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Innovative Food Science and Emerging Technologies 62 (2020) 102346
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Innovative Food Science and Emerging Technologies

journal homepage: www.elsevier.com/locate/ifset
Chitosan-based biodegradable functional films for food packaging T

applications
⁎
Ruchir Priyadarshi, Jong-Whan Rhim
Department of Food and Nutrition, BioNanocomposite Research Center, Kyung Hee University, 26 Kyungheedae-ro, Dongdaemun-gu, Seoul 02447, Republic of Korea
A R T I C LE I N FO A B S T R A C T
Keywords: Chitin is the structural material of crustaceans, insects, and fungi, and is the second most abundant biopolymer
Chitin after cellulose on earth. Chitosan, a deacetylated derivative of chitin, can be obtained by deacetylation of chitin.
Chitosan It is a functionally versatile biopolymer due to the presence of amino groups responsible for the various prop-
Biodegradable erties of the polymer. Although it has been used for various industrial applications, the recent one is its use as a
Functional
biodegradable antimicrobial food packaging material. Much research has been focused on chitosan-based flex-
Nanocomposite
Food packaging
ible food packaging and edible food coatings to compete with conventional non-biodegradable plastic-based
food packaging materials. Various strategies have been used to improve the properties of chitosan - using
plasticizers and cross-linkers, embedding the polymer with fillers such as nanoparticles, fibers, and whiskers, and
blending the polymer with natural extracts and essential oils and also with other natural and synthetic polymers.
However, much research is still needed to bring this biopolymer to industrial levels for the food packaging
applications.
Industrial relevance: As a major by-product of the seafood industry, a massive amount of crustacean shell waste is
generated each year, which can be used to produce value-added chitin, which can be converted to chitosan using
a relatively simple deacetylation process. Being extracted from a bio-waste product using many energy-efficient
methods, chitosan is much cheaper as compared to other biopolymers. Nevertheless, the exceptional properties
of chitosan make it a relatively stronger candidate for food packaging applications. Chitosan has already been
used in various industries, such as biomedical, agriculture, water treatment, cosmetics, textile, photography,
chromatography, electronics, paper industry, and food industry. This review article compiles all the essential
literature up to the latest developments of chitosan as a potential food packaging material and the outcomes of
its practical utilization for this purpose.
1. Introduction betterment and innovations kept on growing, which led to the begin-
ning of research in the field of food packaging. The far-reaching con-
The packaging is the art, science, and technology to deliver goods to sequences of this beginning led to the formation of packaging materials
end consumers at economical prices securely. The food packaging that are being used in contemporary times.
process is of immense importance when preservation of fresh or pro- Paper has been considered as one of the most economical packaging
cessed food is concerned. It plays a major role in making the food, materials. In 1844, first commercial paper bags were manufactured in
which is prepared in one place, available to consumers at some other Bristol, England, which initiated the use of papers in packaging for the
place after days, weeks, or even months after initial harvest or manu- first time on an industrial scale (Risch, 2009). From the early 1900s to
facture. The nomadic humans used to gather food only at the time of the late 1960s, glass containers dominated the food packaging market
consumption. With the increased need for homes and shelters, and with primarily for the liquid products. Glass packaging is still the most ef-
the development in agriculture, it became necessary to store food for fective and preferred one for the products which require strong flavor
future use. That was the time when the need for storage and packaging and aroma protection (Berger, 2002; Risch, 2009). In 1809, Nicholas
arose. The materials that served the purpose were leaves, shells, woven Appert discovered that boiling a sealed container containing food helps
grasses, hollowed logs, animal organs, etc. (Berger, 2002). However, to preserve food longer. This discovery opened the door for food ster-
these storage forms were rather unsophisticated, and the need for ilization using metal containers for food packaging. In 1830, the selling
⁎
Corresponding author.
E-mail address: jwrhim@khu.ac.kr (J.-W. Rhim).
https://doi.org/10.1016/j.ifset.2020.102346
Received 5 December 2019; Received in revised form 25 March 2020; Accepted 30 March 2020
Available online 01 April 2020
1466-8564/ © 2020 Elsevier Ltd. All rights reserved.
R. Priyadarshi and J.-W. Rhim Innovative Food Science and Emerging Technologies 62 (2020) 102346
of cookies in sealed metal containers started (Berger, 2002; Risch, Chitin is a cellulose-like biopolymer (Fig. 1) that is present in the
2009). With the invention of various kinds of synthetic polymers in the exoskeletons of crustaceans and insects and cell walls of fungi and
19th century, plastic-based food packaging materials entered the yeast. Chitin acts as a structural material, just like cellulose plays a role
market. Vinyl chloride and polyethylene terephthalate (PET) are used in plant cells. It is a polymer composed mainly of (1-4)-linked 2-acet-
today to make bottles for packaging water and other beverages. PET is amido-2-deoxy-β-D-glucose monomers and is produced at an enormous
also used to pack foods and hot-fill products like jam (Berger, 2002; quantity of 1011 metric tons annually (Elieh-Ali-Komi & Hamblin,
Risch, 2009). 2016). However, only 150,000 tons out of this are made available for
An ideal food packaging material is the one that protects the food commercial use (Guan et al., 2019). With this figure, it is the second
quality over time; it is easily portable and convenient to use; it is in- most abundant biopolymer on earth after cellulose and the most
expensive; and is renewable or biodegradable, resulting in no municipal abundant biopolymer of animal origin. It is different from other poly-
solid waste accumulation. However, there is no such ideal material saccharides because of its nitrogen content. Chitosan is the deacetylated
produced to date, and each material has its pros and cons, which are not derivative of chitin and is majorly composed of (1-4)-linked 2-amino-2-
absolute but relative depending on these factors. deoxy-β-D-glucose monomers. Properties such as the degree of acet-
Depending on material properties and food packaging needs, the ylation (DA), nitrogen composition, N/C ratio, molecular size, and
glass may be the best choice of packaging materials because it has high polydispersity are dependent on the source of chitin. In general, chitin
barrier properties, is inert and transparent. However, it lags due to its has a DA above 90%, the nitrogen content of 7% and N/C ratio of
brittleness, heaviness, and poor portability. Metal and metal sheets give 0.146. Chitosan has a DA of less than 40% and nitrogen content greater
good competition to glass, but it also lags due to low inertness, no than 7%. The presence of the amino group in chitosan renders it with
transparency, and low portability due to weight. This is where paper functional properties; the electronegative amino group takes up protons
and plastics come in, which are inexpensive, flexible and inert. Plastics and develops a positive charge providing chitosan with various che-
also have good tensile and barrier properties, good transparency, and mical, physical, and biological properties. Solubility, film formation,
lightweight along with functional advantages like moldability, heat viscosity, ion binding, antimicrobial properties, etc. make chitosan a
sealability, etc. (Marsh & Bugusu, 2007). versatile polymer capable of applications in different areas of research,
Plastics and polymers are by far the most utilized and best-con- including food packaging (Wang, Qian, & Ding, 2018).
sidered resources for food packaging applications. They have in-
tensively been used in rigid as well as flexible packaging. However, 2. Sources of chitosan
plastics are non-biodegradable, and according to the report of the
United States Environmental Protection Agency regarding the muni- The most popular and most economical way for the isolation of
cipal waste generation (MSW) in 2014, the contribution of plastics is chitosan is the deacetylation of chitin. However, it is also possible to
worth pondering. The total MSW generated in 2014 was 258 million extract chitosan directly from some fungi (Rane & Hoover, 1993; White,
tons, of which 32.28 million tons were plastic wastes, accounting for Farina, & Fulton, 1979). Chitin serves as the structural material for a
12.9% of the total MSW. Plastic packaging, which accounts for 43% of wide range of organisms of the animal and fungal kingdom. Some
this plastic waste, generated 14.32 million tons (US EPA, 2014). common sources of chitin with their chitin content are given in Table 1.
Ever since the use of plastics started in food packaging, MSW gen-
eration has been a big environmental issue. This has led to the initiation 3. Chitosan production and processing
of research on Biopolymers, which are not only biodegradable but show
properties like being antimicrobial and antioxidative, which can be The process of chitosan production from animal waste is a two-step
beneficial in enhancing the shelf life of food products. protocol involving the extraction of chitin from biological waste and
The industrial production of any product brings with it the un- then conversion of the extracted chitin into chitosan.
avoidable production of a lot of by-products, which are most of the
time, discarded as wastes. Although a lot of byproducts have the po- 3.1. Extraction of chitin
tential to be used in some alternate way, they usually go unutilized. The
sea-food processing industry produces a lot of animal wastes like skins Chitin can be extracted from animal waste in two ways; Chemical
and shells of organisms. Approximately 75% of the weight of crusta- method and Biological method. The chemical method, despite its sev-
ceans contributes to the byproduct waste, and due to the inefficient eral disadvantages like being uneconomical and non-eco-friendly, it is
waste management protocols, they lead to serious environmental ha- the commonly used commercial method due to its shorter processing
zards; they are drowned in sea, land-filled, burned, or left for spoiling time (Arbia et al., 2013). However, in a comparative study to extract
naturally. These animal wastes are rich in proteins and are excellent chitin from prawn and shrimp shells by both chemical as well as bio-
sources of chitin. The utilization of this waste for chitin extraction may logical method, it was observed that the biological method resulted in a
provide a way to minimize environmental hazards while facilitating higher yield of chitin (Beaney, Lizardi-Mendoza, & Healy, 2005;
value addition. Khanafari, Marandi, & Sanatei, 2008). Although biological extraction,
Fig. 1. Structure comparison of chitosan monomer (glucosamine) and cellulose monomer (glucose) (illai, Paul, & Sharma, 2009).
2
Table 1 Chitosan was found to be present naturally in the cell walls of some
Various sources for chitin extraction with percentage content in each by dry fungi, and it was observed that the presence of chitin deacetylase en-
mass (Hamed, Özogul, & Regenstein, 2016). zyme in those fungal strains which was responsible for the conversion
Organism Chitin References of cell wall chitin to chitosan. However, the fungal strains used to
content (%) produce chitin deacetylase give a very low enzyme yield and have a
complicated fermentation requirement. Furthermore, the enzyme iso-
Crustaceans Arbia, Arbia, Adour, & Amrane,
lated from a fungal source shows the lower activity as compared to the
Nephro (lobster) 69.8 2013; Synowiecki and Al-Khateeb
Euphausia superb (krill) 24.0 (2003) ones isolated from bacterial strains. Also, bacteria grow faster and ea-
Homarus (lobster) 60.0 - 75.0 sier on large scale fermentation systems. Therefore, the isolation of
Crangon crangon 17.8 chitin deacetylase from bacterial strains is gaining popularity (Kaur,
(Shrimp)
Dattajirao, Shrivastava, & Bhardwaj, 2012; Teng, 2011; Zhou, Zhang,
Lepas (goose barnacle) 58.3
Chionoecetes opilio 26.6
He, & He, 2010).
(Crab)
Insects Kaur and Dhillon (2015) 3.3. Industrial scenario of chitosan
Blatella (cockroach) 18.4
Coleoptera (ladybird) 27.0 -35.0
With over 30 years of research on chitin and chitosan, the re-
Diptera 54.8
Pieris (butterfly) 64.0 searchers have found that the biopolymer possesses some unique
Bombyx (silkworm) 44.2 characteristics and properties (Section 4), which have made it a ver-
Galleria (wax worm) 33.7 satile material to be used in numerous applications. The last few years
Fungi Synowiecki and Al-Khateeb (2003)
have witnessed a rise in the number of chitosan manufacturers world-
Aspergillus niger 42.0
Penicillium notatum 18.5
wide that producing different grades of chitosan suitable for different
Penicillium chrysogenum 19.5 - 42.0 applications. Iceland based Primex commercializes chitosan-based
Saccharomyces gutulata 2.3 skincare and water purification products with a brand name Chit-
Mucorrouxii 9.4 oClear® and SeaKlear®, respectively. The products are claimed as an-
Siboglinidae 33.0 Crini, Guibal, Morcellet, Torri, and
timicrobial and non-toxic (Primex EHF, 2020). Norwegian Chitosan
Badot (2009)
Cnidaria 3.0 - 30.0 Crini et al. (2009) produces chitosan-based KitoFlokk® for water and wastewater treat-
Brachiopod 4.0 - 29.0 Crini et al. (2009) ment applications (Norwegian Chitosan, 2020). China-based Chibio
Mollusks Crini et al. (2009) Biotech produces chitosan from fungal sources like Aspergillus and
Squid, cuttlefish, 6.0 - 40.0 mushrooms for use in food, beverages, cosmetics, pharma, and agri-
octopus
culture (Chibio Biotech, 2019). Canada based ChinovaBioworks is an-
other company that produces high purity food grade chitosan from
being safe and economical, is a rapidly growing topic of interest, its use mushrooms exclusively for use as a food preservative (Chinova
is still limited to laboratory research (Kaur & Dhillon, 2015). Bioworks, 2019). Meron Biopolymers, India manufactures chitosan
The basic protocol for the extraction of chitin involves washing, using the locally abundant crustacean resources like crabs and shrimps
drying, and crushing of the raw material obtained from various sources for biomedical, cosmetic, agricultural, and food applications (Meron
to get a fine powder. This powder then undergoes a series of the three- Biopolymers, 2016). SFlyGreentech, based in France, utilized insect
step process - demineralization, deproteinization, and decoloration. Black Soldierfly as the source for chitosan extraction. The product is
These three steps are common for both the chemical as well as biolo- endorsed primarily for use in cosmetics and pharmaceuticals (SFly
gical methods (Hamed et al., 2016). Greentech, 2017). Heppe Medical Chitosan GmbH, Halle, Germany, and
Advanced Biopolymers, Trondheim Norway manufactures chitosan-
based products for biomedical applications (Heppe Medical Chitosan
3.2. Conversion of chitin to chitosan
GmbH, 2020). The grade and purity of the chitin and chitosan govern
their market price. G.T.C. BioCorporation, Quingdao, China commer-
The chitin obtained, as described in Section 3.1, is then converted to
cializes chitin at 20 € per kg and chitosan between 18 € and 45 € per kg
chitosan by the process known as deacetylation, which involves the
(Ferreira, Alves, & Coelhoso, 2016).
removal of acetyl groups (COCH3) from the chitin structure. This N-
deacetylation is never complete, and hence chitosan with different
4. Properties of chitosan
degrees of deacetylation is obtained (Abdulkarim, Isa, Abdulsalam,
Muhammad, & Ameh, 2013) as shown in Fig. 2. The deacetylation of
4.1. Ion binding
chitin is done by two methods - alkaline method and enzymatic method.
Chitosan can form complexes with transition metals and post-tran-
3.2.1. Alkaline method sition ions. Several studies suggest that chitosan can bind to ions such as
The chitin powder is added to a 40-50 % NaOH solution. This Cu2+, Hg2+, Zn2+, Cd2+, Ni2+, etc. According to Varma et al., the
process leads to hydrolysis of acetyl groups and conversion of N-acetyl- binding of Cu2+, Hg2+, Zn2+, Cd2+, and Ni2+ to chitosan is maximum
D-glucosamine units into D-glucosamine units having free amine groups. 222, 815, 164, 558, and 75 mg/g chitosan, respectively (Varma,
The degree of deacetylation (DD) of the chitosan produced by this Deshpande, & Kennedy, 2004). However, Vold et al. reported that from
method depends on the reaction time, temperature, and concentration a mixture of binary ions Cu2+, Zn2+, Cd2+, and Ni2+, the probability
of the alkali solution used. This method consumes a lot of energy and that chitosan will bind to Cu2+ was much higher than other ions (Vold,
causes environmental pollution, and hence the trend is shifting towards Vårum, Guibal, & Smidsrød, 2003). This ion binding property is the
a more eco-friendly enzymatic method (Jung & Zhao, 2011; Raval, most important one in the context of active food packaging as it pro-
Raval, & Moerschbacher, 2013; Teng, 2011; Yuan, Chesnutt, Haggard, vides the biopolymer with the ability to bind metallic nanoparticles and
& Bumgardner, 2011). their controlled release leading to prolonged antimicrobial activity.
Wang et al. (2012) demonstrated the synthesis of chitosan-Ag/poly-
3.2.2. Enzymatic method vinylpyrrolidone films through ion complexation and their behavior
Chitin deacetylase enzyme is employed to convert chitin to chit- regarding the controlled release of silver ions to show prolonged anti-
osan. This enzyme was first isolated in 1974 from fungi Mucorrouxii. microbial activity.
3
Fig. 2. Structure of chitin and chitosan-based on varying degrees of deacetylation (Pillai et al., 2009)
4.2. Solubility
Three important parameters determine the solubility of chitosan in

water
• the pH of the solution

• the ionic strength of the solvent which guides salting-out effect
• ions in the solvent interacting with chitosan and limiting its solu-
bility
Chitosan has a characteristic property of being soluble at acidic and

insoluble at basic pH values. All varieties of chitosan are soluble below
a pH value of 6 (Vårum, Ottøy, & Smidsrød, 1994). However, chitosans
with medium molecular weight (MW) and DA around 0.5 can be soluble
at pH 7. The variation in the solubility values with pH for chitosans
with different DA is shown in Fig. 3. The difference in solubility values
can greatly affect the biological properties of chitosan due to the al-
teration of its availability to enzymes (Stephen, Phillips, & Williams,
2006; Tømmeraas, Vårum, Christensen, & Smidsrød, 2001).
4.3. Charge density

Fig. 3. Solubility vs pH curves for chitosans with DA = 0.01 (■), 0.14 (◊), 0.37
The charge density of chitosan depends on the degree of protonation
(+), 0.60 (○) (Vårum et al., 1994).
of the amino group. It is determined by DA of chitosan as well as pH and
ionic strength. The dissociation constant of chitosan varies in the range
of 6.2 to 7.0 depending on chitosan type and ambient conditions. 4.4. Chain conformation
Chitosan samples with different DA values were tested for their mean
electrophoretic mobility as a function of pH at constant ionic strength. Like all polysaccharides, chitosan is also a stiff molecule as com-
The electrophoretic light scattering curve for all the samples followed pared to many of the synthetic water-soluble polymers. The reason for
the same trend when the pH value was increased. At all pH-values, this stiffness is the presence of diequatorial glycosidic linkages in the
chitosans with higher DA, and therefore lower charge density, had chitosan structure due to the 1C4 ring conformation in the chain. This
lower mobility, as expected (Anthonsen & Smidsrød, 1995; Domard, leads to restricted rotation in the chain resulting in extension in chain
1987; Rinaudo & Domard, 1989; Sorlier, Denuzière, Viton, & Domard, length and high intrinsic viscosity. Apart from this, when chitosan is in
2001; Strand, Tømmeraas, Vårum, & Østgaard, 2001). protonated form, an additional ionic strength dependent chain exten-
sion occurs resulting from the repulsion between the charged groups on
4
the polymer chain (Anthonsen, Vårum, & Smidsrød, 1993). with temperature to physically gel chitosans giving rise to transparent
products (Chenite, Buschmann, Wang, Chaput, & Kandani, 2001). The
4.5. Chemical stability reason for this is the temperature-dependent proton transfer from
chitosan to glycerol phosphate leading to reduced electrostatic repul-
The chitosan chain structure has glycosidic linkages susceptible to sion and hence aggregation of chitosan (Lavertu, Filion, & Buschmann,
degradation by both alkalis and acids and also to oxidation by free 2008). Gelling of chitosan can also be done by controlled exposure to
radicals. However, since most applications of chitosan are concerned alkali under well-defined conditions (Ladet, David, & Domard, 2008;
with its stability at low pH, the stability of chitosan against oxidation is Montembault, Viton, & Domard, 2005). Different gel morphologies can
very less studied as compared to the pH stability. The DA value of be obtained depending on the nature of the solvent and DA of chitosan;
chitosan seems to govern its degradation catalyzed by acids, and the soft and easily degradable gels were obtained with chitosan with higher
degradation rate constant increases with the increase in DA value. It DA, while solid gels were obtained with chitosan with low DA values
was found that the cleavage of A-A and A-D glycosidic linkages was (Rami et al., 2014). A variety of ionic cross-linkers and complexing
specifically susceptible to acid hydrolysis. For these linkages, the hy- agents can also be used to obtain physically gelled chitosan. Anionic
drolysis was three times faster than that for D-D and D-A linkages. The molecules like phosphate and citrate (Shu & Zhu, 2002) and transition
reason for this was the presence of a positively charged amino group in metal ions like molybdate (Draget, Värum, Moen, Gynnild, & Smidsrød,
close proximity to the glycosidic linkage to be hydrolyzed, which de- 1992) have also been used for chitosan cross-linking.
creases the rate of hydrolysis (Piszkiewicz & Bruice, 1968; Vårum,
Ottøy, & Smidsrød, 2001). 4.9. Film-forming properties
4.6. Enzymatic degradation From particularly the food packaging perspective, film formation is
considered the most important property of chitosan, which has been
The hydrolases enzymes, which are widely distributed in nature, are extensively studied by many researchers. The capability of the films to
responsible for enzymatic degradation of chitosan (Amano & Ito, 1978). enhance the food quality has been examined, which is not only due to
One such hydrolases enzyme is the lysozyme, which is a presence in the barrier properties of chitosan but also due to its innate anti-
abundance in the human body itself; it is not only found in body fluids microbial activity. A lot of work has been done to fabricate pure chit-
like tears, saliva, urine, and serum but also the tissues. The con- osan or chitosan blend films, membranes, and edible coatings, which
centration of lysozyme varies from 4-13 mg/L in serum (Henry & will be discussed in detail in later sections with a food packaging per-
Henry, 1996) to 450-1230 mg/L in tears (Temel, Kazokoglu, & Taga, spective.
1991). Studies show that the rate of lysozyme degradation of chitosan
increases in direct proportion to its DA value (Nordtveit, Vårum, & 4.10. Antimicrobial properties
Smidsrød, 1996). Also, the maximum initial degradation rates are ob-
tained when a minimum of four acetylated units are contained in the Chitosan is perhaps the only natural polysaccharide that shows
lysozyme binding site. These studies indicate that it may also be pos- antimicrobial activity against bacteria, fungi, and yeast. Its anti-
sible to tailor chitosan as per its application area and degradation re- microbial activity depends on its cationic nature, DA, concentration, the
quirements (Vårum, Holme, Izume, Stokke, & Smidsrød, 1996). Another period of exposure, and the test organism. The exact mechanism of the
study shows that it is possible to fractionate chitosan chains without antimicrobial activity of chitosan is still obscure, but several mechan-
acetyl groups from those consisting of acetyl groups by utilizing im- isms have been proposed for the same:
mobilized chitosan (Sasaki, Kristiansen, Fukamizo, & Vårum, 2003).
(1) Formation of a cellophane-like protective layer by chitosan on the
4.7. Flocculation food surface, thus physically preventing it from microbial attack
(Kumari, Barman, Patel, Siddiqui, & Kole, 2015).
Chitosan displays the potential to replace synthetic polycationic (2) The chitosan layer on the food limits the gas exchange between the
flocculants. It has a high efficiency in removing toxic compounds, heavy food and the outer environment. This leads to hindrance in oxygen
metals, dyes, etc. suspended in water. Although not much has been transportation, thus making the gas unavailable to aerobic microbes
studied on the structure-function relationship of the chitosan as a in the food, causing their elimination (Dotto, Vieira, & Pinto, 2015).
flocculant, it is known that both DD and MW of chitosan affect its (3) The interaction between the cationic group of chitosan with the
functional properties. The number of free amino groups increases as the anionic bacterial cell wall peptidoglycans leads to the dissolution of
DD of chitosan is increased, which in turn enhances flocculation effi- the bacterial cell wall and leakage of the intracellular fluids and,
ciency by increasing the charge neutralization effects. The positive consequently, cell death (Latou, Mexis, Badeka, Kontakos, &
charge density of the chitosan backbone is also affected by external Kontominas, 2014).
factors such as pH and ionic strength, which alter the protonation (4) Chitosan forms an impermeable layer around the bacterial cell and
equilibrium of chitosan in solution. Therefore the effect of DD is not an blocking the exchange of essential solutes between the interior and
independent effect but a combination of several effects. Also, the floc- exterior, resulting in failure of metabolic machinery and ultimately
culation effect increases in direct proportion to the MW of chitosan. cell death (Diez-Pascual & Diez-Vicente, 2015).
Hence, it can be said that the effect of DD and MW on the flocculation (5) The low MW chitosan can enter the cell nucleus and inhibit DNA
efficiency of chitosan is a synergistic effect of various factors (Yang, Li, translation into RNA by binding to the DNA molecule (Youssef,
Huang, Yang, & Li, 2016). Abou-Yousef, El-Sayed, & Kamel, 2015).
(6) The quaternized chitosan can also chelate the essential trace metals
4.8. Gelling properties required for microbial metabolism and hence decrease their avail-
ability to the microbes leading to inhibition of their metabolic
Several methods have been proposed for preparing chitosan gels. machinery and ultimately their death (Hafsa et al., 2016).
One such method is using acetic anhydride to reacetylate chitosan and (7) The synthesis of Chitinase in fruits is enhanced by Chitosan, which
shifting the monomer composition towards the acetylated units. The increases the corresponding gene expression. Since the fungal cell
hydrophobic intermolecular interactions favor the sol-gel transitions wall is made up of chitin, it gets degraded by the action of chitinase
leading to the formation of turbid chitin gels (Vachoud, Zydowicz, & enzyme (Ochoa-Velasco & Guerrero-Beltrán, 2014).
Domard, 2000). Another method uses β-glycerol phosphate combined
5
4.11. Importance of chitosan properties in relation to food packaging methods for film fabrication have been developed. The current fabri-
cation techniques for chitosan films include direct casting, coating,
The properties of chitosan, as discussed in the previous sections, are layer-by-layer assembly, and extrusion. The methods can be used for
not independent but interdependent and give chitosan its unique neat chitosan films without any other polymers, or the chitosan films
nature. The properties are primarily attributed to the presence of the blended with other polymers.
amine groups in the chitosan chain that are formed as a result of the
deacetylation of the chitin structure. Therefore, the degree of deace- 5.1.1. Direct casting
tylation is an important aspect where the properties originate. This The direct casting method has been widely used since it is the most
degree of protonation of the amine groups governs the charge density simple method for the preparation of biopolymer films. The preparation
on the chitosan chain, which directly affects the solubility of chitosan in steps are (1) dissolving chitosan in a weak acidic solution having de-
the aqueous media. Also, varying the pH can vary the protonation of the finite pH value; (2) addition of active or functional compounds or
amine groups leading to a variation in the charge density and solubility materials like cross-linkers, plasticizers, fillers, etc.; (3) blending with
of the chitosan. Besides, the chain conformation of the chitosan also other polymers if required; (4) stirring to obtain a viscous and homo-
gets affected by the charge density as repulsive forces are created in the geneous solution; (5) filtration or centrifugation to get rid of insoluble
chain due to similar charged groups leading to chain extension. The particles and air bubbles; (6) pouring the solution on a flat and leveled
chain conformation, hence, affects the mechanical as well as barrier surface of definite size and shape, usually coated with an inert non-
properties of the chitosan films that have a significant value when food sticky material; (7) drying under set conditions like temperature and
packaging is concerned (Kerch & Korkhov, 2011). At higher degrees of humidity for a certain time; (8) peeling off; (9) curing and storing
deacetylation, the inter-chain interactions are more favorable leading (Sakurai, Takagi, & Takahashi, 1984; Shankar & Rhim, 2018). In some
to enhancement in dynamic rheology of the solution. Therefore, the cases, specialized equipment like vacuum oven, hot air oven, climatic
degree of deacetylation, and hence the solubility, are important factors chamber, humidity controller, etc. may be required for drying. How-
that determine the rheology of the film-forming solution on which the ever, the conditions are milder generally, and simple operations are
film morphology depends (Grant, Cho, & Allen, 2006). The anti- required. The solution cast films show great potential as a food
microbial activity of the chitosan films is also dependent on the degree packaging material. They have been applied to various food products,
of deacetylation – the presence of charged amino groups of chitosan and it was observed that they ensured food safety by maintaining the
responsible for interacting with negatively charged microbial cell quality and enhancing shelf life. Although the casted films show some
membranes leading to its degradation followed by microbial cell death important advantages and exceptional properties, they often have some
(Latou et al., 2014). Also, these charged groups give chitosan its char- defects and disadvantages. Also, slight variations in the formulation can
acteristic ion-binding property which helps the polymer to bind and greatly affect film properties. Hence, care must be taken in formulation
form complexes with metallic ions or nanoparticles as active in- designing while using a direct casting method. Besides, the solution
gredients (Tankhiwale & Bajpai, 2010). Hence, the presence of charged casting method is not currently being employed on the industrial scale,
amine groups makes chitosan a suitable candidate for active and anti- being uneconomical and time-consuming (Mujtaba et al., 2019; Zhang
microbial food packaging. Gelling or cross-linking property is also an- et al., 2019).
other important property of chitosan in the context of food packaging.
For the gelling to take place, either the partial acetylation of the dea- 5.1.2. Coating
cetylated groups is carried out, or a suitable cross-linker is added which Chitosan-based thin films have often been coated either directly on
is responsible for forming cross-links between the chitosan chains. The the surface of the food material (mostly fruits, vegetables, and meat
gelled chitosan, when dried to form films, displays decent water re- products) or indirectly on the surface of the packaging material (El
sistance and moisture barrier properties, which are desirable from the Ghaouth, Arul, Ponampalam, & Boulet, 1991; Fang, Lin, Warner, & Ha,
food packaging perspective (Priyadarshi, Sauraj, Kumar, & Negi, 2018a, 2018; González-Saucedo et al., 2019; Khwaldia, Basta, Aloui, & El-
2018b). Another important property of a food packaging material that Saied, 2014; Ortiz-Duarte, Pérez-Cabrera, Artés-Hernández, &
is looked-for is biodegradation. When the role of the food packaging Martínez-Hernández, 2019). When a direct coating is undertaken, the
material is over, it is required that it goes back to nature and is not purpose is to immobilize the polymer, which contains active substances,
accumulated as municipal solid waste. Enzymatic degradation property directly on the food surface to achieve protection from the environ-
of chitosan makes it a suitable material for food packaging purposes. mental factors like microbes. Also, the coating can alter the gas per-
The hydrolyze enzymes which degrade chitosan are widely distributed meability coefficients of the fruits and vegetables, thereby altering the
in nature. Also, this property of chitosan can be exploited in the case of respiration rate and hence shelf life. These coatings are environmentally
edible coatings and films. The lysozyme present in body fluids can ea- friendly, biodegradable, and, in most cases, edible. Overall, three dif-
sily digest chitosan, making it suitable for ingestion with food (Freier, ferent types of coating methods have been explored: Spread coating,
Koh, Kazazian, & Shoichet, 2005). spray coating, and dip coating.
Spread coating often involves the use of tools like a brush or spatula
5. Chitosan-based films for direct coating on the food or indirect coating on the packaging
material. The direct coating is an important method to limit the mi-
For food packaging applications, the chitosan can be used either in crobial growth on the food surface. The procedure involves (1) pre-
the form of packaging films or coatings directly on food material. Both paration of chitosan formulation containing active ingredients like an-
approaches have been studied by various researchers across the globe. tioxidants, antimicrobials, or strengthening agents; (2) preparing the
In this section, the fabrication methods of chitosan films and coatings food material for coating via treatments like washing, cutting, pas-
have been discussed. Also, some research works on formulations and teurizing, irradiation, or heating; (3) using sterile tool for spreading the
applications of neat chitosan films without any other polymer/biopo- chitosan formulation evenly onto the food material surface; (4) drying
lymer, and chitosan films blended with other polymers/biopolymers under specified conditions in sterile environment like laminar air-flow
such as polysaccharides, proteins, and synthetic plastics, have been cabinet; (5) packaging and storing in refrigerator or vacuum. The in-
conversed. direct coating, on the surface of the packaging material, is aimed at
fabricating multi-layered functional packaging material which has
5.1. Fabrication methods better properties as compared to single-layer packaging. Chitosan
coating on the paper surface can not only impart antimicrobial activity
Depending on the final application in food packaging, various to it but also reduces the moisture permeability without having much
6
effect on the mechanical attributes (Khwaldia et al., 2014). 5.1.4. Extrusion

Spray coating involves the use of several spray tools like compressed Melt extrusion is another widely developed method to process
air-assisted sprayers. This method has been proven more useful when chitosan-based biodegradable active-packaging films. It involves (1)
combined with other treatment processes. Green beans coated with formulation of materials with several compositions; (2) mixing and
chitosan-based formulation when treated with gamma irradiation and blending the materials; (3) extrusion of the blend material under spe-
packed under modified atmosphere caused a reduction in the microbial cified conditions; (4) using a pelletizer to cut the extrudates into pellets;
population and enhanced safety (Severino et al., 2015). Also, spray (5) drying the pellets; (6) using a twin-screw extruder attached with a
coating chitosan on wolfberries followed by heat treatment improved flat die to extrude the pellets into flat sheets or using a blown film
their postharvest quality and acceptability (Ban et al., 2015). extruder with an annular die to make a blown film (Goskonda &
The dip-coating technique requires the immersing of the food ma- Upadrashta, 1993; Steckel & Mindermann-Nogly, 2004).
terial into chitosan-based solutions leading to uniform film formation Films made through the extrusion process generally show good
on the surface. The film formulation depends greatly on the wetting mechanical properties and thermal stability (Martinez-Camacho et al.,
capacity of the surface, processing time, and draining time. The process 2013). For blown film fabrication, the two-step melt compounding
involves: (1) preparation of chitosan solution of definite pH; (2) selec- process was also used. However, with the increase in chitosan content,
tion and processing of food sample to be coated; (3) immersion of the the film properties dropped – an increase was observed in the water
food sample in the solution for designated time ranging from 30 s to 30 vapor permeability (WVP) while the elongation and breaking strength
min depending on the food type; (4) emersion of food, draining excess of the films decreased (Wang, Chen, & Chuai, 2015). The compression
solution, and drying; (5) placing the food in trays or plastic support; (6) molding method has also been used for film fabrication. It was used to
packing and storing. The food samples that were dip-coated were found fabricate a blend film of polyethylene glycol chitosan and low-density
to be more resistant to chilling injury, had lower color variation and polyethylene (LDPE), which showed excellent transparency. However,
decay, consequently better quality and extended shelf life (Kanatt, Rao, the thermal stability of the films was dependent mainly on LDPE. Also,
Chawla, & Sharma, 2013; Zhang, Zhang, & Yang, 2015). Some reports the addition of polyethylene glycol decreased Young's modulus
provide a higher recommendation for the double-dipping process. It (Davidovich-Pinhas, Danin-Poleg, Kashi, & Bianco-Peled, 2014). In
was observed that cheese that was double-layer dip-coated had retained another study, blending and extrusion of polylactic acid (PLA)/starch/
the best aroma and flavor (Cano Embuena et al., 2017). Overall, the chitosan were carried out, resulting in the fabrication of antimicrobial
dip-coating process is relatively easy, time-saving, and avoids the re- films with thickness ranging from 0.15 mm to 0.18 mm. The films were
quirement of large equipment while being highly effective in preserving successful in protecting high water activity food material - like fresh
the food and maintaining its quality (Liu et al., 2016; Souza et al., meat (Bie et al., 2013). It was also observed that for the extruded
2010). polyamide/chitosan films, the poor interaction between chitosan and
polyamide matrix led to inferior mechanical properties and brittle
failure for the films (Lago et al., 2014).
5.1.3. Layer-by-layer assembly Overall, extrusion is an effective process to fabricate films with good
The layer-by-layer (LBL) is an electrostatic deposition technique mechanical, thermal, and antimicrobial properties. However, to achieve
that effectively controls the material properties and functionality of the best performance, it is necessary to take the processing temperature
biomaterial films. This technique can be applied for the fabrication of and the interaction between the polymers into account (Alix et al.,
multi-component films without requiring any sophisticated instru- 2013; Lago et al., 2014; Woranuch & Yoksan, 2013; Woranuch, Yoksan,
ments. Moreover, in this process, the films prepared are not dependent & Akashi, 2015).
on substrate shape (Li, Biagioni, Finazzi, Tavazzi, & Piergiovanni, 2013;
Xiao, Xu, Liu, Hu, & Huang, 2013). 5.2. Pure chitosan films
The role of pH is very imperative in the formation of LBL films. The
electrical charge on each layer is governed by the pH resulting in a Neat chitosan films, without the use of any other polymer with
significant alteration in the biopolymer deposition on the previous chitosan, have been fabricated and studied for packaging applications
layer. The charge on the former layer will determine the number of by several researchers. However, these films include other ingredients
biopolymer molecules required to neutralize it (Acevedo-Fani, Salvia- like dilute acids, plasticizers, and emulsifiers, which assist film forma-
Trujillo, Soliva-Fortuny, & Martin-Belloso, 2015). With the decrease in tion. The films have been fabricated by dissolving chitosan in dilute
pH and increase in layer number, an increment in the thickness, acidic solutions, like acetic acid and hydrochloric acid, and have been
roughness, and modulus of the films was observed. Besides this, the successfully used for the preservation of various food items like fruits,
films also displayed a high degree of photo-orientation (Fernández, vegetables, berries, fish, and meat. The sub-micron size of chitosan film-
Ocando, Fernandes, Eceiza, & Tercjak, 2014). forming dispersions affects the properties of the film. It was observed
The combination of the LBL assembly technique with the immersing that dispersions with 600 nm diameter displayed better preservation as
method is often used. The films are formed by oppositely charged compared to that with 1000 nm diameter (Ali, Zahid, Manickam,
materials viz., polyanion materials and polycation chitosan and the Siddiqui, & Alderson, 2014). Plasticizers like glycerol are generally used
objects are immersed into the respective solutions of these charged to improve film flexibility, chain mobility and strength (Gol, Patel, &
materials for numerous times (da Silva, Iamanaka, Taniwaki, & Rao, 2013; Treviño-Garza, Garcia, del Socorro Flores-González, &
Kieckbusch, 2013; Huang et al., 2019; Maciel, Yoshida, & Franco, 2015; Arévalo-Niño, 2015; Vimaladevi, Panda, Xavier, & Bindu, 2015).
Martiñon, Moreira, Castell-Perez, & Gomes, 2014; Poverenov et al., Emulsifiers like Tween 20, Tween 80, Span 20, and Brij 56 have also
2014; Serizawa, Yamaguchi, Matsuyama, & Akashi, 2000; Souza et al., been used for property enhancement (Rui et al., 2017; Vieira et al.,
2015). The microbial quality, sensory analysis, pH, color, moisture, 2016; Zahid, Ali, Manickam, Siddiqui, & Maqbool, 2012). Although
texture, and growth of material were examined to infer that the pre- pure chitosan films show food packaging attributes, they are far behind
pared film could elongate the shelf life of the food. This combination the synthetic polymeric films and hence are not suitable for mass ap-
method is relatively simple and offers an easy way for functional ma- plications in food packaging.
terials fabrication that could effectively prolong the shelf life and retain
the quality of the treated food product. Besides this, in conditions with 5.3. Chitosan blend films
high humidity, some films also show the best barrier performances
(Tzeng, Stevens, Devlaming, & Grunlan, 2015). There are several reports on chitosan films blended with other
polymers such as polysaccharides, proteins, synthetic polymers, etc. as
7
explained in the following subsections. a purified form of collagen, and when blended with chitosan, the re-
sultant films displayed improved mechanical, moisture barrier, and UV-
5.3.1. Chitosan-polysaccharide blend films resistant properties (Hosseini, Rezaei, Zandi, & Ghavi, 2013;
Several studies report the blending of chitosan with polysaccharides Kowalczyk, Kordowska-Wiater, Nowak, & Baraniak, 2015;
for property improvement. Starch is an important renewable poly- Prateepchanachai, Thakhiew, Devahastin, & Soponronnarit, 2019).
saccharide obtained from plants, being abundant, economical, and Plant-based and microbial proteins have also been used in chitosan
biodegradable. It displays very good film-forming properties and when films. However, they have not been blended with chitosan but have
blended with chitosan, and the resulting film shows improved water been used as active substances in films. These are usually added to
barrier properties along with enhanced antioxidant and antimicrobial chitosan as antimicrobial agents or natural extracts and will be covered
activity (Alix et al., 2013; Talón et al., 2017; Woranuch & Yoksan, briefly in the later sections of this review (Sections 6.1.4 and 6.1.6).
2013).
Cellulose is another abundant biopolymer of plant origin. Chitosan- 5.3.3. Chitosan-synthetic polymer blend films
cellulose blend films showed improved mechanical properties that were Besides blending chitosan with other biopolymers, several re-
attributed to the electrostatic interactions between the two polymers searchers report its blending with synthetic polymers. Although the
(Xiao et al., 2013). The films successfully enhanced the shelf life of resulting polymer is not completely biodegradable, blending chitosan
cheese and wheat bread (Noshirvani, Ghanbarzadeh, Mokarram, & with synthetic polymers can enhance the disintegration of the other-
Hashemi, 2017; Youssef, El-Sayed, El-Sayed, Salama, & Dufresne, wise hard-to-degrade plastics. Polyvinyl alcohol (PVA), being non-toxic
2016). Other cellulosic materials like methylcellulose, hydroxypropyl and water-soluble, is one of the most commonly used synthetic poly-
methylcellulose, quaternized hemicellulose, and microfibrillated cellu- mers blended with chitosan. The blend films not only show highly
lose have also been blended with chitosan to form composite films improved mechanical properties but enhanced barrier performances
(Chen et al., 2016; Gol et al., 2013; Liu et al., 2013). The obtained films towards water and oxygen (Giannakas et al., 2016; Giannakas,
possessed attributes like high moisture resistance, transparency, elas- Stathopoulou, Tsiamis, & Salmas, 2020). Synthetic acids like PLA and
ticity, etc. that are important from the food packaging context. salicylic acid have also been used with chitosan. Chitosan/starch/PLA
Alginate, a chemically versatile and non-toxic biopolymer of algal blend films displayed enhanced flexibility and better thermal proper-
origin, is anionic and interacts with the cationic biopolymer chitosan ties. The films were also found to show sustained antimicrobial activity
via electrostatic interactions to form stable films. These interactions of against common foodborne pathogenic bacteria (Pal & Katiyar, 2016).
charges had a significant influence on properties such as contact angle, The chitosan grafted salicylic acid films were found to be successful in
microstructure and thermal performances (Acevedo-Fani et al., 2015). protecting cucumber from chilling injury, maintaining its quality, and
The chitosan-alginate films showed excellent gas exchange and water enhancing its shelf life (Zhang et al., 2015). Co-extrusion of low-density
vapor transmission properties that prevented the treated foods from polyethylene and chitosan was carried out to obtain chitosan/LDPE
developing off-flavors and protected their appearance (Poverenov et al., blend films, which showed improved moisture barrier properties,
2014; Souza et al., 2015). making it a promising material for food packaging applications
Pectin is another polysaccharide of plant origin. It is present in (Martinez-Camacho et al., 2013; Wang et al., 2015). The ameliorated
many parts of the plant as a structural polysaccharide. Like alginate, it flexibility of the films was also obtained when blended with poly
is also anionic and forms stable intermolecular interaction with chit- (ethylene oxide) (Kohsari, Shariatinia, & Pourmortazavi, 2016). Re-
osan to form uniform films (Maciel et al., 2015). The composite films cently, PET/PP films assembled with chitosan layers containing various
showed desirable characteristics and were found to maintain the phy- preservatives were fabricated and studied for food packaging applica-
sicochemical and sensory values of fresh-cut cantaloupe (Martiñon tions. The films were found to possess antimicrobial activity against
et al., 2014). foodborne bacteria E. coli and B. subtilis (Lei et al., 2014).
Cyclodextrin, konjac glucomannan, glucose, xylan, kefiran, etc. are
some other polysaccharides that have been studied in combination with 6. Chitosan film in food packaging
chitosan for film fabrication. The obtained films can be applied to food
packaging, and their potential can be exploited to extend the food shelf The utilization of chitosan in food packaging is either in the form of
life (Gao, Zhu, & Zhang, 2013; Higueras, López-Carballo, Hernández- flexible packaging films or coatings. The flexible films may either be
Muñoz, Catalá, & Gavara, 2014; Kamdem, Shen, & Nabinejad, 2019; edible or inedible, unlike the coatings, which are edible in almost all
Kosaraju, Weerakkody, & Augustin, 2010; Salmanian, Khodaiyan, & cases since they form a layer directly on the top surface of the food-
Hosseini, 2019). stuff. Several additives have been incorporated in each type of packa-
ging to impart several properties. However, the edible coatings are
5.3.2. Chitosan-protein blend films composed of minimum additives, or sometimes no additives, to avoid
Proteins obtained from several plants, animal, and microbial compromising the food-grade status of chitosan. The flexible films or
sources have been used with chitosan to tailor its properties. Since coatings based on different formulations and functions are as sum-
different proteins have different amino acid monomeric compositions marized in Table 2.
leading to different functional groups (Gravel & Doyen, 2020), the
chitosan-protein films have abundant functions that promote their ap- 6.1. Flexible packaging films
plications in food packaging.
Proteins obtained from animals are highly biocompatible and have Flexible chitosan films are prepared, preferably by the solvent
high nutritive value. Caseinate is an excellent thermoplastic protein casting method in which chitosan is dissolved in suitable solvents, in
with excellent film-forming properties that combines well with chitosan most cases slightly acidified water, and is poured on a flat surface for
through ionic interactions and improves the water vapor barrier prop- allowing the solvent to evaporate (Kim, Son, Kim, Weller, & Hanna,
erties (Khwaldia et al., 2014). Collagen has also attracted the interest of 2006). The properties of the films depend on various factors like DD,
several researchers. The chitosan-collagen composite films show high solvent pH, type of acid used, MW, etc. Lower WVP was observed for
compatibility and better thermal properties (Ahmad, Nirmal, Danish, the films prepared by chitosan having a lower DD value. Also, the WVP
Chuprom, & Jafarzedeh, 2016). Lysozyme chitosan blend films were decreased with a decrease in the pH of the film-forming solution. Also,
used for eggs, and the effect on shelf life was studied. The stored eggs the type of acid used significantly affects the WVP values. It is observed
not only had increased freshness and improved shell strength, but the that the films obtained by using acetic acid and propionic acid show
internal quality was also maintained (Yuceer & Caner, 2014). Gelatin is comparatively lower WVP values that for the films obtained using lactic
8
Table 2
Food packaging applications of chitosan.
Material type Additive Property References
Films with nanomaterials

Chitosan film Silver Antimicrobial activity Tripathi, Mehrotra, and Dutta (2011)
Chitosan film Silver Mechanical strength Vimala et al. (2011)
Chitosan film Zinc oxide Moisture barrier Priyadarshi & Negi (2017)
Mechanical strength
Antimicrobial activity
Chitosan film Silver/Zinc oxide Antimicrobial activity Youssef et al. (2015)
Chitosan film Titanium dioxide Antimicrobial activity Zhang et al. (2017), Zhang et al. (2017)
Chitosan film Copper Moisture barrier Cárdenas, Díaz, Meléndrez, Cruzat, and García Cancino (2009)
Mechanical strength
Chitosan film Graphene oxide Moisture barrier Ahmed, Mulla, Arfat, Thai, and L. A. (2017)
Mechanical strength
Thermal stability
Chitosan film Multiwalled CNTs Mechanical strength Sun et al. (2011)
Chitosan film Reduced graphene oxide Mechanical strength Barra et al. (2019)
Water resistance
Antioxidant activity
Electrical conductivity
Chitosan film Sulfur Antimicrobial activity Shankar & Rhim (2018)
Hydrophobicity
Mechanical strength
Moisture barrier
Films with clays

Chitosan film Micro/nanoclay Moisture barrier Casariego et al. (2009)
Chitosan-PVA film Montmorillonite Mechanical strength Zhang, Wang, et al. (2017), Zhang, Xiao, et al. (2017)
Moisture barrier
Chitosan film Cloisite 30B Mechanical strength Rhim (2011)
Moisture barrier
Chitosan film Cloisite 15A Thermal stability Swain, Kisku, and Sahoo (2014)
Oxygen barrier
Films with polysaccharide particles, fibers, and whiskers

Chitosan film Chitin nanowhiskers Mechanical strength Sriupayo, Supaphol, Blackwell, and Rujiravanit (2005)
Chitosan film Chitin nanowhiskers Mechanical strength Ma et al. (2014)
Chitosan film Chitin nanowhiskers Mechanical strength Li et al. (2009)
Thermal stability
Moisture barrier
Chitosan film Nanocrystalline cellulose Mechanical strength Khan et al. (2012)
Chitosan-PVA film Lignin nanoparticles Mechanical strength Yang et al. (2016)
Thermal stability
Films with natural oils and extracts

Chitosan film Apricot kernel essential oils Moisture barrier Priyadarshi et al. (2018)
Food shelf life extension
Chitosan film Rosemary extract Food shelf life extension Xiao, Zhu, Luo, Song, and Deng (2010)
Chitosan film Cinnamon essential oils Antimicrobial activity Ojagh, Rezaei, Razavi, and Hosseini (2010a, 2010b)
Mechanical strength
Moisture barrier
Food shelf life extension
Chitosan film Bergamot essential oils Moisture barrier Fisher & Phillips (2008)
Sanchez-Gonzalez, Chafer, Chiralt, and Gonzalez-Martinez (2010)
Svoboda & Greenaway (2003)
Chitosan film Ganagal extract Antimicrobial activity Gómez-Estaca, López de Lacey, López-Caballero, Gómez-Guillén, and
Montero (2010)
Holey & Patel (2005)
Mohammed & Al-Bayati (2009)
Chitosan film Curcumin Antimicrobial activity Roy & Rhim (2020)
Chitosan film Mango leaf extract Antioxidant activity Rambabu, Bharath, Banat, Show, and Cocoletzi (2019)
Mechanical strength
Moisture barrier
Chitosan film Purple-fleshed sweet potato extract pH sensing Yong et al. (2019)
Films with cross-linkers and plasticizers

Chitosan film Citric acid Cross-linking Priyadarshi, Sauraj, Kumar, Deeba, et al. (2018)
Moisture resistance
Thermal stability
(continued on next page)
9
Table 2 (continued)
Material type Additive Property References
Chitosan film Citric acid Cross-linking Cui, Beach, and Anastas (2011)
Chitosan film Glycerol Plasticizing Priyadarshi, Sauraj, Kumar, Deeba, et al. (2018)
Flexibility Cui et al. (2011)
Chitosan film Dialdehyde starch Cross-linking Tang et al. (2003)
Mechanical strength
Chitosan film Genipin Mechanical strength Jin, Song, and Hourston (2004)
Water resistance
Chitosan-PVA film Glutaraldehyde Cross-linking Tripathi, Mehrotra, and Dutta (2009)
Chitosan film Tannic acid Water resistance Rubentheren, Ward, Chee, and Tang (2015)
Cross-linking
Chitosan film Polyethylene glycol Plasticizing Kelnar et al. (2015)
Flexibility
Films with antimicrobial agents

Chitosan film Amikacin Antimicrobial activity Mu, Zhang, Zhang, Niu, and Duan (2014)
Vancomycin
Clindamycin
Erythromycin
Chitosan film Ԑ-Polylysine Antimicrobial activity Wu et al. (2019)
Edible coatings
Chitosan coating – Retention of sensory attributes Han, Lederer, McDaniel, and Zhao (2006)
Extended shelf life
Chitosan coating Oleic acid Retention of texture and color Vargas, Albors, Chiralt, and González-Martínez (2006)
Extended shelf life
Chitosan coating – Retention of sensory attributes Chien, Sheu, and Yang (2007)
Extended shelf life
Reduced moisture loss
Chitosan coating – Retention of sensory attributes Moreira, Roura, and Ponce (2011)
Extended shelf life
Chitosan + Guar gum coating – Retention of sensory attributes Huang, Cheng, et al. (2019), Huang, Qian, Jiang, and Zheng (2019)
Chitosan coating Silver-chitosan nanocomposite particles Extended shelf life Ortiz-Duarte et al. (2019)
Chitosan coating Chitosan nanoparticles Antimicrobial activity Gomes et al. (2019)
and formic acid. Another study suggests that with the increase in sto- commercially used as antimicrobial agents (Sharma et al., 2013). Silver
rage time and MW, and with the decrease in storage temperature, the ions cause inactivation of respiratory enzymes leading to the formation
mechanical properties and WVP of the chitosan films tend to increase of reactive oxygen species resulting in cell damage (Priyadarshi & Negi,
(Kerch & Korkhov, 2011). The tensile properties of the films are very 2019). Tripathi et al. (2011) studied the effect of silver nanoparticle
important from the food packaging perspective. Chitosan films show addition on the antimicrobial activity of chitosan films. It was observed
tensile strength (TS) and elongation at break (EB) values comparable to that the silver nanoparticle embedded chitosan films showed enhanced
that of high-density polyethylene (HDPE), LDPE, and cellophane antimicrobial activity against gram-positive bacteria S. aureus and B.
(Butler, Vergano, Testin, Bunn, & Wiles, 1996). However, the applica- subtilis as well as gram-negative bacteria P. aeruginosa and E. coli. An-
tion of chitosan in food packaging is limited due to its high sensitivity to other study carried out by Vimala et al. (2011) suggested an increased
humidity and moisture. Also, since chitosan films are not thermoplastic mechanical strength of chitosan films embedded with silver nano-
and degrade before the melting temperature, they could not be pro- particles along with enhanced antimicrobial effect. Zinc oxide is an-
cessed by industrial methods like extrusion and molding and also can't other important nanomaterial that is recognized as safe and is being
be stretched or heat sealed (Pelissari, Yamashita, & Grossmann, 2011; increasingly used in various human use products. Priyadarshi & Negi,
Van Den Broek, Knoop, Kappen, & Boeriu, 2015). To overcome these 2017 synthesized zinc oxide nanoparticles and used them as fillers in
limitations of this otherwise excellent polysaccharide biomaterial, the chitosan matrix. The resulting films show synergistic antimicrobial
several attempts have been made, such as blending the films with other activity against B. subtilis and E. coli due to chitosan and zinc oxide.
natural or synthetic polymers, or the addition of several active and Besides, a substantial increment in mechanical properties due to the
functional substances like fillers, plasticizers, crosslinkers, natural oils, reinforcing effect of zinc oxide nanoparticles (Fig. 4). In a very recent
etc. study, similar incremental effects of zinc oxide nanoparticles on the
mechanical properties of chitosan films were also reported by
6.1.1. Films embedded with nanomaterials Rodrigues et al. (2020).
Metallic ions like silver, zinc, copper, titanium, etc. have been found Youssef et al. (2015) prepared chitosan-silver-zinc oxide composite
in abundance in nature. Most of these ions serve as essential minerals in films that were effective against a wide range of food pathogens like S.
certain eukaryotes, including humans. Also, these do not possess any aureus, E. coli, S. typhimurium, B. cereus, and L. monocytogenes. A lot of
threat to eukaryotic cells below a defined concentration. Hence, metal literature is available on the photocatalytic antimicrobial activity of
and metal oxide nanoparticles find application as fillers in the food titanium dioxide nanoparticles. However, a limited is explored using
packaging polymers to enhance their mechanical, barrier, thermal, and these nanoparticles in food packaging. Zhang, Wang, et al. (2017) in-
photodegradation properties. Additionally, these nanoparticles address corporated titanium dioxide nanoparticles in chitosan film and used it
other properties relevant to food packaging as well. They show out- for packing red grapes for six days. The films performed exceptionally
standing antimicrobial and antioxidant activities and consequently lead well and protected the grapes from microbial attack while extending
to enhanced shelf life of the products. Silver is the most widely used their shelf life (Fig. 5). Besides, the microorganisms were completely
nanoparticle due to its noble nature, and silver salts have been destroyed at 12 h of exposure time.
10
Fig. 4. Stress vs. strain curve for neat chitosan (a), chitosan nanocomposite films with ZnO nanoparticle concentration 1% (b), and 2% (c) (Priyadarshi & Negi, 2017).
Copper nanoparticle incorporation in the chitosan matrix has also increased mechanical properties with about 40 % increment as com-
been a research focus. The amount of copper required to give anti- pared to pure chitosan films. Chitosan films incorporated with different
bacterial properties to chitosan films is very small and much lower than kinds of sulfur nanoparticles were studied by Shankar and Rhim (2018).
the level of toxicity, but it is desirable to avoid it if there are other It was observed that the films incorporated with chitosan-coated sulfur
relatively safe options. However, there is some published data on the nanoparticles displayed enhanced mechanical and moisture barrier
use of copper nanoparticles in food packaging chitosan films. Cárdenas properties. They also found that the sulfur nanoparticles incorporated
et al. (2009) compared the properties of plain chitosan film with that of chitosan films exhibited significantly higher antimicrobial activity than
the one embedded with copper nanoparticles and observed an en- the neat chitosan films. It was suggested that the chitosan coating on
hanced mechanical, barrier and antimicrobial activity of the latter. sulfur nanoparticles enhanced their compatibility with the chitosan
Apart from the inorganic metallic nanoparticles, carbon-based nano- matrix, thus resulting in enhanced properties of the composite films. In
materials like graphene oxide and carbon nanotubes have also been a very recent study, Barra et al. (2019) embedded reduced graphene
used for incorporation in food packaging chitosan films. Ahmed et al. oxide (rGO) nanoparticles in chitosan films at various concentrations.
(2017) prepared graphene oxide embedded chitosan film and studied Besides enhancing the mechanical and moisture barrier properties as
its properties from the food packaging perspective. It was observed that expected, the nanocomposite films developed good electrical con-
the more compact network structure was formed, leading to the in- ductivity and displayed a very high antioxidant activity as well. Other
creased tensile strength of the film. An increase in barrier properties than metal and metal oxide nanoparticles, nanoclays and biopolymeric
was also observed due to the enhanced tortuosity of the diffusion path polysaccharide nanoparticles like nanosized cellulose, chitin and lignin
for oxygen and water vapor through the film. The glass transition have also been used for this purpose.
temperature also increased, showing betterment in the thermal prop-
erties of the film as well (Table 3).
Sun et al. (2011) used multi-walled carbon nanotubes as a re- 6.1.2. Films embedded with clays
inforcement material for chitosan films. The composite films showed Polymer/clay composites have been increasingly gaining popularity
as a food packaging material due to the extraordinary property of clays
Fig. 5. Preservation of red grape packed in different materials at 37oC for six days: (a) plastic wrap; (b) pure chitosan film; (c) chitosan-TiO2 film (Zhang, Wang, et al.,
2017).
11
Table 3
Mechanical and thermal properties of CS films incorporated with GO (Ahmed et al., 2017).
Sample Tensile strength (MPa) Elongation at break (%) Tg
c a
CS 6.99 ± 0.95 72.70 ± 6.76 200.44 ± 2.1d
CS/0.5% GO 8.97 ± 1.82b 69.88 ± 2.72a 204.84 ± 1.04c
CS/1% GO 13.46 ± 0.82a 61.89 ± 2.21b 219.88 ± 3.38b
CS/2% GO 15.32 ± 2.47a 57.34 ± 4.60c 226.56 ± 2.14a
Values are given as mean ± SD (n = 3).

Different lowercase letters in the same column indicate significant differences (p < 0.05).
to disperse evenly in the polymer matrix, even in very small con- concentration in chitosan films not only increased the tensile strength
centrations, to fill up the spaces in the polymer matrix giving rise to a about three times but also showed strong antimicrobial activity against
very compact structure. This compact structure shows exceptional foodborne pathogenic bacteria. A similar study was performed by Li,
barrier properties as the intercalation of clays make diffusion path Zhou, & Zhang (2009) using cellulose whiskers of approximately the
tortuous for the water vapor and gases, hence making it difficult for same dimensions and observed an improvement not only in the me-
them to pass. The mechanical properties are also largely improved due chanical properties of composite films but also in thermal stability and
to the reinforcement effect of clays. The scientific literature consists of a water resistance. Khan et al. (2012) produced nanocrystalline cellulose
fairly good collection of works where clays of different types have been (NCC), having a dimension of 25 % of the chitosan whisker used in the
used as fillers to ameliorate the properties of plain chitosan as well as above study by Li et al. (2009) and used to strengthen the chitosan film.
chitosan blend films. Casariego et al. (2009) incorporated micro/nano The reason for the strengthening effect of NCC in the chitosan matrix
clay particles in chitosan films in varying concentrations and studied was the formation of a percolation network due to hydrogen bonding
the properties of the composite films. A good decrease in the WVP and between well dispersed NCC and chitosan matrix. It was observed that
water solubility was observed with increasing clay concentration. In the improvement in the mechanical property was comparable to that
another study, Zhang, Xiao, et al. (2017) prepared chitosan-PVA/ reported by Li et al., but was attained at a lower concentration of NCC
montmorillonite composite films where intercalation was achieved due as compared to cellulose nano-whiskers. Yadav, Behera, Chang, and
to the presence of interfacial interaction in the form of hydrogen bonds Chiu (2020) also studied the effect of cellulose nanocrystals on chitosan
between the constituents. It was observed that the films showed better films and observed improved mechanical and barrier properties at 4%
mechanical properties along with improved barrier properties. An in- nanocrystals concentration. Lignin nanoparticles were used by Yang
crease in the MMT content significantly increased Young's modulus and et al. (2016) as fillers for chitosan/PVA blend films for food packaging
tensile strength of the films while also improving the gas and moisture applications. According to their report, not only an increment in me-
barrier. The addition of Cloisite 30B clay to chitosan films led to an chanical properties and thermal stability was observed, but also anti-
increase in the tensile strength and elasticity up to a certain extent microbial and antioxidant properties were imparted to the films making
before decreasing again with the increasing clay concentration (Rhim, them a potential active food packaging material.
2011). On the other hand, there was an exponential increase in the
moisture barrier properties of the film, while a slight decrease in water 6.1.4. Films embedded with natural oils and extracts
solubility and the swelling ratio was observed. Similar trends for their The increased interest of the packaging researchers towards active
mechanical properties were also observed by Kusmono and Abdurrahim food packaging materials, more focus is shifted to the search for ma-
(2019) in a relatively recent study. Another similar study conducted by terials which, apart from improving the film properties, impart biolo-
Swain et al. (2014) focused on thermal stability and gas permeability of gical activity to the films. Increasing market competition to improve
Cloisite 15A incorporated chitosan films. The increase in clay con- food shelf life, while maintaining eco-friendliness and economics, the
centration in the films led to increased thermal stability and decreased discovery of biodegradable, biocompatible, non-toxic, and economic
oxygen permeability of the nanocomposite films, hence making them a biologically active substances are becoming more important. Natural
suitable candidate for food packaging applications. essential oils, which are extracted from renewable plant parts or agri-
culture wastes, are proving to be a good candidate for this purpose.
6.1.3. Films embedded with polysaccharide particles, fibers, and whiskers Other than fatty acids and lipids, they are composed of several plant
The addition of fillers for reinforcement to achieve improved me- metabolites, which are antimicrobial as well as antioxidant in nature.
chanical performances in polymeric films and sheets is gaining a lot of Moreover, they are non-toxic and do not have any effect on human
importance as a simple and effective method. Although many inorganic health if consumed in small quantities. Many researchers have tried to
materials like clays and carbon nanotubes have been used for this incorporate these essential oils in the chitosan films and have so far
purpose, they are considered less suitable from the environmental point achieved the results as expected. Apart from imparting the films with
of view as they are not completely biodegradable and limit the biode- antimicrobial and antioxidant activities, these essential oils can have
gradability of the composite material as a whole. This led to the plasticizing and water-resisting effects on the film, which is a major
opening of passage for the polysaccharide-based biodegradable re- advantage. The components, structures, and functional groups of es-
inforcing materials like chitin and cellulose in the form of powders, sential oils play an important role in determining antimicrobial activity
crystals, fibers, etc. Another reason for the growing use of these bio- (Holley & Patel, 2005). The most consistently effective natural essential
degradable reinforcing materials is their potential antimicrobial ac- oils against the foodborne microorganisms are thyme, cinnamon, ro-
tivity, unlike clays. These natural reinforcing materials have been used semary, sage, basil, vanillin, oregano, and clove oils (Dorman & Deans,
by many researchers to ameliorate the properties of chitosan films for 2000). Besides antimicrobial activity (Canillac & Mourey, 2001; Kanatt,
food packaging applications. Sriupayo et al. (2005) prepared chitin Chander, & Sharma, 2008), these oils also possess antifungal (Mari,
nano-whiskers with average length and width of about 400 nm and 30 Bertolini, & Pratella, 2003), antiparasitic (Pessoa, Morais, Bevilaqua, &
nm, respectively, and used them to reinforce chitosan films. The chitin Luciano, 2002) and antioxygenic (Juglal, Govinden, & Odhav, 2002)
whisker reinforced chitosan films displayed an initial increase in the effects.
tensile strength up to a certain nano-whisker concentration and thereby Rosemary is a good source of antioxidant compounds and hence, is
a decrease with a further increment in nano-whisker concentration. Ma widely used in the food industry to prevent oxidative degradation of
et al. (2014) reported that the use of rod-shaped chitin whiskers at a 3% foods (Georgantelis, Ambrosiadis, Katikou, Blekas, & Georgakis, 2007;
12
Ponce, Roura, del Valle, & Moreira, 2008; Zivanovic, Chi, & Draughon, recent study, plant based flavanoid Quercetin was complexed with
2005). The chitosan films incorporated with rosemary extract were starch and the food packaging properties of Quercetin-starch based
found to extend the shelf life of fresh-cut pears (Xiao et al., 2010). Shelf films were investigated. The films show enhanced antimicrobial activity
life extension of poultry meat was also observed when packaged in and high antioxidant activity making them suitable for food packaging
chitosan films incorporated with rosemary essential oils (Souza et al., (Yadav, Mehrotra, Bhartiya, Singh, & Dutta, 2020).
2019). Ojagh et al. (2010a) reported the effect of incorporation of
cinnamon essential oils (CEO) on the packaging properties of chitosan 6.1.5. Films embedded with plasticizers and cross-linkers
films. Cinnamaldehyde is the known active compound in CEO with a A cross-linker is a molecule that helps in the establishment of a
total share of around 60% in oil (Matan et al., 2006). Besides improving cross-link between two polymer chains. Crosslinking can be either ionic
the antimicrobial activity, the CEO also helped in lowering the water or covalent, which bonds two polymer chains together and alters their
solubility, moisture content, WVP, and elongation at the break of the properties, making them ideal for many applications. Excellent di-
chitosan films. It also helped in enhancing the tensile strength of the mensional stability, lower creep rates, enhanced solvent resistance,
film by providing cross-linking between chitosan chains. CEO in- enhanced glass transition temperature, etc. are some of the important
corporated chitosan films were used to preserve trout fish filet, and the properties achieved by the cross-linked polymers (Nielsen, 2008). The
packet achieved an exceptional shelf life of 16 days without any mi- most common example of the cross-linked polymers is the vulcanization
crobial growth and significant color, odor, and texture loss (Ojagh et al., of rubber. The cross-link present, in this case, is the disulfide bonds.
2010b). Bergamot essential oil (BEO), extracted from Citrus bergamia, is Chemical cross-linking of chitosan is a versatile method for improving
another important essential oil used in chitosan-based active food its properties. Although there are a variety of crosslinkers such as glu-
packaging films. It contains limonene and linalool as the major che- taraldehyde, formaldehyde, dialdehyde starch, citric acid, tannic acid,
mical compounds responsible for antimicrobial activity and the BEO glyoxal, genipin, quinone, etc. which are being used for altering the
incorporated chitosan film is found to be effective against most of the properties of chitosan suited to different applications, a limited number
common foodborne pathogens (Fisher & Phillips, 2008; Moufida & of cross-linkers have been used for food packaging applications due to
Marzouk, 2003; Sanchez-Gonzalez et al., 2010; Svoboda & Greenaway, consumer health concerns. Tang, Du, & Fan (2003) used dialdehyde
2003). The results also show that BEO helps in decreasing the WVP of starch as a cross-linking agent in chitosan films and observed improved
the chitosan films. Recently the effect of lemongrass essential oil (LEO) mechanical properties. The neat chitosan film showed a tensile strength
on properties of chitosan films for active packaging was also studied by of around 47 MPa and elongation of 17 % at the break, while the cross-
Lyn and Hanani (2020). The films incorporated with 9% LEO were linked films displayed better tensile strength and breaking elongation of
found to inhibit the growth of most food borne microbes. The Ganagal 113 MPa and 27 %, respectively. No effect of cross-linking was ob-
is a member of the rhizome family, similar to ginger and turmeric. served on the antimicrobial activity of the chitosan films. Genipin cross-
Ganagal extract (GE) consists of terpenes which are common natural linked chitosan films were prepared by Jin et al. (2004), and their
antimicrobial agents (Gómez-Estaca et al., 2010; Holley & Patel, 2005; mechanical properties and solubility were studied. The results suggest
Mohammed & Al-Bayati, 2009). The GE containing chitosan films were improved elasticity and mechanical properties. Besides this, the films
found to be active against food poisoning bacteria like S. aureus were insoluble in acidic and alkaline media. However, the films tend to
(Mayachiew & Devahastin, 2008). Another member of the rhizome fa- swell in aqueous media. Tripathi et al. studied the physicochemical and
mily is turmeric. Curcumin, a metabolite found in the turmeric extract, antimicrobial properties of glutaraldehyde cross-linked chitosan-PVA
is a good antioxidant and antimicrobial agent. Roy and Rhim (2020) blend films (Tripathi et al., 2009). The cross-linked films displayed a
studied the effect of curcumin dosing in chitosan films. It was observed better antimicrobial effect and enhanced the shelf life of processed to-
that the composite films displayed excellent antioxidant activity. Also, matoes. Tannic acid was used as a cross-linker in chitosan-chitin
the synergistic effect of chitosan and curcumin makes it an efficient whisker nanocomposite films by Rubentheren et al. and observed a
antimicrobial packaging material (Roy & Rhim, 2020). Bitter apricot decreased solubility and water uptake behavior (Rubentheren et al.,
Prunus armeniaca is also an agro-waste resource, and the essential oils 2015). However, the crosslinked films were more rigid as compared to
extracted from its seeds are also good antioxidants and antimicrobial neat chitosan films. Other researchers also observed this increase in
agents. The properties of chitosan films incorporated with apricot rigidity and brittleness in chitosan films due to cross-linking. Cui et al.
kernel essential oils were studied, and it was found that the oil helped in (2011) reported that the citric acid cross-linked chitosan films were
enhancing the antimicrobial and antioxidant properties of the films rigid and brittle and were more prone to cracking. To avoid this, they
leading to enhanced shelf life of packed bread slices by preventing the used glycerol as a plasticizer, which not only retained the properties
mold growth for almost ten days (Priyadarshi, Sauraj, Kumar, Deeba, developed due to cross-linking but also make them flexible. A similar
et al., 2018). Among plant extracts, plant-based proteins are emerging combination was studied by Priyadarshi, Sauraj, Kumar, and Negi
as promising active agents being economical and highly abundant. (2018a, 2018b), and it was observed that besides possessing thermal
Kidney bean protein isolates imparted the chitosan films with flexibility stability and water resistance, the films also showed decent antioxidant
and low surface free energy leading to high hydrophobicity, which is an activity which was attributed to the presence of citric acid (Fig. 6).
important requirement for the packaging of dry, moisture-sensitive food The addition of plasticizers helps in imparting flexibility to the host
products (Ma, Tang, Yang, & Yin, 2013). In another study, the quinoa polymer. Also, they help in making the biopolymeric films suitable for
protein blended chitosan film showed improved mechanical and water processing via conventional routes like extrusion and compression
barrier properties (Abugoch et al., 2016; Caro et al., 2016). Rambabu molding. Chitosan films are generally prepared by the solution casting
et al. used Mango leaf extract as an active agent in the chitosan films – it method and not by these conventional plastic processing routes. The
imparted antioxidant properties to the film and the shelf life of the reason is that chitosan, like cellulose, possesses a degradation tem-
packed cashew nuts remained preserved for 28 days. It was suggested perature below its melting temperature. Plasticization leads to the
that the mango leaf extract incorporation led to 56 % higher oxidation lowering of the glass transition temperature of chitosan by decreasing
resistance in the nuts (Table 4). Besides this, the mechanical, as well as the number of intermolecular bonds between polymer chains, hence
moisture barrier properties of the films, were also enhanced (Rambabu making it more processable (Matet, Heuzey, Ajji, & Sarazin, 2015).
et al., 2019). In an intelligent chitosan film prepared using purple-fle- Glycerol is the most commonly used plasticizer for chitosan. However,
shed sweet potato extract as the pH-responsive agent of the film, the other plasticizers like ethylene glycol, polyethylene glycol, propylene
color of the film changed from yellow at pH 3 to dark brown at pH 10 glycol, and sorbitol have also been used. Kelnar et al. (2015) prepared
(Yong et al., 2019). In addition, the purple sweet potato extract, an chitosan/chitin nanowhisker composite films plasticized with glycerol
anthocyanin pigment, gave antioxidant activity to the film. In a more and polyethylene glycol and studied their properties. The results show
13
Table 4
Day-wise peroxide values for oil derived from cashew nuts stored in various control and MLE films (Rambabu et al., 2019).
Film samples Peroxide index values (mEq kg−1)
Day 1 Day 7 Day 14 Day 21 Day 28
a a a a
Open control 1.78 ± 0.00 1.94 ± 0.00 4.03 ± 0.27 4.75 ± 0.47 8.82 ± 0.59a
PA/PE control 1.77 ± 0.00b 1.80 ± 0.01b 2.68 ± 0.14b 3.45 ± 0.36b 6.59 ± 0.47b
Chitosan control 1.76 ± 0.00c 1.86 ± 0.01c 2.73 ± 0.08b 3.44 ± 0.27b 6.34 ± 0.41b
1% MLE film 1.76 ± 0.00c 1.85 ± 0.02c 2.71 ± 0.07b 3.42 ± 0.22b 6.28 ± 0.39b
3% MLE film 1.75 ± 0.00d 1.78 ± 0.01d 1.47 ± 0.09c 3.08 ± 0.19c 3.65 ± 0.16c
5% MLE film 1.73 ± 0.00e 1.75 ± 0.00e 1.24 ± 0.05d 2.69 ± 0.03d 2.88 ± 0.11d
Different alphabets in a given column, indicate significant change (p < 0.05) of value as established by Tukey's test.
on the use of conventional antibiotics as antimicrobial agents in the

films. Mu et al. (2014) reported the incorporation of commonly used
antibiotics like amikacin, clindamycin, vancomycin, and erythromycin
in the chitosan food packaging films and the effectiveness of the films in
retarding the growth of common food pathogen Listeria monocytogenes
and also other Listeria species was estimated. As expected, the results
showed eradication of Listeria biofilms due to synergistic action of
chitosan and antibiotics, particularly amikacin. Microbial proteins are
also considered important functional material, and chitosan films
blended with them show various properties in the context of food
packaging. Lactococcus lactis is a source of amphiphilic cationic peptide
Nisin. Chitosan-nisin blend films show antimicrobial activity resulting
in an enhanced shelf life of the packaged food product (Imran, Klouj,
Revol-Junelles, & Desobry, 2014; Zimet et al., 2019). In another re-
search, Wu et al. (2019) studied the antimicrobial activity of chitosan
films incorporated with ℇ-polylysine against gram-positive S. aureus
and gram-negative E. coli bacteria. It was found that the films with
chitosan to ℇ-polylysine ratio 9:1 displayed a 2-fold antimicrobial ac-
tivity against S. aureus and a 1.5 fold activity against E. coli bacterial
strains as compared to neat chitosan films. Although this strategy of
using antibiotics in active packaging proves to be better for microbial
inhibition than that discussed in previous sections, the use of antibiotics
in food contact materials can cause serious health hazards. It is sug-
gested that the antibiotics might diffuse in the packaged food material
with time resulting in their indirect consumption by humans. As a
consequence of this, the human pathogens may develop multiple drug
resistance, and the human body may become highly susceptible to
microbial infections (Phillips et al., 2004).
6.2. Edible coatings
It is not a necessity that all biodegradable materials fall into the

Fig. 6. Scheme of cross-linking between chitosan and citric acid (Priyadarshi, category of edible materials. However, the vice-versa is true. Edible
Sauraj, Kumar, & Negi, 2018a, 2018b). films are the ones which are composed wholly of the food-grade com-
ponents whether they are film-forming polymer matrix, the solvent in
that the elasticity and flexibility were imparted to both the chitosan which it is dissolved, or the additives such as fillers, plasticizers, cross-
matrix as well as the chitin nanowhiskers resulting in stretchable filler linkers, or biologically active agents which are incorporated in the films
particles leading to improved mechanical properties. The decrease in (Otoni et al., 2017).
glass transition temperature was also observed for both the components Flexible films, unlike edible coatings, are free-standing structures
indicating enhanced processability. that are produced separately and then applied on the food surface or are
sealable pouches in which the packaging of food is done. On the other
hand, edible coatings are formed on the food surface directly (Fig. 7) by
6.1.6. Films embedded with antimicrobial agents several methods like dipping, spraying, and panning, or by mixing both
The increasing research inclination towards the active food packa- food material and coating solution in a rotating container and drying
ging materials is leading to the emergence of packaging materials that after that. Although edible coatings have limitations and are not sui-
show biological activity like antimicrobial, antioxygenic, antioxidative, table as a complete replacement of conventional packaging materials,
etc. For this purpose, a lot of literature is present, as already described they have the potential to extend the shelf life of food by regulating the
in previous sections, on the addition of functional agents in food moisture, aroma and gas exchange between food and environment and
packaging biopolymeric films. Natural essential oils and extracts, by protecting the surface of food from contamination and discoloration
modified clay fillers, metallic and biological nanoparticles, etc. have (Dutta, Tripathi, Mehrotra, & Dutta, 2009).
been increasingly used to impart biological activity, especially anti- The edible coatings are usually supposed to be transparent and
microbial activity to the films. However, very limited has been reported devoid of any odor or flavor. They should not interfere with food
14
Fig. 7. Schematic representation of the production of films and coatings (Otoni et al., 2017).
sensory properties. McHugh, Huxsoll, & Robertson (1997), for the first % relative humidity. Furthermore, no change in the sensory attributes
time, produced edible films based on fruit purees, and this provided a of the berries was observed. A similar study was performed by Vargas
whole new dimension to food packaging research. Most of the research et al. (2006), who used chitosan combined with oleic acid to coat cold-
reports combined film-forming biopolymeric materials like cellulose, stored strawberries and it was observed that the coatings preserved the
chitosan, starch, pectin, etc. with fruit and vegetable purees. The purees mechanical properties and color of the berries. It also protected the
used were themselves a good source of nutrients and antioxidants and strawberries against fungal pathogens and hence extending its shelf life.
were found to be healthy for ingesting. Gradually, the focus shifted A reduced water loss and higher retention of sensory attributes of sliced
from edible films to edible coatings, primarily for the lightly processed mango coated with chitosan were reported by Chien et al. (2007). The
food products. Cutting, slicing, peeling, and sectioning processes fall edible chitosan coating also inhibited microbial growth leading to en-
under light processing. Lightly processed food products have an in- hancement in the shelf life of the sliced fruit. Tripathi, Mehrotra, &
creased rate of respiration and ethylene production, which leads to Dutta et al. (2009) studied the effect of chitosan-PVA coating on the
faster deterioration. Besides this, they have a high surface water activity minimally processed tomatoes. It was observed that the coating in-
that invites microbial attack. Edible coatings on these lightly processed hibited the growth of E. coli, B. subtilis and S. aureus on the tomatoes.
food products have been a successful method to protect them and en- Moreira et al. (2011) studied the effectiveness of chitosan edible coat-
hance their shelf life (Otoni et al., 2017). ings on the sensory quality of fresh-cut broccoli. The coating led to a
Chitosan has been widely used in edible coatings, either alone or in decreased microbial attack and inhibited the yellowing and opening
combination with other biopolymers or additives. Many reports have florets hence, promoting the retention of its sensory attributes (Moreira
been published which present research work on chitosan-based edible et al., 2011). Ortiz-Duarte et al. (2019) studied the effect of chitosan
coatings on unprocessed or lightly processed fruits, vegetables, and fish coatings embedded with Ag-chitosan composite nanostructures on the
and meat products. Han et al. (2006) studied the effect of chitosan quality of fresh-cut melons. It was observed that the coating treatments
coating on the shelf life of fresh strawberries and found that the coated reduced the respiration rates of the fruit, thereby extending the shelf
berries, unlike non-coated ones, were accepted by consumers even after life. Also, the coated fruits were more resistant to microbe mediated
one week when stored near-freezing temperatures and slightly below 90 deterioration. Gomes et al. (2019) prepared edible chitosan coatings
15
incorporated with chitosan nanoparticles. The coatings displayed an Acevedo-Fani, A., Salvia-Trujillo, L., Soliva-Fortuny, R., & Martin-Belloso, O. (2015).
antimicrobial effect towards both Gram-positive as well as Gram-ne- Modulating biopolymer electrical charge to optimize the assembly of edible multi-
layer nanofilms by the layer-by-layer technique. Biomacromolecules, 16(9),
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was studied which was found to be suitable for their increased pre- tural and barrier properties of crab shell chitosan/graphene oxide composite films.
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Declaration of competing interest Casariego, A., Souza, B. W. S., Cerqueira, M. A., Teixeira, J. A., Cruz, L., Díaz, R., &
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The authors declare no conflict of interests. Chen, G.-G., Qi, X.-M., Guan, Y., Peng, F., Yao, C.-L., & Sun, R.-C. (2016). High strength
hemicellulose-based nanocomposite film for food packaging applications. ACS
Sustainable Chemistry & Engineering, 4(4), 1985–1993.
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Innovative Food Science and Emerging Technologies 62 (2020) 102346

Contents lists available at ScienceDirect

Innovative Food Science and Emerging Technologies

Chitosan-based biodegradable functional ﬁlms for food packaging T

Three important parameters determine the solubility of chitosan in

• the pH of the solution

Chitosan has a characteristic property of being soluble at acidic and

4.3. Charge density

eﬀect on the mechanical attributes (Khwaldia et al., 2014). 5.1.4. Extrusion

Films with nanomaterials

Films with clays

Films with polysaccharide particles, ﬁbers, and whiskers

Films with natural oils and extracts

Films with cross-linkers and plasticizers

Material type Additive Property References

Films with antimicrobial agents

Values are given as mean ± SD (n = 3).

Day 1 Day 7 Day 14 Day 21 Day 28

on the use of conventional antibiotics as antimicrobial agents in the

6.2. Edible coatings

It is not a necessity that all biodegradable materials fall into the

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