“Liposomes and Niosomes

Drug Delivery Systems”

Presented by :
MUTHIA FADHILA
1821012003

Lecturer
Dr. Muslim Suardi, MS, Apt
 Studi biofarmasetika
Sifat fisika dan Rute pemakaian
kimia bahan obat obat

Efek Sifat toksikologi

farmakodinamik obat

Pengaruh eksipien
Keamanan eksipien
dan bentuk sediaan

Shargel, 2004
 LIPOSOMES
• A Liposome is a spherical vesicle with a membrane
composed of a phospholipid & cholesterol bilayer.
• Liposomes can be produced from cholesterols, non toxic
surfactants,sphingolipids,glycolipids,long chain fatty
acids & even membrane proteins.
• Liposomes are the drug carrier loaded with great variety
of molecules such as small drug molecules, proteins,
nucleotides & even plasmids.
• Liposomes can be made in a particular size range that
makes them viable targets for natural macrophage
phagocytosis.
Hu, et al., 2016
 STRUCTURAL COMPONENTS
The main components of liposomes are :

1. Phospholipids

2. Cholesterol

Hu, et al., 2016

 PHOSPHOLIPIDS
• Phospholipids are the major structural components
of biological membranes such as the cell
membrane.

Two types of
Phospholipids
• Phosphoglycerides
(along with their • Sphingolipids
hydrolysis
products)
Yadav, et al., 2017
• Each phospholipid molecule has 3 major parts, 1
head & 2 tails. Head is made from 3 molecular
components: choline , phosphate & glycerol which
is hydrophilic. Each tail with a long chain which are
hydrophobic.

Yadav, et al., 2017

Commonly Used Phospholipids

Yadav, et al., 2017

 CHOLESTROL
• Cholesterol is generally used steroid in the
formulation of liposomes.
• It improves the fluidity of the bilayer membrane and
reduces the permeability of bilayer membrane in
the presence of biological fluids such as blood /
plasma.
• Cholesterol appers to reduce the interactions with
blood proteins.

Yadav, et al., 2017

 CLASSIFICATION
Liposomes can be
classified based on three
different criteria

Based on structural Based on method of Based on composition and

parameters preparation applications

Yadav, et al., 2017

Yadav, et al., 2017
Yadav, et al., 2017
Yadav, et al., 2017
 Transferosomes

Archaeosomes

NOVEL
Proteosomes
LIPOSOMES

Virosomes

Ethosomes
Yadav, et al., 2017
Yadav, et al., 2017
MECHANICAL DISPERSION Yadav, et al., 2017

• Hand-shaken Multi-lamellar vesicles :

• Non-Shaking Vesicles : Yadav, et al., 2017
• Microemulsification Liposomes :

Yadav, et al., 2017

• Sonicated Uni-lamellar Vesicles :

Yadav, et al., 2017

SOLVENT DISPERSION
• Ethanol Injection :

Yadav, et al., 2017

• Ether Injection :

Yadav, et al., 2017

• Reverse Phase Evaporation Vesicles :

Yadav, et al., 2017

 DETERGENT SOLUBILIZATION

• Their shape & size depends on the chemical nature of

detergent , the concentration & other lipids involved.
• The concentration of detergent in water at which micelles just
start to form is known as critical micelle concentration.Yadav, et al., 2017
A three stage model of interaction for detergents with
lipid bilayers:

Stage2: After reaching a

Stage1: At low
critical detergent
concentration detergents
concentration, membrane Stage3: All lipid exists in
equilibrates between
structure tends to unstable mixed micelle form.
vesicular lipid and water
and transforms gradually
phase.
in to micelles.

Yadav, et al., 2017

Some methods are applied for removal of detergent
and transition of mixed micelles to concentric
bilayered form.

DIALYSIS:
• The molecules of detergent are removed from mixed micelle
by dialysis by lowering the concentration of detergent in bulk
aqueous phase.
• eg: sodium cholate,octylglucoside.

COLUMN CHROMATOGRAPHY:
• Removal of detergent is achieved by by passing the
dispersion over a sephadexg-25 column pre-saturated with
constitutive lipids and pre-equilibrated with hydrating buffer.
• eg: deoxycholate.
Yadav, et al., 2017
 EVALUATION
1) Particle size
Microscopic method
• Light microscopy has been utilized to examine the gross size distribution of
large vesicles produced from single chain amphiphiles.
Laser light scattering
• Proton correlation spectroscopy is the analysis of time dependence of
intensity fluctuation in scattered laser light due to the Brownian motion of
particles in solution/suspension.
• Translational diffusion coefficient can be measured here to determine the
mean hydrodynamic radius (Rh) of the particles using stokes-Einstein
equation
Gel permeation
• Exclusion chromatography on large pure gels enable to separate small
uni-lamellar vesicles from radial multi-lamellar vesicles.

Kalepu, et al., 2013

2) Surface Charge
• Free-flow electrophoresis on a cellulose acetate plate in
a sodium borate buffer pH 8.8 & Zeta potential
measurement can also be done.
• The samples are applied to plate & electrophoresis is
carried out at 4˚C on a flat bed apparatus for 30 min.
• The plate is dried and phospholipids are visualised by the
molybdenum blue reagent
• The surface charge can be calculated by estimating the
mobility of the liposomal dispersion in a suitable buffer.
Kalepu, et al., 2013
3) Percent Capture (Entrapment)
• This can be determined by ‘PROTAMINE AGGREGATE’ &
‘MINICOLUMN CENTRIFUGATION method .
• In Mini-column centrifugation , the hydrated gel is filled in a
barrel of 1ml syringe.
• This barrel is rested in a centrifuge tube spun at 2000rpm for 3
min to remove excess saline solution from gel.
• After centrifugation , gel column is dried.
• Liposome suspension is added dropwise to top of gel bed, &
column is spun at 2000rpm for 3min to expel void volumes
containing liposomes into centrifuge tube.
• The elute is then removed & set aside for assay.
Kalepu, et al., 2013
4) Lamellarity
• The average number of bilayers present in liposome can be
found by Freeze electron microscopy & P NMR method.

5) Phase Behaviour
• Liposomes at transition temperature undergo reversible phase
transition i.e the polar head groups in gel state become
disordered to form the liquid crystalline state which can be
determined by DSC.
• The phase transition temperature (Tc) is a function of
phospholipid content of bilayers.
• Tc can give good clues regarding liposomal stability ,
permeability or whether the drug is entrapped in bilayers or in
aqueous compartments.
Kalepu, et al., 2013
6) Drug Release

• The mechanism of drug release from liposome can

be assessed by the use of a well calibrated in-vitro
diffusion cell.
• The rate of drug release from the liposomes can be
determined by in vitro assays which helps to predict
the pharmacokinetics and bioavailability of the
drug.
Kalepu, et al., 2013
7) Quantitative determination of phospholipids
• (i) Initially the phosphorous present in the lipid bilayer of the sample is
hydrolyzed to inorganic phosphate.
• (ii) Then ammonium molybdate is added to convert inorganic phosphate
to phosphomolybdic acid(PMA).
• (iii) The sample is then treated with aminonaphthylsulphonic acid to
quantitatively reduce the PMA to a blue-
• coloured compound.
• (iv) The intensity of the blue colour produced can be measured by
spectrophotometric means and the value is plotted on the standard
curve to obtain the content of phospholipids

8) Quantitative determination of cholesterol

• The sample is reacted with a reagent (containing ferric perchlorate, ethyl

acetate and H₂SO₄) and the absorbance of purple coloured complex is
measured at 610 nm.

Kalepu, et al., 2013

 APPLICATIONS
Liposomes in cancer
therapy
Liposomes as carrier of
drug in oral treatment
Drug targeting
Liposomes in delivery of Liposomes in
anti-microbial agents ophthalmic delivery
Protection against
Liposomes as Vaccine
enzyme degradation of
adjuvants
drugs
Topical drug delivery
Kalepu, et al., 2013
Wagh, et al., 2012
 Introduction
• Niosomes have potential applications in topical
drug delivery system.
• The objective of the study was to formulate and
evaluate the niosome of Itraconazole. Surfactant :
cholesterol ratio and quantity of ethanol used were
studied by applying factorial design.

Wagh, et al., 2012

 Evaluation
• Formulated niosomes were evaluated for vesicle
size, entrapment efficiency, drug release, skin
permeation, and antimycotic activity.

Wagh, et al., 2012

Wagh, et al., 2012
Wagh, et al., 2012
Wagh, et al., 2012
Wagh, et al., 2012
Wagh, et al., 2012
Wagh, et al., 2012
Wagh, et al., 2012
Wagh, et al., 2012
Wagh, et al., 2012
Wagh, et al., 2012
 Conclusion
• Vesicle size, entrapment efficiency, and drug release
were markedly dependent on surfactant : cholesterol
ratio and quantity of ethanol used.
• Permeation of the drug through the skin was affected by
cholesterol content in formulation.
• Itraconazole niosome were having larger zone of
inhibition than marketed formulation when activity was
checked against C. albicans.
• Niosomes may be a promising carrier for topical delivery
of Itraconazole especially due to their simple production.

Wagh, et al., 2012

 DAFTAR PUSTAKA
• Hu, Y.J., Fan Z., Rui-Jun J., & Wan-Liang L. 2016. Advances in
Liposomal Drug Delivery System in the Field of Chemotherapy.
Clinics in Oncology, Volume 1, Article 1092
• Kalepu, S., Sunilkumar K.T., Sudheer B., & Mohanvarma M. 2013.
Liposomal drug delivery system - A Comprehensive Review. Int. J.
Drug Dev. & Res, Vol. 5, Issue 4, ISSN 0975-9344
• Shargel, L., Andrew, B.C & Sussanna, W.U. 2004. Apllied
Biopharmaceutics and Biopharmakokinetics 5th Ed. Boston:
Appleton Century Croft.
• Wagh V.D., & Onkar J.D. 2012. Itraconazole Niosomes Drug
Delivery System and Its Antimycotic Activity against Candida
albicans. International Scholarly Research Network, Volume 2012,
Article ID 653465, 7 pages
• Yadav, D., Kumar S., Deepak P., & Ranu K.D. 2017. Liposomes for
Drug Delivery. Journal of Biotechnology & Biomaterials, 7(4)