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Int. J. LifeSc. Bt & Pharm. Res. 2012 K L Senthilkumar et al., 2012
PREPARATION AND CHARACTERIZATIONOF NABUMETONE LIPOSOMES
K L Senthilkumar1*, R P Ezhilmuthu2, Praveen P2
Research Paper
Nabumetone is the effective drug for the treatment of reumatoid arthritis and osteoarthritis. Thedrug has a half life of 23 hrs and its oral bioavailability is only 30%. The aim of the study is toprepare 6 formulations of liposomal carrier for Nabumetone for the treatment of arthritis that iscapable of delivering the drug to the specific target site by topical route by using different ratiosof phospho lipid and cholesterol with a desired amount of drug by thin film hydration techniqueand to find out the drug release from the liposome's of different ratios, mechanism kinetics ofdrug release pattern and also to find out the size distribution of liposome's of different ratios andalso to increase the bioavailability and efficacy of the drug.
Keywords: Liposome's, Thin Film Hydration Method, Nabumetone.
*Corresponding Author: K L Senthilkumar,[email protected]
INTRODUCTIONLiposomes are colloidal particles formed as
concentric bimolecular layers that are capable of
encapsulating drugs. They are lipid vesicles that
fully enclose an aqueous volume. These lipid
molecules are usually phospholipids with or
without some additives. Cholesterol may be
included to improve bilayer characteristics of
Liposome’s; increasing micro viscosity of the
bilayer, reducing permeability of the membrane
to water soluble molecules, stabilizing the
membrane and increasing rigidity of the vesicle.
Liposomes are micro vesicles, whose
membranes are made of phospholipids, which
ISSN 2250-3137 www.ijlbpr.comVol.1, Issue. 1, January 2012
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Int. J. LifeSc. Bt & Pharm. Res. 2012
1 Department of Pharmaceutics, Padmavathi college of pharmacy and Research institute, Dharmapuri.2 Department of Pharmaceutics, Padmavathi college of pharmacy and Research Institute, Dharmapuri.
are bilayer having head and tail. The head group
is hydrophilic and the tail group which is made of
long hydro carbon chain is hydrophobic. The drug
molecules can be encapsulated in aqueous
space or into lipid bilayer. The exact location of
the drug will depend upon its physicochemical
properties and composition of lipids. The water
soluble substances can be trapped inside the
water sphere while fat soluble substances can
be trapped inside the fat soluble opposite end of
the molecule. Liposome’s give a unique delivery
system for nutrients, vaccines, enzymes, or
drugs. Liposomes are effective in treating
diseases that affect the phagocytes of the immune
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Int. J. LifeSc. Bt & Pharm. Res. 2012 K L Senthilkumar et al., 2012
system because they tend to accumulate in the
phagocytes, which recognize them as foreign
invaders. They have also been used to carry
normal genes into a cell in order to replace
defective, disease-causing genes. Because of
their moisturizing qualities Liposome’s are also
used in cosmetics.
Liposome’s, which are vesicles consisting of
one or more concentrically ordered assemblies
of phospholipids bilayer, range in size from a
nanometer to several micrometers. Phospholipids
such as egg phosphatidylcholine, phosphatidyl-
serine, synthetic dipalmitoyl-DL-alpha-
phosphatidylcholine or phosphatidylinositol, have
been used in conjunction with cholesterol and
positively or negatively charged amphiphiles such
as stearylamine or phosphatidic acid. Because
of the multifold characteristics as drug carriers,
Liposome’s have been investigated extensively
as carriers of anticancer agents for the past
several years. Liposomal entrapments include a
variety of pharmacologically active compounds
such as antimalarial, antiviral, anti-inflammatory
and anti-fungal agents as well as antibiotics,
prostaglandins, steroids and bronchodilators.
MATERIALS AND METHODSNabumetone drug, Soya lecithin 30% Cholesterol,
Chloroform, Methanol Potassium dihydrogen
phosphate, Disodium hydrogen phosphate,
Sodium chloride, Acetone were used for this study.
Methods
Formulation of Liposomes
Drug: Soya lecithin: Cholesterol in different ratios
was dissolved in 5 ml chloroform. This solution
was taken in a 250 ml round bottom flask. The
flask was rotated in rotary flash evaporator at 80
rpm for 15 minutes in thermostatically controlled
water bath at 40°C under vacuum 900 mmHg.
The organic solvent was slowly removed by this
process such that a very thin film of dry lipids
was formed on the inner surface of the flask. The
dry lipid film was slowly hydrated with 5 ml of
phosphate buffer ph 7.4.the fluid is allow to
hydrate with phosphate buffer pH 7.4 for 2 hr
Swirling the contents to yield milky white
suspension. The formulation is subjected to
centrifugation. The unentrapped drug is removed
by centrifugation at 3000 rpm for a period of 30
minutes. The pellets are dispersed in phosphate
buffer.
CHARACTERIZATION OFLIPOSOMESDrug Entrapment Efficiency
Drug entrapment efficiency was calculated by
using centrifugation method. The liposome
suspension of 1ml was taken and centrifuged at
3500rpm for 15 min. The sediment obtained from
the centrifugation was suspended in 100 ml of
phosphate buffer pH 7.4. Then the absorbance
was taken at 261 nm. From that the amount
present in 1 ml of suspension was obtained. The
drug entrapment efficiency was calculated from
the following formula.
Amount of Drug in the sediment% Drug Entrapped (PDE) = ×100
Total amount of Drug
Particle Size Analysis
The particle size of Liposome’s was
determined by optical microscopy by using Perkin
Elmer. All the prepared batches of Liposome’s
were viewed under microscope to study their size.
Size of liposomal vesicles was measured at
different location on slide by taking a small drop
of liposomal dispersion on it and average size of
liposomal vesicles were determined
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Int. J. LifeSc. Bt & Pharm. Res. 2012 K L Senthilkumar et al., 2012
In Vitro Drug Release Study
The release studies were carried out in diffusioncell having 10 ml capacity. 10 ml Phosphate bufferpH 7.4 was placed in diffusion cell. The diffusioncell contained a magnetic bed and the mediumwas equilibrated at 37±50C. Dialysis membranewas taken and placed on the diffusion cell. Afterseparation of non-entrapped Nabumetone,liposome dispersion was filled in the dialysismembrane. The dialysis membrane containingthe sample was suspended in the medium.Aliquots were withdrawn (1 ml) at specificintervals, f iltered, diluted with phosphatebuffer and the absorbance was taken at 261 nm.Then the apparatus was immediately replenishedwith same quantity of fresh phosphate buffer pH
7.4 medium.
RESULTS AND DISCUSSIONFTIR of pure drug and physical mixture.
The drug release kinetics of prepared
Nabumetone Liposomes were conducted using
Higuch's model and Poppa's model. Higuch's plot
was done with square root of time in X axis and
%cumulative drug in Y axis (Figure 1). The
Poppa's plot was done with log time in X axis and
log %cumulative drug release in Y axis. The study
of drug release kinetics showed that majority of
the formulations governed by peppas model.
Results showed that F1 have a particle size
of 7.994 micron and F6 have a highest value com-
pared to other formulations. From the results of
particle size analysis F1 shows satisfactory
particle size (Figure 2).
According to the drug entrapment study
conducted the maximum drug entrapment was
shown by F1 (52±0.5).
Figure 1: FTIR of Pure Drug and Physical Mixture
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Int. J. LifeSc. Bt & Pharm. Res. 2012 K L Senthilkumar et al., 2012
Figure 2: In Vitro Drug Release Study of F1 to F6
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Int. J. LifeSc. Bt & Pharm. Res. 2012 K L Senthilkumar et al., 2012
Figure 3: SEM Photographs of Liposome’s
CONCLUSIONNabumetone Liposome's were prepared using
soya lecithin, cholesterol and chloroform as
solvent by thin film hydration method using rotary
evaporator. The prepared Liposomes were
evaluated by drug entrapment study, particle size
analysis. In vitro drug release study and
mechanism of release kinetics using Higuchi's
plot and Korsemeyer Peppas plot.
The present study demonstrated the
successful preparation of Nabumetone Liposomes
and its evaluation. Formulation F1 showed high
encapsulation efficiency with minimum particle
size and drug release over a 4 hr, hence suppose
to give greater bioavailability and considered as
good liposomal formulation. The difference in
drug entrapment and drug release was due to
the different ratios of lipid to cholesterol. The drug
entrapment shows decreasing when the
concentration of lipid and cholesterol changes.F1
have high entrapment and satisfactory drug
release within 4 hr. F1 shows least particle size
from the results F1 selected as a good formulation
of Nabumetone Liposomes.
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