Trends in Food Science & Technology 112 (2021) 651–666

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Trends in Food Science & Technology

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

Plant extract mediated silver nanoparticles and their applications as

antimicrobials and in sustainable food packaging: A state-of-the-art review
Santosh Kumar a, *, Indra Bhusan Basumatary a, Hemanth P.K. Sudhani b, Vivek K. Bajpai c, **,
Lei Chen d, Shruti Shukla e, ***, Avik Mukherjee a, ****
a
Department of Food Engineering and Technology, Central Institute of Technology Kokrajhar, Kokrajhar, 783370, Assam, India
b
Sree Vidyanikethan Engineering College, Tirupati, 517502, India
c
Department of Energy and Materials Engineering, Dongguk University-Seoul, 30 Pildong-ro 1-gil, Seoul, 04620, Republic of Korea
d
College of Food Science, Fujian Agriculture and Forestry University, Fuzhou, Fujian, 350002, China
e
TERI-Deakin Nanobiotechnology Centre, The Energy and Resources Institute, Gwal Pahari, Gurugram, Haryana 122003, India

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

Keywords: Background: Nanotechnology is an emerging, novel area of applications in antimicrobial and eco-friendly food
Green synthesis packaging that have tremendous potential benefits. Silver nanoparticles (AgNPs) have been extensively studied
AgNPs and frequently used in food packaging applications due to their superior physicochemical, biological and anti­
Bio-nanocomposite
microbial properties.
Antimicrobial food package
Edible films and coatings
Scope and approach: Recently, plant extract-mediated synthesis of AgNPs has gained enormous popularity due to
Food toxicology environment-friendliness, relatively economical synthesis, and effective functionality in food packaging appli­
cations. Numerous medicinal plant extract(s) are used in the synthesis of AgNPs, which are most commonly
spherical in shape and within 2–80 nm range in size. Amount and composition of plant extract, metal ion
concentration, reaction temperature and time, and pH of the reaction mixture are the key factors affecting shape,
size, morphology and crystallinity of the synthesized AgNPs.
Key findings and conclusions: Compared to other synthesis methods, including microbial synthesis, plant-
mediated synthesis of AgNPs is faster, more convenient, and can be effective in utilization of wastes gener­
ated by fruit and vegetable producers and handlers. Plant-mediated AgNPs are effective against both Gram-
positive and Gram-negative foodborne pathogens and spoilage bacteria. Silver (Ag+) ions of AgNPs bind with
negatively charged functional groups present in the microbial cell membrane, thereby increase permeability of
the membrane disturbing the cellular transport system leading to cell death. In addition, entry of Ag+ ions into
the cell cytoplasm may damage DNA, RNA, peptide forming compounds and metabolic enzymes resulting in the
inhibition or hampering of cell division and microbial respiration. Scientific efforts are being made on the
application of plant-mediated AgNPs in bionanocomposite films and coatings, as effective food preservation
strategies. This review highlights innovative applications of plant-mediated AgNPs in sustainable antimicrobial
food packaging.

1. Introduction exhibit significantly different physical and chemical properties

compared to that of macroscale or bulk materials composed of the same
Nanotechnology involves fabrication, production, manipulation, substance. Researches in the field of nanotechnology have skyrocketed,
characterization, and application of structures, devices and systems by particularly for specialized applications in the field of healthcare, energy
controlling shape and size at nano-scale (1–100 nm). Nanoparticles production, environment, molecular computing, water treatment, and

* Corresponding author.
** Corresponding author.
*** Corresponding author.
**** Corresponding author.
E-mail addresses: s.kumar@cit.ac.in (S. Kumar), vbiotech04@dongguk.edu (V.K. Bajpai), shruti.shukla@teri.res.in (S. Shukla), ak.mukherjee@cit.ac.in
(A. Mukherjee).

https://doi.org/10.1016/j.tifs.2021.04.031
Received 3 January 2021; Received in revised form 21 March 2021; Accepted 15 April 2021
Available online 21 April 2021
0924-2244/© 2021 Published by Elsevier Ltd.
S. Kumar et al.
Fig. 1. (A) Top-down and bottom-up approach, and (b) various methods of synthesis of nanoparticles.
food packaging. In 2017, the global market value of nanotechnology was
about 49 billion US dollars, and is projected to increase to about 76
billion US dollars by 2020 (Garside, 2020).
Synthesis of nanomaterials can be classified according to the strategy
used; bottom-up approach and top-down approach (Fig. 1a). In the top-
down approach, bulk material/particles are systematically converted
into smaller particles by the use of physical processes such as crushing,
milling, grinding, lithography, ion, and plasma etching. The primary
shortcomings of this approach are high energy consumption, inability to
synthesize smaller particles, and imperfect surface morphology that may
significantly impact on physical and surface properties of nanomaterials
(Khalaj, Kamali, Costa, & Capela, 2020). The bottom-up approach in­
volves assembling of atoms and molecules of a substance to produce
nanoparticles of various sizes and morphology. This approach is mostly
used to prepare nanomaterials having uniform size, shape and distri­
bution. In the bottom-up approach, self-assembly of monomer/polymer
molecules, supercritical hydrothermal processing, sol-gel synthesis,
laser pyrolysis, microwave heating, chemical vapor deposition (CVD),
and bio-assisted synthesis, including plant-mediated green synthesis are
commonly used. Based on the nature of the synthesis, it is classified into
physical, chemical, and biological processes (Ahmed, 2020).
Physical methods apply mechanical pressure, thermal energy, elec­
trical energy, and high energy radiations to cause material abrasion,
melting, evaporation or condensation to generate nanoparticles (NPs).
Chemical methods are the most suitable methods for synthesis of metal
and metal oxide nanoparticles, where metal ions are reduced by chem­
ical reducing agents and capping agents are used to stabilize NPs. So­
dium borohydride, hydrazine hydrate, and potassium aurochlorate are
most commonly used reducing agents, whereas, poly-vinyl pyrolidone
(PVP), poly-ethylene glycol (PEG), poly-methacrylic acid (PMAA), poly-
methylmethacrylate (PMMA), and gelatin agar are frequently used as
capping agents (Sharma, Yngard, & Lin, 2009). Biological methods
employed mainly plant extracts, such as, (leaf, fruit, root etc.), micro­
organisms (bacteria, fungi, algae, etc.), and biomolecules as templates,
as reducing and stabilizing agents (Huang et al., 2015; Kharissova, Dias,
Kharisov, Pérez, & Pérez, 2013; Sharma et al., 2009). Green synthesis
method is environmentally-friendly, economical, convenient, and an
efficient process (Bhattarai, Zaker, & Bigioni, 2018; Chandra, Kumari,
Bontempi, & Yadav, 2020; Kharissova et al., 2013). Numerous re­
searchers have approached on the green synthesis of NPs, and their
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Trends in Food Science & Technology 112 (2021) 651–666
applications are widely used in the fields of biomedical, pharmaceutical,
cosmetic, environmental, and food sectors (Chandra et al., 2020). The
following figure (Fig. 1b) summarizes physical, chemical, and biological
routes of their synthesis.
Silver is a fascinating material and has been utilized for various day-
to-day applications since ancient time. Silver nanoparticles (AgNPs)
have been extensively studied due to their superior physicochemical,
biological and antimicrobial properties (Das et al., 2020). Properties of
AgNPs primarily depend on their size, shape, distribution, and surface
characteristics which can be tuned during green synthesis via modu­
lating the concentration of reducing agents, stabilizers, metal pre­
cursors, and reaction conditions (Pryshchepa, Pomastowski, &
Buszewski, 2020). Green synthesis of AgNPs has growing demands
especially in food, biomedical, and therapeutic applications as the pro­
cess avoids use of toxic and hazardous synthetic chemicals (Kumar,
Rajendran, Houreld, & Abrahamse, 2018; Pryshchepa et al., 2020).
Among green synthesis methods, plant-mediated synthesis of AgNPs is
convenient, cost-effective and single-step process that uses plant extracts
as natural resources (Duan, Wang, & Li, 2015). The plant extracts mainly
contain polysaccharides, polyphenols, alkaloids, aldehydes, proteins,
and amino acids that can act as reducing and stabilizing agents
(Rajeshkumar & Bharath, 2017).
Food quality and safety are major concerns for consumers, food in­
dustries, and regulatory bodies worldwide. Demands for fresh, clean,
and hygienically manufactured food having prolonged shelf-life without
use of synthetic chemical preservatives are rapidly growing, which has
become the necessity in this era. Food businesses and trades have
become global, and require novel mitigation strategies to inhibit mi­
crobial food contamination. Consumers are increasingly relying on
processed and ready-to-eat foods that are convenient for on-the-go
consumption. Therefore, antimicrobial packaging technology is one of
the most suitable tools to provide consumers with safe and healthy foods
(Fahmy, Salah Eldin, Abu Serea, Gomaa, AboElmagd, Salem, et al.,
2020; Mustafa & Andreescu, 2020). In this review, we present a
comprehensive and contemporaneous account on plant-mediated syn­
thesis of AgNPs, their characterizations and the antimicrobial activities
as natural food packaging alternatives. In addition, this review also
explores various factors affecting synthesis of AgNPs and their antimi­
crobial efficiency. The review also focuses on applications of
plant-mediated synthesized AgNPs as potential antimicrobials in
S. Kumar et al.
Fig. 2. (A) Schematic presentation of plant-mediated synthesis of AgNPs, their characterization and their food packaging applications, and (b) Steps involved and
mechanisms of plant-mediated synthesis of AgNPs.
sustainable food packaging.
2. Synthesis of silver nanoparticles (AgNPs)
2.1. Synthesis of AgNPs using plant extracts
The plant-mediated biosynthesis is predominantly utilized to syn­
thesize metal, metal oxide, and bi-metallic alloys nanoparticles. Plant-
mediated biosynthesis of AgNPs is one of the effective, convenient,
rapid, non-toxic and environment-friendly technique (Das et al., 2020;
Keat, Aziz, Eid, & Elmarzugi, 2015). The use of plants and their different
parts as reducing as well as stabilizing agents has drawn increased
research attention in synthesis of nanomaterials, especially for
food-safety and biomedical applications (Alomar, AlMasoud, Awad,
El-Tohamy, & Soliman, 2020). Number of different plants and their
various parts such as leaf, latex, seed, flower, fruit, peel, bark, root, gum,
etc. have been used for the synthesis of AgNPs (Das et al., 2020;
Rajeshkumar & Bharath, 2017). In this context, numerous medicinal
plants such as Azadirachta indica, Rosmarinus officinalis, Aloe vera,
Jatropha curcas, Cinnamon zeylancium, Euphorbia hirta, Acalypha indica,
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Trends in Food Science & Technology 112 (2021) 651–666
Tinospora cordifolia, Vitex Negundo, Catharanthus roseus, Ananas comosus,
Malva parviflora, etc. were also used for synthesis of AgNPs (Chandra
et al., 2020). In a recent study, Alomar et al. (2020) reported the syn­
thesis of highly antibacterial AgNPs using leaf extract of Peganum har­
mala, a glabrous shrub. The synthesized AgNPs showed inhibition zone
of 65 mm against Gram-negative pathogenic bacterium E. coli and 50
mm against Gram-positive bacteria S. aureus and Bacillus cereus (Alomar
et al., 2020). In another study, fruit pulp extract of Aegle marmelos was
used to synthesize AgNPs for enhanced medicinal purposes. The syn­
thesized AgNPs were used to prepare an antiseptic formulation that
showed better antimicrobial activity than commonly marketed anti­
septic creams (Devi et al., 2020; S.; Patil & Muthusamy, 2020).
In a typical plant-mediated synthesis of AgNPs, following steps are
primarily involved (Fig. 2a). Initially, collection of plant materials fol­
lowed by thorough washing 2–3 times with double distilled water is
performed. There are two different ways to prepare extracts from the
washed plant materials. Firstly, these are dried, ground into powder, and
stored in airtight container for further use. The extract is prepared from
the dried powder by aqueous or alcoholic extraction. In aqueous extract,
weighed plant material is heated in double distilled water usually at
S. Kumar et al. Trends in Food Science & Technology 112 (2021) 651–666

2009). In another study, it was reported the functional groups of alka­

loids, flavones, and anthracenes, such as –C═C–, –C–O–, –C–O–C–, and
–C═O– facilitate reduction of metal ions resulting in synthesis of nano­
particles (Kesharwani et al., 2009).
The physical and chemical factors that affect the synthesis of plant-
mediated AgNPs include amount of plant extract, composition of plant
extract, concentration of metal ions, pH of the reaction mixture, reaction
temperature, reaction time, and agitation speed (Rajeshkumar & Bhar­
ath, 2017). The size, shape and morphology of the AgNPs mainly depend
on the composition of the plant extract, metal ion concentration, pH, and
reaction time (Rajeshkumar & Bharath, 2017; Raza et al., 2016). A
change in pH results in change in the ability of phytochemicals to bind
with or to reduce metal cations during nanoparticle synthesis, and it
may affect the shape, size, morphology and yield of the nanoparticles
(Gahlawat & Choudhury, 2019). Temperature is another important
factor that affects the synthesis of AgNPs, as elevated temperature in­
creases the rate of reaction and efficiency of the synthesis (Liu, Zhang,
Wang, & Wei, 2020). Table 1 summarizes names of the plants and their
different parts used in plant-mediated synthesis of AgNPs, including
phytochemicals responsible as reducing and stabilizing agents and
characteristics of the synthesized NPs.
Fig. 3. Phytochemicals involve in bio-reduction of Ag+ ions and in the stabi­
lization of AgNPs.
2.2. Advantages and disadvantages of plant-mediated synthesis

Physical and chemical methods use heavy equipments, large amount

50–80 ◦ C for 5–15 min. The resulting extract is filtered and the filtrate is
of energy, toxic chemical compounds that are harmful for the environ­
stored at 4–6 ◦ C for synthesis of AgNPs. In second method, the washed
ments and living beings (Keat et al., 2015; Rasheed et al., 2017). The
plant materials are crushed in a crucible, water is added into it, and the
chemicals, such as sodium borohydride, alcohols, hydrazines which are
resulting extract is filtered, and the filtrate is stored in a similar way as
toxic, flammable, and non-degradable are required in physical and
stated above. For the synthesis, aqueous solution of metal precursor,
chemical synthesis methods (Virkutyte & Varma, 2013, pp. 7–33).
such as, AgNO3 (usually 1 mM) is prepared, predetermined amounts of
Plant-mediated synthesis of AgNPs is very rapid, convenient, easy,
the plant extract are added into it, and the mixture is either heated at
cost-effective, eco-friendly, non-toxic, and requires natural plant-based
around 60–80 ◦ C or kept at room temperature or in sun-light for the
resources, which are abundantly available (Fig. 4). Compared to
formation of nanoparticles, as indicated by the subsequent color change
plant-mediated synthesis, other green synthesis methods, such as mi­
in the reaction mixture (Kumari, Brahma, Rajak, Singh, & Kumar, 2016;
crobial synthesis involve very difficult and time-consuming process, as
Saha, Begum, Mukherjee, & Kumar, 2017; Swargiary, Mitra, Halder, &
bio-reduction is faster in case of plant-mediated synthesis. In addition,
Kumar, 2019).
waste generated in the microbial synthesis may be harmful to the
In case of AgNPs, the colorless solution of AgNO3 changes into yellow
environment based on kinds of microbes used in the synthesis.
or yellowish brown (S. Kumar, Singh, Halder, & Mitra, 2016; Swargiary
Plant-based fabrication of AgNPs is suitable for large-scale commercial
et al., 2019). The formation of AgNPs is further confirmed by UV–visible
production that can be safe for food, medical and therapeutic applica­
spectrophotometer. The prepared nanocolloids of AgNPs are lyophi­
tions. Utilization of wastes from fruit and vegetable process industries
lized, and the dried powder is used for the characterization of the syn­
can be achieved in the AgNPs synthesis that makes plant-mediated
thesized AgNPs using various analytical methods, such as transmission
synthesis more environment-friendly (Jalilian et al., 2020). Thus,
electron microscopy (TEM), scanning electron microscopy (SEM), X-ray
plant-mediated synthesis is a simple, effective with zero-contamination
diffraction (XRD), dynamic light scattering (DLS), Zeta potential,
process that could a good alternative to conventional chemical and
Fourier-transform infrared spectroscopy (FTIR), etc. There are three
physical methods, and comparable to microbial synthesis. However,
primary steps involved in the mechanism of plant-mediated synthesis of
there are a few drawbacks and a concern associated with plant-mediated
AgNPs (Fig. 2b). The initial activation phase involves reduction of silver
synthesis of AgNPs. Destruction of plant and plant parts is a major
ions by plant biomolecules and nucleation of the reduced metal atoms.
drawback while using fresh plant resources for nanoparticle synthesis
During second step, growth of the nucleus occurs as spontaneous ag­
(Herlekar, Barve, & Kumar, 2014). This can be addressed by utilizing
gregation of adjacent nanoparticles to form larger size particles. In the
plant-based wastes or agro-wastes, which otherwise contribute to
final step, capping of the aggregated particles takes place giving the final
environmental pollution. The other concern is the subjectivity of the
shape to the nanoparticles. In the plant-mediated synthesis, phytoche­
plant resources whose chemical compositions significantly vary
micals/biomolecules play the key role in reduction of Ag+ ions. The
depending upon their geographical locations in different parts of the
primary phytochemicals involved are terpenoids, polyphenols, carbo­
world. This variation may lead to different results in different labora­
hydrates, alkaloids, phenolic acids, and proteins (Fig. 3) (Rajeshkumar
tories. This problem can be resolve by identifying the biomolecules
& Bharath, 2017). Commonly, these reducing agents and other metab­
present in the plant extract that are responsible for the nanoparticles’
olites present in the plant extract also work as capping and stabilizing
synthesis (Ali, Ahmed, Wu, Hossain, Hafeez, Islam Masum et al., 2020).
agents. This gives the additional advantage to the plant-mediated syn­
thesis, as no added stabilizing agents are required. Proteins, carbohy­
3. Antimicrobial activities of AgNPs
drates and polyphenolic compounds are important biomolecules of plant
extracts that is responsible for the reduction of Ag+ ions and formation
3.1. Antimicrobial studies and action mechanism
of NPs (Jena, Singh, Panigrahi, Suar, & Mandal, 2016). Kesharwani
et al., reported that phenolic biomolecules such as quinones and
Silver has been extensively used as an antimicrobial agent for food
plasto-hydroquinone in plant extract assist in reduction of metal ions,
preservation and storage for medical and therapeutic applications since
and thus facilitate NPs synthesis (Kesharwani, Yoon, Hwang, & Rai,
ancient time. It has been used as an antiseptic, and for treating burns and

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Table 1
Plant-mediated synthesis of AgNPs, and the phytochemicals involved in reduction of Ag+ ions.
Reducing agents Phytochemicals involved AgNPs characteristics (shape, References
and size)
Plant name Plant parts
used

Musa Flower Protein Spherical; 12.6–15.7 Valsalam et al. (2019)

Acuminate
Moringa oleifera Flower Proteins, phenolic Spherical; 8 nm Bindhu, Umadevi, Esmail, Al-Dhabi, and
Arasu (2020)
Caesalpinia Flower Alcohol groups, aliphatic amine, alkane, ester, carbonyl, nitro Spherical; 2–22 nm Moteriya and Chanda (2017)
pulcherrima compounds
Datura inoxia Flower Amine groups, ketones groups, nitro-compounds, aromatic Polygonal; 15–73 nm Gajendran, Durai, Varier, Liu, Li,
compound Rajendran, et al. (2019)
Ipomoea digitata Flower Aromatic amines, amides, carbonyl groups, polyphenols, Roughly spherical; >100 nm Varadavenkatesan, Selvaraj, and
Linn. alcohols and proteins Vinayagam (2019)
Madhuca longifolia Flower Alcohol, phenols, alkanes, Spherical and oval; (M. P. Patil et al., 2018)
Aromatic, amines 30–50 nm
Fritillaria sp. Flower Hydroxyl groups, amine groups, carbonyl groups Spherical, 5–10 nm Hemmati, Heravi, Karmakar, and Veisi
(2020)
Lythrum salicaria Aerial and Phenolic, flavonoid, polyphenol, anthocyanin, tannin, gallic Spherical; 45–65 nm Mohammadalinejhad, Almasi, and Esmaiili
flower acid (2019)
Allium Aerial Secondary plant metabolites (alcohol, phenol, methoxy Spherical; 2.3–27 nm Jalilian, Chahardoli, Sadrjavadi, Fattahi,
ampeloprasum parts groups, aromatic compounds, ester, ether etc.), protein, and Shokoohinia (2020)
carboxylic acid,
Equisetum arvense Leaf and Phenolic, carboxylic acid, ester and several other plant ~ Spherical; 170.5 Miljković, Lazić, Davidović, Milivojević,
stem metabolites of E. arvense extract. hydrodynamic diameter (d. Papan, Fernandes, et al., (2020)
nm)
Anthemis atropatana Aerial Amide of protein, amine, ether, ester, cyanide Spherical; 10–80 nm (mean - Dehghanizade, Arasteh, and Mirzaie
parts 38.8 nm) (2018)
Swertia paniculata Aerial Flavonoids, xanthones, carboxylic group of protein, amino Spherical; 31.2–43.3 nm Ahluwalia, Elumalai, Kumar, Kumar, and
parts acids Sangwan (2018)
Glaucium Aerial Hydroxyl carboxyl, amine Spherical; 45–53.7 nm Allafchian, Jalali, Aghaei, and Farhang
corniculatum L. parts (2018)
Nepeta leucophylla Root Phenol, protein Spherical and elongated; 20 Singh and Dhaliwal (2019)
nm (TEM)
Phoenix dactylifera Root Amide-I of protein, C–H bonds, amine groups, hydroxyl Spherical; 15–40 nm Oves, Aslam, Rauf, Qayyum, Qari, Khan,
groups et al., (2018)
Colocasia esculenta Bulb Polyphenol, amide, protein Spherical; 20 nm Inamuddin and Kanchi (2020)
Alpinia nigra Fruit Polyphenol, protein, flavonoides Spherical; 6 nm Baruah, Yadav, Yadav, and Das (2019)
Phyllanthus emblica Fruit Carbpxylic acid, alcohol/phenol, nitro compound, carbonyl Spherical; 19–45 nm Renuka et al. (2020)
groups, amide, alkynyl groups
Cleome viscosa L. Fruit Protein, phenolics compounds, carbonyl groups, aromatic Spherical; 20–50 nm (TEM) (G, A, P.T, & K, 2018)
groups, tannins, alkaloids
Garcinia indica Fruit Amine, amide, carbonyl groups Spherical and hexagonal; Sangaonkar and Pawar (2018)
(Kokum) 5–30 nm (TEM)
Piper longum L. Leaf Hydroxyl, carbonyl, amide, flavonoids, terpenoids Spherical; 25–32 nm Yadav, Saini, Kumar, Pasi, and Agrawal
(2019)
Prosopis Juliflora Leaf Alcoholic, Phenolic, Aromatic compound Spherical; 10–20 nm Arya et al. (2018)
Withania coagulans Leaf Amides, flavonoides, phenols, amino acids Spherical; ~14 nm Tripathi, Modi, Narayan, and Rai (2019)
Dillenia indica L. Leaf Ester, carboxylic acid, Spherical; 10–23 nm Nayak et al. (2020)
Solanum Leaf Phenolic compound (aromatic groups and carboxylate Spherical and Oval; 35–50 (Santiago et al., 2019)
lycopersicum groups) nm (TEM)
Allium saralicum R. Leaf Polyphenolics, flavonoidic group Spherical; 20–40 nm (TEM) Zangeneh, Bovandi, Gharehyakheh,
M. Fritsch Zangeneh, and Irani (2019)
Veronica amygdalina Leaf Phenol, saponin, and alkaloid Spherical; 2–18 nm Aisida, Ugwu, Akpa, Nwanya, Nwankwo,
Botha, et al., (2019)
Artemisia vulgaris Leaf Phenols, aromatic amine, carbonyl groups Rounded; 27–53 nm (SEM), Rasheed, Bilal, Iqbal, and Li (2017)
25 nm (TEM)
Aesculus Leaf Saponin, protein, alkanes, ester Spherical; 50 ± 5 nm Küp, Çoşkunçay, and Duman (2020)
hippocastanum
Satureja hortensis Leaf Aromatic compounds, carbohydrates, alcohols, carboxylic Spherical; 2.9–3.4 nm (SEM) Rasaee, Ghannadnia, and Baghshahi
acids, esters, ethers (2018)
Telfairia Leaf Protein (hydroxyl, carboxyl, alkane) Spherical; 16–19 nm (Aisida et al., 2020)
Occidentalis
Azadirachta Indica Leaf Terpenoids, Spherical; 20–50 nm Asimuddin et al. (2020)
Flavonoids, Polyphenols
Melissa officinalis Leaf Phenolic/flavonoids, carbonyl, aliphatic compound Spherical; 5–30 nm Ahmeda, Zangeneh, and Zangeneh (2020)
Gundelia tournefortii Leaf Antioxidant compounds containing O–H, C–H, C– –C, C–
–O, Spherical; 16.5 nm (Han et al., )
L. C–O, C–O–C groups
Rumex acetosa Leaf N–H and O–H groups Spherical; 5–80 nm Kota, Dumpala, Anantha, Verma, and
Kandepu (2017)
Ocimum Sanctum Leaf Quercetin Spherical; 11.35–14.6 nm Jain and Mehata (2017)
Delonix regia Leaf Phenols, organic molecules, aliphatic amines Non-uniform; 43.5 nm Nogueira et al. (2014)
Oriza sativa Leaf Sugars, proteins, phenolics Spherical; 3.7–29 nm Kora, Mounika, and Jagadeeshwar (2020)
Cannabis sativa Leaf Phenols, proteins, amino groups, alkanes, alkenes, flavonoids Spherical; 26.52 nm Chouhan and Guleria (2020)

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S. Kumar et al.
Fig. 4. Advantages of plant-mediated synthesis of AgNPs.
wounds (Duan et al., 2015; Jalilian et al., 2020). Recent studies have
established the facts that AgNPs are effective against bacteria, fungi and
viruses. The effectiveness of AgNPs is commonly more prominent
against Gram-negative bacteria compared to that of Gram-positive due
to thick peptidoglycan layers in the cell wall structure of the latter
resulting in rigid cell wall, which causes difficulties in AgNPs penetra­
tion (El-Seedi, El-Shabasy, Khalifa, Saeed, Shah, Shah, et al., 2019). For
example, AgNPs synthesized using banana peel extract showed higher
effectiveness against E. coli and P. aeruginosa (Gram-negative bacteria)
compared to B. subtilis and S. aureus (Gram-positive) bacteria. However,
the synthesized AgNPs showed less effectiveness against a fungal path­
ogen C. albicans (Fig. 5a) (Ibrahim, 2015). Recent studies on
green-synthesized AgNPs reported that plant extract(s) or phytochemi­
cals used in the synthesis may have some contribution to enhance the
antimicrobial activities of the nanoparticles, compared to those
Fig. 5. (A) Effectiveness of plant-mediated AgNPs against various pathogenic microorganisms (Reproduced with permission (Ibrahim, 2015) Copyright 2015,
Elsevier), (b) Schematic representation of mechanism of action of AgNPs against bacteria.
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Trends in Food Science & Technology 112 (2021) 651–666
synthesized through non-biological routes (Fard et al., 2018; Oliver,
Wagh, Liang, Yang, & Boyer, 2018). Panpaliya et al. (2019) reported
that the green-synthesized AgNPs had higher antibacterial activity
against certain human pathogens and food spoilage lactic acid bacteria
compared to that of synthetic/chemical antibacterial compounds such as
chlorohexidine (Panpaliya et al., 2019). Table 2 summarizes the
plant-mediated AgNPs and their antimicrobial properties against
different hazardous microorganisms.
AgNPs can be growth inhibitors or biocides. The antibacterial ac­
tivity of AgNPs depends on various factors such as pH, temperature, test
microorganisms, concentration, size and shape of nanoparticles (Tang &
Zheng, 2018). The antimicrobial activity of plant-mediated AgNPs
shows various mechanisms (Fig. 5b). Firstly, the antimicrobial proper­
ties of AgNPs are associated with release of Ag+ ion, Ag+ ions bind with
electron-donating negatively charged functional groups like sulfhydryl,
carbonyl, imidazole, amino, and phosphate, abundantly present in
membrane protein, which alter membrane structure, and thus increases
permeability of the membrane. Ag+ ions cause disturbance in the
cellular transport system leading to cell death (S. Kumar, Singh, Halder,
& Mitra, 2014; Ramalingam, Parandhaman, & Das, 2016; Yan et al.,
2018). In addition, Ag+ ions can enter into microbial cell and damage
DNA, RNA, peptide forming compounds and metabolic enzymes
resulting in inhibition of transcription, translation i.e., cell division, and
microbial respiration. Second mechanism is based on accumulation of
AgNPs on the cell wall and membrane (Dakal, Kumar, Majumdar, &
Yadav, 2016; Slavin, Asnis, Häfeli, & Bach, 2017). AgNPs cause
morphological changes, leading to cytoplasmic shrinkage, membrane
detachment, and finally ruptures of membrane. Similar to Ag+ ion,
AgNPs can also penetrate into bacterial cell, and can interact with bio­
molecules such as lipids, proteins, and DNA, leading to inhibition of cell
division, and disruption of cellular functioning (Dakal et al., 2016;
Goswami, Sahareen, Singh, & Kumar, 2015). The third mechanism is
based on generation of reactive oxygen species (ROS). In this mecha­
nism, AgNPs and silver ions increase oxidative stress by generation of
reactive oxygen species (ROS) and free radicals such as hydroxyl radi­
cals (OH●), H2O2, superoxide anions (O2− ), hydroxyl radical, singlet
oxygen, and hypochlorous acid, which are strong oxidizing agents that
can damage cellular components, enzymes, and genetic materials
(Quinteros, Cano Aristizábal, Dalmasso, Paraje, & Páez, 2016; Zou et al.,
2018).
S. Kumar et al. Trends in Food Science & Technology 112 (2021) 651–666

Table 2
Antimicrobial activity of plant-mediated synthesized silver nanoparticles (AgNPs).
Plant used in AgNPs Size and shape of AgNPs Test microorganism Concentration of ZoI (mm) MIC References
synthesis nanohybrid

Moringa oleifera Spherical; K. pneumoniae (1 mM AgNO3/8 17 - Bindhu et al. (2020)

8 nm S. aureus mL extract) 29
Peganum harmala Spherical; E. coli 500 μL (for 65 - Alomar et al. (2020)
184 nm (average) S. aurous bacteria) 50
B. cereus 50
E. faecalis No
inhibition
F. oxysporum 250 μL (for fungi) 14
A. alternate 7
Tricoderma 20
Pistacia atlantica Spherical; E. coli 31 μg/mL 10 3 μg/mL Hamelian, Zangeneh,
40–60 nm P. aeruginosa 11 31 μg/mL Shahmohammadi, Varmira, and Veisi
S. aureus 10 7 μg/mL (2020)
B. subtilis 11 7 μg/mL
Caesalpinia pulcherrima Spherical; B. cereus 10–0.019 mg/mL - 5 mg/mL Moteriya and Chanda (2017)
2–22 nm B. subtilis 5 mg/mL
S. aureus 2.5 mg/
mL
Corallium rubrum 10 mg/mL
E. coli 5 mg/mL
P. aeruginosa 5 mg/mL
S. typhimurium Resistant
K. pneumoniae Resistant
C. albicans 5 mg/mL
C. glabrata 2 mg/mL
Cryptococcus 5 mg/mL
neoformans
Madhuca longifolia Spherical and Oval; E. coli 400 μg/mL ~15.5 90 μg/mL (M. P. Patil et al., 2018)
30–50 nm S. typhimurium ~15.7 40 μg/mL
B. cereus ~18.5 60 μg/mL
S. saprophyticus ~20.5 80 μg/mL
Lythrum salicaria Spherical; E. coli 100 μL 14.19 ± - Mohammadalinejhad et al. (2019)
45–65 nm 0.092
S. aureus 14.94 ±
0.416
Allium ampeloprasum Spherical; E. coli (ATCC25922) 300 μg/mL 23 37.5 μg/ Jalilian et al. (2020)
2.3–27 nm mL
MDR E. coli (MDR- 13 37.5 μg/
BL21) mL
S. aureus (ATCC25923) 8 18.75 μg/
mL
MDR P. aeruginosa 27 18.75 μg/
mL
Equisetum arvense Spherical; 170.5 S. enterica 100 μL 11.64 ± - Miljković et al., (2020)
hydrodynamic diameter 0.29
(d.nm) B. cereus 10.75 ±
0.18
L. monocytogenes 12.46 ±
0.37
E. faecium 9.68 ±
0.0.16
S. aureus 12.53 ±
0.44
A. hydrophila 10.80 ±
0.12
Anthemis tropatana Spherical; S. aureus (ATCC 6538) 12.5–100 μg/mL - 12.5 μg/ Dehghanizade et al. (2018)
10–80 nm mL
Streptococcus pyogenes 50 μg/mL
(ATCC 19615)
P. aeruginosa (ATCC 100 μg/
15442) mL
E. coli (ATCC 25922) 25 μg/mL
Swertia paniculata Spherical; 31.2–43.3 nm P. aeruginosa 20 μg/mL 18.80 ± - Ahluwalia et al. (2018)
0.72
K. pneumoneae 17.80 ±
0.20
S. aureus 14.67 ±
0.35
Glaucium corniculatum Spherical; E. coli 100 mg/L 9.6 ± 0.3 - Allafchian et al. (2018)
45–53.7 nm S. typhimurium 8.6 ± 0.2
S. aureus 8.3 ± 0.4
B. cereus 7.6 ± 0.2
Phoenix dactylifera E. coli 80 μg/mL 22 - Oves et al., (2018)
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Table 2 (continued )
Plant used in AgNPs Size and shape of AgNPs Test microorganism Concentration of ZoI (mm) MIC References
synthesis nanohybrid

Spherical; C. albicans 20
15–40 nm
Alpinia nigra Spherical; S. aureus 100 μg/mL 17.67 ± 30 μg/mL Baruah et al. (2019)
6 nm 0.57
K. pneumoniae 19.67 ± 20 μg/mL
0.57
Candida albicans 12.33 ± 50 μg/mL
0.57
Madhuca latifolia Spherical; S. aureus, 10 μL 7.89 - Biswal and Misra (2020)
2–30 nm S. faecalis 14.45
L. monocytogenesi 15.98
E. Coli 8.23
S. Typhimurium 12.83
Cleome viscosa Spherical; B. subtilis 40 mg/mL 14 ± 1.5 - (G, A, P.T, and; K, 2018)
20–50 nm S. aureus 17 ± 0.8
E. coli 16 ± 0.8
K. pneumoniae 14 ± 1.52
Garcinia indica Spherical and hexagonal; E. coli 20 mg/mL 14 ± 0.3 20 mg/mL Sangaonkar and Pawar (2018)
5–30 nm B. subtilis 12 ± 0.2 20 mg/mL
S. aureus 15 ± 0.2 20 mg/mL
P. aeruginosa 12 ± 0.5 40 mg/mL
S. entericatyphi No -
inhibition
Proteus vulgaris No -
inhibition
Serratia marcescens No -
inhibition
Prosopis juliflora Spherical; E. coli 1 μg ~14.00 - Arya et al. (2018)
10–20 nm P. aeruginosa ~11.50
Withania coagulans Spherical; E. coli 20 μg/mL 4 ± 0.57 - Tripathi et al. (2019)
~14 nm K. pneumoniae 8.34 ±
0.33
S. aureus 10.66 ±
0.88
Dillenia indica Spherical; 10.01–23.24 E. faecalis 0.25 mg/mL 27 - Nayak et al. (2020)
nm E. coli 16
Allium saralicum Spherical; P. mirabilis 500 μg/mL 19.2 ± 1.3 250 μg/ Zangeneh et al. (2019)
20–40 nm mL
S. flexneri 23.6 ± 125 μg/
1.34 mL
P. aeruginosa 28.6 ± 31 μg/mL
0.89
E. coli 24 ± 0.7 125 μg/
mL
E. faecalis 30.6 ± 31 μg/mL
0.89
B. subtilis 30.2 ± 1.3 31 μg/mL
S. aureus 25.6 ± 125 μg/
1.14 mL
S. saprophyticus 32 ± 0.7 31 μg/mL
S. pneumoniae 26.8 ± 62 μg/mL
0.44
S. typhimurium 20.8 ± 250 μg/
0.44 mL
C. albicans 23.2 ± 62 μg/mL
0.44
C. glabrata 24.6 ± 62 μg/mL
1.14
C. parapsilosis 24.4 ± 62 μg/mL
1.34
C. krusei 26.2 ± 1.3 31 μg/mL
C. guilliermondii 33.6 ± 31 μg/mL
1.34
Veronica amygdalina Spherical; S. aureus 80 μg/mL 13.0 - Aisida, Ugwu, Akpa, Nwanya,
2–18 nm Coliform 18.8 Ejikeme, Botha, et al., (2019)
Aesculushippocastanum Spherical; S. aureus 100 μg/mL 17.5 ± 1.56 mg/ Küp et al. (2020)
(Horse chestnut) 50 ± 5 nm 2.12 mL
S. epidermidis 13 ± 0.00 3.12 mg/
mL
L. monocytogenes 13 ± 0.00 1.56 mg/
mL
C. renale 15 ± 2.64 12.5 mg/
mL
M. luteus 12 ± 0.00
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Table 2 (continued )
Plant used in AgNPs Size and shape of AgNPs Test microorganism Concentration of ZoI (mm) MIC References
synthesis nanohybrid

12.5 mg/
mL
B. subtilis 17 ± 0.00 6.25 mg/
mL
B. cereus 10.5 ± 3.12 mg/
1.41 mL
E. faecalis 10.5 ± 0.19 mg/
1.41 mL
P. aeruginosa 20.0 ± 1.56 mg/
0.00 mL
P. fluorescens 8 ± 0.00 1.56 mg/
mL
E. coli 10 ± 0.00 1.56 mg/
mL
E. aerogenes 11 ± 0.00 12.5 mg/
mL
K. pneumonia 12.5 ± 6.25 mg/
0.70 mL
P. mirabilis 10 ± 0.00 6.25 mg/
mL
C. albicans No -
inhibition
C. tropicalis No -
inhibition
C. krusei No -
inhibition
Satureja hortensis Spherical; B. subtilis, 100 μg/mL 8.00 - Rasaee et al. (2018)
2.9–3.4 nm B. vallismortis 7.50
E. coli 7.00
Telfairia occidentalis Spherical; 15.84–19.2 nm K. pneumonia 50 μg/mL 21.30 0.2 mg Aisida et al., (2020)
Azadirachta indica Spherical; E. coli - - 0.5 μg/mL Asimuddin et al., (2020)
20–50 nm S. aureus 1.0 μg/mL
Gundelia tournefortii Spherical; P. aeruginosa 64 mg/mL 34.6 ± 4 ± 0 mg/ (Han et al., )
16.5 nm 1.14 mL
S. typhimurium 33.6 ± 4 ± 0 mg/
0.89 mL
E. coli 34.8 ± 4 ± 0 mg/
1.09 mL
S. aureus 35.8 ± 4 ± 0 mg/
1.09 mL
S. pneumoniae 37 ± 1 2 ± 0 mg/
mL
B. subtilis 37.8 ± 2 ± 0 mg/
1.09 mL
C. krusei 38.6 ± 2 ± 0 mg/
0.89 mL
C. guilliermondii 37.6 ± 2 ± 0 mg/
1.14 mL
C. glabrata 35.2 ± 4 ± 0 mg/
0.44 mL
C. albicans 34.2 ± 1.3 4 ± 0 mg/
mL
Rumex acetosa Spherical; 5–80 nm B. cereus 20 μl 16 ± 0 - Kota et al. (2017)
Enterococcus sp. 15.3 ±
1.25
S. aureus 18 ± 0
S. epidermidis 18 ± 0
Vibrio cholera 15.5 ± 0.5
E. coli 17 ± 0
Klebsiella sp. 15 ± 0
Proteus mirabilis 15 ± 0
P. vulgaris 15 ± 0
Pseudomonas sp. 12.7 ± 0.5
Salmonella typhi 14.3 ± 0.5
S. paratyphi A 12.4 ±
2.15
S. typhimurium 15.5 ± 0.5
Shigellaflexneri 17.3 ±
1.25
S. dysentery A 17 ± 0
Cannabis sativa Spherical; 26.52 nm S. aureus 20 μL 23.56 10 μL Chouhan and Guleria (2020)
(average) M. luteus 24.66 5 μL
B. subtilis 22.80 10 μL
E. coli 28.00 5 μL
K. pneumoniae 20.50 5 μL
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Table 2 (continued )
Plant used in AgNPs Size and shape of AgNPs Test microorganism Concentration of ZoI (mm) MIC References
synthesis nanohybrid

C. albicans 21.70 10 μL
S.cerevisiae 22.80 5 μL
Prosopis farcta Sphercal; S. aureus 5 mg 14 ± 2.5 - (Salari, Esmaeilzadeh Bahabadi,
10.26–14.65 nm S. pneumonia 13.33 ± Samzadeh-Kermani, & Yosefzaei,
0.58 2019)
E. coli 15 ± 2.65
S. typhi 17.33 ±
0.58
Artemisia vulgaris Round shaped; 25 nm K. pneumonia 20 μg NPs/disc 11 - Rasheed et al. (2017)
(average) H. influenza 12
P. aeruginosa 9
E. coli 10
S. aureus 8

3.2. Parameters affecting antimicrobial activity 3.2.2. Shape of AgNPs

Several factors, either individually or synergistically, influence
antimicrobial efficiency of AgNPs. These include size, shape, surface
charge(s), morphology, concentration of nanoparticles, and/or target
microorganism(s) (Dakal et al., 2016; Hamouda, Hussein, Abo-elmagd,
& Bawazir, 2019). Some of the most important factors among these
are discussed below.
3.2.1. Size of AgNPs
The size and shape of the nanoparticles greatly influence their anti­
microbial activity. Numerous researchers reported that smaller particle
size showed greater antibacterial activity, and vice versa (Raza et al.,
2016; Sotiriou & Pratsinis, 2010). Smaller sizes of nanoparticles have
larger surface-to-volume ratio resulting in higher concentration of silver
atoms or ions, which increases their reactivity with microbial cell
wall/membrane, and can also increase ROS production (Quinteros et al.,
2016). Additionally, smaller particles have higher tendency to enter into
microbial cells leading to interactions with intra-cellular components.
Oliver et al. (2018) reported that antimicrobial activities AgNPs having
more than 20 nm size depend on Ag+ ion release, whereas those having
less than 10 nm size are more effective antimicrobial, as they directly
enter into microbial cells and interact with cellular components and
enzymes (Oliver et al., 2018). Mei et al. (2017) also reported superior
antimicrobial activity of ultra-small silver nanocluster (AgNCs)
compared to that of AgNPs larger than 10 nm due to high surface to
volume ratio, high local surface concentration, and high mobility (Mei,
Teng, Zhu, Liu, Zhang, Zhang, et al., 2017).
In addition to size, shape of the Ag NPs also affects antimicrobial
activity, as it is associated with their interaction to the cell wall/mem­
brane of microorganisms. In general, different shapes of AgNPs such as
spherical, oval, rectangular, rod-shaped, and polyhedral have been
studied for their antimicrobial activities. Spherical AgNPs were more
effective antimicrobial than the rod shaped ones (Zhang, Liu, Shen, &
Gurunathan, 2016). It has also been reported that triangular shaped
AgNPs had higher microbial activity against E. coli compared to the
spherical and rod-shaped NPs (Acharya et al., 2018). On the other hand,
dos Santos et al. (2017) reported that antimicrobial efficacy of spherical
AgNPs against Streptococcus mutans was not significantly different from
that of triangular AgNPs (dos Santos Junior, Targino, Flores, Rodrí­
guez-Díaz, Teixeira, Heimer, et al., 2017). Although, report have sug­
gested that triangular AgNPs are more effective antimicrobial agents,
but there is no conclusive data on effect of shape of AgNPs on antimi­
crobial efficacy.
3.2.3. Concentration of AgNPs
Concentration of AgNPs also strongly correlates with their antimi­
crobial efficiency. Studies reported that higher the concentration of
nanoparticles, greater is the antimicrobial effect, and vice-versa (S.
Kumar et al., 2014, 2016). Recently, MICs of AgNPs against human
pathogens such as S. mutans, and S. orlis were reported to be 45–60
μg/mL, and those against food spoilage bacteria such as Lactobacillus
acidophilus and L. fermentum ranged from 15 to 90 μg/mL (Panpaliya
et al., 2019). MIC for AgNPs against biofilms of foodborne pathogens
like Staphylococcus aureus and Pseudomonas aeruginosa were about 62.5
μg/mL, whereas 85 and 90% of the biofilm inhibition were achieved at

Fig. 6. Sustainable food packaging applications of AgNPs.

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S. Kumar et al.
AgNPs concentrations of 125 and 250 μg/mL, respectively (Gupta et al.,
2019).
4. Antimicrobial and sustainable food packaging applications of
AgNPs
AgNPs are promising antimicrobial nanomaterials that are effective
against a broad range of microorganisms, including bacteria, yeasts,
fungi, and viruses. In recent times, scientists have garnered huge interest
in green synthesis of AgNPs owing to their applications in food in­
dustries, including sustainable food packaging (Fig. 6) (Mei & Wang,
2020). Nanomaterial-based food packaging can be categorized into two
groups; (a) improved packaging in which nanomaterials are incorpo­
rated into the polymer matrix to enhance its mechanical and barrier
properties; and (b) nanocomposite-based antimicrobial packaging that
can protect the food from food spoilage microorganisms, and improve
the shelf-life of the food.
Sustainable food packaging (SFP) is an innovative development so­
lution to the food packaging industry (also global market demands for its
cost and performance), which is beneficial, safe, and healthy for in­
dividuals, communities, and the environment throughout its life cycle
(Chemat et al., 2017). SFP products should be produced from recycled
material, and it generates zero-landfill, minimizes the use of water and
efficient enough to utilize renewable resources. Besides, due to these key
characteristics, SFP must also fulfill its purpose and meet relevant in­
ternational standards to protect packaged food items through effective
communication strategies with stakeholders across the entire food
supply chain (Chemat et al., 2017).
4.1. Bio-plastic based antimicrobial food packages/films
According to the United Nations, over 8.3 billion metric tons of
plastic was created as of 2017 and that number is rising alarmingly. The
United Nations Environment Programme (UNEP) urges researchers and
entrepreneurs to seek alternatives to plastics because of their non-
biodegradable properties that pose a serious threat to the environment
(Löhr et al., 2017). Biodegradable plastics made from renewable mate­
rials (plants, seaweed, bacteria, etc.) are valued to be an alternative
source to petroleum-based plastics, and they not only contribute to
sustainability but also provide feasible solutions to the growing need for
an eco-friendly food packaging market.
Research on developing food packaging materials using bio-
nanocomposites has seen a sharp rise over the past decade. Accord­
ingly, these bio-plastics can be classified into three categories, namely
(1) natural biopolymers derived from natural resources, such as poly­
saccharide (starch, chitosan, carrageenan, cellulose, and alginate),
protein (gelatin, soy protein, wheat gluten and collagen), and lipids
(carnauba wax, and, shellac wax); (2) synthetic polymers derived from
natural monomers; such as polylactic acid (PLA); and (3) microbial
polymers produced by natural or genetically-modified microorganisms,
such as bacterial cellulose, polyhydroxyalkanoates (PHA), including
poly (3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) and poly
(β-hydroxybutyrate) (PHB) (Fabra, López-Rubio, & Lagaron, 2014).
AgNPs are used as active antimicrobial agents in food packaging
systems, which can extend the shelf-life of food for prolonged periods by
inhibiting the growth of foodborne pathogenic and spoilage microor­
ganisms. There has been a considerable number of studies on green
synthesis of AgNPs and their applications in food packaging systems to
control foodborne pathogens. On that account, polyethylene oxide
(PEO) nanocomposite films together with AgNPs and Acca sellowiana
extract exhibited antibacterial activity against S. aureus and E. coli
(Sganzerla, Longo, de Oliveira, da Rosa, de Lima Veeck, de Aquino et al.,
2020). In a similar study, methylcellulose films carrying plant-mediated
AgNPs were designed using Lippia alba plant extract, which also
demonstrated antimicrobial activity against S. aureus and E. coli (Nunes,
de Souza Maguerroski Castilho, de Lima Veeck, da Rosa, Noronha,
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Trends in Food Science & Technology 112 (2021) 651–666
Maciel, et al., 2018). Evidently, it is understood that the presence of
methylcellulose and PEO protects phenolic compounds from degrada­
tion and thereby establishing the ideal conditions for biodegradable
food packaging material. Antibacterial activity against E. coli and
L. monocytogenes was also noticed when green synthesized AgNPs using
organosolv lignin (a reducing agent) were imbedded into polylactic acid
(PLA) to prepare composite films (Shankar, Rhim, & Won, 2018).
Recently, green synthesis of ginger extract generated AgNPs incorpo­
rating biodegradable polyvinyl alcohol (PVA) and clay (montmor­
illonite-MMT) seems to be an effective approach for packaging of food
materials, such as chicken sausage. This study also indicates that due to
presence of AgNPs imbedded within MMT and PVA, it took 110 days to
completely degrade the packaging film in comparison with different
blend-based composites (films) and also efficient to inhibit the growth of
microbial pathogens (Mathew, S, Mathew, & E.K, 2019).
More recently, AgNPs were synthesized using leaf broth of Capparis
zeylanica and incorporated with PVA and polyethylene glycol (PEG)
biopolymer, that displayed significant antimicrobial activity (Ernest
Ravindran, Subha, & Ilangovan, 2020). Moreover, polylactic acid (PLA)
films render a new sort of biodegradable material, which is produced
from plant-based natural resources (corn, starch, and sugar beets), have
acquired FDA accreditation authorizing food-contact applications (Zhu,
Tang, Yin, & Yang, 2018). These films show a greater level of tensile
strength and water-resistance that can be employed to improve food
safety standards and to extend the shelf-life of fresh produce (Chi et al.,
2019; Xing, Li, Wang, Li, Xu, Guo, et al., 2019). Additionally, integration
of AgNPs in agar-banana powder mix films exhibited unique antimi­
crobial activity against foodborne pathogens, such as E. coli and
L. monocytogenes (Orsuwan, Shankar, Wang, Sothornvit, & Rhim, 2016).
Encapsulation of probiotic cells in edible films is not new to the scientific
world. However, due to the instability during food processing and poor
performance in gastrointestinal tract, its relevance is of not great value.
Henceforth, incorporation of AgNPs into natural polymers may improve
the quality of the films with better physico-chemical and mechanical
properties (Kraśniewska, Galus, & Gniewosz, 2020). In a recent study,
furcellaran, a low-sulfated polysaccharide obtained from Furcellaria
lumbricalis (red algae) was used to synthesize AgNPs, which can form a
stable nanocomposite film, and fight against pathogenic microorganisms
like C. albicans, E. coli, Enterococcus facealis, S. aureus, and Pseudomonas
aeruginosa. These nanocomposite films have not only potential antimi­
crobial activities but also considered to be an eco-friendly approach
exhibiting good mechanical and barrier properties, including UV radi­
ation resistance and water vapor abilities (Jamróz, Khachatryan, Kopel,
Juszczak, Kawecka, Krzyściak, et al., 2020).
4.2. Biopolymer-based antimicrobial edible coatings
Applications of edible coatings on perishable foods have a long his­
tory. The first edible coating to be used on fruits and vegetables during
twelfth and thirteen centuries is “wax” that arrests respiratory gas ex­
change (Hardenburg, 1967). Number of studies on the application of
plant-mediated AgNPs on perishable food surfaces have been reported.
Edible coatings are also used as a matrix for introducing antimicrobial
nanoparticles that can extend the shelf-life and storage quality of fruits
and vegetables. For instance, calcium-alginate coating filled with silver-
montmorillonite nanoparticles applied on fresh-cut carrots was able to
reduce respiration rate under controlled atmosphere storage (low oxy­
gen concentration) that can double the shelf-life of the end-product
(Costa, Conte, Buonocore, Lavorgna, & Del Nobile, 2012). Similarly,
the effect of carboxy methyl cellulose (CMC) and guargum-based coat­
ings derived AgNPs significantly enhanced antioxidant activity and
flavor stability of Kinnow fruit (Citrus reticulata) for nearly 4 months at
4 ◦ C (Shah, Jahangir, Qaisar, Khan, Mahmood, Saeed, et al., 2015). In
another study, fresh-cut melon coated with Ag-chitosan nanocomposites
not only inhibited microbial growth but also exhibited lower respiration
rate together with enhanced vitamin C levels and prolonged shelf-life
S. Kumar et al. Trends in Food Science & Technology 112 (2021) 651–666

Fig. 7. TEM images of duodenum of mice fed with

300 mg/kg/day of AgNPs for 3 days; (A) Duo­
denum of control mouse, (B) Duodenum of female
mice showing AgNPs along microvilli (circled) and
in the cell cytoplasm (arrow), (C) Duodenum of
female mice showing bulk amount of AgNPs over
and under the nucleus (N, nucleus), (D) Enlarged
squared area of image C, (E) Duodenum of male
mice showing an aggregation of AgNPs within a
membrane bound structure, and (F) Enlarged
squared area of image E. Reproduced with
permission from (Narciso et al., 2020) Copyright
2020, Elsevier.

after 13 days at 5 ◦ C. This suggests that Ag-chitosan biopolymer nano­ nanofillers can induce the release of Ag+ ions from AgNPs in food con­
composites might be one of the most promising approaches for fresh-cut tact materials (FCMs) (Carbone, Donia, Sabbatella, & Antiochia, 2016).
active food packaging systems (Ortiz-Duarte, Pérez-Cabrera, Arté­ Release of Ag+ ions in dissolved form and in nanoparticles form among
s-Hernández, & Martínez-Hernández, 2019). FCMs can be quantified and analyzed using ICP–MS and SEM–EDX
analysis for testing their toxicity. Not many countries around the globe
were able to implement certain standard safety guidelines and regula­
4.3. Release studies of AgNPs from film/package to food
tions regarding the release of AgNPs in Food and FCMs. For instance, the
European Food Safety Authority (EFSA) recommends permissible limit
There is a growing demand to increase the shelf-life of packaged
of Ag+ ions release from food packaging, which should not exceed 0.05
foods and protect them from foodborne pathogens. AgNPs demonstrate
mg/L in water and 0.05 mg/kg in food, whereas USFDA and Canada are
excellent antimicrobial activity towards many foodborne pathogens and
yet to set proper guidelines for safer utilization of AgNPs in food and
economical among other nanomaterials that have become the finest
FCMs (Carbone et al., 2016). Therefore, it is very important to examine
choice for many researchers and food industries to embed them in
new packaged food material combined with AgNPs release from food
various biodegradable packaging materials. These AgNPs can be coa­
packaged materials. However, due to the slow release of Ag+ ions in
lesced with edible (organic materials) and non-degradable polymers
FCMs, AgNPs provide great strength in stored food materials, and they
(inorganic materials) to improve gas-barrier, mechanical properties,
can be ideal candidates in food packaging industry. When a detailed
thermal features, antimicrobial properties for sustainable food pack­
study was conducted on the release of Ag+ ions from various types of
aging. The antimicrobial activity becomes competently effective when
nanomaterials, such as low-density polyethylene (LDPE) and poly­
foodborne pathogens come in contact with well-designed nanoparticles
propylene into the food, the release of Ag+ ions was way more than the
in polymeric matrices. This approach will maintain the food quality and
specified permissible limits due to acidic nature and conventional oven
also enhance the shelf-life of the food (Nafchi, Alias, Mahmud, & Robal,
conditions (Echegoyen & Nerín, 2013). Release of a few low molecular
2012).
weight materials from polymeric matrices can destroy or degrade the
Despite having many advantages of AgNPs, there is a rising concern
quality of FCMs, and for this reason, it is not recommended to utilize
amongst researchers about their unusual kind of physicochemical
such materials in food packaging (Fahmy et al., 2020). Additionally,
properties (types of AgNPs, characteristics, concentrations and surface
when AgNPs combined with starch were proved to be effective
chemistry). Furthermore, acidic foods, microwave heating and

662
S. Kumar et al.
antibacterials and improved the mechanical strength of the packaging
film. Besides, the release of Ag+ ions from nanocomposite-film was
within the recommended limit (Talegaonkar, Sharma, Pandey, Mishra,
& Wimmer, 2017). In order to trace the release of AgNPs to FCMs,
dedicated set of guidelines and regulations such as “ESPA” has to be
implemented by all the countries to maintain the safety and quality of
food and feed (Chaudhary, Fatima, & Kumar, 2020).
5. Toxicological and hazardous aspects of AgNPs
AgNPs are effective biocides against various pathogenic and spoilage
microorganism and can be used as antimicrobial agents to enhance the
shelf-life of food. However, uncontrolled migration of Ag+ ions from
packaging to food may increase the risk of consumer health hazard. The
European Food Safety Authority (EFSA) mandates the upper limits of
Ag+ ion release from package not more than 0.05 mg/L in water and
0.05 mg/kg in food. Determination of Ag+ ion release from AgNP-based
nanocomposite package and their accumulation in the packaged food is
essential to ensure consumer safety. Although there are several reports
on toxicity of nano-silver beyond the recommended limit, long term
exposure and resulting disease need further research to fulfill the
knowledge gap in this area. Gliga et al. (2000) reported toxicological
effect of AgNPs and revealed that intracellular release of AgNPs was
responsible for the toxicity to human lung cells. Several studies have
reported that AgNPs may accumulate in several organs such as kidneys,
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Trends in Food Science & Technology 112 (2021) 651–666
Fig. 8. TEM images of liver of mice fed with 300
mg/kg/day of AgNPs for 3 days; (A) Liver of con­
trol mouse (N, nucleus), (B) Liver of male mice
showing an accumulation of nanoparticles next to
mitochondria (arrow) (P, peroxisomes), (C)
Micrograph of liver of female mice showing many
AgNPs within a membrane bound structure, (D)
Enlarged squared area of image C, (E) Liver of male
mice showing AgNPs in two membrane-bound
structures, (F), Enlarged squared area of E.
(Reproduced with permission from (Narciso et al.,
2020) Copyright 2020, Elsevier).
liver, brain, and testicles (Bagheri-Abassi, Alavi, Mohammadipour,
Motejaded, & Ebrahimzadeh-Bideskan, 2015) (Jorge, José, & Miguel,
2011). An experiment carried out on Sprague Dawley adult rats showed
that the oral dose of 50, 100 and 200 mg/kg/day AgNPs resulted in
accumulation of Ag+ ions in different tissues. High doses of AgNPs may
cause hepatotoxic (El Mahdy, Eldin, Aly, Mohammed, & Shaalan, 2015),
neurotoxic (Bagheri-Abassi et al., 2015), and genotoxic effects (El
Mahdy et al., 2015) on animals. Toxicological and bio-distribution
assessment of AgNPs conducted on male and female mice with 50,
150, 300 mg/kg/day body weight oral administration for three days
showed that AgNPs were mostly accumulated in duodenum tissues
(Fig. 7) followed by subsequently migration to liver (Fig. 8), spleen and
kidney (Narciso, Coppola, Lori, Andreoli, Zjino, Bocca, et al., 2020). It
was evident from TEM analysis that AgNPs were present in the cyto­
plasm or remained membrane bound, and never entered the nuclei,
resulting in lack of genotoxic damage in any of the internal organs and
blood (Figs. 7 and 8) (Narciso et al., 2020). However, such high toxic
levels of Ag+ ion migration from active packaging to foods are very rare,
and risk of disease as a consequence of Ag+ ion migration from packages
has not been fully assessed so far. Additionally, plant-mediated synthesis
renders the resulting AgNPs less harmful to the environment due to
adsorption of natural organic matter on their surfaces preventing
exposure to oxidation and formation of harmful Ag+ ions (Jorge de
Souza, Rosa Souza, & Franchi, 2019), and to animal and plant tissues
due to their biocompatibility. AgNPs synthesized by phyto-extracts of
S. Kumar et al.
three Indian medicinal plants were non-toxic to seeds of common Indian
pulses like mung bean and chickpea (Banerjee, Satapathy, Mukhopa­
hayay, & Das, 2014). Rónavári et al. (2017) reported that coffee
extract-mediated AgNPs have no harmful effect of on human and mouse
cells, suggesting that plant-mediated AgNPs can be a better alternative
for antimicrobial applications in sustainable food packaging (Rónavári,
Kovács, Igaz, Vágvölgyi, Boros, Kónya, et al., 2017).
6. Conclusion and future perspectives
Silver nanoparticles (AgNPs) are well-studied nanofillers for food
packaging applications due to their superior physicochemical, optical,
thermal and antimicrobial properties. The plant-based methods for
synthesis of AgNPs seems to be promising because they require non-
toxic chemicals for the reduction of silver ions. These methods are
cost-effective, biocompatible, environment-friendly, and can be carried
out at a large-scale for commercial production. Incorporation of plant-
mediated AgNPs into biopolymer-based nanocomposite films and coat­
ings may lead to improve mechanical, barrier, thermal, and antimicro­
bial properties of these films. Such improved films and coatings have
been evaluated for food packaging applications, and they have been
reported to be effective in prolonging the post-harvest life of numerous
perishable foods such as fresh produce, dairy, fish and meat products.
These bionanocomposite-based packagings are antimicrobial in nature
and contribute significantly to sustainable food packaging, thus sug­
gesting their potential application in food safety and shelf-life extension
of perishable food. Though future research is needed, to evaluate the
concentration Ag+ ions released from various biopolymer-based nano­
composite primary packagings or food contact materials, in order to
ensure prevention of any toxicity in the packaged food. This review
provides a wide spectrum on the use of plant-based natural resources for
the production of AgNPs, and their applications in food packaging and
preservation.
Acknowledgments
Shruti Shukla would like to thank Department of Biotechnology
(DBT) - Ramalingaswamy Fellowship Grant (D.O. NO. BT/HRD/35/02/
2006), Government of India for collaborative support. SK is very much
thankful to Dr. Dipankar Halder, Associate Professor, Department of
Food Technology and Biochemical Engineering (FTBE), Jadavpur Uni­
versity, Kolkata and Dr. Atanu Mitra, Associate Professor, Sree Chaita­
nya College, Habra for their continuous support and encouragement to
complete this review article.
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