Trends in Food Science & Technology 124 (2022) 259–277

Contents lists available at ScienceDirect

Trends in Food Science & Technology

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

Make your packaging colorful and multifunctional: The molecular

interaction and properties characterization of natural colorant-based films
and their applications in food industry
Jiayin Huang a, b, Zhiheng Hu a, c, Gaoshang Li a, b, Lingping Hu a, b, Jianchu Chen a, Yaqin Hu b, *
a
Institute of Food Engineering, College of Biosystems Engineering and Food Science, Zhejiang University, Hangzhou, Zhejiang, 310058, China
b
College of Food Science and Engineering, Hainan Tropical Ocean University, Marine Food Engineering Technology Research Center of Hainan Province, Collaborative
Innovation Center of Marine Food Deep Processing, Sanya, Hainan, 572022, China
c
Hainan Key Laboratory of Herpetological Research, College of Fisheries and Life Science, Hainan Tropical Ocean University, Sanya 572022, Hainan, China

A R T I C L E I N F O A B S T R A C T

Keywords: Background: Great attentions have been paid on the safety and quality of food products, leading to emerging
Natural colorants innovations in food packaging industry. The natural colorants with eco-friendly and multi-functional properties
Food packaging are widely integrated into packaging films. And the natural colorant-based film can not only protect food from
Molecular interaction
surrounding environment, but also behave indicator and active functions as a promising component for novel
Physical properties
Functional properties
food packaging.
Scope and approach: This review provides a thoroughly insight into recent advances in natural colorant-based
food packaging. Main 7 kinds of natural pigments were critically summarized. The molecular interaction be­
tween natural colorants and bio-polymers was investigated. Meanwhile, the influences of molecular interaction
on physical and functional properties of natural colorant-based films were clearly discussed. Additionally, the
applications of natural colorant-based film in terms of indicator and active food packaging were systematically
summed. Furthermore, the major bottlenecks and future prospects were also illustrated for the further appli­
cation of natural colorant-based film.
Key findings and conclusions: The molecular interactions which were varied due to different charged natures of
natural colorants, exerted significant influences on film physical properties, including structural, mechanical,
hydrophobic, thermal and oxygen permeability properties. The natural colorant-based film which exhibited great
pH-responsive, active and barrier characteristics, was suitable for the freshness monitoring and maintenance of
food products, being regarded as indicator and active food packaging. Nevertheless, further studies focusing on
the stabilization modification of colorants as well as the commercial production method of natural colorant-
based films are critically needed.

1. Introduction recent years.

Food packaging which performs the responsibility of protection,
convenience, containment and communication, plays a critical role in
the storage, distribution and retail chain of food products. Nowadays,
large amounts of conventional petroleum-based packaging have posed
huge burdens on environment issue. Furthermore, several harmful
chemical compounds contained in the packaging system have the risk to
cause severe health diseases. Therefore, emerging novel food packaging,
which not only reacts as effective barriers but also behaves specific
functions complying with the social development, has been fabricated in
The synthetic colorants with high stability and low cost have been
integrated with the packaging matrix for perceiving food spoilage pro­
cess by color variations. Nevertheless, people are worried about safety
problems resulted from their biological and toxicity characteristics. To
this end, natural colorant has become an ideal alternative towards
synthetic dye owing to its non-toxicity, biodegradability and eco-
friendly characteristics. Several reviews on the natural colorants (like
anthocyanin, curcumin, betalain) have been reported in terms of their
sources, extraction methods, stability or biological activities such as
cardiovascular protection, antioxidant, neuroprotection

* Corresponding author.
E-mail address: 1004346262@qq.com (Y. Hu).

https://doi.org/10.1016/j.tifs.2022.04.028
Received 2 March 2022; Received in revised form 9 April 2022; Accepted 25 April 2022
Available online 29 April 2022
0924-2244/© 2022 Elsevier Ltd. All rights reserved.
J. Huang et al.
(Carreón-Hidalgo, Franco-Vásquez, Gómez-Linton, & Pérez-Floresb,
2022; Yang, Guo, et al., 2022; Aliabbasi, Fathi, & Emam-Djomeh, 2021).
Therefore, in addition to behaving similar functions as synthetic
colorant to sense the deterioration procedure of packaged food, the
natural colorants can also maintain food quality due to their outstanding
active functions. As a result, natural colorants play vital parts in the
development of packaging industry, which broaden the promising
application of food packaging. Till now, critical reviews about natural
colorant-based films have mainly focused on their preparation proced­
ures, properties characterization or their performances in freshness
monitoring, behaving as real-time freshness indicator, while paid little
attention on their roles in food preservation (Alizadeh-Sani, Moham­
madian, Rhim, & Jafari, 2020; Zheng, Liu, Yu, & Shao, 2022). Mean­
while, only anthocyanin and curcumin these two natural colorants have
been widely reviewed as potential functional components in food
packaging system. For example, Roy, Priyadarshi, Ezati, and Rhim
(2022) discussed the uses of curcumin in the food packaging regarding
with the structure, biological activity, application and commercial
prospects of curcumin-based film. Similarly, the recent advances in the
preparation, properties and potential application of anthocyanin-based
packaging films have been reviewed by Bacerril, Nerín, and Silva
(2021), Filho et al. (2021) and Yong and Liu (2020). Nevertheless, there
are many natural colorants existed in the world, the reviews about other
natural colorant-based packaging films as well as their applications in
both freshness monitoring and food preservation are deficient yet.
Hence, a systematic review on the indicator and active packaging films
incorporating all kinds of common natural colorants is necessary for the
development of food packaging industry.
Nowadays, there are two main topics of the researches on natural
colorant-based films. One is the properties characterization, evaluating
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Trends in Food Science & Technology 124 (2022) 259–277
the influences of different colorant contents or preparation conditions
on the film characteristics (Ge et al., 2020; Jiang et al., 2020; Mei et al.,
2020; Zheng, Li, et al., 2022). The other one is preliminary application,
behaving as indicator or active packaging for freshness monitoring as
well as shelf-life extension (Guo, Ge, et al., 2021; He et al., 2022; Qin
et al., 2021; Sganzerla et al., 2021; Yang et al., 2022; Zhang, Li, et al.,
2021). Interestingly, these two areas can be both influenced by molec­
ular interactions contained in the packaging system. Generally, in order
to immobilize natural colorants for stability enhancement, biodegrad­
able macromolecular polymers have been used as suitable substitutes to
petroleum-based derivatives, which will interact tightly with natural
colorants. It is believed that the molecular interactions between natural
colorant and polymer matrix will be varied because of various structural
conformation, exerting different influences on the packaging properties.
However, the systemic reviews on the types and formation mechanism
of molecular interactions, as well as their related effects on film char­
acteristics are limited.
Therefore, this review provides a throughout insight into recent
advances of novel food packaging incorporating natural colorants. There
are 7 kinds of natural colorants characterized in this review regarding
their classification and differences in chemical structures. Additionally,
the molecular interactions and their effects on film physical and func­
tional properties are summarized systematically. Furthermore, the
application in terms of freshness monitoring and food preservation,
existing problems and prospects of natural colorant-based food pack­
aging are discussed thoroughly (Fig. 1).
2. Natural colorants
Natural colorants, presenting advantages of free-pollution, free-
Fig. 1. The schematic diagram of the parts discussed
in the current review.
A) The different kinds of natural colorants applied in
the food packaging industry widely are discussed in
the first part, mainly are anthocyanin, curcumin,
betalain, chlorophyll, carotenoid, shikonin and aliz­
arin. B) In the second part, the molecular interaction
between natural colorants and polymers are identi­
fied systematically. C) The physical properties of
natural colorant-based films are characterized in the
third part. D) The functional properties of natural
colorant-based films are characterized in the forth
part. E) In the last part, the application of natural
colorant-based films, including freshness indicator
and active packaging are summarized.
J. Huang et al.
toxic, renewable and biodegradable, can be classified according to their
sources (plant, animal or microorganism) and structures (porphyrin,
isoprene, polyphenol, ketone, quinone-derivatives, etc) (Ghosh, Sarkar,
Das, & Chakraborty, 2022). Recently, they have been used as sensing
and active compounds in the food packaging. Briefly, the most common
natural colorants applied in novel food packaging are anthocyanin,
curcumin, betalain, chlorophyll, carotenoid, shikonin and alizarin
(Fig. 2).
2.1. Anthocyanin
As a kind of water-soluble flavonoid derivative, the anthocyanins are
mainly distributed in the tissue of red or purple fruits and vegetables,
such as blackberry, purple sweet potato, red cabbage, strawberry, etc.
Till now, over 500 kinds of anthocyanins have been identified.
Structurally, anthocyanins are glycosides of anthocyanidins, con­
taining a basic C6-C3-C6 skeleton, which consists of two aromatic rings
separated by a 6-membered heterocyclic ring with an oxygen atom
(Fig. 2). Meanwhile, there is a sugar esterification at 3-position, which
can be attached to saccharides. Notably, the anthocyanins in nature are
divided into 6 forms based on their substituent: cyanidin, pelargonidin,
peonidin, delphinidin, petunidin and malvidin, whose conformations
differ greatly because of the differences in the position and the number
of -OH and -CH3 groups.
After being extracted, the anthocyanins exhibit red, purple or blue
colors originally. While their colors will change under different acid-
base environment, making them become ideal sensing materials for
food freshness monitoring. Besides, researches have demonstrated the
promising effects of anthocyanins on human health, behaving as effec­
tive cardiovascular protector, anti-diabetes and anti-obesity com­
pounds, etc. Apart from outstanding biological properties, active
characteristics like antibacterial and antioxidant capacity are helpful for
the development of food packaging. The blueberry anthocyanin was
reported to exhibit strong antimicrobial activity against Listeria mono­
cytogenes (L.monocytogenes) and Staphylococcus aureus (S.aureus) by
destruction of cell membrane (Albuquerque, Oliveira, Barros, & Fer­
reira, 2020). On the other hand, the positively charged oxygen atom and
phenolic hydroxyl groups which allow the donation of hydrogen,
contribute to its potent antioxidant functions (Yong & Liu, 2020).
2.2. Curcumin
The curcumin (1,7-bis(4-hydroxy-3-methoxyphenyl)-1,6-
Fig. 2. The kinds, sources and structures of the most common natural colorants applied in novel food packaging.
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heptadiene-3,5-dione) can be isolated from the rhizomes of curcuma
longa, belonging to the ginger family. As a derivative of isoprene, cur­
cumin does not dissolve in water, but presenting well lipid-soluble
property (Roy et al., 2022). The curcumin is a rare natural colorant
with a typical crystalline structure. There are phenolic ortho methoxy
groups which give curcumin excellent antioxidant capacity, being
attached to the β-diketone moiety (Fig. 2).
The initial color of curcumin is yellow-orange, which will experience
noticeable change under different pH environment, exhibiting its pos­
sibility in freshness monitoring. In addition to the great antioxidant
capacity mentioned above, the curcumin contributes to the inhibition of
microbial activity such as Salmonella and L. monocytogenes (Gao &
Matthews, 2020). Besides, the curcumin also has a series of physiolog­
ical effects such as anti-cancer and anti-malarial activity, which is
famous for its outstanding therapeutic influences on chronic diseases.
2.3. Betalain
As a kind of heterocyclic compound, betalain can be found in edible
parts of plants of order Caryophyllales, such as the Beta vulgaris L. root
(the main source of betalain), Swiss chard, pear cactus and Opuntia
stricta (Ghosh et al., 2022; Kanatt, 2020). Several functions of betalain
have been investigated, such as its ability to scavenge free radicals,
retard lipid oxidation, inhibit microorganism growth as well as fight
against inflammation (Koop et al., 2022; Yao, Hu, Qin, & Liu, 2020).
The betalain, as the metabolite production derived from L-tyrosine
metabolism, is the ammonium conjugate of betalamic acid with amino
compounds or cyclo-DOPA (l-3,4-dihydroxy-phenylalanine) (Fig. 2).
Therefore, the batalain can be characterized into betacyanin and
betaxanthin, showing reddish-violet and yellowish-orange color,
respectively. The color of betalain depends on the proportion of these
two forms, which enables betalain to be a promising freshness indicator.
2.4. Chlorophyll
The chlorophyll whose breakdown is responsible for the leaf senes­
cence, is well known as the photosynthetic greenish colorant, acting as
an effective energy provider (Ghosh et al., 2022). It can be classified into
5 major classes, in which the chlorophyll a and b are mainly distributed
in green plants, the chlorophyll c and d are originated from algae while
the chlorophyll e can be derived from photosynthetic bacteria.
As shown in Fig. 2, the basic structure of chlorophyll is a porphyrin
ring and a cyclic tetrapyrrole, containing a divalent magnesium (Mg) in
J. Huang et al.
its center. When the central Mg2+ is replaced, the color of chlorophyll
will become darker, leading to the formation of pH-responsive ability.
The unique conformation of chlorophyll is crucial for reduction of free
radical. Meanwhile, the rearrangement of electron in the peripheral
position will modify the characteristics of π-electron system, affecting
several biological properties including regulation of xenobiotic meta­
bolism, antimutagenic, antimicrobial and anti-inflammation activities
(Zepka, Jacob-Lopes, & Roca, 2019).
2.5. Carotenoid
The carotenoid is a lipid-soluble tetratepenoid, which also can be
dissolved in non-polar solvents. The main sources of carotenoid are
fruits, vegetables, algae, insects and fungi.
Chemical structurally, the carotenoid consists of eight isoprenoid
units and their order can be inverted at the center of molecule (Aliza­
deh-Sani et al., 2020) (Fig. 2). According to different functional groups
contained in the molecular chain, the carotenoid can be divided into two
major classes: 1) the carotenoids contain parent hydrocarbon chain
only, like α-carotene, xanthophylls, and lycopene. They are strong
antioxidant agents, which can extinct the singlet oxygen and reduce
oxidative stress as well as react as effective protector to retard the
growth of cancerous cells; 2) the carotenoids contain oxygen, including
lutein and astaxanthin. These two kinds of carotenoid especially lutein,
is the predominant carotenoid in the prevention of sever diseases like
cancer, cardiovascular disease and cataracts, and they also perform
excellent antioxidant, antimicrobial and anti-inflammatory properties
(Koop et al., 2022). Despite of different potentials in biological prop­
erties, all carotenoids have similarities in color formation, that is, the
conjugated C=C chemical bonds are responsible for their red/or­
ange/yellow color, and the colorful compound will be decomposed
under alkaline conditions, which is suitable for its incorporation in in­
dicator film for freshness monitoring.
2.6. Shikonin
Shikonin is a novel and non-toxic natural colorant extracted from the
roots of Oriental medicinal herb, such as Arnebia euchroma and Lith­
ospermum erythrorhizon (Rhim & Roy, 2021). It it a typical naph­
thoquinone colorant, which consists of a naphthazarin group as well as a
chiral six-carbon side chain. Since the naphthazarin moiety is chemi­
cally reactive, the shikonin is sensitive to the environmental alterations.
The shikonin exhibits bright red color originally. Although it is
relatively stable at room temperature compared with anthocyanin, it
presents pronounced color changes at different acid-based conditions. In
addition, as a traditional Chinese medicine, shikonin shows a wide range
of biological properties such as anti-inflammatory, wound healing,
antimicrobial, antioxidant, anti-tumor, etc (Ezati, Bang, & Rhim, 2021).
2.7. Alizarin
The alizarin (1,2-dihydroxyanthraquinone) is another highly appli­
cable natural colorant, which exhibits lower solubility in polar solvent.
Besides dye industry, it has been also widely used in other industries
such as pharmaceutical and tissue chemical industry. Generally, the
alizarin, also known as Turkey Red, can be derived from madder plant
roots (Ezati & Rhim, 2020). It is an anthraquinone compound, con­
taining three benzene rings or conjugated orbitals of hydroxyl groups.
The color of alizarin is orange-red, while the chemical reaction of
hydroxyl groups contained in its molecular chain contributes to its color
variation towards different pH environment. Meanwhile, the alizarin
exhibits several functional properties, including antioxidant, UV
blocking, etc (Ezati, Rhim, Moradi, Tajik, & Molaei, 2020). Raj, Kim,
Kim, Lee, and Lee (2022) reported the efficient biofilm inhibition ability
of alizarin against S. aureus.
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3. Molecular interactions between natural colorant and bio-
polymers in the film system
Generally, the Fourier transform infrared spectroscopy (FT-IR)
spectra of packaging film with or without natural colorants are relatively
similar, with no supplementary peaks presence, suggesting the colorants
can not exert influences on chemical composition of bio-polymers due to
their extremely low concentrations. However, the slight variations of
characteristic peaks can be identified due to the molecular interaction
between colorants and polymers, which are the vital parameters of film
properties. On the one hand, they play important roles in formation of
physical characteristics. On the other hand, the molecular interactions
are closely related to the release of natural colorant from the film matrix,
affecting the functional properties (Fig. 3(a)). The types of molecular
interactions highly depend on the charged nature of film components.
Hence, according to different electronic properties of natural colorants,
we divide them into two groups, 1) charged colorants: anthocyanin,
betalain and chlorophyll; 2) neutral colorants: curcumin, carotenoid,
shikonin and alizarin. Their molecular interactions with polymers are
illustrated in Fig. 3(b).
3.1. The molecular interactions between charged natural colorants and
common polymer matrix
The characteristic peaks in the FT-IR spectra of natural colorants can
represent the chemical bonds, contributing to further definition of their
interaction with polymers. The FT-IR spectra of anthocyanins exhibited
characteristic peak of the aromatic ring C-H deformation band (around
1028 cm− 1), the C=C stretching band (1634-1514 cm− 1) and the O-H
stretching band (3700-3000 cm− 1) (Ge et al., 2020; Qin et al., 2021;
Yong et al., 2019). Meanwhile, several typical peaks were reported in
the FT-IR spectra of betalains, such as the N-H stretching band (around
3274 cm− 1), C=O stretching band (around 1712 cm− 1), C=N stretching
band (around 1643 cm− 1), C=C stretching band (around 1600 cm− 1),
C-O stretching band (around 1071 cm− 1) and the C-O-C band (around
1024 cm− 1) (Qin, Liu, Zhang, & Liu, 2020). As for chlorophyll with a
different chemical structure compared with anthocyanins and betalains,
presented a relatively different FT-IR spectra, in which the bands around
900 cm− 1 and between 1500 and 1800 cm− 1 were responsible for the
Chla-pyrrol ring and the Amide I as well as the amine signals (Cha­
voshizadeh, Pirsa, & Mohtarami, 2020; Rizzi et al., 2016).
After combination, several variations in the position or the intensity
of characteristic peaks will be observed. Qin et al. (2021) developed a
lycicum ruthenicum anthocyanin-based film incorporating starch and
polyvinyl alcohol (PVA), obvious shifts of O-H and C-H stretching vi­
bration were identified, from 3279 cm− 1 to 2927 cm− 1 to 3290 cm− 1
and 2923 cm− 1, respectively. And the O-H bond was intensified after the
addition of anthocyanin, which might be due to the formation of
hydrogen bonds. Similarity, the bands at 2879 cm− 1 and 2924 cm− 1 of
Artemisia sphaerocephala Krasch gum film significantly weakened, which
was associated with the hydrogen bond resulted from the combination
with purple onion peel anthocyanin (Liang, Sun, Cao, Li, & Wang, 2018).
The conversion of characteristic peaks were also investigated in
betalain-based films. Compared with starch/PVA film, the starch/P­
VA/red pitaya (Hylocereus polyrhizus) betalain exhibited broaden O-H
stretching band which shifted to a higher wave number, and they
attributed it to the generation of hydrogen bonds (Qin, Liu, et al., 2020).
Aforementioned, when being incorporated with neutral bio-polymers
(starch, gum, PVA, etc), the hydroxyl structure contained in the natu­
ral colorants were supposed to facilitate the formation of H-H with the
hydrophilic groups in the polymer matrix. Besides, it is worthy to note
that the chlorophyll as a kind of charge colorant, indeed can also interact
with neutral bio-polymer such as poly(methylethacrylate) (Abdelrazek,
Damrawi, Elashmawi, & EI-Shahawy, 2010). Although several changes
of typical stretching bands are identified, the specific kinds of molecular
interaction between chlorophyll and neutral charged polymers have not
J. Huang et al. Trends in Food Science & Technology 124 (2022) 259–277

Fig. 3. The molecular interaction within natural colorant-based films.

In which, (a) the relationship between molecular interaction and film properties Specifically, the molecular interaction can affect film physical properties by
alteration of structure network, as well as affect film functional properties by controlling the colorant release; (b) A brief description of molecular interactions
between neutral colorants and common polymer matrix. The charged colorants (anthocyanin, betalain and chlorophyll) interact with neutral polymer by hydrogen
bonds, whereas with charged polymer by hydrogen bonds and electronic repulsion; while the neutral colorants (curcumin, carotenoid, shikonin and alizarin) interact
with polymer mainly by hydrogen bonds.

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J. Huang et al.
been exactly defined yet.
In addition to neutral polymers, there are also some charged bio-
polymers being applied as film matrix. The stretching bands of sodium
carboxymethyl (CMC)/κ-carrageenan film at 1410 cm− 1 and 845 cm− 1
widened and shifted, respectively, which was due to the incorporation of
mulberry anthocyanin, indicating the generation of electrostatic repul­
sion and hydrogen bonds (Zhang, Sun, Cao, & Wang, 2020). The new
molecular interaction (electrostatic repulsion) was also reported in the
chitosan/oxidized chitin nanocrystal/black rice anthocyanin film (Wu
et al., 2019) and CMC film incorporating red cabbage anthocyanins
(Liang, Sun, Cao, Li, & Wang, 2019), manifesting as N-H band shifts
around 1600 cm− 1. Similar result was obtained in the biodegradable
film based on the charged components chitosan and betalain. Apart from
hydrogen interactions, the amide II bond exhibited a blue shift owing to
the addition of red pitaya betacyanin, which was associated with strong
electrostatic interactions among these film compounds (Yao, Liu, Hu,
Yun, & Liu, 2022). Rizzi et al. (2016) prepared a chitosan/chlorophyll
film, and the FT-IR spectra demonstrated the intensity changes of Amide
I and amine signals (between 1500 and 1800 cm− 1), which were
attributed to the rearranging of hydrogen bonds and formation of elec­
trostatic repulsion. In addition, the shifts in 3248 cm− 1 corresponding to
the -OH stretching vibration of CMC/silica/pectin film incorporated
with chlorophyll also indicated the chlorophyll could be immobilized
through electrostatic interactions (Sharifi & Pirsa, 2021). Therefore, it is
supposed that the charged oxygen atom, carboxyl groups and macro­
cycle contained in the anthocyanin, betalain and chlorophyll have the
ability to interact with functional groups (carboxyl, amino, etc) in the
charged polymers such as CMC, chitosan and κ-carrageenan via the
electrostatic repulsion and hydrogen bonds.
3.2. The molecular interactions between neutral natural colorants and
common polymer matrix
In the FT-IR spectra of curcumin, an obvious peak was observed at
1623 cm− 1, attributing to the C=O stretching, the typical keto-enol
tautomersim structure of curcumin. The characteristic peaks were also
identified such as the -OH stretching (3507 cm− 1), C=C groups (1503
cm− 1), C-O stretching (1275 cm− 1) and C-H stretching (below 1000
cm− 1) (Liu, Dang, Zhang, Munsop, & Li, 2021). In terms of carotenoid,
considering its oxygen-containing forms were the main contributors in
food packaging, the characteristic peaks in the region between 1633 and
1646 cm− 1 as well as at the 3600 cm− 1 were responsible for the C=O
sand the -OH stretching vibration, respectively. In addition, the CH2
asymmetrical and symmetrical stretching could also induce the ab­
sorption peaks around 2900 cm− 1 (Hari, Francis, Nair, & Nair, 2018;
Łupina, Kowalczyk, Lis, Raszkowska-Kaczor, & Drzlowska, 2022). As for
shikonin, it presents several characteristic peaks around 1612, 1714,
2925 and 3500 cm− 1, corresponding to the C=C aromatic ring stretch­
ing, carbonyl groups, -C-H stretching band and -OH group, respectively
(Rhim & Roy, 2021). While there are also several typical peaks can be
identified in the FT-IR spectra of alizarin, such as the -OH stretching
vibration (3412 cm− 1), C=O (1710 cm− 1) and C=C (1572-1380 cm− 1)
aromatic stretching vibration (Ezati, Tajik, Moradi, & Molaei, 2019).
Compared with charged natural colorants, the molecular interaction
of neutral colorants with bio-polymers is simpler, in which the hydrogen
bond is the major interaction mode, whatever the charge natures of
polymers are. The combination of curcumin and whey protein isolate
(charged polymer) contributed to the blue shift of hydroxyl stretching
vibrations from 3508 cm− 1 to 3276 cm− 1, which was resulted from the
formation of hydrogen bonds (Kevij, Salami, Mohammadian, & Khoda­
dadi, 2020). An active film on the basis of curcumin and neutral poly­
mers (PVA and starch) presented the stronger intensity of -OH and -CH
stretching vibration around 3260 cm− 1 and 2916 cm− 1 which accounted
for new hydrogen bonds generated (Chen, Zhang, Bhandari, and Yang
(2020). In addition, Hari et al. (2018) observed a slight shift in the po­
sition of characteristic peak in chitosan-gelatin film after the
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incorporation of β-carotene. Similar results were obtained in which the
addition of alizarin could intensify the peak around 3300-2800 cm− 1 in
the CMC/alizarin film, which was due to the hydrogen interactions
(Ezati et al., 2020). Thus, the hydroxyl groups in neutral natural col­
orants (curcumin and carotenoid) are concluded to facilitate the gen­
eration of hydrogen bonds with the hydrophilic groups (such as amino
and hydroxyl groups) contained in the bio-polymers.
3.3. The influencing factors of molecular interactions
There are several factors affecting molecular interactions. Firstly, the
molecular interactions are highly dependent on the kinds and sources of
natural colorants. The pyranose ring of glucose residues intensified more
significantly in starch/PVA film combination with red pitaya peel
betalain than L. ruthenicum anthocyanin, being attributed to the stronger
inter-molecular bond force (Qin et al., 2020). Yao et al. (2020) prepared
a chitosan/PVA/batalain film, and an obvious blue shift of O-H
stretching vibration was observed by the addition of betalain extracted
from cactus pears. However, on the contrary, the O-H stretching band
was reported to shift to a higher wave number in the starch/PVA/red
pitaya peel betalain film (Qin et al., 2019). Similar results were obtained
in the chitosan/starch-based film incorporating purple or black eggplant
anthocyanins, which were manifested as different shift degrees of -OH
stretching (Yong et al., 2019). Secondly, the concentration of natural
colorants is also the important factor of molecular interaction strength.
For instance, In the FT-IR spectra of agar/PVA/curcumin film, the
typical peak of -OH stretching shifted from 3365 cm− 1 to 3358, 3347
and 3342 cm− 1 with the increasing amounts of curcumin (0–3.0%),
followed by the decreased intensity (Zhang, Li, et al., 2021). The
changes of peak position and intensity could be also observed in other
films with different colorant contents (Chen, Yan, Huang, Zhou, & Hu,
2021; Tran, Athanassiou, Basit, & Bayer, 2017; Yong et al., 2019),
indicating the amounts of natural colorant exerted significant effects on
molecular interaction. Furthermore, with regard to charged compo­
nents, the molecular interaction is also related to the pH value of the
film-forming solution, due to the close relationship between charged
nature and pH status (Yong & Liu, 2020).
4. Physical properties of natural colorant-based films
The physical properties can be determined by several factors such as
conformation of film components, film preparation methods, molecular
interactions, etc. The film with better physical properties will exhibit
more excellent functional properties, leading to its greater application in
food industry. Hence, investing the physical properties of natural
colorant-based film is of vital importance (Table 1).
4.1. Structural properties
4.1.1. Microstructural properties
The microstructural properties usually analyzed by scanning elec­
tron microscopy (SEM) and atom force microscopy (AFM), can reflect
arrangement rules, compatibility and miscibility of film components.
In general, the addition of anthocyanin (Wu, Sun, Zheng, Kang, &
Chen, 2019), curcumin (Roy & Rhim, 2020a), betalain (Etxabide, Maté,
& Kilmartin, 2021), chlorophyll (Chavoshizadeh et al., 2020), carot­
enoid (Hari et al., 2018; Łupina et al., 2022), shikonin (Dong et al.,
2020; Rhim & Roy, 2021) and alizarin (Ezati & Rhim, 2020; Ezati et al.,
2019) effectively resulted in the more uniform, homogeneous and
denser cross-section, indicating natural colorants can be distributed
evenly, which was accounted for the formation of hydrogen bonds and
the well compatibility. However, the existence of electrostatic repulsion
may exert negative influences on film microstructure. Singh, Nwabor,
Syukri, and Voravuthikunchai (2021) developed an anthocyanin-based
film incorporating chitosan and PVA, in which the generation of elec­
trostatic interaction resulted in significantly higher roughness value (Sa
J. Huang et al.
Table 1
The physical properties of natural colorant-based films.
Natural colorants
Clitoria ternatea or Carissa
carandas anthocyanin
Black rice bran anthocyanin
(BACN)
Black wolfberry anthocyanins
Roselle anthocyanin
Torch ginger anthocyanin
purple sweet potato (Ipomoea
batatas (L.) lam)
anthocyanin
Black plum peel anthocyanin
Curcumin
Curcumin
Curcumin
Curcumin
Amaranth (Amaranthus tricolor
L.) betalain
Red pitaya betalain
Polymers
Chitosan, polyvinyl alcohol (PVA)
Gelatin, oxidized chitin nanocrystals
Chitin whiskers, sodium alginate/
gelatin
PVA, hydroxypropyl methylcellulose
(HPMC)
Sago starch
Sodium carboxymethyl (CMC), starch
Chitosan, TiO2 nanoparticles
Poly(lactic acid) (PLA)
CMC, zinc oxide
κ-carrageenan
Rice starch
Chitosan, fish gelatin
Different polysaccharides (chitosan,
κ-carrageenan or locust bean gum),
PVA
Trends in Food Science & Technology 124 (2022) 259–277
Physical properties References
Mechanical properties:anthocyanin reduced tensile strength (TS) from Singh et al. (2021)
43.71 to 14.72 and 16.67 MPa; increased elongation at break (EB) from
92.25% to 227.41% and 167.17%
Hydrophobic properties: anthocyanin reduced water solubility (WS) from
7.22% to 6.89% and 6.99%, whereas increased moisture content (MC)
from 12.57% to 12.63% and 12.71%
Mechanical properties: anthocyanin decreased TS value from 9.44 MPa Ge et al. (2020)
(BACNs-Ch0) to 2.53 MPa (BACNs-Ch100); while increased EB value from
115.33% to 141.67% with increasing anthocyanin contents
Oxygen permeability (OP) properties: anthocyanin decreased OP from
2.035 to 1.323 cm3 m− 2⋅d− 1⋅atm− 1
Hydrophobic properties: anthocyanin increased MC significantly; the WS Zheng, Liu, et al. (2022)
decreased at first and then increased with increasing anthocyanin
contents; The water contact angle (WCA) increased when anthocyanin
was incorporated with hydrophilic polymers (Chitin whinskers/sodium
alginate, CS) while decreased when being incorporated with hydrophobic
polymers (Chitin whiskers/gelatin, CG)
Thermal properties: the introduction of anthocyanin shifted the
endothermc peak of CS (192 ◦ C) or CG (197 ◦ C) film to a lower frequency
(196–210 ◦ C; 199–207 ◦ C), indicating the decreased in thermal stability
Mechanical properties: with the increasing of anthocyanin contents, the Huang et al. (2021)
TS value significantly increased (45.66–56.98 MPa), and the EB value
decreased (17.50–13.75%)
Hydrophobic properties: The addition of roselle anthocyanin contributed
to lower MC (17.50%~13.75%), WCA (50.00◦ ~ 34.65◦ ) and higher WS
values (14.71%~26.88%)
Hydrophobic properties: Larger MC (27%) and WS (37.92%) values were Mei et al. (2020)
obtained with the increasing anthocyanins
Mechanical properties: the TS increased from 13.47 to 23.69 MPa and the Jiang et al. (2020)
EB decreased from 26.9% to 14.1% when the anthocyanin contents
ranged from 0.1 to 0.9 g/100g
Hydrophobic properties: The MC decreased significantly from 22.41% to
13.91% after the incorporation of anthocyanin
Thermal properties: when anthocyanin up to 0.5 g/100g, the Tm
increased significantly (127.11 ◦ C–128.81 ◦ C).
Mechanical properties: anthocyanin significantly enhanced mechanical Zhang et al. (2019)
properties by increasing TS from 14.93 to 19.72 MPa
Hydrophobic properties: anthocyanin reduced MC from 31.49% to
27.77% and increased WS from 18.14% to 29.23%
Thermal properties: anthocyanins decreased thermal stability of CS film
Mechanical properties: TS increased firstly (46.9–49.4 MPa) when the Roy and Rhim (2020b)
curcumin contents increased to 0.5%, then decreased to 40.8 MPa when
content reached 1.5%; Similarly, EB declined at first (7.8%–6.2%) and
increased to 7.9% (curcumin at 1.5%)
Hydrophobic properties: WCA ranged from 71.2◦ to 74.3◦ as Cur
increased
Thermal properties: curcumin increased the final residue from 4.2% to
5.6%
Mechanical properties: TS increased to 43.7 MPa (Cur 0.5%) and then Roy and Rhim (2020a)
decreased to 38.0 MPa (Cur 1.0%); EB increased from 16.7% to 20.6%
(Cur increased to 1.0%)
Hydrophobic properties: the incorporation of Cur increased WCA
significantly (from 43.2◦ to 49.1◦ )
Thermal properties: the introduction of curcumin decreased the onset/
endset temperature and final residue (34.4%~32%) slightly
Mechanical properties: TS significantly enhanced when increasing Cur Liu et al. (2018)
contents up to 3%, and then decreased when Cur contents exceeded 3%
Thermal stability: lower content of curcumin improved thermal stability,
while when the Cur content reached 7%, the thermal stability decreased
OP properties: lower content of curcumin decreased OP from around 1.25
to 0.2 cm3 m− 2⋅d− 1⋅atm− 1, while then increased to 1.75 cm3
m− 2⋅d− 1⋅atm− 1 finally
Mechanical properties: TS increased significantly to 0.032 MPa from Erna et al. (2022)
0.013 MPa, whereas the EB reduced from 0.0025% to 0.00082%
Hydrophobic properties: increasing Cur contents led to lower MC (3.58%)
and WS (71.30%) values compared with starch film (MC = 4.99%, WS =
79.2%)
Mechanical properties: TS increased from 10.4 to 12.5 MPa, and then Hu et al. (2020)
decreased to 9.3 MPa when the betalain content reached its maximum
value; EB increased gradually from 66.7% to 88.8%
Mechanical properties: Betalains increased the TS of chitosan/ Yao et al. (2022)
κ-carrageenan-based film, while decreased the TS of gum-based film; in
addition, betalin increased the EB value of κ-carrageenan/gum-based film
(continued on next page)
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J. Huang et al.
Table 1 (continued )
Natural colorants
Red pitaya betalain
Beet root betalain
Chlorophyll
Chlorophyll extracted from
black mulberry leaf
Chlorophyll extracted from
spinach leaves
Chlorophyll a
β-carotene
Astaxanthin
β-carotene
Commercial β-carotene,
β-carotene extracted from
shrimp or tomato
Carotenoid extracted from
tomato, carrot and annatto
seed
Astaxanthin
Shikonin extracted from
Lithospermum erythrorhizon
root
Shikonin extracted from
Lithospermum erythrorhizon
root
Alizarin
= 21.16 ± 0.44 nm) of films contained Clitoria ternatea anthocyanin
extracts compared with films without anthocyanin (Sa = 4.94 ± 0.86
nm). Meanwhile, Campos et al. (2019) reported an uneven cross-section
of starch/curcumin film, which might be because of the restriction
behavior of curcumin toward the citric acid as a kind of plasticizer,
limiting the dissolution process of starch during the film preparation.
Trends in Food Science & Technology 124 (2022) 259–277
Polymers Physical properties References
Thermal stability: melting temperature increased with the increasing CEO
contents
Starch, PVA Mechanical properties: Increasing betalain contents led to elevated TS Qin, Liu, Zhang, and Liu
(27.41–35.02 MPa) and EB values (35.15%~41.99%) (2020)
Hydrophobic properties: the introduction of betalain significantly reduced
the MC from 13.14% to 10.25%
Thermal stability: the Tg and Tm increased from 56.62, 167.48 to 57.9,
169.09 as the increasing CNC contents (0–7%)
Furcellaran Mechanical properties: The incorporation of betalains increased TS (from Jamróz et al. (2019)
25.05 to 33.00 MPa) and EB values (from 57.25% to 93.57%)
Hydrophobic properties: the combination of beet root betalain decreased
MC and WS from 18.06% to 100%–16.57% and 90.59%, respectively
Wheat gluten, polypyrrole Mechanical properties: chlorophyll increased TS and EB values Chavoshizadeh et al.
nanocomposite significantly (2020)
Hydrophobic properties: with increasing chlorophyll content, the WS
increased significantly
Black mulberry pulp pectin, CMC, silica Hydrophobic properties: chlorophyll decreased MC and WS values Sharifi and Pirsa (2021)
nanoparticles Thermal properties: chlorophyll had not effects on thermal stability of
pectin-based film
Poly(methylmethacrylate) (PMMA) Thermal properties: chlorophyll enhanced the onset temperature from Abdelrazek et al. (2010)
147 ◦ C to around 160 ◦ C, and the temperature at certain weight loss values
also increased with increasing chlorophyll
Chitosan, 2-HP-β-cyclodextrin Thermal properties: the exothermic peaks of film incorporated with Rizzi et al. (2016)
chlorophyll shifted to a higher temperature, indicating higher thermal
stability
Chitosan, gelatin, Starch Hydrophobic properties: decreased WS values were observed (from 79.1% Hari et al. (2018)
to 49.1%) after the incorporation of β-carotene
CMC/octenyl succinic anhydride Mechanical properties: astaxanthin decreased TS from 11.8 MPa to 8.75 Łupina et al. (2022)
(OSA), gelatin MPa, and increased EB values with the increasing contents of astaxanthin
Hydrophobic properties: the introduction of astaxanthin reduced WCA of
CMC/GEL film; while increased WCA of OSA/GEL film gradually
OP properties: the astaxanthin significantly reduced OP values from
174.12 to 34.06 cm3 μm/(m2⋅d)
Pectin, nanoclay, Carum copticum Mechanical properties: β-carotene reduced TS value at minimum EO Adsagh and Pirsa (2020)
essential oils content and increased TS value at maximum content; increasing
β-carotene also increased EB values
Shrimp muscle protein Hydrophobic properties: the β-carotene (commercial or extracted from Gómez-Estaca et al.
shrimp) reduced the WS values from 88.6% to 83.4% and 83.9%, while (2015)
the β-carotene (extracted from tomato) improved the WS value to 89.5%
PLA Mechanical properties: carotenoid significantly increased TS and EB value Stoll, Rech, Flôres,
from 22.4 MPa to 246% to 29.9 MPa and 374%, respectively Nachtigall, and Rios
Thermal properties: the carotenoid would not affect the thermal stability (2018)
of PLA film
Chitosan Mechanical properties: after the incorporation of astaxanthin, TS Roy et al. (2021)
increased from 22.25 to 28.09 MPa while the EB decreased from 35.35%
to 32.17%
Hydrophobic properties: the hydrophobic properties were improved after
the addition of a, the MC and WS decreased significantly from 16.24% to
53.75%–14.82% and 48.00%, respectively.
Thermal properties: the thermal stability of chitosan-based film was
improved by the addition of astaxanthin
Cellulose nanofiber (CNF) Mechanical properties: the introduction of shikonin increased TS and EB, Rhim and Roy (2021)
respectively
Hydrophobic properties: after the addition of shikonin, the MC slightly
decreased, while the WS and WVP increased
Thermal properties: the addition of shikonin improved the film thermal
stability
CMC or CNF Mechanical properties: after the introduction of shikonin, TS increased Ezati, Priyadarshi, et al.
from 78.9 to 79.4 MPa (2021)
Hydrophobic properties: the shikonin reduced the MC, WS and WVP
values significantly
Thermal properties: the addition of shikonin increased the final residue
contents of packaging film under high temperature
Chitosan Mechanical properties: the alizarin decreased TS from 30.6 to 29.1 MPa, Ezati and Rhim (2020)
while increased EB from 48.3% to 54.4%
Hydrophobic properties: the alizarin contributed to larger WCA
Thermal properties: the incorporation of alizarin modified the film
thermal stability
The microstructural characteristics also depend on the contents of
natural colorants. On the one hand, with relatively low concentration,
the colorant-based film kept uniform; on the other hand, extremely large
amounts of natural colorants contributed to agglomerates and hetero­
geneity, resulting in slight cracks and rough cross-section (Hu, Yao, Qin,
Yong, & Liu, 2020; Roy & Rhim, 2020b; Zheng, Li, et al., 2022).
266
J. Huang et al.
Nevertheless, there were also some researchers demonstrating totally
opposite phenomenons, in which the films combined with higher
amounts of anthocyanin were smoother and more immaculate when
compared with lower concentrations. While they considered that the
large numbers of -OH in the colorants could bind with polymers tightly,
generating more hydrogen bonds to overcome the defects of electro­
static repulsion. Consequently, the compact and rigid micro-structures
were obtained (Chen et al., 2021; Ge et al., 2020).
4.1.2. Crystalline
The crystalline which can be determined by X-ray diffractometer
(XRD) is an important structural parameter of film characterization. The
intensity of the peaks in XRD spectra can represent the crystalline level,
the sharper and wider peaks are responsible for the higher crystalline
and amorphous region of film material, respectively.
Generally, the crystalline degree mainly depends on the polymers
rather than natural colorants, which is owing to low contents of color­
ants, imposing relatively limited effects on crystalline (Sharifi & Pirsa,
2021; Yao et al., 2022). However, several studies have revealed that the
addition of natural pigments altered the diffraction peak intensity of
food packaging, which was related to the crystalline or amorphous
conformation of natural colorants (Chavoshizadeh et al., 2020; Liu et al.,
2018; Qin et al., 2021; Yao et al., 2022; Zhang, Li, et al., 2021). As kinds
of crystalline compounds, the curcumin and carotenoid can lead to
accumulation of crystallization. For instance, the
chitosan-gelatin-carotenoid film exhibited an increase in the intensity of
peak at 2θ = 17.79◦ /28.71◦ , suggesting the enhancement of crystalline
structure after the incorporation of carotenoid (Hari et al., 2018; Liu
et al., 2018). Similar results were found in curcumin-based film (Liu
et al., 2018; Zhang, Li, et al., 2021), which was attributed to the crys­
talline of colorants and the interweaving between colorants and poly­
mers, resulting in the formation of more ordered structure. On the other
hand, the amorphous compounds, anthocyanin, betalain and chloro­
phyll which present a broad peak around 20◦ (2θ), usually decrease
crystalline degrees of films (Chavoshizadeh et al., 2020; Huang, Liu,
Chen, Yao, & Hu, 2021; Qin et al., 2021; Qin, Liu, et al., 2020; Yao et al.,
2022). For example, owing to the disruption of polymer rearrangement
via the generation of strong interactions, the peak at 20◦ whose intensity
decreased significantly became wider after the addition of chlorophyll
into wheat gluten film (Chavoshizadeh et al., 2020). Nevertheless, the
anthocyanins extracted from eggplant (Wang et al., 2019) and purple
corn (Qin, Liu, Yuan, et al., 2019) could improve the diffraction peak
intensity of films contrarily, signifying that the crystalline degree might
be also associated with the sources of natural pigments.
4.2. Mechanical properties
The mechanical properties which are mainly characterized by tensile
strength (TS) and elongation at break (EB), are the symbols of film
ability to maintain its good integrity. The food packaging with better
mechanical properties are able to protect food products from physical
stress and environmental conditions.
The TS stands for the maximum capacity to withstand external stress.
The microstructural network and molecular interaction can have sig­
nificant influences on film mechanical properties (Adsagh & Pirsa, 2020;
Jamróz, Kulawik, Guzik, & Duda, 2019). Many researchers reported that
the molecular interaction enhanced TS values after incorporation with
natural colorants, the elevated TS levels were attributed to formation of
hydrogen bonds and electrostatic repulsion, contributing to the greater
interfacial adhesion and more compact and stable film network (Roy, &
Rhim, 2022a; Ezati, Priyadarshi, Bang, & Rhim, 2021; Rhim & Roy,
2021; Adsagh & Pirsa, 2020; Qin, Liu, et al., 2020; Jamróz et al., 2019).
However, in some cases, the addition of natural colorants might lead to
interruption of molecular interactions among film components and
disruption of uniform spatial structure. As a result, the TS was signifi­
cantly declined (Ezati et al., 2019; Ge et al., 2020; Singh et al., 2021;
267
Trends in Food Science & Technology 124 (2022) 259–277
Łupina et al., 2022). Notably, the mechanical properties can be also
affected by concentrations of natural colorants. Hu et al. (2020) fabri­
cated an Amaranth betalain film based on the chitosan and fish gelatin,
and they found the lower contents of betalain (5% and 10%) contributed
to higher TS values, while weak TS was obtained when betalain reached
its maximum values. Similar results were also reported by Roy and Rhim
(2020b), Liu et al. (2018), etc, in which the film TS could be enhanced
due to the well dispersion and effective external load transformation of
colorants as well as their suitable interactions with bio-polymers when
an optimum content of natural pigment was added. Nevertheless,
excessive amounts of colorants would reduce the TS value, which was
accounted for the aggregation of those colorants, resulting in the for­
mation of intermittent areas and discontinuous microstructures.
The EB value can represent film flexibility degree. On the one hand,
the introduction of alizarin increased flexibility of chitosan/alizarin film
by the reduction of intramolecular forces and the increase of polymer
structure mobility (Ezati et al., 2019). Similarly, when the astaxanthin
was incorporated with the CMC-gelatin film, the flexibility was signifi­
cantly improved due to the increasing of free volume among polymers
and astaxanthin. Consequently, the chain-chain movement increased
(Łupina et al., 2022). Hence, the natural colorants would behave as
plasticizers to enhance the EB by generation of molecular interaction,
contributing to the facilitation of chain motions (Adsagh & Pirsa, 2020;
Ge et al., 2020; Hu et al., 2020; Jamróz et al., 2019; Qin, Liu, et al., 2020;
Yao et al., 2022). On the other hand, the rigid properties or extremely
high amounts of natural colorants could cause the reduction of film
flexibility by restriction of chain-chain interactions and movements
(Huang et al., 2021).
4.3. Hydrophobic properties
The hydrophobic properties of packaging film are important for its
application, due to their strong correlation with film stability, the
release of natural colorants, functional performances, etc. Many in­
dicators are applied to evaluate film hydrophobic characteristics, such
as moisture content (MC), water solubility (WS) and water contact angle
(WCA).
The MC represent the total volume of water molecules contained in
the film matrix, while different kinds of natural colorants will exert
different influences on MC. The incorporation of anthocyanin (Huang
et al., 2021), curcumin (Erna, Felicia, Rovina, Vonnie, & Huda, 2022),
betalain (Qin, Liu, et al., 2020), chlorophyll (Sharifi & Pirsa, 2021) or
shikonin (Ezati, Priyadarshi, et al., 2021) contributed to tighter struc­
tural network formed by the hydrogen bonds and electrostatic in­
teractions, limiting film water absorption and binding capacity. As a
result, the moisture transfer was retarded, MC was reduced and the film
hydrophobic property was improved. However, some studies reported
that the MC value would be enhanced after the addition of natural
colorants (Mei et al., 2020; Singh et al., 2021; Zheng, Li, et al., 2022),
which might be owing to the abundant of hydroxyl groups contained in
the molecular chains, interacting with functional groups of bio-polymers
to bind with moisture.
The film with lower WS value is believed to maintain integrity better.
The WS is highly dependent on water solubility of natural colorants. The
entire dissolution of colorants led to an increase in the hydrophilic sites
for water absorption, hence, the WS was enhanced significantly (Cha­
voshizadeh et al., 2020; Huang et al., 2021; Mei et al., 2020). On the
contrary, when being incorporated with curcumin, which was insoluble
in polar water molecules, the WS of starch-based film decreased from
79.2% to 71.30% due to the strong hydrophobic properties of curcumin
(Erna et al., 2022). Similar results were obtained in shikonin (Rhim &
Roy, 2021) and alizarin-based (Ezati et al., 2019) films, owing to their
low solubility in water. Secondly, the molecular interaction reduces WS
in general. For examples, the formation of hydrogen bonds was able to
limit film matrix-water interactions, contributing to the decrease in film
solubility (Hari et al., 2018; Jamróz et al., 2019). Meanwhile, the
J. Huang et al.
generation of electrostatic repulsion could prevent water molecules
from entering film matrix, and the lower WS values as well as stronger
hydrophobic properties would be obtained (Sharifi & Pirsa, 2021).
Thirdly, the WS is also related to natural colorants concentrations.
Zheng, Liu, Yu, and Shao (2022) developed a black wolfberry antho­
cyanin film based on chitin whiskers and sodium alginate, the WS
decreased initially resulted from the enhancement of film dimensional
stability after the incorporation of anthocyanin, whereas the WS
increased when large amounts of anthocyanin added, which might be
owing to the introduction of excessive -OH, contributing to the strong
ability to bind water. Furthermore, the addition of carotenoid extracted
from shrimp exhibited a reduction effect on WS, while the carotenoid
extracted from tomato would increase WS, indicating that the WS was
also determined by different sources of natural colorants (Gómez-Estaca,
Calvo, Sánchez-Faure, Montero, & Gómez-Guillén, 2015).
The WCA measures film surface wettability directly, and the larger
WCA represents stronger hydrophobic properties. The WCA is signifi­
cantly affected by the hydrophobic or hydrophilic nature of film com­
ponents. For instance, the hydrophilic colorants (such as roselle
anthocyanin) were documented to decrease the WCA from 50.00◦ to
34.65◦ , indicating the enhancement of film hydrophilicity (Huang et al.,
2021). By contrast, the hydrophobic colorants (like curcumin) caused
the increase in WCA (Roy & Rhim, 2020a, 2020b). Notably, the
hydrophobic/hydrophilic natures of polymers might also impose
different influences on WCA. The incorporation of astaxanthin with
CMC/gelatin (hydrophilic) film contributed to a decreasing WCA, due to
the formation of porous structure, allowing large amounts of water to be
immobilized in the film matrix. However, when being incorporated with
octenyl succinic anhydride (OSA)/gelatin (hydrophobic) film, the WCA
increased significantly owing to interruption effects of astaxanthin to
retard the water movement access, contributing to higher hydrophobic
properties (Łupina et al., 2022).
4.4. Thermal properties
The packaging films with better thermal stability are in high de­
mand. Generally, the differential scanning calorimetry (DSC) and ther­
mogravimetric analysis (TGA) are applied to evaluate film thermal
properties.
Generally, it can not be denied that the natural colorants exhibit the
capacity to alter thermal stability of packaging film due to their complex
molecular structure, etc. The formation of molecular interaction after
the incorporation of natural colorants led to more stable thermal prop­
erties, as expressed as higher melting temperature (Tm), larger enthalpy
energy (ΔHm) value or more residue amounts at extremely high tem­
peratures, resulted from the induction effects of colorants in molecular
chain reorganization, contributing to the limited chain mobility.
Consequently, the cohesive energy increased, and the film thermal sta­
bility was enhanced (Ezati, Bang, & Rhim, 2021; Ezati et al., 2020; Jiang
et al., 2020; Rizzi et al., 2016; Roy et al., 2021; Roy & Rhim, 2020b).
Conversely, the addition of natural colorants was also reported to result
in the reduction of thermal stability, which was attributed to the inter­
ruption influences exerted by those colorants, leading to the weaken of
molecular interactions (Roy & Rhim, 2020a; Zhang et al., 2019; Zheng,
Li, et al., 2022).
In addition to molecular interactions, thermal properties could be
also influenced by concentrations of natural colorants. Liu et al. (2018)
developed a curcumin/κ-carrageenan active film, and the typical peaks
in DSC pattern shifted from 93 ◦ C to 102 ◦ C with increasing contents of
curcumin (0%–5%), owing to the formation of hydrogen bonds; How­
ever, when the curcumin concentrations were greater than 7%, the
thermal stability decreased obviously, which might be resulted from the
serious agglomeration of curcumin, leading to the disruption of ordered
structural network. Similar results were also obtained in other natural
colorant-based film (Liu et al., 2019), indicating the important roles the
colorant contents played in the thermal properties.
268
Trends in Food Science & Technology 124 (2022) 259–277
4.5. Oxygen permeability properties
The oxygen permeability (OP) properties are the essential physical
characteristics for food packaging material, since the migration and
diffusion of oxygen is crucial for the shelf-life of food products. It is
believed that the food packaged by film with weaker oxygen perme­
ability properties will experience less oxidation reactions.
Generally, the natural colorants have the ability to enhance film
oxygen barrier properties (Stoll, Rech, Flôres, Nachtigall, & Rios, 2019;
Xiao, Liu, Kang, Cui, & Xu, 2021). For example, Łupina et al. (2022)
found the combination of astaxanthin into CMC-gelatin film resulted in a
5-folds reduction in OP, which decreased from 174.12 to 34.06 cm3
μm/m2⋅d. The promising oxygen barrier characteristics of natural
colorant films could be explained by intermolecular crosslinking in the
film matrix. The hydrogen bonds or the unique molecular geometry of
colorant skeleton could increase the tortuous degrees of O2 diffusion
way, contributing to the hindrance of oxygen free movement (Ge et al.,
2020; Xiao et al., 2021). However, when the contents beyond ideal
range, the large amounts of agglomerated natural colorants would be
distributed unevenly in the film matrix, resulting in a significant in­
crease in OP (Ge et al., 2020; Liu et al., 2018).
5. Functional properties of natural colorant-based films
As Fig. 4 shown, the functional properties characterized as indicator,
active and barrier characteristics, are closely related to the application
of food packaging. They can be affected by properties of both natural
colorants and polymers, storage condition, film development methods,
etc.
5.1. pH-responsive properties
The pH-responsive properties are the fundamental functions of nat­
ural colorants-based packaging to monitor food freshness in real time.
During the spoilage process, the food products will release certain gases,
such as H2S, CO2, ethylene or the nitrogen compounds. Those gases
whose contents increase with the extension of storage time, will alter the
pH environment. Hence, the packaging with the characteristics of pH-
response have capacities to convey freshness information to con­
sumers directly and efficiently.
Among all the kinds of pigments, the anthocyanins exhibit the most
complex discoloration behaviors. The indicator films based on antho­
cyanins extracted from black rice, butterfly pea, purple sweet potato,
roselle, etc, present similar color variations, whose color are purple or
dark red initially, then change to pink (pH around 4.0–6.0) and colorless
(pH = 7.0). Furthermore, the anthocyanin-based film colors turn from
green/blue to yellow when the pH values up to 11.0 (Jiang et al., 2020;
Wu et al., 2019). Compared with anthocyanins, the curcumins, beta­
lains, carotenoids, shikonin and alizarin are more stable in the pH values
ranging within acid condition, which means that the film incorporating
colorants will present colors of bright yellow, fuchsia, bold orange/­
yellow, bright red and yellow, respectively. While when being immersed
into alkaline environment (pH > 7), the film colors turn to reddish
brown, yellow, light yellow, blue and reddish purple, respectively
(Alizadeh-Sani et al., 2020; Ezati, Priyadarshi, et al., 2021; Ezati &
Rhim, 2020; Liu et al., 2018; Liu et al., 2022; Wu et al., 2021). In
contrast, chlorophyll behaves totally opposite discoloration perfor­
mances, which was found more unstable when pH values are between
2.0 and 7.0, facilitating color alteration to olive green from green (Qu
et al., 2020).
The conformation conversion and the degradation of natural color­
ants can be accounted for the pH-responsive properties of natural
colorants-based films. For instance, the original red color of
anthocyanin-based film is attributed to structure of flavylivm cations
(under acid environment), which will turn into colorless carbinol psendo
based under the effects of deprotonation and hydrotion (pH at 7.0) and
J. Huang et al. Trends in Food Science & Technology 124 (2022) 259–277

Fig. 4. The functional properties of natural colorant-based films, including pH-responsive, active and barrier properties.

269
J. Huang et al.
purple/blue/green quinoidal base (pH between 7.0 and 11.0). Further­
more, the anthocyanins are degraded partially at extremely alkaline
conditions, leading to the formation of yellow chalcone (pH > 11.0.
Similarly, the structure transformation from betalains (purple red) and
chlorophyll (green) to betalamic acid (yellow), cyclo-Dopa glucoside
(colorless) and pheophytin (olive green), also results in the pH-
responsive behaviors of colorant films (Jamróz et al., 2019; Yao et al.,
2022; Zheng, Liu, et al., 2022). The conformation changes of betalains
are due to the hydrolysis degradation under alkaline conditions, while
the reason for alterations of chlorophyll structure is the replacement of
central Mg2+ by H+ (Zheng, Liu, et al., 2022). Similarly, the chromo­
phore conformation variation of shikonin under different acid-base
conditions results in its color changes (Dong et al., 2020). Meanwhile,
because of the conjugated and unsaturated double bonds contained in
the molecular structure, the discoloration principle of carotenoid-based
film is related to its oxidation, contributing to the color degradation
(Gómez-Estaca et al., 2015). In addition, as a weak brosted acid con­
taining three unstable protons, the protonation process plays an
important role in the discoloration of curcumin-based films, which in­
duces the conformation alteration from keto form (acid conditions) to
enol form (alkaline conditions), owing to the generation of phenoxide
anion (Erna et al., 2022; Xiao et al., 2021). The protons transfer is also
responsible for the color variation of alizarin-based films. Specifically,
the hydroxyl groups in the alizarin can be combined with carbonyl ox­
ygen atoms, leading to the formation of intramolecular hydrogen bonds
and the film discoloration (Ezati & Rhim, 2020).
The pH-responsive properties are affected by several factors. For
example, the colorant films stored at higher temperatures presented
worse pH-responsive ability, which might be due to the instability of
natural colorants (Zhai et al., 2017). In addition, the polymers incor­
porated can also affect the pH-responsive properties. Huang, Chen,
Zhou, Li, and Hu (2020) developed a series of roselle anthocyanin-based
film based on different ratio of PVA and hydroxypropyl methylcellulose
(HPMC), and the film with PVA:HPMC ratio of 1:3 exhibited the best
pH-responsive properties, owing to the better physical characteristics
obtained via the formation of tighter network, immobilizing anthocy­
anin well and maintaining its original discoloration properties better.
Similar results were also obtained by Yao et al. (2022), in which the
betalain-based films presented different pH-responsive capacities with
regards to various polymers combined with.
5.2. Active properties
5.2.1. Antioxidant properties
Food products are easy to oxidize, leading to the quality deteriora­
tion. Hence, natural colorant films with strong oxidative resistance ca­
pacities can be applied as the feasible ways to protect food from
oxidation.
Owing to the excellent antioxidant properties of natural colorants,
the film antioxidant characteristics are improved significantly after the
addition of anthocyanin (Singh et al., 2021; Wu et al., 2019; You, Wang,
Zhou, Yang, & Pang, 2022; Zhang et al., 2019), curcumin (Roy & Rhim,
2020a, 2020b; Zhang, Li, et al., 2021), betalains (Qin, Liu, et al., 2020;
Yao et al., 2022), chlorophyll (Chavoshizadeh et al., 2020), carotenoid
(Roy et al., 2021; Łupina et al., 2022), shikonin (Ezati, Bang, & Rhim,
2021; Rhim & Roy, 2021) and alizarin (Ezati et al., 2020). You et al.
(2022) fabricated a konjac glucomannan/CMC film incorporating black
currant anthocyanin, which presented significantly higher 2,2-Diphe­
nyl-1-picrylhydrazyl (DPPH) scavenging activity of 49.23% compared
with control film (32.65%), while the film antioxidant properties could
be attributed to the phenolic hydroxyl groups contained in anthocyanin,
behaving as an effective reduction agent. Meanwhile, the combination
of curcumin enhanced the DPPH and 2,2’ - Azinobis - (3- ethyl­
benzthiazoline - 6-sulphonate) (ABTS) radical scavenging activities of
poly(lactic acid) (PLA) films, from 1.8% to 3.1%–76.6% and 94.7%,
respectively, due to the H-donation from hydroxyl groups in the
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curcumin structure (Roy & Rhim, 2020b). Similarly, the -OH groups
were also responsible for the modification of antioxidant properties in
betalain, carotenoid, shikonin and alizarin based films, which reacted as
the singlet oxygen quencher, resulting in the massive increase of DPPH
scavenging activity (Ezati & Rhim, 2020; Gómez-Estaca et al., 2015;
Rhim & Roy, 2021; Wu et al., 2021). Notably, although the incorpora­
tion of chlorophyll could increase the antioxidant ability of gluten film,
the enhancement was not attribute to the capacity to donate H atom. The
mechanism of chlorophyll antioxidant performances was believed to
relate to the prevention of hydroperoxide decomposition (Chavoshiza­
deh et al., 2020).
The film antioxidant properties depend on sources of natural color­
ants. The chitosan-PVA films incorporated with anthocyanin from Clit­
oria ternatea and Carissa carandas exhibited relatively different DPPH
scavenging capacities (Singh et al., 2021). In addition, many studies
demonstrated the noticeable concentration-dependent principle of film
antioxidant properties, suggesting that the antioxidant performances
would be enhanced with the increasing contents of natural colorants
(Hari et al., 2018; Liu et al., 2022; Wang, Zhang, & Zhang, 2022; Wen
et al., 2021; Wu et al., 2021).
Furthermore, the film antioxidant characteristics are also deter­
mined by release of colorants, which are closely associated with the
nature of polymers, the molecular interactions, etc. The red pitaya
betacyanin/PVA/chitosan films presented lower DPPH scavenging
ability compared with betacyanin films incorporating PVA/κ-carra­
geenan or PVA/gum, owing to the generation of hydrogen bonds and
electrostatic repulsion within the chitosan, κ-carrageenan and locust
bean bum, leading to the restriction of the betalain release (Yao et al.,
2022). On the other hand, Qin et al. (2021) found that the addition of
nano loaded anthocyanin reduced the film antioxidant capacities by
nearly 15%, which was resulted from the slowly release efficiency of the
encapsulated anthocyanin. However, by contrast, the antioxidant
properties of cellulose nanocrystals film were improved by the addition
of nano-encapsulation curcumin when compared with free curcumin,
due to the protection effects of encapsulation, facilitating the mainte­
nance of the curcumin properties and structural integrity (Xiao et al.,
2021).
5.2.2. Antibacterial properties
The film antibacterial properties are important to food quality
maintenance since the spoilage resulted from foodborne pathogens
contributes to several food security problems. Because of the great ca­
pacity of natural colorants in microorganism activity inhibition, the
packaging films incorporating natural pigments present excellent anti­
microbial activities (Hu et al., 2020; Qin, Liu, et al., 2020; Rhim & Roy,
2021; Roy & Rhim, 2020a; Sharifi & Pirsa, 2021; Soltaninejad & Maleki,
2021; Stoll et al., 2019; Wen et al., 2021; Zhang et al., 2019, 2021). For
instance, the PLA/curcumin film exhibited distinctively slower growth
of Escherichia coli (E.coli) and L. monocytogenes, whose viable colony
forming counts decreased by about 1–2 log cycle compared with pure
PLA film (Roy & Rhim, 2020b). The 2,2,6,6,-tetramethylpiperidi­
ne-1-oxyl radical (TEMPO)-oxidized bacterial cellulose film containing
purple sweet potato anthocyanin showed larger inhibition diameters
with the values of 14.5 and 18.4 mm for E. coli and S. aureus compared to
film without anthocyanin (14.2 and 17.4 mm) (Wen et al., 2021).
Similarly, The larger areas of inhibition zones for both S. aureus and
E. coli were observed in PVA/TiO2/chitosan film after the addition of
chlorophyll (Soltaninejad & Maleki, 2021).
Generally, the antimicrobial properties of natural colorants-based
films can be attributed to their disruption behaviors toward bacteria
cell membranes. On the one hand, natural colorants decrease the sta­
bility of cytoplasmic membrane and increase the permeability of cell
membrane. On the other hand, they also perform important roles in cell
metabolism, contributing to the death of bacteria (Qin, Liu, et al., 2020;
Soltaninejad & Maleki, 2021; Stoll et al., 2019; Zhang et al., 2019).
Meanwhile, in addition to perform interaction behaviors with cell
J. Huang et al.
membrane, the curcumin can also inhibit the function of filamentous
temperature-sensitive protein (FtsZ), the essential protein for cell divi­
sion, contributing to its outstanding antibacterial capacity (Liu et al.,
2022).
In most studies, the natural colorant-based film exhibit stronger
antimicrobial capacity against Gram-positive than Gram-negative bac­
teria, owing to the differences in cell wall structure, cell metabolism and
cell physiology. Specifically, the Gram-negative bacteria consist of a
complex cell wall structure surrounded by a barrier outer membrane,
which do not contain in the Gram-positive bacteria. As a result, it is more
difficult for colorants to destroy the cell conformation integrity (Hu
et al., 2020; Roy & Rhim, 2020a; Stoll et al., 2019; Zhang et al., 2019).
Apart from the strain-dependence property, the film antibacterial
properties also depend on the contents of colorants, which will increase
gradually with the increasing colorant amounts (Liu et al., 2022; You
et al., 2022). In addition, similar with antioxidant properties, the anti­
microbial capacities are affected by bio-polymer matrix, too, due to their
influences on molecular interaction within the film system, resulting in
the alteration of colorant release (Roy & Rhim, 2020c).
5.3. Barrier properties
The natural colorant-based films can be regarded as potent barrier,
which protect food products from light, water and oxygen, resulting in
the quality maintenance.
The light barrier properties determined by light transmittance or
opacity, are necessary for food packaging because the ultraviolet (UV)-
light will facilitate a series of photochemical reactions, contributing to
the food deterioration. As colorful pigments, the introduction of natural
colorants significantly improves light barrier properties of bio-polymer
films, as manifested by the lower light transmittance or higher opacity
(Chavoshizadeh et al., 2020; Ezati, Priyadarshi, et al., 2021; Liu et al.,
2022; Qin et al., 2021; Stoll et al., 2019; Wu et al., 2021; You et al., 2022;
Zhang, Li, et al., 2021). On the one hand, the natural colorants contain
certain chromophores including C=C, C=O, C=N, aromatic ring, etc,
exhibiting capacities for light absorption (Hu et al., 2020; Wen et al.,
2021; Xiao et al., 2021; Yao et al., 2022). As mentioned above, the
colorants can act as compatibilizer to alter crystalline degree and reduce
phase separation within film matrix, leading to the light scattering and
reflection (Chavoshizadeh et al., 2020; Łupina et al., 2022).
Moisture transformation is responsible for the food deterioration too.
Hence, the water vapor barrier is the essential function for food pack­
aging. Water vapor permeability (WVP) is usually applied to evaluate
the water barrier properties, which reflects the water diffusion behav­
iors. The water barrier properties were improved by the addition of
natural colorants such as black plum peel anthocyanin (Zhang et al.,
2019), sweet purple potato anthocyanin (Zhang, Li, et al., 2021), cur­
cumin (Roy & Rhim, 2020a), red pitaya betalain (Qin, Liu, et al., 2020),
chlorophyll (Chavoshizadeh et al., 2020), alizarin (Ezati & Rhim, 2020),
etc, which could be attributed to the prolongation of tortuous pathway
for water after the colorant incorporation. Meanwhile, the colorants
could also contribute to the generation of new molecular interactions,
leading to the densified structural network. As a result, the film water
affinity was reduced correspondingly (Qin et al., 2021; Zhang et al.,
2019). However, on the contrary, the natural colorants would decline
water barrier properties in some studies. You et al. (2022) prepared a
black rice anthocyanin-based film, and the WVP increased from 0.67 to
2.53 g mm/m2⋅d⋅kPa with the increasing anthocyanin contents, which
was accounted for the hydrophilicity of anthocyanin as well as its
disruption effects on alteration of initial molecular interaction, leading
to the disordered of compact film structure. Similar results were also
observed by Wang et al. (2022) and Roy and Rhim (2020b).
The film barrier properties are highly correlated with concentration
of natural colorants. Generally, the light barrier capacity will be
improved dramatically as the increasing amounts of colorants (Wu et al.,
2021; Xiao et al., 2021). However, different contents of natural pigments
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Trends in Food Science & Technology 124 (2022) 259–277
can exert different influences on water vapor and oxygen barrier prop­
erties. For instance, low contents of cockscomb flower betalains (4%–
8%) contributed to the more uniform and compact film network,
resulting in the decrease of WVP from 11.65 to 10.34 × 10− 11 g
m− 1⋅s− 1⋅Pa− 1. Whereas, when being incorporated with excessive beta­
lains (12%), the WVP increased to 12.43 × 10− 11 g m− 1⋅s− 1⋅Pa− 1
significantly, owing to the formation of agglomeration of betalains and
reduction of film homogeneity (Wu et al., 2021). Similarly, Liu et al.
(2018) observed that the lower WVP values were obtained in the
κ-carrageenan film containing lower contents of curcumin, while the
water vapor barrier properties would be destroyed after the addition of
relatively large amounts of curcumin, suggesting the important role the
natural colorant contents played in the film barrier functions. In addi­
tion, the film barrier properties also depend on nature of polymers
because of the differences in molecular interaction. Yao et al. (2022)
compared the WVP of PVA/red pitaya betalain film based on three types
of bio-polymers, and the results demonstrated that the chitosan and
locust bean gum-based film exhibited greater water vapor barrier ca­
pacities, due to the stronger molecular interaction caused the estab­
lishment of tighter inner structural network, so the moisture
transmission was limited.
6. Application of natural colorant-based films in food industry
6.1. Indication packaging
During spoilage process, food products will release certain sub­
stances, contributing to the alteration of packaged environment. The
natural colorant-based film can act as smart indicator, reflecting the
quality deterioration by sensing the changes of environment. Notably,
the indication behaviors of natural colorant films are non-destructive
and efficient, which are in accordance with the urgent demands of
consumers for food quality and safety. The recent applications of natural
colorant-based film in freshness monitoring are listed in Table 2.
The natural colorant-based films have been widely applied in the
freshness monitoring of meat products including poultry and aquatic
products (Dong et al., 2020; Ezati et al., 2019; Gasti et al., 2021; Guo
et al., 2021; Wu et al., 2021; Yildiz, Sumnu, & Kahyaoglu, 2021; You,
Wang, Zhou, Nang, & Pang, 2022). The abundant protein contained in
the meat products will be degraded to nitrogen compounds with the
extension of storage time, leading to the significant rise of pH value. As a
result, these films can monitor the variation of total volatile basic ni­
trogen (TVB-N) contents to reflect the freshness degree. Guo, Ge, et al.
(2021) developed a watermelon peel beetroot betalain indicator film
based on pectin for beef freshness monitoring. The TVB-N values
increased from 6.73 to 18.71 mg/100g at 8 d. A significant decrease in
film a* value was observed from 45.27 to 29.26, and the film color
shifted from pink to brownness correspondingly. The measurement of
film color differences (ΔE) can evaluate the color variations, and when
ΔE>5, the color change can be identified by naked eyes. A high value of
ΔE (36.5) was observed when the curcumin-based film was applied to
shrimp freshness monitoring, accompanied by the obvious color changes
from yellow to orange-red, indicating its feasibility in freshness detec­
tion (Zhang, Li, et al., 2021).
The natural colorant film can monitor the freshness of dairy product
(Weston, Phan, Arcot, & Chandrawati, 2020; Zheng, Li, et al., 2022). The
deterioration of milk is highly associated with the accumulation of lactic
acid, which is produced under the effects of microorganisms, resulting in
a sharp decline in pH. Weston et al. (2020) reported a clear color
response from purple-blue to light red of red cabbage anthocyanin/a­
garose film when the pH of milk decreased from 6.8 to 4.0. In addition to
milk, the natural pigment film can also be applied to the freshness
monitoring of other dairy products like cheese. Adsagh and Pirsa (2020)
prepared a β-carotene smart film incorporating pectin and nanoclay, and
results demonstrated that as increasing oxidation degree of butter, the
film color turned from orange to light yellow, owing to the reduction of
J. Huang et al. Trends in Food Science & Technology 124 (2022) 259–277

Table 2
The recent advances in the application of natural colorant-based films.
Natural colorants based Indication or Applica-tion Application test Application effects References
packaging active packaging object

Phyllanthus reticulatus Freshness Fish fillet The film color changed from dark gray to light Gasti et al. (2021)
anthocyanin/chitos-an/ indication yellow when the fish was totally spoiled
methylcellulos-e film

Blueberry anthocyanin/ Freshness Litchis The film changed from purple to pink; Zhou et al. (2021)
cellulose nanofiber/ indication and Prolonged shelf-life of litchis by 6 days
chitosan film active packaging

Black wolfberry Freshness Milk The color of the indicator film changed from blue- Zheng, Liu, et al.
anthocyanin/chitin indication purple to light red, and ΔE reached 5.39–8.25 (2022)
whiskers/sodium within 12 h
alginate film

Purple cabbage Freshness Mutton Film color changed from mauve to baby blue. Guo et al. (2021)
anthocyanin/ indication Specifically, with an extended storage time, the L*
watermelon peel pectin and b* increased from 34.75 to 5.86 to 47.95 and
film 10.80. While the a* and △E values decreased from
12.65 to 60.82 to 7.61 and 47.52, respectively.

Blackcurrant anthocyanin/ Freshness Crucian carp The film color turned from pink-purple to yellow You, Wang, Zhou,
konjac glucomannan/ indication Nang, & Pang
carboxymethyl cellulose (2022)
film

Red cabbage anthocyanin/ Freshness Milk The film underwent a visible blue-purple-pink Weston et al.
agarose film indication color change to discriminate between fresh, (2020)
spoiling, and spoiled milk

Anthocyanin/methyl Freshness Ricotta cheese – The film changed color from colorless to deep Krithenkov et al.
cellulose/polyethylene indication purple at the given temperature (8 ◦ C) for the given (2021)
glycol/chitosan film period of time (70 min).
Brassica oleracea L. Leak indication CO2 The film changed from green to purple with the Saliu and Pergola
anthocyanin/L-Lysine/ increasing CO2 contents (2018)
ε-poly-L-lysine film
Red cabbage anthocyanin/ Freshness Rutab The film color turned from violet to purple Maftoonazad and
PVA film indication Ramaswamy
(2019)

Radish anthocyanin/agar/ Active Banana – Improved the appearance of banana; inhibited the Yang et al. (2022)
TiO2/essential oil film packaging mould and yeast activity; reduced weight loss and
maintained the firmness
Lycium ruthenicum Active Bass fillets – Extended shelf-life to 7 days as manifested by lower Qin et al. (2021)
anthocyanins/starch/ packaging total viable counts (TVC), total volatile basic
PVA film nitrogen (TVB-N), thiobarbituric acid (TBARS) and
weight loss
Black berry anthocyanin/ Active Cherry tomato – Improved physicochemical characteristics of Sganzerla et al.
CMC film packaging cherry tomato, such as increasing the firmness, pH (2021)
while decreasing the weight loss; Higher sensorial
properties were obtained
Black rice bran Freshness Shrimp and The film color shifted from purple to gray blue or Ge et al. (2020)
anthocyanin/gelatin/ indication and hairtail brown;
oxidized chitin active packaging Inhibited the generation of nitrogen compounds
nanocrystal film and reduced the pH values
Prunus maackii Freshness Pork and lard The film color changed from pink to dark red (10 h) Sun et al. (2020)
anthocyanin/ indication and and purple (20 h) when being applied in pork
κ-carrageenan/HPMC active packaging freshness monitoring;
film Reduced the POV and acid price of lard
significantly

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Table 2 (continued )
Natural colorants based Indication or Applica-tion Application test Application effects References
packaging active packaging object

Purple sweet potato Freshness Pork The initial pink color turned into light brown at the He et al. (2022)
anthocyanin/agarose/ indication and start point of spoilage, and to dark green finally;
PVA film active packaging Inhibited the increasing in pH, TVB-N and TVC
values

Cranberry anthocyanin/ Active Olive oil – Inhibited the increasing of peroxide values Wang et al. (2019)
chitosan/gelatin/CMC packaging obviously
film
Curcumin/PVA film Freshness Shrimp The film gradually changed from yellow to orange- Zhang, Li, et al.
indication (Macrobr- red with the decline of shrimp freshness, with the (2021)
achium increasing ΔE values to 36.5 at 60 h
rosenberg-ii)
Curcumin/chitosan/ Freshness Chicken breast Yellowness of the films decreased while the redness Yildiz et al. (2021)
polyethylene oxidie indication of the indicator increased and became dominant
(PEO) nanofiber film over time
Curcumin/chitosan/CNF Active Pineapple – Extended shelf-life by maintaining the weight loss Ghosh et al.
film packaging and reducing the respiration rate and mesophilic (2021)
count
Curcumin nanoparticles/ Active Chilled lamb – Effectively reduced the number of total mesophilic Fallah et al.
ajowan essential oil/ packaging loins bacteria, psychrotrophic bacteria, lactic acid (2022)
pectin film bacteria and Enterobacteriaceae; reduced pH, TVB-
N, TBARS values, carbonyl and MetMb contents;
improved sensory attributes including appearance,
odor, etc
Celosia cristata L. flower Freshness Shrimp Film color changed from reddish-purple to brown/ Wu et al. (2021)
betalain/locust bean indication yellow
gum/PVA film

Watermelon peel beetroot Freshness Beef Film changed from pink at beginning to brownness Guo, Ge, et al.
betalain/pectin film indication after 8 days (2021)

Betalain/gelatin/chitosan Active Beef – Decreased pH, TBARS, S.aureus and E.coli count to Amjadi et al.
nanofiber/ZnO packaging 6.27, 0.71 mg MDA/kg, 2.63 and 3.83 log CFU/g, (2020)
nanoparticle film respectively (the values of film without betalain
extract were 6.77, 0.82, 4.56 and 4.10,
respectively; Higher L* and a* values were
obtained
Amaranthus leaf betalain/ Active Fish – Extended shelf-life of fish by reducing the pH, TVB- Kanatt (2020)
PVA/gelatin film packaging N and TVC values
β-carotene/pectin/ Freshness Butter The film color changed from orange to light yellow; Adsagh and Pirsa
nanoclay/Carum indication and Retarded lipid oxidation by lowering the peroxide (2020)
copticum essential oils active packaging content to 1.07 mEq/kg at 90 d (the CK was 4.3
film mEq/kg); Reduced microbial counts significantly

Carotenoid/PLA film Active Sunflower oil – Retarded oxidation by inhibiting the formation of Stoll et al. (2019)
packaging conjugated dines, whose content (5.77) was lower
than CK group (7.52)
Shikonin/cellulose film Freshness Shrimp The film color changed from bright red to brown Dong et al. (2020)
indication red

Shikonin/CNF film Freshness Mackerel fillet The film color changed from pink to dark purple Ezati, Priyadarshi,
indication et al. (2021)

Alizarin/chitosan film Freshness Fish fillet The film color changed from khaki to yellow Ezati and Rhim
indication (2020)

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273
J. Huang et al.
Table 2 (continued )
Natural colorants based Indication or Applica-tion Application test
packaging active packaging object
Alizarin/cellulose/strach Freshness Rainbow trout
indication fillet
double bands in β-carotene molecular structure.
Meanwhile, the natural colorant films are capable to monitor the
freshness of fruits and vegetables too, because of their abilities to sense
certain gases volatile compounds released by the respiration and
metabolism behaviors during the ripening procedure. For example, with
the decrease in litchis freshness, the blueberry anthocyanin/chitosan/
cellulose nanofiber film exhibited an obvious color alteration from
purple to pink (Zhou et al., 2021). Similarly, the production of CO2 as
well as the accumulation of organic acid contributed to the color vari­
ation from violet to purple of anthocyanin-based film when it was
applied to rutab freshness monitoring (Maftoonazad & Ramaswamy,
2019).
Apart from freshness indicator, the natural colorant-based film have
also been reported to behave as time-temperature indicator for con­
trolling the storage temperature of food products (Krithenkov et al.,
2021; Rahimah et al., 2020), or as leak indicator which is of vital
importance for the regulation of atmosphere modification packaging
(Saliu & Pergola, 2018).
6.2. Active packaging
Owing to the outstanding active functions mentioned above, the
natural colorant-based film which is considered to be a promising way in
food preservation, has been widely applied in the quality maintenance of
meat products (Amjadi, Nazari, Alizadeh, & Hamishehkar, 2020; Fallah
et al., 2022; Ge et al., 2020; He et al., 2022; Kanatt, 2020; Qin et al.,
2021; Sun, Chi, Xu, & Wang, 2020), dairy products (Adsagh & Pirsa,
2020), fruits and vegetables (Ghosh, Nakano, Mulchandani, & Katiyar,
2021; Sganzerla et al., 2021; Yang et al., 2022).
Shelf-life is a comprehensive indicator for freshness evaluation. The
litchis wrapped by black berry anthocyanin film incorporating cellulose
nanofiber and chitosan exhibited a slower spoilage process, showing the
initial mildew spots on 9 d, whereas the unpackaged groups started to
spoil on the 3rd day. The extension of shelf-life could be attributed to the
release of active compounds to retard the ripening procedure (Zhou
et al., 2021). Similarly, when being packaged by Amaranthus leaf beta­
lain film, the shelf-life of fish was prolonged by 5 days at least, owing to
the antibacterial property of betalain in extension of lag phase and
reduction of microorganism growth phase (Kanatt, 2020).
Sensory attributes including flavor, appearance, color, etc, show the
strong correlation with consumer acceptability, and several studies have
reported that the application of natural colorant-based film had the
ability to improve sensory profiles of food products. An obvious decline
in a* value of beef samples was observed (from 17.24 to 11.24), which
was effectively controlled by the betalain active film (from 18.24 to
15.91), resulted from the functions of betalain in limiting the accumu­
lation of myoglobin (Amjadi et al., 2020). In addition, the active prop­
erties of anthocyanin film were responsible for the modification effects
on cherry tomato flavor, which exhibited a higher acceptance score of
3.86 compared with control group (Sganzerla et al., 2021). Similarly,
the lamb loins packaged by curcumin/pectin film also presented rela­
tively pleasant odor, owing to the film capacities to mask the unfavor­
able flavors (Fallah et al., 2022).
Oxidation is a critical issue in food preservation. The butter packaged
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Trends in Food Science & Technology 124 (2022) 259–277
Application effects References
The film color changed from orange to bright Ezati et al. (2019)
brown
by β-carotene active film presented a steady slope in peroxide content
increasing, with a final value of 1.07 mEq/kg compared with control
group (4.3 mEq/kg), due to the migration of β-carotene into the butter,
resulting in the scavenge of free radicals (Adsagh & Pirsa, 2020). Stoll
et al. (2019) developed a series of carotenoid-based film for sunflower
oil preservation, and the results revealed that the bixin-film demon­
strated the best antioxidant effect on sunflower oil, which might be
accounted for its higher hydrophilicity, interacting with hydroperoxides
and behaving protection against oxidation better. Meanwhile, the lower
lipid oxidation degree was also observed in beef packaged by bela­
tin/ZnO/gelatin/chitosan film, manifesting as the significantly lower
thiobarbituric acid (TBA) values.
Protein degradation and denaturation is the severe problem espe­
cially for meat product preservation. TVB-N can be used to evaluate the
protein degradation degree, and researches have demonstrated that
lower TVB-N values were obtained in bass fillet (Qin et al., 2021),
shrimp and hairtail (Ge et al., 2020) packaged by Lycium ruthenicum
anthocyanin and black rice bran anthocyanin-based film, respectively.
Besides, Fallah et al. (2022) measured the carbonyl levels in lamb loins
packaged by active film with or without curcumin nanoparticles, and
samples packaged by curcumin film showed lower carbonyl contents,
ascribed to the protection influences of curcumin in protein degradation.
Apart from aforementioned issues, the microorganism growth which
can contribute to food deterioration is also an important problem in food
preservation. The total mould counts of banana packaged by radish
anthocyanin/agar film increased to 2.00 log CFU/g at the end of storage
time, significantly lower than control group (4.50 log CFU/g), suggest­
ing the excellent antimicrobial capacity of natural colorant film to
inhibit the microbial activities (Yang et al., 2022). The inhibition effects
of pigment-based film in microorganism growth have been also reported
by Ghosh et al. (2021), in which the curcumin film could effectively
reduce the mesophilic count of pineapple, due to its outstanding anti­
bacterial properties.
Meanwhile, the natural colorant-based films can also preserve food
products by performing other functions, such as the reduction of weight
loss (Ghosh et al., 2021; Qin et al., 2021; Sganzerla et al., 2021; Yang
et al., 2022), restriction of respiration actions (Ghosh et al., 2021) or the
maintenance of textural profiles (Sganzerla et al., 2021; Yang et al.,
2022). And other recent studies about the application of natural
pigment-based film in food preservation are listed in Table 2.
7. Conclusion and future prospects
The natural colorant-based film has become a promising alternative
for conventional petroleum-based food packaging owing to its eco-
friendly, multifunctional properties and widespread sources. The most
common natural colorants applied in novel food packaging are antho­
cyanin, curcumin, betalain, chlorophyll, carotenoid, shikonin and aliz­
arin. They can interact with macromolecular polymer matrix through
different ways due to their different charge natures. On the one hand,
the charged colorants (anthocyanin, betalain and chlorophyll) can
combine with neutral polymers through hydrogen bonds, while interact
with charged polymers via hydrogen bonds and electronic repulsion. On
the other hand, the neutral colorants (curcumin, carotenoid, shikonin
J. Huang et al.
and alizarin) can interact with polymers mainly by hydrogen bonds. The
molecular interactions contained in the film matrix can not only affect
film physical properties (mechanical, hydrophobic, thermal and oxygen
permeability characteristics) by formation of structural network, but
also affect film functional properties (pH-responsive, active and barrier
characteristics) via controlling of colorant release. Furthermore, the
natural colorant-based film with desired properties, can be applied as
potential indicator and active packaging for real-time freshness moni­
toring and efficient food preservation.
According to the market dynamics, the natural colorant-based food
packaging has a promising prospect due to the rising awareness of
environmental protection. The global bio-plastics production, in which
the packaging industry is accounted for 47%, is estimated to reach 2.87
million tonnes by 2025 (Hadidi et al., 2022), suggesting the inevitable
trend of natural colorant-based film in the future commercial world.
Although the production cost is higher than conventional
petroleum-based food packaging now (maybe is 10–50 times), it will be
cheaper by utilization of inexpensive or recycled by-products, devel­
opment of large-scale production and adoption of more feasible film
development routes. Meanwhile, there are still several limitations of
natural colorant-based films need to be overcome for better commercial
application. The biggest challenge is the instability of natural colorants,
which are easy to degrade under the effects of external environment,
leading to the invalid of application effects. Secondly, the relationship
between molecular interaction and release behaviors of natural color­
ants has not been defined thoroughly. Nevertheless, its investigation is
necessary since it is highly correlated with film functional properties.
Meanwhile, in recent studies, the application objects of natural
colorant-based film have mainly focused on protein-rich food products
especially for freshness real-time monitoring, and related preservation
mechanism is still unclear. Furthermore, the preparation of natural
colorant-based film is still in the laboratory stage, which can not satisfy
the demands of scale-up production yet.
Therefore, further researches are crucial in following aspects: 1)
Improve the stability of natural colorants by incorporation of co-
pigment compounds, encapsulation, modification of chemical confor­
mation, etc; 2) Investigate the influences of molecular interactions on
release performances of natural colorants to regulate their migration
effectively, resulting in better functional characteristics; 3) Extend the
application of natural colorant-based films into other food products and
study the preservation mechanisms by food metabolomics or other
innovative technology, in order to provide theoretical information for
further applications; 4) Develop technically feasible film fabrication
method to meet requirements at the industrial level for reduction of
production cost and enhancement of commercial feasibility.
Declaration of competing interest
We declare that we have no financial and personal relationships with
other people or organizations that can inappropriately influence our
work, there is no professional or other personal interest of any nature or
kind in any product, service and/or company that could be construed as
influencing the position presented in, or the review of, the manuscript
entitled Make your packaging colorful and multifunctional: the molec­
ular interaction and properties characterization of natural colorant-
based films and their applications in food industry.
Acknowledgement
The authors gratefully acknowledge the funding support of National
Natural Science Foundation of China (NSFC31871868), Hainan Pro­
vincial Natural Science Foundation of China (321CXTD1012), Scientific
Research Foundation of Hainan Tropical Ocean University
(RHDRC202117).
275
Trends in Food Science & Technology 124 (2022) 259–277
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