial activity are
terpinen-4-ol and 1, 8-cineole, TTO has been used successfully in the
management of oral candidiasis in AIDS patients [101] and other oral
fungal infections in patients suffering from advanced cancer [102].
Sanchez-Gonzalez et al. [100] incorporated TTO into chitosan matrix
and investigated the physical and antibacterial behavior of the obtained
composite. The CS films were rough and a reduction of its gloss occurred
after the incorporation of TTO. Water vapor permeability was also re-
duced by 40% when the CS/TTO ratio was 1:2. Likewise, the films' resis-
tance to break was notably reduced by TTO incorporation due to the
presence of discontinuities in the film matrix that affect its mechanical
response. The poor mechanical properties obtained by the addition of
TTO may be related with the structural arrangement of the lipid phase
into the chitosan matrix. Thus, the structural discontinuities provoked
by the incorporation of the oil could explain the lowest resistance to frac-
ture of the composite films. Some of these results are in line with those
reported by other authors when adding oils to a chitosan matrix
[103,29] but differ in some aspects due to the great influence of several,
widely studied factors related to CS preparation. Only the composite
1829
films with TTO:CS ratios higher than 1 showed a limited antifungal effec-
tiveness against Penicillium, which was notably reduced after 3 days of
storage. Nevertheless, CS films presented a significant antimicrobial ac-
tivity against L. monocytogenes and the incorporation of TTO in the CS/
TTO ratio of 1:2 improved the antibacterial properties of these films,
showing a complete inhibition of the microbial growth during the fifth
day at 10 °C.
Synergism and antagonism between components of EOs and food
constituents require more study before these substances can be reli-
ably used in commercial applications [104].
Biodegradable coatings based on chitosan (CS) with and without
bergamot essential oil were applied to table grapes, cv. Muscatel in-
vestigated by Sánchez-González et al. [105] in order to find environ-
mentally friendly, and healthy treatments with which to better
preserve fresh fruit quality and safety during postharvest cold stor-
age. Physicochemical properties (weight loss, °Brix, total phenols, an-
tioxidant activity, color and texture), respiration rates and microbial
counts of samples were determined throughout cold storage when
using CS. CS coatings containing bergamot oil produced the most ef-
fective antimicrobial activity, and showed the greatest inhibition of
the respiration rates in terms of both O2 consumption and CO2 gener-
ation. Although the coatings did not seem to reduce the rate of grape
browning during storage, they inhibited color development, thus im-
proving the product appearance. Taking into account the overall re-
sults obtained, the most recommended coating for Muscatel table
grape is the CS containing bergamot oil.
An intensive study has been conducted by Gómez-Estaca et al. [106]
in which chitosan–gelatin films, containing sorbitol and glycerol as plas-
ticizers were incorporated with several different essential oils and tested
for their antibacterial behavior against 18 different bacterial strains
which included some important food pathogen and spoilage bacteria.
Clove essential oil showed the highest inhibitory effect, followed by rose-
mary and lavender. Clove and thyme essential oils were the most
effective food preservatives, when tested on an extract made of fish.
The gelatin/chitosan-based edible films incorporated with clove essential
oil were tested against six selected microorganisms: Pseudomonas
fluorescens, Shewanella putrefaciens, Photobacterium phosphoreum,
Listeria innocua, E. coli and Lactobacillus acidophilus. The clove-
containing films inhibited all these microorganisms irrespectively of
the film matrix or type of microorganism. When the complex gelatin–
chitosan film incorporating clove essential oil was applied to fish during
chilled storage, the growth of microorganisms was drastically reduced in
Gram-negative bacteria, especially enterobacteria, while lactic acid bac-
teria remained practically constant for much of the storage period [106].
Mayachiew et al. [107] studied the effect of drying methods and
conditions (i.e., ambient drying, hot air drying at 40 °C, vacuum dry-
ing and low-pressure superheated steam drying (LPSSD) within the
temperature range of 70–90 °C at an absolute pressure of 10 kPa) as
well as the concentration of galangal extract on the antimicrobial ac-
tivity of edible chitosan films against S. aureus. Galangal extract was
added to the film forming solution as a natural antimicrobial agent
in the concentration range of 0.3–0.9 g/100 g.
Galangal similar to ginger and turmeric is a member of the rhi-
zome family. Rhizomes are knobby underground stems that are
known for their pungent and flavorful flesh, it is a traditional spice
used extensively for flavoring and medicinal purposes. Galangal ex-
tract has also proved to be an effective natural antimicrobial agent
against some food poisoning bacteria, e.g., S. aureus [108]. The main
compounds of galangal extract are the terpenes, which have potential
antimicrobial activity [78,106,109].
Chitosan films containing galangal extract at 0.6% and 0.9% (w/w)
were effective in inhibiting the growth of S. aureus. No inhibition zone
was observed when the extract concentration of 0.3% (w/w) was
used. This could be ascribed to a limited galangal extract release proba-
bly due to interaction between the extract and chitosan. Another possi-
ble reason could be the limit of detection of antimicrobial activity when
1830 M.Z. Elsabee, E.S. Abdou / Materials Science and Engineering C 33 (2013) 1819–1841
using the disk diffusion method [13,110]. It was noted that drying
methods and conditions had significant effects on the antimicrobial ac-
tivity of chitosan films incorporated with galangal extract. The results
showed that ambient dried film had the highest antimicrobial activity;
this was followed by LPSSD films and vacuum dried films. This may be
due to the fact that the film temperature increased more rapidly and
stayed at higher levels in the case of vacuum drying than in the case
of LPSSD, thus inducing more thermal degradation of the antimicrobial
compound [104,111]. In addition, different intermolecular interactions
also contributed to the observed results. The decrease in bacteria inhibi-
tion might be due to lower diffusion of the active agent into the agar
medium as a result of higher interaction between chitosan and galangal
extract. The antimicrobial films prepared at higher drying temperatures
had lower antimicrobial activity, both in the cases of vacuum drying and
LPSSD. The antimicrobial films prepared by LPSSD at 70 °C had the
highest antimicrobial activity compared with films prepared at other
conditions of vacuum drying.
The results showed that the antimicrobial activity of the films in-
creased with an increase in the extract concentration, as expected.
Chitosan film incorporated with 0.9% (w/w) galangal extract and pre-
pared by ambient drying could reduce the number of S. aureus by
about 3.6 log cycle within the contact time of 24 h. On the other
hand, ambient dried film incorporated with 0.3% (w/w) galangal ex-
tract exhibited lower cell reduction number of around 2.0 log cycle.
Sanchez-Gonzalez et al. [112] prepared antimicrobial films by
incorporating different concentrations of tea tree essential oil (TTO)
into chitosan (CS) films. These films have been tested against
L. monocytogenes and P. italicum. The possible antifungal effect against
P. italicum at 20 °C of CS and composite films was determined on
potato dextrose agar (PDA) medium and shown in Figs. 19 and 20. This
effectiveness was evaluated through the analysis of the growth (or sur-
vival) of a determined infection level of P. italicum (105 spores/ml), the
growth of fungus was followed by counts immediately after the inocula-
tion and periodically during the storage period of (PDA) plates.
CS films did not show antifungal effect for the assayed times. Pre-
vious studies have demonstrated that the antimicrobial effect of chi-
tosan depends on the type of microorganism, being mainly effective
against bacteria and also against some molds and yeast [113].
The TTO composite films delayed the fungal growth of P. italicum (in
comparison to the control), which was dependent on the TTO concen-
tration. At low TTO levels, no antimicrobial effect was observed. Only
when TTO–CS ratio was higher than 1, a moderate inhibition of fungus
Fig. 19. Effect of CS and CS–TTO composite films on the growth and survival of Penicillium
italicum on PDA medium stored at 20 °C. Mean values and 95% LSD intervals for each
sample time [112].
Fig. 20. Effect of CS and CS–TTO composite films on the growth and survival of Listeria
monocytogenes on TSA NaCl medium stored at 10 °C. Mean values and 95% LSD inter-
vals for each sample time [112].
growth was detected. The level of reduction of the Penicillium popula-
tion in CS–2TTO films observed during the first 3 days of storage was es-
pecially remarkable, reaching a fungal reduction of 3 logs in comparison
with the control plates. Nevertheless, the inhibition level of the compos-
ite films decreased throughout storage time. Considering that the anti-
microbial activity of TTO has been probed with very low concentrations
in the liquid phase, the observed behavior could be explained by the
availability level of active antimicrobial compounds against the fungi
agent. Numerous studies have demonstrated that these compounds
are more effective in reducing microbial growth when incorporated
into a film or gel and applied to the product surface than when applied
on the surface via spray solution or directly added to the product
[114–117] because of the active substances ability to evaporate or dif-
fuse into the medium.
4. Chitosan and clay
Natural polymers suffer from lower mechanical strength compared
to synthetic polymers and high moister barrier because of their hydro-
philic nature. Many strategies have been explored to improve these
problems of chitosan based biodegradable packaging films. These in-
clude the addition of plasticizers such as glycerol which increases the
flexibility of the final product [103]. The addition of other biodegradable
aliphatic polyesters, such as poly-caprolactone (PCL), poly(butylene
succinate) (PBS), poly(lactic acid) (PLA), poly(butylene terephthalate
adipate) (PBTA), and poly(butylene succinate adipate) (PBSA), has
also been investigated to produce materials with properties intermedi-
ate between the two components [118].
Other methods included the addition of layered silicates nanoparticles
(e.g. sodium montmorillonite) to chitosan to improve its end-use proper-
ties such as barrier and mechanical properties [119]. Montmorillonite
(MMT) is the most studied nanoscale clays. It is hydrated alumina-
silicate layered clay made up of two silica tetrahedral sheets fused to an
edge-shared octahedral sheet of aluminum hydroxide. Its advantages of
high surface area and platelet thickness of 10 Å make it suitable for rein-
forcement purposes.
When nanoclay is mixed with a polymer, three types of composites
(tactoids, intercalation, and exfoliation) can be obtained. In the case of
tactoids, complete clay particles are dispersed within the polymer matrix
and the layers do not separate. Mixing a polymer and organoclay forms a
micro-scale composite, with the clay serving only as a conventional filler.
Intercalation and exfoliation are two ideal nano-scale composites. Inter-
calation occurs when a small amount of polymer is inserted between the
layers of the clay, thus expanding the interlayer spacing and forming a
 M.Z. Elsabee, E.S. Abdou / Materials Science and Engineering C 33 (2013) 1819–1841 1831

and MMT, hindering the flocculation and leaves the MMT stacks random-
ly orientated in space. A higher reduction of permeability (50%) is
obtained in chitosan films not containing glycerol due to the alignment
and flocculation of MMT stacks. This system could have potential applica-
tions in the packaging field.

5. Gas permeation properties of edible coatings

Permeability is a steady-state property that describes the extent to

which a permeating substance dissolves and then the rate at which it dif-
fuses through a film, with a driving force related to the difference in con-
centration of that substance between the two sides of the film [128].
Gas permeability of edible films and coatings depend on several
factors such as the integrity of the film, the ratio between crystalline
and amorphous zones, the hydrophilic–hydrophobic ratio and the
Fig. 21. Influence of medium pH on the effects of chitosan, ♦ without chitosan, ■ polymeric chain mobility; the interaction between the film-forming
0.005% chitosan, and ▲ 0.01% chitosan. Open symbols are pH 4 and closed symbols
polymer and the presence of a plasticizer or other additives are also
are pH 6 (T = 7 °C). The lines represent the fitted Baranyi-model [154].
important factors in film permeability [129].
The measurement of permeability of oxygen and carbon dioxide of
well-ordered multilayer structure. In exfoliation, the layers of the clay are the edible films provides important information for their further devel-
separated completely and the individual layers are distributed through- opment. Oxygen is the key factor that might cause oxidation, inducing
out the polymer matrix. The formation of intercalation or exfoliation several unwanted food changes such as odor, color and flavor, as well
depends on the types and amounts of nanoclay used [119]. as nutrients deterioration. Therefore, films providing a proper oxygen
Several reports deal with the preparation characterization of MMT/ barrier can help in improving food quality and extending food shelf life
chitosan composites and films [120–126]. It has been shown that chito- [130]. Carbon dioxide that is formed in some foods due to deterioration
san interact with MMT and forms a homogenous film. Chitosan/MMT and respiration reactions should be removed from the package to avoid
nanocomposites were prepared by an ion exchange reaction between further food deterioration and/or package destruction [131]. Such films
water soluble oligomeric chitosan and Na MMT. Chitosan showed can maintain food quality and improve stability and shelf life by
high affinity to MMT clay host. According to thermogravimetric analysis retarding unwanted mass transfer (O2 and CO2) in food products migra-
(TG) and powder X-ray diffraction analysis the thermal stability of chi- tion of moisture for dried and intermediate moisture foods, and migra-
tosan was remarkably improved due to the strong electrostatic interac- tion of solutes for frozen foods.
tion of cationic chitosan molecules with anionic silicate layers. It was There are several possible edible coatings for fruits, such as cellulose,
also found that the nanocomposites showed a synergistic effect in the casein, zein, soy protein, and chitosan. These were chosen since they
antimicrobial activity against to E. coli and S. aureus [127]. Delaminated have the desirable characteristics of generally being odorless, tasteless
MMT were found to be enriched on the surface of the nanocomposites and transparent. It is not easy to measure the gas permeation properties
when the amount of MMT was >103 ppm. This was accompanied by of the coatings after being placed on fruits. Therefore, separate flat films
a decrease of the contact angle [121]. The proliferation of fibroblasts need to be prepared and tested.
on MMT/CS 103 ppm was significantly greater than on other materials.
The antimicrobial activity was enhanced markedly with the increased
amount of MMT. The inflammatory responses of MMT in vitro and in
subcutaneous rats were not obvious until the concentration of MMT
was >103 ppm. The biocompatibility of MMT/CS at 103 ppm was
even better than that of CS. The biodegradation rate of CS of the MMT/
CS nanocomposite was much faster than that of the pure CS polymer.
These results suggested potential antimicrobial applications for MMT/
CS nanocomposites, especially for those containing 10 3 ppm (0.1%) of
MMT [121].
Montmorillonite (MMT) nanoclay and rosemary essential oil (REO)
were incorporated into chitosan film [120]. The MMT weight percent rel-
ative to chitosan was varied from 1 to 5 and was activated by three REO
levels (0.5%, 1%, and 1.5% v/v), and their impact on physical, mechanical,
and barrier properties of the chitosan was investigated. The results of
these investigation showed that the combined effect of adding MMT
and REO improved the tensile strength and the antibacterial properties
of the chitosan composites (films). It was also found that the MMT/chito-
san biocomposite particles exhibited a higher thermal decomposition
temperature compared to pure chitosan particles. The dual effect of
adding glycerol and MMT to chitosan has been investigated by Lavorgna
et al. [124]. The authors found that the mechanical properties of
nanocomposite containing glycerol are improved as clay loading in-
creases. This is due to a combined effect of clays and plasticizer. Glycerol
modifies the hydrogen-bonding network within the material and allows
better interaction between filler and matrix, thus facilitating the stress
transfer to the reinforcement phase and improving its mechanical Fig. 22. The influence of soluble starch on the antimicrobial activity of chitosan. Lag
properties. The addition of glycerol lowers the water permeability by phase (A) and growth rate (B) of Candida lambica with different chitosan and starch
(30%). Glycerol reduces the hydrogen interactions between chitosan concentrations (T = 7 °C and pH = 5). , 0% starch; , 1% starch; ■, 30% starch [154].
1832 M.Z. Elsabee, E.S. Abdou / Materials Science and Engineering C 33 (2013) 1819–1841
Fig. 23. The change of antibacterial activities of starch/chitosan blend films with radi-
ation dose (80% starch, 20% chitosan) [155].
Formulations based on polysaccharides and proteins have proved
their excellent selective permeability to O2 and CO2. However, because
of their hydrophilic nature they exhibit poor moisture barrier properties,
which can be improved by adding hydrophobic materials such as natural
waxes, acetylated monoglycerides and surfactants through emulsion or
lamination technology. That is why edible films usually are heteroge-
neous in nature.
Literature data indicate that O2, CO2 and water vapor permeability
of edible coatings are lower than the conventional plastic films. Starch
films ability for food protection is by controlling and reducing oxygen
transport, thus extending the shelf life of the food.
However, the incorporation of antimicrobial agents to starch films in-
creases O2 gas permeability thus incorporation of potassium sorbate to
sweet potato starch films, led to higher oxygen permeability [52]. The
mass transfer of oxygen in a semi-crystalline polymer is primarily a func-
tion of the amorphous phase, because the crystalline phase is usually
assumed to be impermeable. Fama et al. [53,55] showed that the incorpo-
ration of potassium sorbate decreased the crystallinity of tapioca-starch
edible films causing an increase in oxygen permeation (OP) of the starch
films. The incorporation of potassium sorbate weakened inter-molecular
forces between adjacent starch polymeric chains, facilitated chain mobil-
ity and increased the free volume between starch molecules, which
promoted oxygen permeability [132]. However, a better oxygen transpor-
tation barrier property was obtained with the starch films when chitosan
was incorporated. With 15% of chitosan, sweet potato starch film had a
significantly lower OP (1.94±0.95×10−6 cm3 ×m/(m2 ×d×kPa)) than
that of the control starch film. Chitosan could form inter-molecular hydro-
gen bonds with starch, which limited the inter-molecular chain mobility
and decreased its free volume, contributing to the decrease of OP. More-
over, Xu et al. [31] reported that chitosan film possessed low permeability
to oxygen.
Mathew and Abraham [43] determined the water vapor transmis-
sion (WVT) of films gravimetrically. The films were fixed on to the
circular opening of permeation cell-containing anhydrous calcium
chloride (0% RH) using melted paraffin. The cups were then weighed
and placed at 92% relative humidity and 37 °C in a humidity chamber.
The cups were weighed at 1-h intervals until the change in weight be-
came constant. The water vapor transferred through the films at dif-
ferent time intervals were determined from the weight gain of the
cups. Changes in the weight of the permeation cell were recorded
and plotted as a function of time. The slope of each line was calculated
by linear regression and WVT rate was calculated from the slope of
the straight line (g/h) divided by the transfer area (m 2).
Oxidized ferulic acid incorporated films were found to significantly
decrease the WVT compared to the control blend films, probably due
to the higher degree of cross linking that results from the formation of
quinones and free radicals which tend to promote the cross linking
[133]. However, the permeability of the ferulic acid incorporated film
at a concentration of 100 mg increased slightly in comparison to the
blend film containing 75 mg of ferulic acid, but was lower than that of
the control film indicating 75 mg of ferulic acid as the optimal concen-
tration of the cross-linking agent. Blend films with higher TS had lower
WVT rates probably owing to the better degree of organization of the
polysaccharide network up to the optimum concentration of ferulic
acid. They also studied oxygen transmission rates of chitosan/starch
films with different amount of ferulic acid. In general, polysaccharide
films are expected to be good oxygen barriers, due to their tightly
packed and ordered hydrogen-bonded network structure and low solu-
bility. The results showed that the oxygen transmission rate reduced
with the increase in the level of cross-linking agent. Kucuk and Caner
[134] have reported better stability and quality for sunflower oil
samples stored under packaging conditions free of air. High levels of ox-
ygen in food packages have been reported to cause the development of
off-flavors, off-odors and nutritional loss in food stuffs [135].
Since a main function of a food packaging is often to avoid or at least
to decrease moisture transfer between the food and the surrounding at-
mosphere, or between two components of a heterogeneous food prod-
uct, WVP should be as low as possible.
Shen et al. [52] showed that the WVP of sweet potato starch films de-
creased significantly with the addition of over 10% chitosan. This was
also the case for tapioca-starch-based edible film examined by Chillo
et al. [47]. They explained the decreasing WVP transmission rate at
higher concentrations of chitosan as a result of reducing the available hy-
drophilic group [31,47]. In addition the hydrogen bonding interaction
between chitosan and starch decreased the free sorption sites for
water, which is responsible for decreasing the WVP of the film [136].
WVP of chitosan films (CS) contain different concentrations of tea
tree essential oil (TTO) had been studied by Sanchez-Gonzalez et al.
[100]. The room temperature conditions used for measuring the WVP
(100/54.4) of the films were established to simulate the environmental
conditions when the films are applied as a coating for vegetables. WVP
values were in the range of those reported by other authors working
with films based on chitosan [29].
The WVP values showed a significant decrease in line with the in-
crease in TTO concentration, reaching a maximum WVP reduction of
about 40% with incorporation of 2% TTO in the film-forming disper-
sions. This behavior is expected as an increase in the hydrophobic
compound fraction usually leads to an improvement in the water bar-
rier properties of films, as was previously reported for essential oils
addition in CS films [96].
Fig. 24. The change of antibacterial activities of starch/chitosan blend films with the
content of chitosan (50 kGy) [155].
 M.Z. Elsabee, E.S. Abdou / Materials Science and Engineering C 33 (2013) 1819–1841
Fig. 25. Inhibition area of (A) control film and (B) AM incorporated film [167].
6. Effect of electric field on film formation
Preliminary works have shown that the presence of a moderate elec-
tric field during the preparation of chitosan coating solutions may influ-
ence their transport properties. If such effect is confirmed, moderate
electric fields could be used to tailor edible films and coatings for specif-
ic applications. Ohmic heating is based on the passage of electrical cur-
rent through a sample where the electrical energy is directly converted
to heat and instant heating occurs. The use of electric fields in the food
area has gained a new interest [137,138]. The application of electric
fields has also been an important instrument among researchers in
the area of edible films and coatings, and there are works showing
that the application of electric fields promotes a significant improve-
ment of several properties. Garcia et al. [139] analyzed the effect of an
electrical field applied during drying on the microstructure and macro-
scopic properties of films obtained with different mixtures of chitosan
(CS) and methylcellulose (MC). The analysis indicated that CS electri-
cally treated film exhibited a more ordered structure lower WVP and
higher Young's modulus values leading to stronger films. The authors
concluded that electrical field treatment would be a good alternative
to improve film flexibility and water vapor barrier properties.
Lei et al. [140] reported that using ohmic heating for the production
of protein–lipid films, improves the yield, of the film formation rate and
the rehydration capacity of the films. Souza et al. [141] determined the
effect of field strength on the functional properties of chitosan coatings.
Four different field strengths were tested, for each electric field treat-
ment, the water vapor, oxygen and carbon dioxide permeability of the
films formed were determined, together with their color, opacity and
solubility in water. Chitosan films formed from solutions subjected to
electric fields at 100 V cm−1 or higher were found to have lower values
of O2P and CO2P (oxygen and CO2 permeability). Atomic force micros-
copy AFM observation of chitosan films surface treated at 100 V cm−1
or above (confirmed by the Ra values) showed smoother surface as op-
posed to a rougher surface of untreated films.
AFM imaging modes can potentially provide structural information
for a sample in its more natural state (without dehydration or coatings).
Nanoscale measurements by AFM allow the influence of different fac-
tors on the hardness, elasticity and permeability of the film surface to
be quantified, which is extremely useful for the design of high perfor-
mance edible food packaging systems [142].
7. Antibacterial activity of chitosan and chitosan blends
The greatest losses in food are due to microbiological alterations.
Many chemical and physical processes have been developed to preserve
1833
food quality. Among such processes, adequate packaging is a funda-
mental factor in the conservation and marketing phases. Thus, packag-
ing plays a prominent role in maintaining food quality. Antimicrobial
films and coatings have vitalized the concept of active packaging and
have been developed to reduce, inhibit or delay the growth of microor-
ganisms on the surface of foods in contact with the packaged product
[143,144].
In most fresh or processed foods, microbial contamination occurs at a
higher intensity on the food surface, thus requiring an effective microbial
growth control. Traditionally, antimicrobial agents are added directly to
the foods, but their activity may be inhibited by many substances in
the food itself, diminishing their efficiency. In such cases, the use of anti-
microbial films or coatings can be more efficient than adding antimicro-
bial agents directly to the food since these may selectively and gradually
migrate from the package onto the surface of the food, thereby high con-
centrations being maintained when most necessary [114].
L. monocytogenes, a Gram-positive rod, is a bacterium that can
cause illness in a variety of food products [145]. One food product of
great concern is the refrigerated, ready-to-eat (RTE) foods contami-
nated with L. monocytogenes [146]. Eating foods contaminated with
L. monocytogenes normally causes the disease listeriosis which is
more serious for elderly adults and adults with compromised im-
mune systems and can cause meningitis [147]. In pregnant women,
the disease may cause spontaneous abortions or stillborn babies.
Greenwood [147] has studied the antimicrobial effect of chitosan, as
an edible film, that was dissolved in lactic acid or acetic acid against
L. monocytogenes on RTE roast beef. Chitosan with low and high mo-
lecular weight (MW) of (4.7 × 10 5 g/mol) and (1.1 × 10 6 g/mol)
were used to test the molecular weight effect on the antimicrobial ca-
pacity. This study showed that the acetic acid chitosan coating was
more effective in reducing L. monocytogenes counts than the lactic
acid chitosan coating. The study indicated also that chitosan coatings
could be used to control L. monocytogenes on the surface of RTE roast
beef. However, it has been found that L. monocytogenes was able to
grow on the surface of the RTE roast beef regardless of the chitosan
treatments used. Coma et al. [13] also observed the ability of
L. monocytogenes to grow on the surface of cheese regardless of chito-
san treatments. This could be due to the decreased antimicrobial ac-
tivity of chitosan films over time as a consequence of the decreased
availability of amino groups on chitosan [147]. The L. monocytogenes
strain used during this study was able to grow on the surface of the
RTE roast beef product. The initial level of 6.50 log CFU/g increased
to over 10 log CFU/g on the control samples at day 21 while samples
dipped in chitosan have a count of 7.4–7.9 for high and low molecular
weight chitosan respectively. The data indicated an improvement of
1834 M.Z. Elsabee, E.S. Abdou / Materials Science and Engineering C 33 (2013) 1819–1841
Fig. 26. Tomato fruit at zero time, (A) similar fruit kept for 13 days into a polypropylene bag fabricated from PP film covered by 12 alternating chitosan/pectin layers in comparison
to similar fruit kept for the same period of time in untreated PP bag (B), and another one kept in open air (C).
the antibacterial capacity of the roast beef when treated with chitosan
coating and also that higher molecular weight polymers have slightly
lower antibacterial properties.
The molecular weight and amino group of chitosan have strong
influence on its antibacterial activity [148,149]. In the Gram-positive
bacteria, the major constituent of its cell wall is peptidoglycan and
very little protein. The cell wall of Gram-negative bacteria on the
other hand is thinner but more complex and contains various poly-
saccharides, proteins and lipids beside peptidoglycan.
It has been found that chitosan activity against fungus to be less ef-
ficient as compared with its activity against bacteria [110,150]. On the
other hand, results from Bautista-Banos et al. [151] were much different
than those of Guo-Jane et al. [152] who emphasized on the efficiency of
Fig. 27. Effects of chitosan coating on browning (A), eating quality (B) and disease in-
cidence (C) of fresh-cut Chinese water chestnut during storage at 4 °C. Each value is
the means for three replicates, and the vertical bars indicated the standard errors. ●,
0 g (control); ■ 0.5 g/100 ml; ▲, 1 g/100 ml; ▼, 2 g/100 ml [173].
chitosan against fungi. Nevertheless, all these studies indicated that the
polycationic nature of chitosan is the key to its antifungal properties in
addition to the possible effect that chitosan might have on the synthesis
of certain fungal enzymes and that the length of the polymer chain en-
hances that activity. Bautista-Banos et al. [151] have shown that not
only chitosan is effective in stopping the growth of the pathogen, but
it also induces marked morphological changes, structural alterations
and molecular disorganization of the fungal cells.
No et al. [153] studied the antibacterial activities of six chitosan
samples and six chitosan oligomers with different molecular weights
(Mws) against four Gram-negative (E. coli, P. fluorescens, Salmonella
typhimurium, and Vibrio parahaemolyticus) and seven Gram-positive
bacteria (L. monocytogenes, Bacillus megaterium, B. cereus, S. aureus,
Lactobacillus plantarum, Lactobacillus brevis, and Lactobacillus bulgaricus).
They found that chitosan markedly inhibited growth of most bacteria
tested; however, the inhibitory effects differed with regard to the molec-
ular weight of chitosan and the type of bacterium. Chitosan oligomers
also inhibited bacterial growth by 1–5 log cycles at a 1.0% concentration;
however, effects were more limited than those of the chitosan. Chitosan
generally showed stronger bactericidal effects for Gram-positive bacteria
than Gram-negative bacteria.
Devlieghere and Debevere [154] investigated the antimicrobial ef-
fect of chitosan, with high deacetylation degree and low molecular
weight against several psychrotrophic food-borne pathogens and
spoilage micro-organisms and compared to those known from the lit-
erature. They found that Gram-negative bacteria were more suscepti-
ble, while the sensitivity of the Gram-positive bacteria was highly
variable: Brochotrix thermosphacta and B. cereus were very sensitive
to the applied chitosan while L. monocytogenes and different lactic
acid bacteria were less susceptible. Yeasts, represented by Candida
lambica and Cryptococcus humicolus, showed an intermediate sensi-
tivity, the effect of pH on the antimicrobial activity of chitosan on
C. lambica is illustrated in Fig. 21. The growth of C. lambica in
Sabouraud medium (Oxoid, CM147) was not influenced by the pH
in the absence of chitosan. In a medium containing 0.005% (w/v) chi-
tosan the growth was completely inhibited at pH 4.0, while at pH 6.0,
the same chitosan concentration led to a significant extension of the
lag phase and a rather small decrease in growth rate.
Devlieghere and Debevere [154] used soluble starch, which caused a
pH decrease in the growth medium. Three different chitosan concentra-
tions (0%, 0.005% and 0.01% (w/v)) and starch concentrations (0%, 1%
and 30% (w/v)) were tested. Without starch, 0.005% chitosan was
enough to cause a significant retardation of the growth of C. lambica at
pH 5.0 and a higher concentration of chitosan even caused inactivation
of the yeast. Neither the lag phase nor the growth rate of C. lambica was
influenced by low amounts of starch (1% (w/v)).
On the other hand, the activity of chitosan was strongly decreased by
high amounts (30% (w/v)) of starch, leading to a significantly shorter lag
phase and a significantly higher growth rate (Fig. 22), demonstrating the
 M.Z. Elsabee, E.S. Abdou / Materials Science and Engineering C 33 (2013) 1819–1841
Fig. 28. Percentage of infected strawberries as a function of storage time at 10 °C for
control and 1% CS coated samples. Vertical bars indicate standard deviation [178].
negative effect of the investigated type of starch on the antimicrobial
activity of chitosan. It should be mentioned that only the gelatinized
starch was investigated, it is possible that native starch interacts in a dif-
ferent way.
Zhai et al. [155] investigated the effect of radiation on the antibacterial
efficiency of chitosan using starch/chitosan blends irradiated at room
temperature by electron beam (EB) with beam current of 1 mA and
acceleration energy of 2 MeV. In a similar work Liu et al. and Zhai et al.
[110,156] have found that the antibacterial activity of chitosan could be
enhanced by irradiation. The starch/chitosan blend films made by irradi-
ation were tested and Figs. 23 and 24 demonstrate the curves of optical
density (OD) versus culture time for the blend films against E. coli. The
blend films exhibited significant antibacterial activities after irradiation
by 30 kGy compared to the pure or un-irradiated blend films (Fig. 24).
OD of the blend films decreased with irradiation does, which indicate
that the antibacterial activity of blend films improved with increasing
the dose. Fig. 24 showed that even at 5 wt.% chitosan content in the
blend films an obvious antibacterial activity could be observed. Further-
more, it was also found that the antibacterial activity of blend films
enhanced by increasing the content of chitosan in blend systems.
The antimicrobial activity of chitosan edible films incorporated with
garlic oil (GO), (KS) and nisin (N) against E. coli, S. aureus, S. typhimurium,
Fig. 29. Appearance of strawberries coated with modified chitosan-based formulation
containing limonene and emulsifier [179].
1835
L. monocytogenes and B. cereus (common meat product contaminants)
has shown that incorporating GO into chitosan film produced high inhib-
itory zones for S. aureus, L. monocytogenes and B. cereus. It also reduced the
bacterial growth underneath film of E. coli and S. typhimurium. Inhibitory
zone increased by the increasing the GO concentration. L. monocytogenes
was the most sensitive against GO incorporated film followed by S. aureus
and B. cereus [157]. Chitosan film incorporated with potassium sorbate
(KS) showed antimicrobial activity against S. aureus, L. monocytogenes
and B. cereus. There was no effect on E. coli and S. typhimurium whether
in its inhibitory zone or underneath film. Increasing KS level higher than
100 mg/g of chitosan did not significantly improve the antimicrobial
effect.
Similar to GO and KS, incorporating nisin (N) did not show inhib-
itory zone on E. coli and S. typhimurium. However, N-incorporated
chitosan film revealed growth inhibition underneath film disks on
these organisms. Among inhibited microorganisms, L. monocytogenes
was the most sensitive and susceptible to N.
The mechanical, physical and antimicrobial properties of Konjac
glucomannan–chitosan–nisin ternary blend film were studied by Li
and Xie [158] Konjac glucomannan, a hetero polysaccharide derived
from the konjac tuber, consists of 1,4-linked β-D-mannose and
β-D-glucose units in a molar ratio of 1.6:1 with a low degree of acetyl
groups at the side chain C-6 position and having an average molecular
weight of 0.67–1.9 million. However, most of the films do not have
practicable mechanical properties or antimicrobial activity. The results
of mechanical and physical properties tests showed that the blend
film KC2 (mixing ratio konjac glucomannan 80/chitosan 20) had the
maximum tensile strength and a higher transparency greater water sol-
ubility and a lower water vapor transmission ratio among other blend
ratios. This means that incorporating the chitosan to the konjac
glucomannan film enhanced the mechanical and physical properties
remarkably.
The antifungal activity of coatings and films of chitosan/PLA (poly (lac-
tic acid)) based bio-packaging for potential food applications has been in-
vestigated by Fimbeau Sebastien et al. [159]. Three mycotoxinogen fungal
strains, Fusarium proliferatum, Fusarium moniliforme and Aspergillus
ochraceus were tested. Exposure to mycotoxins can produce both acute
and chronic toxicities ranging from death to deleterious effects upon, for
example, the central nervous, cardiovascular and pulmonary systems.
Their general teratogenicity, cancerogenicity and their toxicological prop-
erties constitute a high human and animal health risk. The mycotoxins
also attract attention because of the significant economic losses associated
with their impact on human health and animal productivity. PLA belongs
to the family of aliphatic polyester commonly made from lactic acid,
which can be produced from renewable resources such as starch via fer-
mentation processes [160]. It is a thermoplastic, high strength, high
Fig. 30. Changes in total viable counts (TVC) of fish sample during frozen storage.
Chitosan-coated fish sample (lot I), control sample (lot II) [181].
1836 M.Z. Elsabee, E.S. Abdou / Materials Science and Engineering C 33 (2013) 1819–1841
Fig. 31. L-value of minimally processed Fuji apples during storage at 5 and 20 °C (◆ control sample, ■ 1% chitosan, ▲ 2% AA + 0.5% CaCl2, △ 2% AA + 0.5% CaCl2 + 1% chitosan). Values
are means of three replicates. Vertical bars represent standard deviation [187].
modulus polymer and is considered biodegradable and compostable
[161]. Moreover, it is possible to use it for food contact. Since it is relatively
hydrophobic it can reduce the hydrophilic nature of chitosan-based films
and consequently improve their moisture barrier properties and decrease
the water/matrix interactions. However the films produced although had
antifungal capacity they lacked homogeneity and good mechanical char-
acteristics which made them unpractical as packing materials.
The antifungal properties of films and solutions based on chitosan
with different molecular weight at different concentrations have been
investigated by Ziani et al. [162]. Surfactants were added to the for-
mulation to assess their impact on treatment efficiency. The antifun-
gal activity was conducted against three fungi, Aspergillus niger,
Alternaria alternata and Rhizopus oryzae. Results indicated important
and significant differences of the antifungal activity between chitosan
based solutions and chitosan based films. Furthermore, the antifungal
activity of the different treatment depended on the type of fungus
treated. Thus, chitosan film treatments were significantly more effec-
tive on A. niger than solution treatments. On the other hand, solution
treatments resulted in higher radial inhibition when applied against
A. alternata or R. oryzae. The highest radial inhibition was observed
against A. alternata (97%) using chitosan solution. The influence of
the other parameters (concentration, molecular weight and surfac-
tant type) on treatment efficiency was not as important and their
significance depended on treatment type and fungus nature [163].
Antimicrobial activity of edible coating solutions based on chitosan
and blends of chitosan–tapioca starch with or without potassium
sorbate (KS) addition against Lactobacillus spp. and Zygosaccharomyces
bailii was studied by Vasconez et al. [1]. They found that the chitosan
antibacterial action suffered the antagonist effect of the presence of KS
and/or tapioca starch. Interactions between chitosan–starch and
chitosan–KS, might inhibit chitosan amino groups bonding to the cell
Fig. 32. a-value of minimally processed Fuji apples during storage at 5 and 20 °C (◆control sample, ■ 1% chitosan, ▲ 2% AA + 0.5% CaCl2, △ 2% AA + 0.5% CaCl2 + 1% chitosan). Values
are means of three replicates. Vertical bars represent standard deviation.
membrane, depressing its antimicrobial activity. The films studied
were not effective against Lactobacillus spp. Therefore, among the
films studied, the one formulated with chitosan and tapioca starch
could have possible applications as a barrier against Z. bailii contamina-
tion in a high water product of reduced pH. Their results suggested that
the chitosan antibacterial action depended on the technique of applica-
tion. Chitosan was more effective in the coating solution than in the film
matrix.
The antimicrobial activities of biodegradable films of sweet potato
starch containing KS or chitosan on E. coli and S. aureus have been
studied by Shen et al. [52]. The sweet potato film control did not
show any inhibitory effect on E. coli or S. aureus. Films incorporated
with potassium sorbate (KS) ≥ 15% or chitosan ≥ 5% were found to
have an anti-E. coli effect. S. aureus could be effectively suppressed
by incorporation of chitosan at ≥ 10%. While KS lowers the tensile
strength and elongation at break, and raises the oxygen permeability,
WVP and water solubility, chitosan has the opposite effect. Among
the food-borne bacteria, E. coli and S. aureus are observed in a wide
range of food products. Furthermore, they are the human pathogens
that cause the most economically important food-borne diseases
throughout the world [164] E. coli is the most common bacteria
from human feces, and S. aureus is one of the indigenous microbiota
on human skin [165].
The antibacterial activity of chitosan–starch film using microwave
treatment has been carried out using agar plate diffusion method
[166]. The antibacterial activity of the film and their same solution has
been evaluated against three different test cultures viz. Gram-negative
bacteria E. coli, Gram-positive bacteria S. aureus and Gram-positive bac-
teria Bacillus subtilis. It was found that the solution of chitosan–starch
showed inhibitory effect against above said test cultures but film proved
to be negative.
 M.Z. Elsabee, E.S. Abdou / Materials Science and Engineering C 33 (2013) 1819–1841
Fig. 33. Mean values and standard deviations of firmness, variation with storage time
at 5 °C (◆ control sample, ■ 1% chitosan, ▲ 2% AA + 0.5% CaCl2, △ 2% AA + 0.5%
CaCl2 + 1 % chitosan). Values are means of three replicates [187].
Incorporating chitosan and lauric acid into starch based film showed
more effective antimicrobial ability against B. subtilis and E. coli [167],
and the film had synergistic antimicrobial effect when chitosan and
lauric acid were combined. The starch-based film incorporated with
lauric acid and chitosan showed better flexibility than when purely
starch-based film was formulated. The solution of starch and chitosan
with different mixing ratios (w/w) 8:2 and 9:1 were the most effective
ratio with greater inhibition on both B. subtilis and E. coli than other
solution in agar plate and liquid culture test. The control (pure wheat
starch) and antimicrobial (AM) film (incorporated with chitosan and
lauric acid) were produced by casting method. The antimicrobial effec-
tiveness of control (pure wheat starch) and AM film incorporated with
chitosan and lauric acid are shown in Fig. 25A and B. A wide clear zone
on solid media was observed for B. subtilis growth inhibition whereas
inhibition for E. coli was not as effective as B. subtilis. From the liquid cul-
ture test, the AM films clearly demonstrated a better inhibition against
B. subtilis than E. coli.
8. Increasing the shelf life of foods
Edible coatings could reduce moisture transfer, restrict oxygen up-
take, lower respiration, retard ethylene production, seal in flavor vola-
tiles and carry additional functional ingredients (such as antioxidants
and antimicrobial agents) that retard microbial growth and potential
discoloration. Many authors have investigated chitosan coatings for
their potential to enhance the quality and extend the storage life of
food products [12,17,168–172].
Elsabee et al. [172] used the modified polypropylene films by chito-
san and chitosan/pectin multilayer as a packaging device for storing and
increasing the shelf life of tomato they fabricated bags 20× 20 cm using
12 multi layers (composed of chitosan/pectin) over corona treated
Fig. 34. Weight loss from Fuji apple slices stored 8 days at 5 °C and 85%RH (◆ control
sample, ■ 1% chitosan, ▲ 2% AA + 0.5% CaCl2, △ 2% AA + 0.5% CaCl2 + 1% chitosan).
Values are means of three replicates [187].
1837
Fig. 35. Effect of edible coatings on the initial respiration rate of fresh-cut apple slices
stored at 5 °C for 8 days (♦ control sample, ■ 1% chitosan, ▲ 2% AA + 0.5% CaCl2, △ 2%
AA + 0.5% CaCl2 + 1% chitosan). Values are means of three replicates [187].
polypropylene (PP) films. A fresh tomato fruits of regular size (50 g)
were collected cleaned and stored in bag (A). Another similar fruit
was stored in regular PP bag (B) and a third one was kept in air (C).
The three species were kept in a refrigerator at 4 °C. The above experi-
ment was done in triplicate. The samples were investigated at intervals,
and after 13 days the samples were compared and Fig. 26 shows the
state of the three samples. It can be seen that samples B and C deterio-
rated completely while the sample in the treated bag was kept almost
intact with no apparent rotting infection.
Application of chitosan coating to browning control and quality
maintenance of fresh-cut Chinese water chestnut (CWC) was investi-
gated by Pen and Jiang [173]. Fresh-cut CWC were treated with aqueous
solution of 0.5, 1 or 2 g chitosan/100 ml, placed into trays over-
wrapped with plastic films, and then stored at 4 °C. The result of their
work is shown in Fig. 27 surface discoloration of fresh-cut CWC
appeared after 3 days of storage at 4 °C and became more serious
after 6 days, while the eating quality decreased markedly. Treatment
with chitosan coating delayed the development of the discoloration.
Furthermore, increasing the concentrations (from 0 to 2 g) of chito-
san/100 ml coating enhanced the inhibition of the discoloration and
resulted in better appearance maintenance. Application of chitosan
coating also inhibited effectively the disease development of fresh-cut
CWC. In the current study, as the use of chitosan coating in combination
with low temperature storage was effective for control of overgrowth of
spoilage organisms, the surface discoloration of the fresh-cut CWC
appears to be major factor limiting storage life.
Carrot (Daucus carota L.) is one of the most popularly consumed veg-
etables, but marketing is limited by its fast deterioration during storage,
due to physiological changes that reduce its shelf life [174]. The product
loses its firmness and develops odors characteristic of anaerobic catabo-
lism, due to the high respiration rate and microbiological deterioration
[175]. Minimally processed carrot quickly loses their bright orange
color during storage, developing a whitish appearance or white blush
on its surface. Durango et al. [14] developed an edible antimicrobial coat-
ing based on a starch–chitosan matrix to evaluate its effect on minimally
processed carrot by means of microbiological analyses. The results of this
experiment showed that the use of an antimicrobial coating consisting of
chitosan-added yam starch is a viable alternative in controlling the
microbiota present in minimally processed carrot, since the growth of
lactic acid bacteria, total coliforms, psychrotrophs, yeasts and molds
and mesophilic aerobes, was substantially inhibited by the application
of 1.5% chitosan coating. Based on the concept of protection barrier tech-
nology, the use of such coating may contribute to improve safety in min-
imally processed carrot thereby prolonging its shelf life. Coating may be
applied on minimally processed fruits and vegetables, combined to other
types of controls, such as quality raw material, hygienic processing con-
ditions and storage temperatures. The combination of these treatments
1838 M.Z. Elsabee, E.S. Abdou / Materials Science and Engineering C 33 (2013) 1819–1841
as barrier offers a greater potential for shelf-life extension of minimally
processed vegetables.
Mango pulp is very perishable and so has a short shelf life, which both
marketers and consumers would like to be longer. Chien et al. [176] man-
ually treated sliced mango with aqueous solutions of 0%, 0.5%, 1% or 2%
chitosan; Chitosan coating retarded water loss and the drop in sensory
quality, increasing the soluble solid content, the acidity and ascorbic
acid content. It also inhibited the growth of microorganisms. The data
reveal that applying a chitosan coating effectively prolongs the quality
attributes and extends the shelf life of sliced mango fruit.
Chitosan (CS1, 1% w/w solution) was used for producing chitosan
film and used for glazing skinless pink salmon filets by Sathivel et al.
[171] Chitosan glazing delayed lipid oxidation in skinless pink salmon
filets after eight months frozen storage. Chitosan film was a very good
barrier to oxygen, while having low WVP. Chitosan coated filets had a
higher thaw yield than that of the control non-glazed. Their study
demonstrated the potential of chitosan solution as an edible glazing
for pink salmon fillets.
Casariego et al. [26] aimed to determine the effects of the concentra-
tions of glycerol and sorbitol (as hydrophilic plasticizers), Tween 80 (as
surfactant) and chitosan on the wettability of Cuban chitosan-based
edible coatings in view of their application on tomato and carrot and
to develop a model allowing the optimization of coating composition.
Their results indicated that chitosan obtained from lobster of the
Cuban coasts can be recommended as an edible coating to be applied
on fruits and vegetables, possibly contributing to extend their shelf life.
Strawberry (Fragaria x ananassa) is a highly perishable non-
climacteric fruit. It must be harvested at full maturity to achieve maxi-
mum quality in terms of visual appearance (freshness, color and absence
of decay or physiological disorders), texture (firmness, juiciness and
crispness), flavor and nutritional value (vitamins, minerals, dietary
fiber and phytonutrients). Gray mold, caused by Botrytis cinerea Pers.
Fr., is the most economically significant postharvest pathogen of straw-
berry fruits. Strawberry spoilage after harvest can also occur by mechan-
ical injury and desiccation. Low storage temperatures and modified
atmospheres with elevated CO2 levels are common tools for avoiding,
at least partially, mold growth and senescence, and for extending fruit
shelf-life. However, prolonged exposure of berries to high CO2 concen-
trations can cause off-flavor development [177].
Hernandez-Munoz et al. [178] studied the effect of chitosan coatings
combined with calcium gluconate on strawberry (Fragaria x ananassa cv.
Camarosa) quality attributes during refrigerated storage. Strawberries
were treated with 1% or 1.5% chitosan acetate solution, with or without
the addition of calcium gluconate. Assessment of the treatments is
based on their effects on fungal decay, respiration rate, quality attributes,
and the visual appearance of strawberries stored for 6 days at 10 °C.
Strawberries were randomly distributed into five groups. Four groups
were assigned to one of four treatments while the fifth group provided
the untreated control. The treatments consisted in immersing fruits for
5 min in: (a) 1% chitosan acetate; (b) 1.5% chitosan acetate; (c) 1% chito-
san+0.5% calcium gluconate; and (d) 1.5% chitosan +0.75% calcium
gluconate solution. Fruits were allowed to dry for 2 h at 20 °C and
were subsequently stored at 10 °C and 70±5% RH.
Uncoated strawberries showed signs of fungal decay after the third
day of storage at 10 °C (Fig. 28). After 6 days of storage, 33.5% of
uncoated fruit was infected by molds while no sign of fungal decay
could be detected by visual inspection of fruits coated with 1.5% chitosan
or 1.5% chitosan+0.75% calcium gluconate. Of the fruit coated with 1%
chitosan, 12.5% was observed to be infected on the sixth day of storage.
In summary, chitosan coating was seen to delay fruit senescence
and fungal decay of strawberry fruits stored at 10 °C and 70 ± 5% rel-
ative humidity.
Vu et al. [179] developed edible bioactive coating based on modified
chitosan for increasing the shelf life of strawberry during storage. They
found that, the functionalized chitosan-based edible coating could be-
come a promising method to carry specific antifungal agents without
detrimental effects on strawberries or other fruits. In this context, future
research will focus on developing a new coating formulation which con-
sists of modified chitosan, lipid (fatty acids, acetylated monoglycerides,
etc.), plasticizers (polyols) and reinforcement agents (nanocellulose) to
improve the functional properties of coatings applied on strawberries
and increase the shelf life of strawberries during storage.
Fig. 29 shows the appearance of strawberries coated with modi-
fied chitosan.
Duan et al. [180] used acid-soluble chitosan (ACS) and water-soluble
chitosan (WCS) for enhancing the shelf life and retaining the antioxi-
dant properties of pre-washed, ready-to-eat high bush blueberry culti-
vars under commercial storage conditions. Their results from this study
indicate the possibility of using edible coatings to develop ready-to-eat
fresh blueberries with no reduction in shelf life. The key for success is
using an appropriate coating material, container, and method of apply-
ing the coatings. In this study, different coatings showed various effects
on the post-harvest quality of pre-washed fresh blueberries. Both
acid-soluble and water-soluble chitosan coatings showed potential for
reducing rate of decay of blueberry during room temperature storage.
The effects of chitosan coating on quality and shelf life of silver carp
during frozen storage were investigated by Fan et al. [181]. Fish samples
were treated with aqueous solution of 2% chitosan, and then stored at −
3 °C for 30 days. Total viable counts are shown in Fig. 30. Fish samples
were given a dip treatment in 2% chitosan solution (lot I) and in 1% gla-
cial acetic acid (lot II) as a control, respectively for 120 min and then
drained well. After that, they were individually packed in plastic trays
and airproofed with polyvinyl dichloride (PVDC); then all the packs
were kept in a refrigerator maintained at −3 °C for 30 days [182,183].
Fish samples were taken randomly every 5 days for microbiological,
chemical and sensory evaluation.
The results of microbiological (total viable count), chemical (pH,
TBA, TVB-N, and K-value), and sensory evaluation analyses indicate
that chitosan coating on silver carp (Hypophthalmichthys molitrix) can
lead to retention of the good quality characteristics and extension of
the shelf life during frozen storage.
Shelf-life extension of Ricotta cheese was studied by Di Pierro et al.
[184] using chitosan/whey protein film as active coating and stored
under modified atmosphere at 4 °C. The chitosan/whey protein film
had 35% and 21% lower oxygen and carbon dioxide permeability, re-
spectively, and about three times higher WVP than film prepared with
chitosan alone, they found that edible film reduced growth of microbial
contaminants and extended the shelf-life of the product packed under
modified atmosphere. The coating delayed the development of undesir-
able acidity, better maintained the texture and did not seem to modify
sensory characteristics. It is possible that the benefits derived from the
biopolymers can also be realized with other dairy products.
A composite coating has been developed by Maqbool et al. [185] by
combining Arabic gum and chitosan to evaluate its potential to control
anthracnose in banana during and after cold storage. They found that
Arabic gum alone did not show any fungicidal effects while the combi-
nation of 1.0% chitosan with all tested Arabic gum concentrations had
fungicidal effects. However, the potato dextrose agar (PDA) medium
amended with 10% Arabic gum incorporated with 1.0% chitosan showed
the most promising results among all treatments in suppressing the
mycelial growth (100%) and conidial germination inhibition (92.5%).
Xing et al. [186] investigated the effect of chitosan coating containing
anti-browning agents and modified atmosphere packaging (MAP) on the
browning and shelf life of fresh-cut lotus root stored at 4 °C for 10 days.
Their study has industrial impact where fresh-cut vegetables have drawn
the attention of industry as a novel lightly processed product. Both edible
coating and (MAP) treatment cause changes in atmosphere composition
and respiration rate of lotus root slices. This combined treatment could
be used to control the browning and improve the storage life of this
fresh-cut vegetable. This information could be useful for the develop-
ment of novel application to edible coating and (MAP) design for lightly
processed lotus root.
 M.Z. Elsabee, E.S. Abdou / Materials Science and Engineering C 33 (2013) 1819–1841
The effect of coatings in combination with anti-browning agents (1%
chitosan; 2% ascorbic acid+0.5% CaCl2 and 2% ascorbic acid+0.5%
CaCl2 +1% chitosan) on minimally processed apple slices was studied
during storage by Qi et al. [187]. Their results show that chitosan-
coating treatments effectively retarded enzymatic browning on mini-
mally processed apples during storage and they effectively retarded or
avoided tissue softening, apple slices underwent a little loss of firmness.
Chitosan-coating did not perform very well as water vapor barriers in
apple slices. Figs. 31 and 32 show effects of chitosan-coating on L*
value (lightness) and a* value (colorimetric) of apple pieces. In compar-
ison with control sample, chitosan sample almost remained stable L* and
a* values at 20 °C for 24 h. L* values of all sample decreased rapidly after
24 h for 20 °C and decreased slightly after 24 h at 5 °C (Fig. 31B). But the
a* values of all samples increased rapidly after 24 h at 20 °C and in-
creased slightly after 24 h at 5 °C (Fig. 32B). This indicated that low tem-
perature storage was better than high temperature storage to retard
browning of apple pieces.
Their results (Fig. 33) showed that 0.2% AA+ 0.5% CaCl2 + 1% chito-
san treated apples had the highest firmness, since calcium chloride, a
known firming agent, helped apple slices maintain firmness. Chitosan
at 1% content treated apples also showed constant firmness throughout
8 days of storage due to low level of Ca2+ contained in aqueous chito-
san solution. In contrast with chitosan coatings, non-coated apples
showed reduced firmness.
After 2 days of storage, control apples lost around 19% of their
weight, while coated apples lost 15% of their weight (p> 0.05). Chitosan
coatings on apple slices did not prove to work effectively as water vapor
barriers during the entire storage period (Fig. 34).
The physical damage or wounding caused by peeling and cutting in-
creases respiration rate within minutes, thus the need to reduce initial
respiration rate is critical in extending shelf-life of minimally processed
fruits so that Qi et al. [187] studied also the respiration rate as shown in
Fig. 35. The effect of chitosan edible coatings on initial respiration rate,
all formulations tested reduced initial respiration rate, and 2% AA+0.5%
CaCl2 +1% chitosan had a significantly greater effect than others.
Minimally processed broccoli was coated with either chitosan or
carboxymethyl-cellulose (CMC) with or without a previous application
of a mild heat shock of 1.5 min at 50 °C. This work was made by Ansorena
et al. [188]. They found that the edible coatings, chitosan and
carboxymethyl-cellulose, seemed to have a beneficial impact on quality
retention of minimally processed broccoli by retarding their weight loss,
browning and yellowing processes, reducing stem hardening, microbial
growth and improving total chlorophyll and ascorbic acid retention.
Moreover, both edible coatings were able to inhibit the florets opening,
which is an important quality improvement for broccoli. However signif-
icant differences were observed among these coatings: chitosan was
superior during storage maintaining broccoli in higher quality levels rela-
tive to CMC coating by reducing total microbial counts, hardening, O2 con-
sumption and improving ascorbic acid retention.
9. Conclusion
In view of the severe environmental pollution caused by plastic food
packaging, there is a considerable interest in edible and biodegradable
films made from renewable and natural polymers. Chitosan, a natural,
non-toxic, biodegradable polymer, available commercially, has been
employed in a variety of applications in food industry as a new edible
packaging material to control the food quality; it can form transparent
films, which may find application in a variety of packaging needs.
Blending of chitosan with other natural polymers gives films and coat-
ings with good properties, chitosan addition in edible films leads to
good film forming and mechanical properties, no toxicity, biodegrad-
ability, relative more hydrophobic nature that could provide higher
moisture barrier and water resistance.
Many researchers studied different properties of chitosan and chi-
tosan blends (preparation, physical, mechanical, rheological, water
1839
vapor permeability and antimicrobial properties) their results show
that chitosan and chitosan blends are extremely promising materials
for bio-based active films preparation and that chitosan based edible
films and coatings can be useful for preserving and extending the
shelf life of foods.
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