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Ionotropic gelation - A novel method to prepare chitosan nanoparticles

Article  in  Research Journal of Pharmacy and Technology · January 2011

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 Research J. Pharm. and Tech. 4(4): April 2011

ISSN 0974-3618 www.rjptonline.org

REVIEW ARTICLE
Ionotropic Gelation – A Novel Method to Prepare Chitosan Nanoparticles
Subhashis Debnath1*, R. Suresh Kumar2 and M. Niranjan Babu1
1
Seven Hills College of Pharmacy, Tirupati-517561, Andhra Pradesh, India.
2
J.S.S College of Pharmacy, Ooty-643001, Tamilnadu, India.
*Corresponding Author E-mail: subhashis_xyz@yahoo.com

ABSTRACT:
Considerable research efforts have been directed towards developing safe and efficient chitosan-based particulate drug
delivery systems. The present review outlines the ionotropic gelation method for the preparation of chitosan-based
nanoparticulate drug delivery systems published over the past decade. From a literature survey, it is realized that
research activities on chitosan micro/nanoparticulate systems containing various drugs for different therapeutic
applications have increased at the rapid rate. Hence ionotropic gelation method can use to prepare these chitosan
nanoarticles as it is very simple and having lots of advantages then other methods.

KEYWORDS: Chitosan, Nanoparticles, Ionotropic gelation.

INTRODUCTION:
Polymer nanoparticles are particles of less than 1 mm It has been reported that chitosan nanoparticles are having
diameter that are prepared from natural or synthetic little particle size and enhanced zeta potential. Chitosan is a
polymers. Nanoparticles have become an important area of polysaccharide, similar in structure to cellulose. Both are
research in the field of drug delivery because they have made by linear h-(1Y4)-linked monosaccharides2-7.
been extensively used to deliver drugs, genes, diagnostics, However, an important difference to cellulose is that
and vaccines into specific cells or tissues. The strategy of chitosan is composed of 2-amino-2-deoxy-h-d-glucan
using nanoparticles as a carrier system for drug and gene combined with glycosidic linkages. The primary amine
groups render special properties that make chitosan very
delivery has attracted increasing interest. The main target of
many pharmaceutical delivery systems is to deliver the drug useful in pharmaceutical applications. Compared to many
to the specific cell types and is successful only when the other natural polymers, chitosan has a positive charge and is
drug through its delivery vehicle is internalized into cells.mucoadhesive. Therefore, it is used extensively in drug
Owing to their small size, prolonged circulation time, and delivery applications. Chitosan is obtained from the
sustained drug release profile, nano-sized polymeric deacetylation of chitin, a naturally occurring and abundantly
nanoparticles bearing drugs have received an increasing available (in marine crustaceans) biocompatible
amount of attention for their ability to improve the efficacypolysaccharide. However, applications of chitin are limited
of potent drugs. It has been reported that nano-sized drug compared to chitosan because chitin is structurally similar
carriers composed of natural and synthetic polymers to cellulose, but chemically inert. Acetamide group of chitin
maintain a prolonged circulation time in the body by can be converted into amino group to give chitosan, which
avoiding the reticuloendothelial system (RES), as such is carried out by treating chitin with concentrated alkali
reduced liver and spleen uptake has been exploited in solution. Chitin and chitosan represent long-chain polymers
cancer therapies1. having molecular mass up to several million Daltons.
Chitosan is relatively reactive and can be produced in
8-11
Recently, polymer nanoparticles have been widely various forms such as powder, paste, film, fiber, etc .
investigated as a carrier for drug delivery. Among them,
much attention has been paid to the nanoparticles made of Commercially available CS has an average molecular
biodegradable polymers such as chitosan(CS ) due to its weight ranging between 3800 and 20,000 Daltons and is
good biocompatibility, biodegra dability, and novel drug 66% to 95% deacetylated. Chitosan, being a cationic
release behavior. Chitosan nanoparticles are potential polysaccharide in neutral or basic pH conditions, contains
delivery systems for vaccines, genes, and anticancer agents. free amino groups and hence, is insoluble in water. In acidic
pH, amino groups can undergo protonation thus, making it
Received on 10.11.2010 Modified on 04.01.2011 soluble in water. Solubility of CS depends upon the
Accepted on 17.01.2011 © RJPT All right reserved distribution of free amino and N-acetyl groups. Usually 1-3
Research J. Pharm. and Tech. 4(4): April 2011; Page 492-495 % aqueous acetic acid solutions are used to solubilize CS.
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 Research J. Pharm. and Tech. 4(4): April 2011

Chitosan is biocompatible with living tissues since it does
not cause allergic reactions and rejection. It breaks down
slowly to harmless products (amino sugars), which are
completely absorbed by the human body. Chitosan degrades
under the action of ferments, it is nontoxic and easily
removable from the organism without causing concurrent
side reactions12,13. It possesses antimicrobial property and
absorbs toxic metals like mercury, cadmium, lead, etc. In
addition, it has good adhesion, coagulation ability, and
immunostimulating activity. If degree of deacetylation and
molecular weight of CS can be controlled, then it would be
a material of choice for developing micro/nanoparticles.
Chitosan has many advantages, particularly for developing
micro/nanoparticles. These include: its ability to control the
release of active agents, it avoids the use of hazardous
organic solvents while fabricating particles since it is
soluble in aqueous acidic solution, it is a linear polyamine
containing a number of free amine groups that are readily
available for crosslinking, its cationic nature allows for
ionic crosslinking with multivalent anions, it has
mucoadhesive14-17character, which increases residual time at
the site of absorption, and so on. Chitin and CS have very
low toxicity; LD50 of CS in laboratory mice is 16 g/kg
body weight, which is close to sugar or salt. Chitosan is
proven to be safe in rats up 10% in the diet. Various
sterilization methods such as ionizing radiation, heat, steam
and chemical methods can be suitably adopted for
sterilization of CS in clinical applications. In view of the
above-mentioned properties, CS is extensively used in
developing drug delivery systems. Particularly, CS has been
used in the preparation of mucoadhesive formulations
improving the dissolution rate of the poorly soluble drugs
drug targeting and enhancement of peptide absorption.
However, the micro/nanoparticulate drug delivery systems
offer numerous advantages over the conventional dosage
forms. These include improved efficacy, reduced toxicity
and improved patient compliance. The present review
addresses the preparation of chitosan nanoparticles by
ionotropic gelation method18-21.
Methods of preparation of chitosan nanoparticles
Different methods such as ionotropic gelation, emulsion
cross-linking, nanoprecipitation, salting out etc have been
used to prepare CS particulate systems. Selection of any of
the methods depends upon factors such as particle size
requirement, thermal and chemical stability of the active
agent, reproducibility of the release kinetic profiles,
stability of the final product and residual toxicity associated
with the final product. Since we are concerned only with the
ionotropic gelation method, we will restrict our discussions
only on these aspects.
Ionotropic Gelation Method
The use of complexation between oppositely charged
macromolecules to prepare CS nanoparticles has attracted
much attention because the process is very simple and mild.
In addition, reversible physical cross-linking by
electrostatic interaction, instead of chemical cross-linking,
has been applied to avoid the possible toxicity of reagents
and other undesirable effects. Tripolyphosphate (TPP) is a
polyanion, which can interact with the cationic CS by
electrostatic forces. After Bodmeier et al, reported the
preparation of TPP–CS complex by dropping CS droplets
into a TPP solution, many researchers have explored its
potential pharmaceutical usage. In the ionic gelation
method, CS is dissolved in aqueous acidic solution to obtain
the cation of CS. This solution is then added dropwise
under constant stirring to polyanionic TPP solution. The
chitosan molecules has abundant NH3 group which can
react with negatively charged phosphoric ions of TPP to
form cross-linked chitosan nanoparticles. During the
process of cross-linking and hardening process water was
extruded from the particles, which may help in sustaining
the release of drug. Three kinds of phenomena were
observed: solution, aggregation and opalescent suspension
while preparing the nanoparticles. The last stage indicates
the completion of the process. Insulin-loaded CS
nanoparticles have been prepared by mixing insulin with
TPP solution and then adding this to CS solution under
constant stirring. Two types of CS in the form of
hydrochloride salt (SeacureR 210 Cl and ProtasanR 110
Cl), varying in their molecular weight and degree of
deacetylation, were utilized for nanoparticle preparation.
For both types of CS, TPP concentration was adjusted to get
a CS/TPP ratio of 3.6:1. Chitosan nanoparticles thus
obtained were in the size range of 300–390 nm with a
positive surface charge ranging from +34 to +45 mV. Using
this method, insulin loading was modulated reaching the
values up to 55%. Efficiency of the method was dependent
upon the deacetylation of CS, since it involves the gelation
of protonated amino groups of CS1,18,22,23.

There are many ongoing investigations, which demonstrate

the improved oral bioavailability of peptide and protein
formulations. Bioadhesive polysaccharide CS nanoparticles
would seem to further enhance their intestinal absorption.
Yan et al. prepared the insulin-loaded CS nanoparticles by
ionotropic gelation of CS with TPP anions. Particle size
Fig 1. Schematic representation of preparation of chitosan distribution and zeta potential were determined by photon
nanoparticles by ionotropic gelation method correlation spectroscopy. The ability of CS nanoparticles to
493
 Research J. Pharm. and Tech. 4(4): April 2011

enhance the intestinal absorption of insulin and the relative

pharmacological bioavailability of insulin was investigated
by monitoring the plasma glucose level of alloxan-induced
diabetic rats after the oral administration of various doses of
insulin-loaded CS nanoparticles. The positively charged,
stable CS nanoparticles showed particle size in the range of
250–400 nm. Insulin association was up to 80%. The in
vitro release experiments indicated initial burst effect,
which is pH-sensitive. The CS nanoparticles enhanced the
intestinal absorption of insulin to a greater extent than the
aqueous solution of CS in vivo. After administration of 21
I.U./kg insulin in the CS nanoparticles, hypoglycemia was
prolonged over 15 h. The average pharmacological
bioavailability relative to s.c. injection of insulin solution Fig 3. Chitosan nanoparticles prepared by ionotropic gelation
was up to 14.9%2,4,6,8. method
Physicochemical, Biological and Pharmacological Properties of
Chitosan
Physical Properties: Particle Size 30mm
Density 1.35-1.40 g/cc
pH 6.5-7.5
Solubility Insoluble in water but
soluble in acids
Chemical properties
o Cationic Polyamine
o High charge density at pH 6.6
o Adheres to negatively charge surfaces
o Forms gels with polyanions
o High molecular weight, linear
polyelectrolyte
o Viscosity- high to low chelates certain
transitional metals
o Reactive hydroxyl/ amino group
Biological Properties
o Nontoxicity
o Biocompatibility
o Biodegradability

Pharmacological Properties
o Hypocholesterolemic action
o Wound-healing properties
o Antacid and antiulcer activity
Fig 2. Interactions of chitosan with TPP (A) Deprotonation, (B) o Antifungal and antibacterial activity
Ionic cross linking
Table 1. Properties of chitosan

Lifeng Qi et al. have prepared the chitosan nanoparticles by

ionotropic gelation method and also evaluated the
antibacterial activity of chitosan nanoparticles. They have Advantages of ionotropic gelation method1,5,7
also characterized the particles by SEM,AFM and their • The method is very economic and simple
MIC value were less than 0.25 g/ml, and the MBC values • The method requires less equipment and time
of nanoparticles reached 1 g/ml .AFM revealed that the • In addition, reversible physical cross-linking by
exposure of S.choleraesuis to the chitosan nanoparticles electrostatic interaction, instead of chemical cross-
lead to the disruption to the cell membranes and the leakage linking, has been applied to avoid the possible toxicity
of cytoplasm21. of reagents and other undesirable effects.
• No use of organic solvent.
Zengshuan Ma et al. have prepared chitosan insulin
nanoparticles by ionotropic gelation of chitosan with TPP
anions. The ability of chitosan nanoparticles to enhance the Disadvantages of ionotropic gelation method7
intestinal absorption. The relative pharmacological activity The only disadvantage of TPP/CS nanoparticles is their
and bioavailability of insulin were investigated by poor mechanical strength
monitoring the plasma glucose level of administration of
various dose of insulin loaded chitosan nanoparticles24.

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 Research J. Pharm. and Tech. 4(4): April 2011

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