Environ Chem Lett (2018) 16:101–112
https://doi.org/10.1007/s10311-017-0670-y
REVIEW
Chitosan nanoparticles preparation and applications
K. Divya1 · M. S. Jisha1   
Received: 31 May 2017 / Accepted: 11 October 2017 / Published online: 31 October 2017
© Springer International Publishing AG 2017
Abstract  Shell fish processing industry is very common in
coastal areas. While processing, only the meat is taken, the
head and shells are discarded as waste. On an average, the
sea food industry produces 80,000 tons of waste per year.
The sheer amount of waste makes degradation a slow pro-
cess causing accumulation of waste over a period of time.
A very simple and effective solution to this environmental
hazard is the recycling of shell waste to commercially viable
products like chitin. Chitosan is the N-acetyl derivative of
chitin obtained by N-deacetylation. Chitosan is widely used
in food and bioengineering industries for encapsulation of
active food ingredients, enzyme immobilization, as a car-
rier for controlled drug delivery, in agriculture as a plant
growth promoter. Chitosan is also a defense elicitor and an
antimicrobial agent. Chitosan has interesting properties such
as biodegradability, biocompatibility, bioactivity, nontoxic-
ity and polycationic nature. This review presents structural
characteristics and physicochemical properties of chitosan.
The methods of preparation of chitosan nanoparticles are
detailed. Applications of chitosan nanoparticles are dis-
cussed. Applications include drug delivery, encapsulation,
antimicrobial agent, plant growth-promoting agent and plant
protector.
Keywords  Chitin · Chitosan · Chitosan nanoparticles ·
Antimicrobial action · Agriculture
* M. S. Jisha
jishams@mgu.ac.in
1
School of Biosciences, Mahatma Gandhi University, Kerala,
India
Introduction
Nanoparticles range in dimension from 1 to 100 nm. They
have unique properties compared to their bulk equivalents
due to the decrease in dimension to atomic level (Ravis-
hankar Rai and Jamuna Bai 2011). The properties of materi-
als change at the nanoscale. This is because bulk materials
have relatively constant properties regardless of their size,
but as the size decreases, the percentage of surface atoms
compared to bulk material increases. This causes unexpected
properties of nanoparticles (Gupta et al. 2007).
Nanoparticles are synthesized by size reduction using
either top–down methods such as milling, high-pressure
homogenization and sonication or bottom–up processes
like reactive precipitation and solvent displacement (Vau-
thier et al. 2003).
Nanoparticles are grouped into organic and inorganic
nanoparticles. The inorganic nanoparticles have gained sig-
nificant importance due to their ability to withstand adverse
processing conditions (Whitesides 2003). Metal oxide nano-
particles such as titanium oxide, zinc oxide, silver oxides
and magnesium oxides are of great interest among inorganic
materials due to their tunable optical properties and physical
and optical stability (Makhluf et al. 2005). Due to the unique
electronic, metallic and structural characteristics, organic
materials like carbon nanotubes, lipids and polymers have
versatile applications (Hatton et al. 2008).
Polymeric nanoparticles can be synthesized from natural
and synthetic polymers. They are used owing to their stabil-
ity and ease of surface modification. Biopolymeric nanopar-
ticles have added advantages, like availability from marine
(chitin and chitosan) or agricultural (cellulose, starch,
pectin) resources, biodegradability, biocompatibility and
nontoxicity. Biodegradable polymers such as chitosan are
studied mainly as delivery systems for controlled release
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102
of active ingredients, stabilization of biological molecules
like proteins, peptides or genetic material (Ghormade et al.
2011).
Chitosan is a modified biopolymer, derived by partial
deacetylation of chitin. It consists of alternating units of
(1 → 4) linked N-acetyl glucosamine and glucosamine units.
It is a white, hard, inelastic and nitrogenous polysaccha-
ride (Badawy and Rabea 2011). Chitosan finds multifaceted
applications due to its nontoxicity, biodegradability and
antimicrobial properties. It is used in biomedical industries,
agriculture, genetic engineering, food industry, environmen-
tal pollution control, water treatment, paper manufacture,
photography and so on (Cheba 2011).
Chitosan nanoparticles (ChNP) have the characteris-
tics of chitosan and the properties of nanoparticles such as
surface and interface effect, small size and quantum size
effects (Ingle et al. 2008). Owing to the enormous potential
of ChNP, this review explores the structural characteristics
of chitosan and the different preparation methods of ChNP.
Special emphasis will be placed on the application of ChNP.
Chitosan‑structure and physicochemical properties
Chitin was first discovered in 1811 by Henri Braconnot
while conducting research in mushrooms. Later in 1859,
Prof. C. Rouget found that alkali treatment of chitin yielded
a substance that unlike chitin can be dissolved in acids.
Hoppe Seiler called this deacetylated chitin ‘Chitosan’ (Bad-
awy and Rabea 2011).
Chitin is the wide-spread biopolymer in nature after cellu-
lose. It is the major component of cuticles of insects, fungal
cell walls, yeast or green algae (Einbu and Vayrum 2008).
It is also present in crab and shrimp shells (Wang and Xing
2007). Chitosan, on the other hand, is much less abundant in
nature. It has been found only in cell walls of certain fungi
(Muzzarelli and Gooday 1986).
Chitin is a homopolymer of β 1-4 linked N-acetyl D-glu-
cosamine (Glc NAc; A unit) residues (Yen et al. 2009).
There is mainly three classes of chitin-α, β and γ chitin.
α-Chitin has antiparallel chains while β-chitin has intrasheet
Fig. 1  Structure of chitin and chitosan
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Environ Chem Lett (2018) 16:101–112
hydrogen bonding by parallel chains. γ-Chitin is a combi-
nation of α and β chitin and has parallel and antiparallel
structure (Yen and Mau 2007) (Franca et al. 2008). Chitosan,
obtained by deacetylation of chitin, has β 1-4 linked A unit
(chitin monomer) and 2-amino 2-deoxy-β-D-glucopyranose
(Glc N; D units; chitosan monomer) (Park et al. 2011; Puv-
vada et al. 2012; Shahidi et al. 1999).
Chitosan contains at least 60% D units (Kumirska et al.
2011). The molar fraction of D units is expressed as the
degree of deacetylation (DD) (Aranaz et  al. 2009). The
structure of chitin and chitosan is shown in Fig. 1. It is an
important characteristic that influences the performance of
chitosan in many applications (Kumirska et al. 2010). DD
can be determined by potentiometric titration (Zhang et al.
2011), infrared radiation (Baxter et al. 1992), UV–visible
spectrophotometry (Kasaai 2009), gel permeation chroma-
tography (Kumar 1999), 1H-liquid-state NMR and solid-
state 13C NMR (Ottey et al. 1996). The presence of the free
amine groups along the chitosan chain makes it unlike chitin
soluble in diluted acidic solvents.
The molecular weight and viscosity development in aque-
ous solution also play a significant role in the biochemical
and pharmacological application of chitosan. Other major
parameters are crystallinity, ash content, moisture content,
heavy metal content and so on (Rinaudo 2006).
In addition to its many applications, chitosan is also an
eco-friendly solution to the pollution caused by the seafood
processing industry. Every year, 60,000–80,000 tons of shell
waste are produced globally. This sheer amount of waste
makes degradation a slow process and an environmental
concern. Conversion of shell waste to chitin is an effective
solution to this problem. Chitin has many applications and
can also be deacetylated to form chitosan which has a myriad
of applications (Divya et al. 2014).
Structural modifications of chitosan
Chitosan contains three functional groups; an amino group
and primary and secondary hydroxyl groups at C2, C3
and C6 positions. The hydroxyl groups of chitosan make a
Environ Chem Lett (2018) 16:101–112 103
chemical modification by attaching side groups to the reac-
tive hydroxyl groups without altering its biophysical proper-
ties (Rajasree and Rahate 2013). Crosslinking chitosan with
glyoxal, glutaraldehyde and terephthaldehyde results in a
hydrogel that can be used in organ transplants and restor-
ing organ function (Kumar and Koh 2012). N-imidazolyl-
O-carboxymethyl chitosan has been used for high-perfor-
mance gene delivery (Shi et al. 2011a, b). Radionuclides like
Ho-166, Sm-153 and Lu-166 crosslinked with chitosan are
used for targeted therapy (Zolghadri et al. 2010). Chemical
modification of chitosan by adding quaternary ammonium
groups (Thanou et al. 2001), carboxy alkyl groups (Aiping
et al. 2006) and acetic anhydrides (Hirano et al. 2002) has
also been reported.
Chitosan nanoparticle production
Chitosan has the ability to form a gel on contact with anions
and form beads. This property enables its use in drug deliv-
ery. But still, the large size of these beads (1–2 mm) limits
its application (Shiraishi et al. 1993).
Fig. 2  Schematic representation of different modes of ChNP synthesis and its various applications
Chitosan nanoparticles (ChNP) were first described in
1994 by Ohya and co-workers. They used ChNP prepared
by emulsifying and crosslinking for intravenous delivery
of anticancer drug 5-fluorouracil (Grenha 2010). Since
then, many methods have been employed for the synthesis
of ChNP. Five methods are presently available. They are
ionotropic gelation, microemulsion, emulsification solvent
diffusion, polyelectrolyte complex and reverse micellar
method (Tiyaboonchai 2003). Out of this, the most widely
used methods are ionotropic gelation and polyelectrolyte
complex. These methods are simple and do not apply high
shear force or use organic solvents (Sailaja et al. 2011). The
schematic representation of different methods of ChNP syn-
thesis is depicted in Fig. 2.
Ionotropic gelation
This technique was first reported by Calvo et al. (1997) and
has been widely examined and developed. The method uti-
lizes the electrostatic interaction between the amine group
of chitosan and a negatively charged group of polyanion
such as tripolyphosphate. Chitosan can be dissolved in
acetic acid in the absence or presence of the stabilizing
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104
agents, such as poloxamer. Polyanion was then added, and
nanoparticles were formed spontaneously under mechan-
ical stirring at room temperature. The size and surface
charge of particles can be modified by changing the ratio
of chitosan to the stabilizer. A general increase in parti-
cle compactness and size was observed on increasing the
chitosan concentration and on increasing the polymer to
polyanion ratio (Jonassen et al. 2012). They also reported
that nanoparticles dispersed in saline solution were more
stable due to the smaller particle size found in the presence
of sodium chloride. This is because a monovalent salt like
sodium chloride when added to the solvent screens out to
the electrostatic repulsion between the positively charged
amine groups on the chitosan backbone. This will increase
the flexibility of the polymer chains in solution and thus
increase its stability (Ilium 1998).
Microemulsion method
This method was first reported by De et al. (1999). Accord-
ing to this method, a surfactant was dissolved in N-hexane
and chitosan in acetic solution and glutaraldehyde was
added to surfactant/hexane mixture under continuous
stirring at room temperature. Nanoparticles were formed
in the presence of a surfactant. The system was stirred
overnight to complete the crosslinking process, between
the free amine group of chitosan and glutaraldehyde.
The glutaraldehyde in this method acts like a crosslinker
(Fang et al. 2009). The organic solvent is then removed by
evaporation under low pressure, and excess surfactant was
removed by precipitate with ­CaCl2 and then the precipitant
was removed by centrifugation. The major disadvantage of
this method is the use of antigenic agent glutaraldehyde.
Also, the incorporation of protein or peptides to nano-
particles is not possible as they may be damaged by the
covalent crosslinking (Calvo et al. 1997).
Emulsification solvent diffusion method
This method was first reported by El-Shabouri (2002).
It is a modified method developed by Niwa et al. (1993)
employing PLGA. An emulsion is obtained upon injec-
tion of an organic phase into chitosan solution containing
a stabilizing agent such as poloxamer under mechanical
stirring, followed by high-pressure homogenization. The
emulsion is then diluted with a large amount of water. Pol-
ymer precipitation occurs due to the diffusion of organic
solvent into the water, thus forming nanoparticles. The
major disadvantages of this method include the high shear
forces used during nanoparticle preparation and the use of
organic solvents.
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Environ Chem Lett (2018) 16:101–112
Polyelectrolyte complex (PEC)
Polyelectrolyte complexes formed by self-assembly of the
cationic charged polymer and plasmid DNA as a result of
fall in hydrophilicity due to charge neutralization between
cationic polymer and DNA. The ChNP can be spontaneously
formed on addition of DNA solution into chitosan dissolved
in acetic acid solution, under mechanical stirring at room
temperature (Erbacher et al. 1998).
Reverse micellar method
This method was reported by Brunel et al. (2008). The major
highlight is the absence of both crosslinker and toxic organic
solvents. Also, ultrafine nanoparticles within a narrow size
range can be obtained with this method. An aqueous solu-
tion of chitosan is added to the organic solvent containing
surfactant under constant agitation to form reverse micelles
(Zhao et al. 2011).
Preparation of chitosan nanofibers
Chitosan nanofibers are solid particles with a diameter
range of 1–1000 nm. Although there are many methods for
nanofiber synthesis, electrospinning process has attracted
attention since it produces nanofibers with a size range of
micrometers to nanometers (Jayakumar et al. 2010a, b). Due
to its popularity, only electrospinning process is discussed
here.
Chitosan gets protonated in acidic solution changing it
into a polyelectrolyte. When the high electric field is applied
during electrospinning, repulsive forces arise between ionic
groups within the polymer, thus producing beads instead of
continuous fibers. This restricts the fabrication of pure chi-
tosan (Min et al. 2004). This problem was solved by Ohkawa
and team by using trifluoroacetic acid (TFA) as a solvent.
The amine groups of chitosan form salts with TFA thus
eradicating the intramolecular interaction between chitosan
molecules (Ohkawa et al. 2004). Acetic acid has also proved
to be effective in producing chitosan nanofibers (Geng et al.
2005).
Electrospinning of chitosan usually produces beads due to
an inadequate stretch of filaments during the whipping of jet
due to low charge density (Sun and Li 2011). To overcome
this, nanofibers of blends of chitosan and synthetic polymers
such as poly vinyl alcohol (PVA), poly ethyl oxide (PVO)
and poly ethylene terephthalate (PET) have been produced
recently (Jia et al. 2007). PVA and PEO are mainly used
for biomedical applications like bone implant (Allen et al.
2004), artificial organs (Chen et al. 1994), wound dressing
Environ Chem Lett (2018) 16:101–112 105
(Yoshii et al. 1999), cartilage tissue repair (Sims et al. 1996),
and so on. Pet is commonly used in textile and plastic indus-
try (Sims et al. 1996).
Application of chitosan nanoparticles
Chitosan nanoparticles are natural materials with excellent
physicochemical, antimicrobial and biological properties,
which make them a superior environmentally friendly mate-
rial and they possess bioactivity that does not harm humans
(Malmiri et al. 2012). Due to these unique properties, chi-
tosan nanoparticles find a wide array of applications. Some
of them are discussed below.
Tissue engineering
Tissue engineering is the use of living cells that have been
manipulated either by genetically or by their extracellu-
lar environment, for developing biological substitutes for
implantation into the body or for remodeling tissues through
some active mechanism. The purpose of tissue engineering
is to repair, replace, maintain or enhance the function of a
particular tissue or organ (Jayakumar et al. 2010a, b).
Chitosan nanoparticles, due to its biological and mucoad-
hesive properties, can improve transmucosal permeability,
thereby enhancing transport through the paracellular path-
way of the nanoparticles and can induce structural reorgan-
ization of tight junction-associated proteins (Peppas and
Huang 2004).
Cancer diagnosis
Semiconductor nanocrystals (or quantum dots) are the most
promising fluorescent probes for many biomedical applica-
tions (Jayakumar et al. 2010a, b). In spite of the success in
using this nanocrystal, there arises the problem of cytotox-
icity of their heavy metal composition. Chitosan nanoparti-
cles owing to its non-toxic nature gains importance in this
respect. The anticancer activity of chitosan nanoparticles
can be attributed to its small size. The small particle size
increases the specific surface area and surface to volume
ratio which in turn increases the dissolution resulting in bio-
availability of chitosan (Ghadi et al. 2014).
Manjusha et al. (2010) developed folic acid (FA) con-
jugated carboxymethyl chitosan (CMCS) coordinated to
manganese-doped zinc sulfide (ZnS: Mn) quantum dot (FA-
CMCS-ZnS: Mn) nanoparticles which find application in
targeting, controlled drug delivery and imaging of cancer
cells. Anticancer drug 5-fluorouracil used for the breast can-
cer treatment was selected for the study. The nontoxicity,
imaging, specific targeting and cytotoxicity of FA-CMCS-
ZnS: Mn were studied.
Drug delivery
The potential use of chitosan nanoparticles as carriers has
paved way for development of wide variety of colloidal
delivery vehicles (Malmiri et al. 2012). Chitosan nanopar-
ticles can cross biological barriers to protect macromole-
cules from degrading in biological media. It can also deliver
drugs or macromolecules by controlled release to a target
site (Lopez-Leon et al. 2005; Perera and Rajapakse 2013).
The small size of ChNP also makes it efficient in interfacial
interaction with cell membrane because the small particles
will be taken up by cell by endocytosis (Ghadi et al. 2014).
Several studies have been reported regarding the ability
of chitosan nanoparticles to improve the bioavailability of
drugs, modifying its pharmacokinetics and protecting the
encapsulated drugs (Janes et al. 2001; Shi et al. 2011a, b).
Apart from being used as an oral delivery carrier, ChNP
can also be applied to other mucous membrane systems like
pulmonary and nasal routes to deliver peptides and proteins
(Fernandez-Urrusuno et al. 1999).
Enzyme immobilization support
Chitosan is known as an ideal material for enzyme immobi-
lization due to its various properties like improved resistance
to chemical degradation and avoiding disturbance of metal
ions to an enzyme (Vazquez-Duhalt et al. 2001; Yang et al.
2010). The amino functional group of chitosan makes it suit-
able for enzyme immobilization (Ghadi et al. 2015).
Liu et al. (2005) studied trypsin immobilized on lino-
lenic acid-modified chitosan nanoparticles using glutar-
aldehyde (GA) as crosslinker and found that the thermal
stability and optimum temperature of immobilized trypsin
increased. Ghadi et al. (2015) reported that chitosan mag-
netic core shell nanoparticles are capable of immobilizing
lipase enzyme. The strong bond between chitosan and lipase
increases the enzyme adsorption and enzyme loading.
Antioxidant activity
Chitosan is a proved antioxidant agent (Rajalakshmi et al.
2013). It can scavenge free radicals and chelate metal ions
by donating a hydrogen or a lone pair of electrons (Lin et al.
2009). The amino and hydroxyl functional groups of chi-
tosan interact with metal ions triggering many activities such
as adsorption, chelation and ion exchange (Onsosyen and
Skaugrud 1990). The semicrystalline structure of chitosan
and the strong hydrogen bonds ensures that chitosan can not
be dissociated from the metal ions (Xie et al. 2001). Chi-
tosan/fucoidan nanoparticles showed DPPH and ROS radical
scavenging activity (Huang and Li 2014). Yen et al. (2008)
reported that chitosan exhibited hydroxyl radical scavenging
activity and iron chelating ability.
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106 Environ Chem Lett (2018) 16:101–112

Encapsulation of biologically active compounds
Chitosan-based systems have wide and rapidly increasing
applications in the food and biochemical industries. Any
ingredients can be encapsulated, irrespective of it being
hydrophobic, hydrophilic or bacterial (Zhao et al. 2011).
Chitosan retains the bioactivity of macromolecules such as
DNA and proteins during encapsulation. The positive charge
of chitosan helps it to establish a strong interaction with
negatively charged molecules without altering its activity
(Mohammadpour Dounighi et al. 2012).
Jang and Lee (2008) investigated the stability and charac-
teristics of vitamin C-loaded chitosan nanoparticles prepared
by ionic gelation of chitosan with TPP anions during heat
processing in aqueous solutions. The chitosan nanoparti-
cles were found to be heat stable, and there was a continu-
ous release of vitamin C from chitosan nanoparticle. This
indicated applicability of the system in food processing. Hu
et al. (2008) investigated the process of fabricating ChNP
to be used as carriers for delivering tea catechins. Sharma
and Sharma (2013) reported chitosan nanoparticles showed
encapsulation efficiency of 77.8% for terbinafine an anti-
fungal agent.
Water treatment
Water pollution has raised serious concerns lately mainly
due to the inadequacy of conventional water treatment meth-
ods. Even though, activated carbon can be used for adsorb-
ing impurities though effective is not cost or energy efficient.
The low-cost adsorbents like chitosan and cellulose are
interesting options in this context (Olivera et al. 2016). The
functional groups of chitosan, hydroxyl and amino groups
make it an excellent absorbent and enable to be used in water
treatment for removal of functional matrices like pesticides
and metal pollutants. (Dehaghi et al. 2014).
Nanochitosan were tested effectively for adsorptive
capacity of Pb(II) (Qi et al. 2004), Cr(VI) (Sivakami et al.
2013), Cd(II) (Seyedi et al. 2013), arsenate (Kwok et al.
2014), acid Green 27 (AG27) dye of anthraquinone type
(Hu et al. 2006), etc. Chitosan nanofibers owing to their high
porosity and higher surface area per unit mass are potential
adsorbents. They were tested to remove Pb(II) and Cu(II)
while retaining their inherent characteristics (Haider and
Park 2009).
ChNP-coated 4-micron membranes were tested for their
drinking water purification ability in a flow through mem-
brane filtration systems. The ChNP-coated membranes held
good bacterial growth compared to noncoated membranes.
Also, the filtered water showed the maximum removal of
coliforms using multiple tube fermentation (MPN) test
(Rajendran et al. 2015).
Arafat et al. (2015) reported that chitosan zinc oxide nan-
oparticle composites were able to remove 99% of the color
from textile effluent. Magnetic chitosan possesses good dye
adsorbing capacity and can also be easily recovered from the
treated water using magnetic force thus exhibiting excellent
reusability (Hosseini et al. 2016).
Antimicrobial agent
The search for natural antimicrobials to avoid synthetic
chemicals led to chitosan and chitosan nanoparticles. Table 1
gives a summary of different works done on the antimicro-
bial activity of ChNP. ChNP were found to be more effective
against plant pathogens like Fusarium solani (Chowdappa
and Gowda 2013). The antimicrobial activity of chitosan
is caused by three mechanisms. The positively charged

Table 1  Antimicrobial activity of chitosan nanoparticles

Compound Organism References

Chitosan nanoparticles Alternaria alterneta, Macrophomia phaseolina, Rhizoctonia De Paz et al. (2011), Divya et al. (2017), Huang
solani, Nigrospora sphaerica, Botryosphaerica dothidea, N. et al. (2009), Nguyen et al. (2016), Qi et al.
oryzae, A. tenussima, Candida albicans, Fusarium solani, (2004), Saharan et al. (2013), Sarwar et al.
Aspergillus niger, Esherechia coli, Staphylococcus aureus, (2014), Xing et al. (2016), Yien et al. (2012)
Streptococcus pneumoniae, Salmonella choleraesuis, S.
typhimurium, Klebsiella pneumoniae, Pseudomonas aer-
uginosa, Sterptococcus mutans Biofilm
Chitosan microspheres E. coli, Salmonella enterica, K. pneumoniae, V. cholera, Jeon et al. (2014), Kong et al. (2008)
Streptococcus uberis, S. aureus
Chitosan–silver nanoparticles E. coli, R. solani, Aspergillus flavus, A. alterneta, Collec- Ali et al. (2011), Chowdappa et al. (2013), Du
totrichum gloesporiodies, S. aureus, Bacillus subtillus, P. et al. (2009), Honary et al. (2011), Kaur et al.
aerugenosa (2012, 2013), Namasivayam and Roy (2013),
Wei et al. (2009)
Turmeric-chitosan nanoparticle C. albicans, Trychophytol metagrophyte, Fusarium oxyspo- Nguyen et al. (2014)
rum, Penicillium italicum
Chitosan R. solani, S. aureus, S. simulans Liu et al. (2012), Raafat et al. (2008)

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Environ Chem Lett (2018) 16:101–112 107
chitosan interacts with negatively charged phospholipid of
plasma membrane altering the permeability of cell caus-
ing leakage of cell components and cell death. Chitosan has
metal ion chelating property which is a possible cause for its
antimicrobial action. There have also been reported that chi-
tosan could penetrate the cell wall and bind to DNA inhib-
iting mRNA synthesis (Hernandez-Lauzardo et al. 2011;
Sudarshan et al. 1992).
Kaur et al. (2012) reported the fungicidal properties of
nanosize silver/chitosan nanoformulations (NFs) used as an
agent for antifungal treatment of seed-borne plant pathogens
like Aspergillus flavus, Rhizoctonia solani and Alternaria
alterneta.
Ma et  al. (2010) obtained chitosan nanoparticles by
hydrogen peroxide degradation of chitosan. It was incor-
porated into antimicrobial paper by the addition of pulp,
impregnation, dispersion coating on hand sheets and insuf-
flations. It was found that the paper prepared by insuffla-
tions had the greatest activity against Escherichia coli and
Staphylococcus aureus.
Qi et al. (2004) evaluated the in the vitro antibacterial
efficiency of ChNP and copper-loaded ChNP against E. coli,
Staphylococcus choleraesuis, Salmonella typhimurium and
S. aureus. The results showed that chitosan nanoparticles
and copper-loaded nanoparticles inhibited the growth of all
tested bacteria. Their MIC values were less than 0.25 lg/ml,
and the MBC values of nanoparticles reached 1 lg/ml.
Low molecular weight (LMW) ChNP and high molecular
weight (HMW) ChNP have shown activity against Candida
albicans, Aspergillus niger and F. solani (Yien et al. 2012).
The antifungal efficacy of oleoyl-ChNP against Nigrospora
sphaerica, Botryosphaerica dothidea, Nigrospora oryzae,
Alternaria tenussima, Gibberella zeae and Fusarium culmo-
rum was tested by Xing et al. (2016) and four phytopatho-
gens N. sphaerica, B. dothidea, N. oryzae and A. Tenussima
were chitosan sensitive, whereas G. zeae and F. culmorum
were chitosan resistant. Chitosan silver nanoparticle com-
posites were tested positive for its activity against mango
anthracnose pathogen Collectotrichum gleosporides (Chow-
dappa et al. 2014).
Agriculture
Agricultural nanotechnology has acquired a great interest
in recent times due to its ability to provide molecular man-
agement of biotic and abiotic stress, fast and easy detec-
tion of diseases, and delivery systems for fertilizers and
pesticides (Kashyap et al. 2015). In spite of these advan-
tages, selection of nanoparticles for the agricultural pur-
pose should be exercised with caution. Since nanoparticles
owing to its increased surface contact might have toxic
effects absent in its bulk counterpart. It is hence advis-
able to use nontoxic materials for nanoparticle synthesis.
ChNP gain its popularity in this respect (Ghormade et al.
2011). Though there are many reports of the application
of chitosan in agriculture, much work has not been done
using ChNP. Table 2 provides a concise list of various
agricultural applications of ChNP.
Chitosan has been reported to activate more than 20
pathogenesis genes like defensis, lignins, ARnase, phy-
toalexins, chitinase and β-gluconase and plant metab-
olism-related genes (Hadwiger et  al. 2002). Chandra
et al. (2015) reported ChNP produce significantly high
defense response in Camellia sinensis by increasing the
activity of defense enzymes peroxide (PO), polyphenol
oxidase (PPO), phenylalanine ammonia lyase (PAL) and
β-1,3-gluconase.
Chitosan has a significant effect on the growth and
development of various plants like rice, coffee (Van et al.
2013), wheat (Wang et al. 2015), strawberry (Saavedra
et al. 2016), Dendrobium formossum orchid (Kananont
et  al. 2010). Chitosan also had the ability to increase
chlorophyll content and nutrient uptake of plants (Van
et al. 2013). ChNP have shown to impact the biophysical
characteristics of coffee seedlings by increasing pigment
content, the rate of photosynthesis and nutrient uptake, etc.
(Dzung et al. 2011).
Agricultural application of chitosan is mainly in the
form of delivery systems due to its cationic properties and
solubility in acidic solution (Kananont et al. 2010). The
amine group of chitosan forms complex with a wide range
of oppositely charged polymers (Sonia and Sharma 2011).
In addition, chitosan gets easily absorbed to plant surfaces
thus prolonging the contact time between plant surface and
agrochemical (Tiyaboonchai 2003).
Application of nanochitosan-NPK fertilizers to wheat
led to significant increase in its growth performance and
yield (Abdel-Aziz et al. 2016). Microspheres of chitosan and
cashew tree gum were used as a carrier of Lippida sidoides
essential oil to control the proliferation of insect larvae
(Kashyap et al. 2015). ChNP-paraquat herbicide composite
was able to reduce the herbicide toxicity (Grillo et al. 2014).
Corradini et al. (2010) incorporated NPK fertilizers to meth-
acrylic acid polymerized chitosan nanoparticles CS-PMAA.
The elements were found to aggregate on the surface of chi-
tosan nanoparticles which was indicated by the increase in
mean diameter of CS-PMAA.
The main reason for low agricultural productivity is envi-
ronmental factors like temperature, moisture content, pests
and weeds. It is therefore important to constantly monitor
the plant growth. Nanosensors act as an effective evalua-
tion mechanism by transferring nanosized biochemical and
physiological changes to macrolevel (Cicek and Nadaroglu
2015). Nanochitosan biosensors with paramagnetic ­Fe3O4
were able to determine and remove heavy metals (Ahmed
and Fekry 2013).
13

108 Environ Chem Lett (2018) 16:101–112

Table 2  Agricultural applications of chitosan nanoparticles

Compound Crop Use References

Chitosan nanoparticles Robusta coffee Plant growth promotion Van et al. (2013)
Chitosan nanoparticles Rice Defense against Pyricularia grisea Manikandan and Sathiyabama (2016)
Chitosan nanoparticles Apple Post-harvest protection Pilon et al. (2015)
Chitosan nanoparticles Strawberry Post-harvest protection Hajirasouliha et al. (2012)
Chitosan nanoparticles-NPK fertilizer Wheat Plant growth and yield Abdel-Aziz et al. (2016)
Chitosan Cucumber Defense against Pythium aphanidermatum Postma et al. (2009)
Chitosan Tobacco Defense against Phytophthora nicotianae Falcon-Rodriguez et al. (2011)
Hydrolyzed chitosan Rice Defense against Pyricularia grisea Rodriguez et al. (2007)
Chitosan Rice Defense/stress response activation Agrawal et al. (2002)
Chitosan Rice Growth promotion and yield Van Toan and Hanh (2013), Cham-
nanmanoontham et al. (2015)
Radiation degraded chitosan Rice, red chilly, Growth promotion and yield Dahlan et al. (2010), Yacob et al.
potato, carrot (2013), Rekso (2008)
Chitosan Carum copticum Growth promotion and yield Mahdavi and Rahimi (2013)
Chitosan Indian spinach Growth promotion (Mondal et al. 2011)
Chitosan Maize Growth promotion and yield Agbodjato et al. (2016)
Chitosan Tomato Growth promotion and Ralstonia wilt control Algam et al. (2010)
Chitosan Okra Growth promotion and yield Mondal et al. (2012)
Chitosan Sunflower Growth promotion and yield Cho et al. (2008)
Chitosan Freesia Growth promotion and yield Salachna and Zawadzińska (2014)
Chitosan Orchid Growth promotion and yield Nge et al. (2006)
Chitosan Rose apples Post-harvest protection Plainsirichai et al. (2014)

Other applications
The effect of different concentrations of chitosan and chi-
tosan nanoparticles as an active coating on microbiologi-
cal characteristics of fish fingers during frozen storage at
− 18 °C was studied by Abdou et al. (2012). Results indi-
cated that fish fingers coated with either chitosan or chitosan
nanoparticles had much lower total bacterial count (TBC),
psychrophilic bacteria, proteolytic bacteria and coliform
bacteria when compared with uncoated fish fingers and that
coated with a commercial edible coating. In addition, chi-
tosan nanoparticles have been used to improve the strength
and washability of textiles (Panyam and Labhasetwar 2003).
Conclusion
This review summarized the preparation of chitosan nano-
particles and their various applications. From this review,
it is concluded that nanostructured chitosans can be used
as bioactive ingredients carriers. They have the potential to
be encapsulation or immobilization carriers. Due to their
favorable biological properties such as nontoxicity, bio-
compatibility, biodegradability and antibacterial ability,
they are also interesting options as drug delivery carriers
and as cell proliferation enhancers. However, most of these
studies are still at the laboratory level. Additional studies
are necessary before their industrial application. We hope
that more chitosan nanoparticle-based application can be
developed and used in the biochemical and food engineer-
ing fields and also in plant protection the near future.
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