Chitosan nanoparticles preparation and applications

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 process 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 carrier 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, nontoxicity 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 discussed. Applications include drug delivery, encapsulation, antimicrobial agent, plant growth-promoting agent and plant protector.

Full text

Vol.:(0123456789)
1 3
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
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
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

102 Environ Chem Lett (2018) 16:101–112
1 3
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
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
Fig. 1 Structure of chitin and chitosan

103Environ Chem Lett (2018) 16:101–112
1 3
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).
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
Fig. 2 Schematic representation of different modes of ChNP synthesis and its various applications

104 Environ Chem Lett (2018) 16:101–112
1 3
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.
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

105Environ Chem Lett (2018) 16:101–112
1 3
(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.

106 Environ Chem Lett (2018) 16:101–112
1 3
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
solani, Nigrospora sphaerica, Botryosphaerica dothidea, N.
oryzae, A. tenussima, Candida albicans, Fusarium solani,
Aspergillus niger, Esherechia coli, Staphylococcus aureus,
Streptococcus pneumoniae, Salmonella choleraesuis, S.
typhimurium, Klebsiella pneumoniae, Pseudomonas aer-
uginosa, Sterptococcus mutans Biofilm
De Paz etal. (2011), Divya etal. (2017), Huang
etal. (2009), Nguyen etal. (2016), Qi etal.
(2004), Saharan etal. (2013), Sarwar etal.
(2014), Xing etal. (2016), Yien etal. (2012)
Chitosan microspheres E. coli, Salmonella enterica, K. pneumoniae, V. cholera,
Streptococcus uberis, S. aureus Jeon etal. (2014), Kong etal. (2008)
Chitosan–silver nanoparticles E. coli, R. solani, Aspergillus flavus, A. alterneta, Collec-
totrichum gloesporiodies, S. aureus, Bacillus subtillus, P.
aerugenosa
Ali etal. (2011), Chowdappa etal. (2013), Du
etal. (2009), Honary etal. (2011), Kaur etal.
(2012, 2013), Namasivayam and Roy (2013),
Wei etal. (2009)
Turmeric-chitosan nanoparticle C. albicans, Trychophytol metagrophyte, Fusarium oxyspo-
rum, Penicillium italicum Nguyen etal. (2014)
Chitosan R. solani, S. aureus, S. simulans Liu etal. (2012), Raafat etal. (2008)

107Environ Chem Lett (2018) 16:101–112
1 3
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).

108 Environ Chem Lett (2018) 16:101–112
1 3
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.
References
Abdel-Aziz HM, Hasaneen MN, Omer AM (2016) Nano chitosan-
NPK fertilizer enhances the growth and productivity of wheat
plants grown in sandy soil. Span J Agric Res 14(1):0902
Abdou ES, Osheba AS, Sorour MA (2012) Effect of chitosan and chi-
tosan-nanoparticles as active coating on microbiological char-
acteristics of fish fingers. Int J Appl Sci Technol 2(7):158–169
Agbodjato NA, Noumavo PA, Adjanohoun A, Agbessi L, Baba-
Moussa L (2016) Synergistic effects of plant growth promot-
ing rhizobacteria and chitosan on invitro seeds germination,
greenhouse growth, and nutrient uptake of maize (zea mays l.).
Biotechnol Res Int 2016:1–11
Agrawal GK, Rakwal R, Tamogami S, Yonekura M, Kubo A, Saji
H (2002) Chitosan activates defense/stress response (s) in
the leaves of oryza sativa seedlings. Plant Physiol Biochem
40(12):1061–1069
Ahmed RA, Fekry A (2013) Preparation and characterization of
a nanoparticles modified chitosan sensor and its application
for the determination of heavy metals from different aqueous
media. Int J Electrochem Sci 8(3):6692–6708
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,
potato, carrot
Growth promotion and yield Dahlan etal. (2010), Yacob etal.
(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)

109Environ Chem Lett (2018) 16:101–112
1 3
Aiping Z, Jianhong L, Wenhui Y (2006) Effective loading and con-
trolled release of camptothecin by O-carboxymethylchitosan
aggregates. Carbohydr Polym 63(1):89–96
Algam S, Xie G, Li B, Yu S, Su T, Larsen J (2010) Effects of paeniba-
cillus strains and chitosan on plant growth promotion and control
of ralstonia wilt in tomato. J Plant Pathol 92(3):593–600
Ali SW, Rajendran S, Joshi M (2011) Synthesis and characterization
of chitosan and silver loaded chitosan nanoparticles for bioactive
polyester. Carbohydr Polym 83(2):438–446
Allen MJ, Schoonmaker JE, Bauer TW, Williams PF, Higham PA,
Yuan HA (2004) Preclinical evaluation of a poly (vinyl alco-
hol) hydrogel implant as a replacement for the nucleus pulposus.
Spine 29(5):515–523
Arafat A, Samad SA, Huq D, Moniruzzaman M, Masum SM (2015)
Textile dye removal from wastewater effluents using chitosan–
ZnO nanocomposite. J Text Sci Eng 5(3):1
Aranaz I, Mengabar M, Harris R, Is Paos, Miralles B, Acosta N, Heras
A etal (2009) Functional characterization of chitin and chitosan.
Curr Chem Biol 3(2):203–230
Badawy MEI, Rabea EI (2011) A biopolymer chitosan and its deriva-
tives as promising antimicrobial agents against plant pathogens
and their applications in crop protection. Int J Carbohydr Chem
2011:1–29
Baxter A, Dillon M, Anthony Taylor KD, Roberts GAF (1992)
Improved method for ir determination of the degree of n-acety-
lation of chitosan. Int J Biol Macromol 14(3):166–169
Brunel F, Varon L, David L, Domard A, Delair T (2008) A novel syn-
thesis of chitosan nanoparticles in reverse emulsion. Langmuir
24(20):11370–11377
Calvo P, Remunanae-Lopez C, Vila-Jato JL, Alonso MJ (1997) Novel
hydrophilic chitosan polyethylene oxide nanoparticles as protein
carriers. J Appl Polym Sci 63(1):125–132
Chamnanmanoontham N, Pongprayoon W, Pichayangkura R, Roytrakul
S, Chadchawan S (2015) Chitosan enhances rice seedling growth
via gene expression network between nucleus and chloroplast.
Plant Growth Regul 75(1):101–114
Chandra S, Chakraborty N, Dasgupta A, Sarkar J, Panda K, Acharya
K (2015) Chitosan nanoparticles: a positive modulator of innate
immune responses in plants. Sci Rep 5:1–14
Cheba BA (2011) Chitin and chitosan: marine biopoly-mers with
unique properties and versatile applications. Glob J Biotechnol
Biochem 6(3):149–153
Chen DH, Leu JC, Huang TC (1994) Transport and hydrolysis of
urea in a reactor–separator combining an anion-exchange mem-
brane and immobilized urease. J Chem Technol Biotechnol
61(4):351–357
Cho M, No H, Prinyawiwatkul W (2008) Chitosan treatments affect
growth and selected quality of sunflower sprouts. J Food Sci
73(1):70–77
Chowdappa P, Gowda S (2013) Nanotechnology in crop protection:
status and scope. Pest Manag Hortic Ecosyst 19(2):131–151
Chowdappa P, Kumar SM, Lakshmi MJ, Upreti K (2013) Growth stim-
ulation and induction of systemic resistance in tomato against
early and late blight by bacillus subtilis OTPB1 or trichoderma
harzianum OTPB3. Biol Control 65(1):109–117
Chowdappa P, Gowda S, Chethana C, Madhura S (2014) Antifungal
activity of chitosan–silver nanoparticle composite against colle-
totrichum gloeosporioides associated with mango anthracnose.
Afr J Microbiol Res 8(17):1803–1812
Cicek S, Nadaroglu H (2015) The use of nanotechnology in the agri-
culture. Adv Nano Res 3(4):207–223
Corradini E, De Moura MR, Mattoso LHC (2010) A preliminary study
of the incorparation of NPK fertilizer into chitosan nanoparticles.
Express Polym Lett 4(8):509–515
Dahlan KZHM, Hashim K, Bahari K, Mahmod M, Yaacob N, Talip N
etal (2010) Application of radiation degraded chitosan as plant
growth promoter. A pilot scale production and field trial study
of radiation processed chitosan as plant growth promoter for rice
crops. International Atomic Energy Agency
de Paz LEC, Resin A, Howard KA, Sutherland DS, Wejse PL (2011)
Antimicrobial effect of chitosan nanoparticles on streptococcus
mutans biofilms. Appl Environ Microbiol 77(11):3892–3895
De TK, Ghosh PK, Maitra A, Sahoo SK (1999) Process for the prepara-
tion of highly monodispersed polymeric hydrophilic nanoparti-
cles: Google Patents
Dehaghi SM, Rahmanifar B, Moradi AM, Azar PA (2014) Removal
of permethrin pesticide from water by chitosan–zinc oxide
nanoparticles composite as an adsorbent. J Saudi Chem Soc
18(4):348–355
Divya K, Rebello S, Jisha MS (2014) A simple and effective method
for extraction of high purity chitosan from shrimp shell waste.
In: Proceedings of the international conference on advances in
applied science and environmental engineering-ASEE
Divya K, Vijayan S, George TK, Jisha M (2017) Antimicrobial proper-
ties of chitosan nanoparticles: mode of action and factors affect-
ing activity. Fibers Polym 18(2):221–230
Du W-L, Niu S-S, Xu Y-L, Xu Z-R, Fan C-L (2009) Antibacterial
activity of chitosan tripolyphosphate nanoparticles loaded with
various metal ions. Carbohydr Polym 75(3):385–389
Dzung NA, Khanh VTP, Dzung TT (2011) Research on impact of
chitosan oligomers on biophysical characteristics, growth,
development and drought resistance of coffee. Carbohydr Polym
84(2):751–755
Einbu A, Vayrum KM (2008) Characterization of chitin and its hydrol-
ysis to glcnac and glcn. Biomacromol 9(7):1870–1875
El-Shabouri MH (2002) Positively charged nanoparticles for improv-
ing the oral bioavailability of cyclosporin-A. Int J Pharm
249(1):101–108
Erbacher P, Zou S, Bettinger T, Steffan A-M, Remy J-S (1998) Chi-
tosan-based vector/DNA complexes for gene delivery: bio-
physical characteristics and transfection ability. Pharm Res
15(9):1332–1339
Falcon-Rodriguez AB, Costales D, Cabrera JC, Martinez-Tellez
MA (2011) Chitosan physico–chemical properties modulate
defense responses and resistance in tobacco plants against the
oomycete phytophthora nicotianae. Pestic Biochem Physiol
100(3):221–228
Fang H, Huang J, Ding L, Li M, Chen Z (2009) Preparation of mag-
netic chitosan nanoparticles and immobilization of laccase. J
Wuhan Univ Technol Mater Sci Ed 24(1):42–47
Fernandez-Urrusuno R, Calvo P, Remuan-Lopez C, Vila-Jato JL,
Alonso MJ (1999) Enhancement of nasal absorption of insulin
using chitosan nanoparticles. Pharm Res 16(10):1576–1581
Franca EF, Lins RD, Freitas LCG, Straatsma TP (2008) Characteriza-
tion of chitin and chitosan molecular structure in aqueous solu-
tion. J Chem Theory Comput 4(12):2141–2149
Geng X, Kwon O-H, Jang J (2005) Electrospinning of chitosan dis-
solved in concentrated acetic acid solution. Biomaterials
26(27):5427–5432
Ghadi A, Mahjoub S, Tabandeh F, Talebnia F (2014) Synthesis and
optimization of chitosan nanoparticles: potential applications
in nanomedicine and biomedical engineering. Casp J Int Med
5(3):156
Ghadi A, Tabandeh F, Mahjoub S, Mohsenifar A, Roshan FT, Alavije
RS (2015) Fabrication and characterization of core-shell mag-
netic chitosan nanoparticles as a novel carrier for immobilization
of Burkholderia cepacia lipase. J Oleo Sci 64(4):423–430
Ghormade V, Deshpande MV, Paknikar KM (2011) Perspectives for
nano-biotechnology enabled protection and nutrition of plants.
Biotechnol Adv 29(6):792–803
Grenha A (2010) Chitosan nanoparticles: a survey of preparation meth-
ods. J Drug Target 20(4):291–300

110 Environ Chem Lett (2018) 16:101–112
1 3
Grillo R, Pereira AE, Nishisaka CS, de Lima R, Oehlke K, Greiner
R, Fraceto LF (2014) Chitosan/tripolyphosphate nanoparticles
loaded with paraquat herbicide: an environmentally safer alterna-
tive for weed control. J Hazard Mater 278:163–171
Gupta AK, Naregalkar RR, Vaidya VD, Gupta M (2007) Recent
advances on surface engineering of magnetic iron oxide nanopar-
ticles and their biomedical applications. Nanomedicine (Lond)
2(1):23–29
Hadwiger L, Klosterman S, Choi J (2002) The mode of action of chi-
tosan and its oligomers in inducing plant promoters and develop-
ing disease resistance in plants. Adv Chitin Sci 5:452–457
Haider S, Park S-Y (2009) Preparation of the electrospun chitosan
nanofibers and their applications to the adsorption of Cu (II) and
Pb(II) ions from an aqueous solution. J Membr Sci 328(1):90–96
Hajirasouliha MJM, Soheili F, Naja Fabadi (2012) Effect of novel
chitosan nanoparticle coating on pot harvest qualities of straw-
berry. In: Proceedings of the 4th international conference on
nanostructures
Hatton RA, Miller AJ, Silva SRP (2008) Carbon nanotubes: a multi-
functional material for organic optoelectronics. J Mater Chem
18:1183–1192
Hernandez-Lauzardo AN, Velaizquez-del Valle MG, Guerra-
Sanchez MG (2011) Current status of action mode and effect
of chitosan against phytopathogens fungi. Afr J Microbiol Res
5(25):4243–4247
Hirano S, Yamaguchi Y, Kamiya M (2002) Novel N-saturated-fatty-
acyl derivatives of chitosan soluble in water and in aqueous acid
and alkaline solutions. Carbohydr Polym 48(2):203–207
Honary S, Ghajar K, Khazaeli P, Shalchian P (2011) Preparation, char-
acterization and antibacterial properties of silver–chitosan nano-
composites using different molecular weight grades of chitosan.
Trop J Pharm Res 10(1):69–74
Hosseini F, SadighianS Hosseini-Monfared F, Mahmoodi NM (2016)
Dye removal and kinetics of adsorption by magnetic chitosan
nanoparticles. Desalin Water Treat 57(51):24378–24386
Hu Z, Zhang J, Chan W, Szeto Y (2006) The sorption of acid dye onto
chitosan nanoparticles. Polymer 47(16):5838–5842
Hu B, Pan C, Sun Y, Hou Z, Ye H, Zeng X (2008) Optimization of
fabrication parameters to produce chitosan tripolyphosphate
nanoparticles for delivery of tea catechins. J Agric Food Chem
56(16):7451–7458
Huang YC, Li RY (2014) Preparation and characterization of anti-
oxidant nanoparticles composed of chitosan and fucoidan for
antibiotics delivery. Mar Drugs 12(8):4379–4398
Huang L, Cheng X, Liu C, Xing K, Zhang J, Sun G, Chen X etal
(2009) Preparation, characterization, and antibacterial activity of
oleic acid-grafted chitosan oligosaccharide nanoparticles. Front
Biol China 4(3):321–327
Ilium L (1998) Chitosan and its use as a pharmaceutical excipient.
Pharm Res 15(9):1326–1331
Ingle A, Gade A, Pierrat S, Sonnichsen C, Rai M (2008) Mycosynthesis
of silver nanoparticles using the fungus fusarium acuminatum
and its activity against some human pathogenic bacteria. Curr
Nanosci 4(2):141–144
Janes KA, Fresneau MP, Marazuela A, Fabra A, Alonso MJ (2001)
Chitosan nanoparticles as delivery systems for doxorubicin. J
Control Release 73(2):255–267
Jang K-I, Lee HG (2008) Stability of chitosan nanoparticles for l-ascor-
bic acid during heat treatment in aqueous solution. J Agric Food
Chem 56(6):1936–1941
Jayakumar R, Menon D, Manzoor K, Nair SV, Tamura H (2010a) Bio-
medical applications of chitin and chitosan based nanomaterials:
a short review. Carbohydr Polym 82(2):227–232
Jayakumar R, Prabaharan M, Nair S, Tamura H (2010b) Novel chitin
and chitosan nanofibers in biomedical applications. Biotechnol
Adv 28(1):142–150
Jeon SJ, Oh M, Yeo W-S, Galvao KN, Jeong KC (2014) Underlying
mechanism of antimicrobial activity of chitosan microparticles
and implications for the treatment of infectious diseases. PLoS
ONE 9(3):92723
Jia Y-T, Gong J, Gu X-H, Kim H-Y, Dong J, Shen X-Y (2007) Fab-
rication and characterization of poly (vinyl alcohol)/chitosan
blend nanofibers produced by electrospinning method. Carbo-
hydr Polym 67(3):403–409
Jonassen H, Kjoniksen AL, Hiorth M (2012) Stability of chitosan
nanoparticles cross-linked with tripolyphosphate. Biomacro-
mol 13(11):3747–3756
Kananont N, Pichyangkura R, Chanprame S, Chadchawan S, Lim-
panavech P (2010) Chitosan specificity for the in vitro seed
germination of two dendrobium orchids (asparagales: Orchi-
daceae). Sci Hortic 124(2):239–247
Kasaai MR (2009) Various methods for determination of the degree
of n-acetylation of chitin and chitosan: a review. J Agric Food
Chem 57(5):1667–1676
Kashyap PL, Xiang X, Heiden P (2015) Chitosan nanoparticle based
delivery systems for sustainable agriculture. Int J Biol Mac-
romol 77:36–51
Kaur P, Thakur R, Choudhary A (2012) An invitro study of the
antifungal activity of silver/chitosan nanoformulations against
important seed borne pathogens. Int J Sci Technol Res 1:83–86
Kaur P, Choudhary A, Thakur R (2013) Synthesis of chitosan–silver
nanocomposites and their antibacterial activity. Int J Sci Eng
Res 4:869–872
Kong M, Chen X, Xue Y, Liu C, Yu L, Ji Q etal (2008) Preparation
and antibacterial activity of chitosan microshperes in a solid
dispersing system. Front Mater Sci China 2(2):214–220
Kumar MNVR (1999) Chitin and chitosan fibres: a review. Bull
Mater Sci 22(5):905–915
Kumar S, Koh J (2012) Physiochemical and optical study of chi-
tosan–terephthaldehyde derivative for biomedical applications.
Int J Biol Macromol 51(5):1167–1172
Kumirska J, Mg Czerwicka, Kaczyaski Z, Bychowska A, Brzozowski
K, Thaming J, Stepnowski P (2010) Application of spectro-
scopic methods for structural analysis of chitin and chitosan.
Mar Drugs 8(5):1567–1636
Kumirska J, Weinhold MX, Thaming J, Stepnowski P (2011) Bio-
medical activity of chitin/chitosan based materials influence of
physicochemical properties apart from molecular weight and
degree of n-acetylation. Polymers 3(4):1875–1901
Kwok KC, Koong LF, Chen G, McKay G (2014) Mechanism of
arsenic removal using chitosan and nanochitosan. J Colloid
Interface Sci 416:1–10
Lin SB, Chen SH, Peng KC (2009) Preparation of antibacterial
chito-oligosaccharide by altering the degree of deacetylation of
β-chitosan in a Trichoderma harzianum chitinase-hydrolysing
process. J Sci Food Agric 89(2):238–244
Liu C-G, Desai KGH, Chen X-G, Park H-J (2005) Preparation and
characterization of nanoparticles containing trypsin based
on hydrophobically modified chitosan. J Agric Food Chem
53(5):1728–1733
Liu H, Tian W, Li B, Wu G, Ibrahim M, Tao Z, Sun G etal (2012)
Antifungal effect and mechanism of chitosan against the rice
sheath blight pathogen, Rhizoctonia solani. Biotechnol Lett
34(12):2291–2298
Lopez-Leon T, Carvalho ELS, Seijo B, Ortega-Vinuesa JL, Bastos-
Gonzailez D (2005) Physicochemical characterization of chi-
tosan nanoparticles: electrokinetic and stability behavior. J
Colloid Interface Sci 283(2):344–351
Ma Y, Liu P, Si C, Liu Z (2010) Chitosan nanoparticles: preparation
and application in antibacterial paper. J Macromol Sci Part B
49(5):994–1001

111Environ Chem Lett (2018) 16:101–112
1 3
Mahdavi B, Rahimi A (2013) Seed priming with chitosan improves
the germination and growth performance of ajowan (carum cop-
ticum) under salt stress. EurAsia J Biosci 7:69–76
Makhluf S, Dror R, Nitzan Y, Abramovich Y, Jelinek R, Gedanken A
(2005) Microwave assisted synthesis of nanocrystalline mgo and
its use as a bacteriocide. Adv Funct Mater 15(10):1708–1715
Malmiri HJ, Jahanian MAG, Berenjian A (2012) Potential applications
of chitosan nanoparticles as novel support in enzyme immobili-
zation. Am J Biochem Biotechnol 8(4):203–219
Manikandan A, Sathiyabama M (2016) Preparation of chitosan nan-
oparticles and its effect on detached rice leaves infected with
pyricularia grisea. Int J Biol Macromol 84:58–61
Manjusha EM, Mohan JC, Manzoor K, Nair SV, Tamura H, Jayakumar
R (2010) Folate conjugated carboxymethyl chitosan–manganese
doped zinc sulphide nanoparticles for targeted drug delivery and
imaging of cancer cells. Carbohydr Polym 80:414–420
Min B-M, Lee SW, Lim JN, You Y, Lee TS, Kang PH, Park WH (2004)
Chitin and chitosan nanofibers: electrospinning of chitin and dea-
cetylation of chitin nanofibers. Polymer 45(21):7137–7142
Mohammadpour Dounighi N, Eskandari R, Avadi M, Zolfagharian
H, Mir Mohammad Sadeghi A, Rezayat M (2012) Preparation
and invitro characterization of chitosan nanoparticles contain-
ing Mesobuthus eupeus scorpion venom as an antigen delivery
system. J Venom Anim Toxins Incl Trop Dis 18(1):44–52
Mondal M, Rana MIK, Dafader N, Haque M (2011) Effect of foliar
application of chitosan on growth and yield in Indian spinach. J
Agrofor Environ 5(1):99–102
Mondal M, Malek M, Puteh A, Ismail M, Ashrafuzzaman M, Naher
L (2012) Effect of foliar application of chitosan on growth and
yield in okra. Aust J Crop Sci 6(5):918
Muzzarelli RAAJC, Gooday GW (1986) Chitin in nature and technol-
ogy. Plenum Publishing Corporation, New York
Namasivayam SKR, Roy EA (2013) Enhanced antibiofilm activity
of chitosan stabilized chemogenic silver nanoparticles against
Escherichia coli. Int J Sci Res 2013:591
Nge KL, Nwe N, Chandrkrachang S, Stevens WF (2006) Chitosan
as a growth stimulator in orchid tissue culture. Plant Sci
170(6):1185–1190
Nguyen T, Dzung T, Cuong P (2014) Assessment of antifungal activity
of turmeric essential oil-loaded chitosan nanoparticles. J Chem
Biol Phys Sci 4:2347–2356
Nguyen TV, Nguyen TTH, Wang S-L, Vo TPK, Nguyen AD (2016)
Preparation of chitosan nanoparticles by tpp ionic gelation com-
bined with spray drying, and the antibacterial activity of chitosan
nanoparticles and a chitosan nanoparticle–amoxicillin complex.
Res Chem Intermed 2016:1–11
Niwa T, Takeuchi H, Hino T, Kunou N, Kawashima Y (1993) Prepara-
tions of biodegradable nanospheres of water-soluble and insolu-
ble drugs with d, l-lactide/glycolide copolymer by a novel spon-
taneous emulsification solvent diffusion method, and the drug
release behavior. J Control Release 25(1):89–98
Ohkawa K, Cha D, Kim H, Nishida A, Yamamoto H (2004)
Electrospinning of chitosan. Macromol Rapid Commun
25(18):1600–1605
Olivera S, Muralidhara HB, Venkatesh K, Guna VK, Gopalakrishna K,
Kumar Y (2016) Potential applications of cellulose and chitosan
nanoparticles/composites in wastewater treatment: a review. Car-
bohydr Polym 153:600–618
Onsosyen E, Skaugrud O (1990) Metal recovery using chitosan. J
Chem Technol Biotechnol 49(4):395–404
Ottey MH, Varum KM, Smidsra DO (1996) Compositional heterogene-
ity of heterogeneously deacetylated chitosans. Carbohydr Polym
29(1):17–24
Panyam J, Labhasetwar V (2003) Biodegradable nanoparticles for
drug and gene delivery to cells and tissue. Adv Drug Deliv Rev
55(3):329–347
Park JK, Chung MJ, Choi HN, Park YI (2011) Effects of the molecu-
lar weight and the degree of deacetylation of chitosan oligosac-
charides on antitumor activity. Int J Mol Sci 12(1):266–277
Peppas NA, Huang Y (2004) Nanoscale technology of mucoadhesive
interactions. Adv Drug Deliv Rev 56(11):1675–1687
Perera U, Rajapakse N (2013) Chitosan nanoparticles: preparation,
characterization, and applications. In: Kim SK (ed) Seafood
processing by-products: trends and applications. Springer, New
York, pp 371–387
Pilon L, Spricigo PC, Miranda M, Moura MR, Assis OBG, Mat-
toso LHC, Ferreira MD (2015) Chitosan nanoparticle coatings
reduce microbial growth on fresh-cut apples while not affecting
quality attributes. Int J Food Sci Technol 50(2):440–448
Plainsirichai M, Leelaphatthanapanich S, Wongsachai N (2014)
Effect of chitosan on the quality of rose apples (syzygium
agueum alston) cv. Tabtim chan stored at an ambient tempera-
ture. APCBEE Procedia 8:317–322
Postma J, Stevens LH, Wiegers GL, Davelaar E, Nijhuis EH (2009)
Biological control of pythium aphanidermatum in cucumber
with a combined application of lysobacter enzymogenes strain
3.1 T8 and chitosan. Biol Control 48(3):301–309
Puvvada YS, Vankayalapati S, Sukhavasi S (2012) Extraction of chi-
tin from chitosan from exoskeleton of shrimp for application
in the pharmaceutical industry. Int Curr Pharm J 1(9):258–263
Qi L, Xu Z, Jiang X, Hu C, Zou X (2004) Preparation and anti-
bacterial activity of chitosan nanoparticles. Carbohydr Res
339(16):2693–2700
Raafat D, Von Bargen K, Haas A, Sahl H-G (2008) Insights into the
mode of action of chitosan as an antibacterial compound. Appl
Environ Microbiol 74(12):3764–3773
Rajalakshmi A, Krithiga N, Jayachitra A (2013) Antioxidant activity
of the chitosan extracted from shrimp exoskeleton. Middle East
J Sci Res 16(10):1446–1451
Rajasree R, Rahate K (2013) An overview on various modifica-
tions of chitosan and it’s applications. Int J Pharm Sci Res
4(11):4175
Rajendran R, Abirami M, Prabhavathi P, Premasudha P, Kanimozhi
B, Manikandan A (2015) Biological treatment of drinking
water by chitosan based nanocomposites. Afr J Biotechnol
14(11):930–936
Ravishankar Rai V, Jamuna Bai A (2011) Nanoparticles and their
potential application as antimicrobials, science against microbial
pathogens: communicating current research and technological
advances. In: Méndez-Vilas A (ed), Formatex, Microbiology
Series, No. 3, Vol. 1. Spain, pp 197–209
Rekso GT (2008) Development of radiation degraded chitosan as plant
growth promoter and its economic evaluation. JAEA CONF
2008-9
Rinaudo M (2006) Chitin and chitosan: properties and applications.
Prog Polym Sci 31(7):603–632
Rodriguez A, Ramirez M, Cardenas R, Hernandez A, Velazquez M,
Bautista S (2007) Induction of defense response of Oryza sativa
L. against Pyricularia grisea (cooke) Sacc. By treating seeds
with chitosan and hydrolyzed chitosan. Pestic Biochem Physiol
89(3):206–215
Saavedra GM, Figueroa NE, Poblete LA, Cherian S, Figueroa CR
(2016) Effects of preharvest applications of methyl jasmonate
and chitosan on postharvest decay, quality and chemical attrib-
utes of fragaria chiloensis fruit. Food Chem 190:448–453
Saharan V, Mehrotra A, Khatik R, Rawal P, Sharma S, Pal A (2013)
Synthesis of chitosan based nanoparticles and their invitro
evaluation against phytopathogenic fungi. Int J Biol Macromol
62:677–683
Sailaja A, Amareshwar P, Chakravarty P (2011) Different techniques
used for the preparation of nanoparticles using natural polymers
and their application. Int J Pharm Pharm Sci 3(Suppl 2):45–50

112 Environ Chem Lett (2018) 16:101–112
1 3
Salachna P, Zawadzińska A (2014) Effect of chitosan on plant
growth, flowering and corms yield of potted freesia. J Ecol Eng
15(3):97–102
Sarwar A, Katas H, Zin NM (2014) Antibacterial effects of chitosan–
tripolyphosphate nanoparticles: impact of particle size molecular
weight. J Nanopart Res 16(7):2517
Seyedi SM, Anvaripour B, Motavassel M, Jadidi N (2013) Comparative
cadmium adsorption from water by nanochitosan and chitosan.
Int J Eng Innov Technol 2(9):145–148
Shahidi F, Arachchi JKV, Jeon Y-J (1999) Food applications of chitin
and chitosans. Trends Food Sci Technol 10(2):37–51
Sharma S, Sharma S (2013) Synthesis, characterization and determi-
nation of encapsulation efficiency of chitosan nanoparticles for
terbinafine. Indo Am J Pharm Res 3(12):1564–1567
Shi B, Shen Z, Zhang H, Bi J, Dai S (2011a) Exploring N-imidazolyl-
O-carboxymethyl chitosan for high performance gene delivery.
Biomacromol 13(1):146–153
Shi LE, Tang ZX, Yi Y, Chen JS, Xiong WY, Ying GQ (2011b) Immo-
bilization of nuclease p1 on chitosanmicro-spheres. Chem Bio-
chem Eng Q 25(1):83–88
Shiraishi S, Imai T, Otagiri M (1993) Controlled release of indo-
methacin by chitosan-polyelectrolyte complex: optimization
and invivo/in vitro evaluation. J Control Release 25(3):217–225
Sims DC, Butler PE, Casanova R, Lee BT, Randolph MA, Lee WA,
Yaremchuk MJ etal (1996) Injectable cartilage using polyethyl-
ene oxide polymer substrates. Plast Reconstr Surg 98(5):843–850
Sivakami M, Gomathi T, Venkatesan J, Jeong H-S, Kim S-K, Sudha
P (2013) Preparation and characterization of nano chitosan for
treatment wastewaters. Int J Biol Macromol 57:204–212
Sonia T, Sharma CP (2011) Chitosan and its derivatives for drug deliv-
ery perspective. Adv Polym Sci 243:23–53
Sudarshan NR, Hoover DG, Knorr D (1992) Antibacterial action of
chitosan. Food Biotechnol 6(3):257–272
Sun K, Li Z (2011) Preparations, properties and applications of chi-
tosan based nanofibers fabricated by electrospinning. Express
Polym Lett 5(4):342–361
Thanou M, Nihot M, Jansen M, Verhoef JC, Junginger H (2001) Mono-
N-carboxymethyl chitosan (MCC), a polyampholytic chitosan
derivative, enhances the intestinal absorption of low molecular
weight heparin across intestinal epithelia invitro and invivo. J
Pharm Sci 90(1):38–46
Tiyaboonchai W (2003) Chitosan nanoparticles: a promising system
for drug delivery. Naresuan Univ J 11(3):51–66
Van Toan N, Hanh TT (2013) Application of chitosan solutions for rice
production in vietnam. Afr J Biotechnol 12(4):382–384
Van SN, Minh HD, Anh DN (2013) Study on chitosan nanoparticles on
biophysical characteristics and growth of robusta coffee in green
house. Biocatal Agric Biotechnol 2(4):289–294
Vauthier CDC, Chauvierre C, Brigger I, Couvreur P (2003) Drug deliv-
ery to resistant tumors: the potential of poly(alkyl cyanoacrylate)
nanoparticles. J Control Release 93:151–160
Vazquez-Duhalt R, Tinoco R, D’Antonio P, Topoleski LDT, Payne
GF (2001) Enzyme conjugation to the polysaccharide chitosan:
smart biocatalysts and biocatalytic hydrogels. Bioconjug Chem
12(2):301–306
Wang X, Xing B (2007) Importance of structural makeup of biopoly-
mers for organic contaminant sorption. Environ Sci Technol
41(10):3559–3565
Wang M, Chen Y, Zhang R, Wang W, Zhao X, Du Y, Yin H (2015)
Effects of chitosan oligosaccharides on the yield components and
production quality of different wheat cultivars (triticum aestivum
l.) in northwest china. Field Crop Res 172:11–20
Wei D, Sun W, Qian W, Ye Y, Ma X (2009) The synthesis of chitosan-
based silver nanoparticles and their antibacterial activity. Carbo-
hydr Res 344(17):2375–2382
Whitesides GM (2003) The’right’size in nanobiotechnology. Nat Bio-
technol 21(10):1161–1165
Xie W, Xu P, Liu Q (2001) Antioxidant activity of water-soluble chi-
tosan derivatives. Bioorg Med Chem Lett 11(13):1699–1701
Xing K, Shen X, Zhu X, Ju X, Miao X, Tian J, Qin S etal (2016) Syn-
thesis and invitro antifungal efficacy of oleoyl-chitosan nano-
particles against plant pathogenic fungi. Int J Biol Macromol
82:830–836
Yacob N, Mahmud M, Talip N, Hashim K, Harun AR, Zaman K,
Dahlan H (2013) Degradation of chitosan for rice crops applica-
tion. Nucl Sci Tech 24(S1):10301–S010301
Yang K, Xu N-S, Su WW (2010) Co-immobilized enzymes in mag-
netic chitosan beads for improved hydrolysis of macromolecu-
lar substrates under a time-varying magnetic field. J Biotechnol
148(2):119–127
Yen MT, Mau JL (2007) Selected physical properties of chitin prepared
from shiitake stipes. LWT Food Sci Technol 40(3):558–563
Yen MT, Yang JH, Mau JL (2008) Antioxidant properties of chitosan
from crab shells. Carbohydr Polym 74(4):840–844
Yen MT, Yang JH, Mau JL (2009) Physicochemical characteriza-
tion of chitin and chitosan from crab shells. Carbohydr Polym
75(1):15–21
Yien L, Zin NM, Sarwar A, Katas H (2012) Antifungal activity of chi-
tosan nanoparticles and correlation with their physical properties.
Int J Biomater 2012:1–9
Yoshii F, Zhanshan Y, Isobe K, Shinozaki K, Makuuchi K (1999) Elec-
tron beam crosslinked PEO and PEO/PVA hydrogels for wound
dressing. Radiat Phys Chem 55(2):133–138
Zhang Y, Zhang X, Ding R, Zhang J, Liu J (2011) Determination of
the degree of deacetylation of chitosan by potentiometric titra-
tion preceded by enzymatic pretreatment. Carbohydr Polym
83(2):813–817
Zhao LM, Shi LE, Zhang ZL, Chen JM, Shi DD, Yang J, Tang ZX
(2011) Preparation and application of chitosan nanoparticles and
nanofibers. Braz J Chem Eng 28(3):353–362
Zolghadri S, Jalilian AR, Yousefnia H, Bahrami-Samani A, Shirvani-
Arani S, Mazidi M, Akhlaghi M, Ghannadi-Maragheh M (2010)
Production and quality control of 166Ho-Chitosan for therapeutic
applications. Iran J Nucl Med 18(2):1–8