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International Journal of Pharmaceutical Research | July – Sept 2015 | Vol 7 | Issue 3|29
Research Article
Formulation and Characterization of Rutin trihydrate
Liposomes for Topical Delivery
APARAJITA VARSHNEYA
AND PADMINI RAVIKUMAR
Department of Pharmaceutics, SVKM’s Dr. Bhanuben Nanavati College of Pharmacy,
Gate No: 1, Mithibai College Campus, V.M. Road, Vile Parle (W), Mumbai-400 056, India
ABSTRACT
In the present study, liposomes were formulated as drug carriers for enhancing the delivery of an antioxidant Rutin
trihydrate for topical administration. Formulation of liposomes was done by thin film hydration method. Optimization
was achieved by varying the lipid to cholesterol ratio and gradually increasing the amount of drug until maximum
entrapment efficiency was achieved. The formulation and process parameters were optimized to attain multilamellar
liposomes with homogeneous size and good entrapment. The optimized batch gave entrapment efficiency of 88%.
Gels were prepared by using Carbopol 940 as the gelling agent. The in vitro and ex vivo release profile of liposomal gels
was compared with conventional gel formulations. The liposomal gel showed prolonged drug release upto 12 h and
also exhibited anti-elastase activity. The safety of liposomal gels was ensured by conducting skin irritation studies on
albino Wistar rats.
Keywords: Liposomes, Rutin trihydrate, topical delivery, gels.

Address for correspondence 
Aparajita Varshneya, SVKM’s Dr. Bhanuben Nanavati College of Pharmacy, Gate No.1, Mithibai, College Campus, V.M. 
Road, Vile Parle (West), Mumbai ‐ 400 056, India. 
Phone no: +91‐9870687261 e‐mail: aparajita.varshney@yahoo.com 
Received:18/02/15, Revised:02/04/15, Accepted:18/04/15 
INTRODUCTION
Topical drug delivery has long been sought after, as
it is coupled with well-established advantages like
localised drug delivery, avoidance of first pass
metabolism, improved patient compliance owing to
its non-invasive nature and reduced systemic side
effects [1]. Despite these merits, the brick and mortar
structure of stratum corneum i.e. the horny layer
serves as the principal barrier to the percutaneous
absorption of drugs [2,3]. Owing to these barrier
properties of the skin, the delivery of active
substances from conventional formulations is
generally compromised. Thus, there arises the need
for a suitable carrier to enhance drug delivery [4].
Liposomes
Liposomes were first described by British
haematologist Dr. Alec D Bangham in early 1960’s
[5]. Liposomes are highly organized structures,
composed of concentric bi-layered vesicles in which
an aqueous volume is enclosed by a membranous
lipid bilayer [6]. The main advantages that have
made these vesicular systems favourable for drug
delivery are their tissue compatibility,
biodegradability, safety and their ability to entrap
almost any drug in the bilayer core or in the outer
domain. Additionally, liposomal drug formulations
can be used to overcome a drug’s non-ideal
properties and unfavourable pharmacokinetic
profiles [7]. Regarding topical application, liposomes
offer a wide range of advantages including
moisturization and prolonged dermal release [8].
Topical use of antioxidants has been adopted as an
important strategy in cosmeceutical industry to avoid
wrinkling of the skin and protect it from degenerative
effects such as photoaging, sunburn,
photocarcinogenesis etc. [9]. Antioxidants help in
preventing and repairing damage caused due to
oxidative stress. They scavenge free radicals which
are postulated to be the mediators in skin aging [10].
They are found to be useful in delaying the signs of
aging such as fine lines, wrinkles and age spots.
Antioxidants are promising in photoprotection with
negligible side effects at physiological concentrations
[11]. Of the various antioxidants, bioflavonoids are
one of the most powerful antioxidants. They are a
class of plant secondary metabolites possessing a
broad range of pharmacological activities. Rutin
trihydrate was chosen as the active molecule as it is a
powerful antioxidant which inhibits free-radical
mediated cytotoxicity and lipid peroxidation. It is
sparingly soluble in water (0.125 g/L). This limits its
use in topical delivery. Thus liposomes were chosen
as drug carriers as they are amphiphilic in nature
and lipophilic molecules get embedded in their
concentric bilayers. Liposomes being physiologically
similar to cell membrane, are nontoxic in nature and
are easily re-absorbed from the epidermis into the
deepest layers [12]. Thus, the objective of the study
was to enhance the accumulation of drug at the site
of administration and achieve prolonged release by
means of liposomal formulation.
Aparajita Varshneya et al /Formulation and Characterization of Rutin Liposomes
30 | International Journal of Pharmaceutical Research | July – Sept 2015 | Vol 7 | Issue 3
MATERIALS AND METHODS
Phosphatidylcholine (Leciva S-70) was a generous
gift sample from VAV Life Sciences, Mumbai, India.
Rutin trihydrate was purchased from Loba Chemie,
Mumbai and Cholesterol was procured from
Qualigens, Mumbai. Carbopol 940, Glass beads
(2.5 mm-3.5 mm) and all other AR grade reagents
were purchased from S D Fine-Chem Ltd., Mumbai.
Dialysis membrane 150 was purchased from
HiMedia Laboratories Pvt. Ltd., Mumbai and porcine
skin was procured from local market. Healthy male
albino Wistar rats (Rattus norvegicus sp.) were
purchased from Bharat Serum, Mumbai, India.
Liposomes were prepared using Rotary Evaporator,
Roteva, Equitron and UV-visible spectrophotometer,
Shimadzu 1800 was used for evaluation.
I. Preparation of Rutin Trihydrate Liposomes:
Multilamellar vesicles were prepared by thin film
hydration technique as described by Bangham et al.
[13]. Phosphatidylcholine, Cholesterol and Rutin
trihydrate constituted the lipid mixture. This lipid
mixture was dissolved in chloroform and methanol
(2:1) in a round bottom flask (RBF) and 25 g glass
beads were added for homogeneous film formation.
The RBF was attached to the rotary flash evaporator
(Roteva, Equitron) and allowed to rotate at 80 r/min
in a thermostated water bath at 40°. The organic
solvents were then removed by slow application of
vacuum leading to lipid film formation on the walls
of the flask. The film was allowed to dry for 60 min
to ensure complete removal of solvent. The dried
lipid film was hydrated with phosphate buffer (pH:
5.4) at 150 r/min and 46° that is above the transition
temperature of lipid. Rotation was continued for 30
min. The Rutin trihydrate liposomal suspension was
kept at room temperature for 2 h for complete
hydration and for annealing structural defects.
The liposomes produced by this method are large
and heterogeneous in size, thus they were downsized
by bath sonication for 20 min. The suspension was
also vortexed for few minutes. The liposomal
suspension was characterized for physical
appearance, settling time, re-dispersion
time, entrapment efficiency and vesicle size by optical
microscopy. It was then filled in amber colored glass
bottles and stored at 4° until further use.
II. Characterization of Liposomes:
Rutin trihydrate liposomes were characterized for
following parameters:
Optical Microscopy
A drop of the liposomal suspension was placed on a
clean glass slide and observed under high power
45x and 100x of the optical microscope (Motic
microscope). Multilamellar vesicles (MLV) were
clearly identified (Figure 1). Microscopic observations
were used to study presence of aggregates,
precipitation or leakage of drug. Size and shape of
the vesicles was noted for all liposomal batches.
Figure	 1:	 (a)	 Large	 multilamellar	 vesicles	 as	
visible	under	100x	magnification	
Determination of Vesicle Entrapment Efficiency
Entrapment efficiency for the batches prepared
during the optimization process was estimated by the
following procedure:
The liposomal pellet i.e. the drug loaded liposomes
were separated from the aqueous phase by
centrifugation at 15,000 r/min using Microspin
centrifuge for 30-45 min until a clear supernatant
was obtained. The amount of drug loaded was
estimated by both direct and indirect method.
Direct method: After centrifugation, the supernatant
was decanted. The formed pellet was washed with
phosphate buffer and vortexed, in order to remove
the free drug adsorbed on the surface of liposomes.
This step was repeated thrice to completely separate
the unentrapped drug. The resulting pellet was then
dissolved in methanol and sonicated for 20 min.
(This causes rupturing of pellet and release of
entrapped drug in the solution). The resulting
methanolic solution was then analyzed by UV
spectrophotometry using the developed analytical
method.
% Entrapment Efficiency (E.E.) =
 
(Amount of drug quantified in the pellet) x 100
Total amount of drug added
Indirect method: The supernatant of the liposomal
dispersion decanted after centrifugation and the
solutions collected after washing of pellet were
combined and analyzed by developed UV
spectrophotometric method for the amount of drug
present. % Entrapment efficiency was estimated as
follows:
% Entrapment Efficiency (E.E) =
(Total amount of drug added - Amount of drug
detected in the supernatant) x 100_
Total amount of drug added
Determination of vesicle size and size distribution
Vesicle size was determined by dynamic light
scattering using a computerized inspection system
(Malvern particle size analyzer). Freshly prepared
batches of vesicles were diluted ten-fold with filtered
Aparajita Varshneya et al /Formulation and Characterization of Rutin Liposomes
International Journal of Pharmaceutical Research | July – Sept 2015 | Vol 7 | Issue 3|31
distilled water and then analyzed at 25° using ten
runs at each scan. Optimization time was fixed at
120 s. Vesicle size and polydispersity index were
measured for the liposomal formulation. The
measurements were done in triplicate.
Zeta potential measurement
Liposomal suspension was diluted with filtered
distilled water and zeta potential was measured in
triplicate with the help of Malvern zetasizer.
Transmission Electron Microscopy (TEM)
TEM analysis was carried out for optimized drug
loaded liposomal suspension. A drop of the diluted
formulation was placed with the aid of a micropipette
on a coated copper grid 3 mm in diameter. Negative
staining was carried out by uranyl acetate. The grid
was allowed to dry under an IR lamp for about 20
min. The grid was then loaded on a probe which was
inserted in the slot designed for it in the electron
microscope. The grid containing the sample was then
bombarded with electrons accelerated at 200 kV and
the sample visualized over a fluorescent stage.
Vesicle size of the sample was measured and marked
and its morphology observed on the computer
(Figure 2).
Figure	2:	Negative	staining	TEM	imaging	of	Rutin	
trihydrate	liposomesMagnification	100,000x	(size	
approx.		600	nm)	
III. Optimization of Process and Formulation
Parameters:
Many factors influence the formation of liposomes
and its characteristics. Therefore, the following
process parameters were optimized to obtain
liposomes with maximum entrapment efficiency,
homogeneous size and stability.
a. Process Parameters
Speed of the rotary evaporator
It is desirable to obtain a thin and uniform film as it
governs the final output of liposomal preparation.
The speed of rotation was varied from 60 r/min to
150 r/min during film formation as well as during
hydration.
The ratio and volume of solvent system
The solvent system was optimized by taking various
combinations of organic solvents i.e. chloroform and
methanol. The ratios 1:1, 2:1 and 3:1 were tried and
the film was evaluated in terms of its uniformity.
Quantity of glass beads
Thin film hydration technique is readily employed for
its feasibility on a laboratory scale, but problems
related to limitations of surface area exist. Glass
beads are commonly used for this purpose which
helps to increase the surface area to provide a thin
film. Liposomal dispersions were prepared using
different ratios of glass beads (diameter: 2.5 mm-3.5
mm) and solvent volume such as 0:1, 2:1, 3:1 and
4:1. The formulations were evaluated in terms of
uniformity of film and separation of vesicle
aggregates (microscopic observation).
Vesicle sizing technique
Size reduction of the vesicles is done to obtain a
homogeneous population of vesicles with low
polydispersity index. The various techniques studied
for size reduction were bath sonication using
ultrasonic bath, high pressure homogenization and
by using ultra turrax. Bath sonication was carried out
at room temperature for 20-30 min. Size reduction
using Ultra turrax was done at 6000 r/min for 15
min. High pressure homogenization was also tried by
carrying out approximately 10 cycles at a pressure of
5000 psi for 10 min. The effect of these techniques
was studied by estimating entrapment efficiency,
vesicle size and polydispersity index.
pH of the hydrating media
The effect of pH of the phosphate buffer was studied
on the formulation. The effect of pH on the
entrapment of the drug is related to its pKa. Hence
the pH of the hydrating buffer was adjusted at values
closer to the pKa of the drug and entrapment
efficiency was estimated. Distilled water, phosphate
buffer pH 5.4, 6.8 and 7.4 were used as hydrating
media and formulations were studied in terms of
colour, sedimentation, redispersibility and
entrapment efficiency.
Table	1:	Formula	optimization	based	on	lipid	concentration	
Lipid Conc. (%) Consistency Settling rate Redispersibility
Vesicle size
(μ)
0.5 Thin suspension +++ + 1-4
1
Moderate, smooth
suspension
++ +++ 0.25-3
2 Thick suspension + ++ 2-6
Aparajita Varshneya et al /Formulation and Characterization of Rutin Liposomes
32 | International Journal of Pharmaceutical Research | July – Sept 2015 | Vol 7 | Issue 3
Settling rate: + Slow ++ Moderate +++Fast Redispersibility + Poor ++ Good
+++Excellent
Table‐2:	Formula	optimization	based	on	Drug:	Lipid	and	PC:	CH	ratio	
Batch No.
PC:CH
(molar
ratio)
Total lipid: 100 mg
Globule Size
(μ)
EE
(%)Amount of PC
(mg)
Amount of CH
(mg)
B1 1:0 100 0 0.5-5 56
B2 1:0.2 90.75 9.25 0.5-1.5 88
B3 1:0.5 79.68 20.32 0.5-3 79
B4 1:1 66.21 33.79 0.25-3.5 74
B5 1:0.1 95.15 4.85 1-4 49
B6 1:0.2 90.75 9.25 1-3 53
B7 1:0.5 79.68 20.32 2-4.5 58
B8 1:1 66.21 33.79 1-5 51
B9 1:0.1 95.14 4.85 1-5 33
B10 1:0.2 90.75 9.25 2-5 41
B11 1:0.5 79.68 20.32 2-4 35
B12 1:1 66.21 33.79 1-6 39
PC: Phosphatidylcholine, CH: Cholesterol, EE: Entrapment Efficiency
Formulation parameters
The basic components of formulation such as
lipid concentration, drug to lipid ratio and
phosphatidylcholine to cholesterol ratio were
optimized to obtain reproducibility in the quality
of liposomes. The key parameters considered for
optimization of formula were entrapment
efficiency and vesicle size. Percent drug
entrapment i.e. the amount of drug inside the
vesicles is the most important parameter for
liposomal formulations.
Lipid Concentration
Liposomal formulations were prepared with
increasing concentration of phosphatidylcholine
0.5%, 1% and 2% i.e. the quantity of lipid taken
was 50 mg, 100 mg and 200 mg for 10 ml
liposomal batch. Rutin trihydrate at low, fixed
concentration was incorporated in the
formulations. The formulations were studied on
the basis of physical observations such as
consistency, settling rate, redispersibility and
vesicle size.
Drug to Lipid ratio
This is an important factor that determines the
amount of drug entrapped within the liposomes.
It varies as per the drug, its molecular size,
charge and physical properties. For lipid soluble
drugs, larger quantities of drug can be
encapsulated and hence drug to lipid ratio can be
large. Water soluble drugs have limitations of
encapsulation efficiency, as comparatively smaller
volume of aqueous portion is available for
entrapment.
Phosphatidylcholine to cholesterol ratio
Liposomes were formulated in different molar
ratios of phosphatidylcholine and cholesterol, with
increasing concentration of drug and parameters
such as % entrapment efficiency and globule size
were estimated.
IV. Preparation of Liposomal Gels
Rutin trihydrate lipogel was prepared by using
Carbopol 940 gel base. Carbopol is a pH
sensitive polymer and gel formation occurs after
inducing neutralization. Definite amount of
Carbopol 940 was kept for hydration in double
distilled water containing preservatives overnight.
Liposomal pellet (obtained after centrifugation
and separation of unentrapped drug) was
dispersed in distilled water by sonication and
added to hydrated Carbopol solution with stirring.
Gelling was induced by neutralization using
triethanolamine. The gels were evaluated for their
visual appearance, colour, texture, feel upon
application (like greasiness, smoothness), pH,
viscosity, spreadability, drug content and in
vitro/ex vivo drug release.
Aparajita Varshneya et al /Formulation and Characterization of Rutin Liposomes
International Journal of Pharmaceutical Research | July – Sept 2015 | Vol 7 | Issue 3|33
Figure	 3:	 Comparative	 in	 vitro	 drug	 release	
through	dialysis	membrane	
Figure	 4:	 Comparative	 ex	 vivo	 drug	 release	
through	porcine	ear	skin	
	in	vitro	/	ex	vivo	drug	release	from	lipogel	and	
conventional	gel	
in vitro / ex vivo drug release studies were
performed using Franz diffusion cells having
surface area of 3.91 cm2
and receptor
compartment having a capacity of 22 ml.
Phosphate buffer pH 5.4 was used as receptor
fluid. The pH of the buffer approaches the natural
pH value of human skin.
in vitro: Dialysis membrane filters (molecular
weight cut off 12 000 to 14 000) which were pre-
hydrated by soaking in buffer overnight were
mounted on the cells. Receptor compartment was
filled with buffer as diffusion medium (37±0.5°).
Reservoir solution was stirred at 300 r/min using
magnetic bead. 0.5 g gel was placed in the
donor compartment on the membrane. Aliquots
were collected at 1, 2, 3, 4, 5, 6, 7, 8 &
24 h intervals and analyzed by UV-
spectrophotometry (Figure 3).
ex vivo: Porcine ear skin was shaved and carefully
separated. Subcutaneous fat was removed using
a scalpel. Skin sections were soaked in buffer
overnight. Skin sections thus obtained were
mounted on Franz Diffusion cells. Epidermal side
of the skin was exposed to ambient condition
while dermal side was kept facing the receptor
solution. Receptor compartment was filled with
buffer as diffusion medium (37±0.5°). Reservoir
solution was stirred at 300 r/min using magnetic
bead. 0.5 g gel was placed in the donor
compartment on the skin. All bubbles were
carefully removed between the underside of the
skin and solution in the receiver compartment.
Aliquots were collected at 1, 2, 3, 4, 5, 6, 7, 8 &
24 h intervals and analyzed by UV-
spectrophotometry (Figure 4).
Kinetic modeling
The release data was analysed using zero order,
first order, Higuchi, Hixson-Crowell and
Korsmeyer-Peppas models, to evaluate the
mechanism of drug release from the liposomal
gel. Mathematical models aid in predicting the
drug release rate and diffusion behaviour, thus
reducing the number of experiments needed. It
also helps in understanding the physics of a
particular drug transport phenomenon, thus
enabling the development of new formulations
[14,15] .
Figure	5:	Pharmacokinetic	release	profile		
of	lipogel	(ex	vivo)	
Anti-elastase assay
Elastase is the only enzyme that is capable of
breaking down elastin, an insoluble elastic fibrous
protein that, together with collagen, determines
the mechanical properties of connective tissue.
Several studies have demonstrated that both skin-
aging and anti-wrinkle effects are significantly
correlated with decreased elastase activity [16].
Therefore anti-elastase assay was carried out of
Rutin trihydrate liposomal gel to confirm its utility
as an anti-wrinkle product. Porcine pancreatic
elastase enzyme was incubated with the sample in
the cuvette. The amount of enzyme left
uninhibited was detected by reacting it with
substrate N-Succ-Ala-Nitroanilide that gives p-
nitroaniline as the final product. This was read
spectrophotometrically at 405 nm. Higher the
amount of product formed, lower is the inhibitory
effect of the sample on the enzyme.
Aparajita Varshneya et al /Formulation and Characterization of Rutin Liposomes
34 | International Journal of Pharmaceutical Research | July – Sept 2015 | Vol 7 | Issue 3
% Elastase inhibition = (Absorbance of
control –Absorbance of test) x 100
Absorbance of control
Figure	 6:	 Anti‐elastase	 inhibition	 by	 Rutin	
trihydrate	
V. Skin Irritation Studies
Healthy male albino Wistar rats (Rattus norvegicus
sp.), 180-220 g, 3 months old were selected as
test models. Six animals were used for the study,
each animal served as its own control. The
research project protocol was approved by Local
Institutional Animal Ethics Committee (Approval
No. CPCSEA/IAEC/BNCP/P-10/2014).
All the animals were allowed to acclimatize to
laboratory conditions prior to study. Hair was
removed from the back side of rats and an area
of 1 cm2
was marked on both the sides. One side
served as control while the other as test. Control
and test formulations (200 mg/rat) were applied
on each side and the site was covered with cotton
bandage. Observations for sensitivity and
reaction were made and graded as follows
Grade 1: No reaction, Grade 2: Slight, patchy
erythema, Grade 3: Slight but confluent or
moderate but patchy erythema, Grade 4:
Moderate erythema, Grade 5: Severe erythema.
RESULTS AND DISCUSSION
Method of preparation of liposomes
Thin film hydration method was selected for
preparation of multilamellar liposomes. The
reason behind the selection of thin film hydration
method was that Rutin trihydrate is lipophilic in
nature and has low aqueous solubility, so
multilamellar vesicles are more capable of
loading a higher mass of a hydrophobic drug
than are unilamellar vesicles. Thin film hydration
method was found to give excellent drug loading
with efficiency over 80%. Hence, this method was
used for preparation of liposomes.
Optimization of process parameters
a. Speed of rotation
Uneven film was observed at higher speed of
rotation (150 r/min) during film formation. This
may be due to improper distribution of heat at
higher speed. During film formation, lower speed
increases the time of contact of the film with the
hot water in the water bath. At higher speed,
since sufficient time is not available for the lipid to
form film, liquid to gel transitions of the lipid does
not occur. Based upon these observations 80
r/min was considered optimum for formation of
uniform film.
But, during hydration of lipid film sufficient energy
and vigorous shaking is required for the dried
film to get hydrated and form liposomes, thus
100-150 r/min was found adequate to form
uniform liposomal dispersion.
b. The ratio and volume of solvent blend
The choice of solvent is usually chloroform owing
to its low boiling point (61.15°) and easy removal
under vacuum leading to thin film deposition on
the walls of the flask. Rutin trihydrate is insoluble
in chloroform but soluble in methanol (Boiling
point 64.7°) and thus methanol was used in
combination with chloroform. Chloroform:
methanol in a 2:1 ratio was selected. This ratio
gave an even, uniform film. When the volume of
solvents is low (5 ml or less), solvents evaporate
rapidly under vacuum leading to improper film
formation. On the other hand when the volume
of solvent is high (10 ml or more), inadequate
film drying was observed. The presence of
organic solvent often leads to emulsion
formation. Volume of the solvent system used was
7.5 ml (5 ml Chloroform and 2.5 ml methanol).
c. Quantity of glass beads
Optimum quantity of glass beads is required to
obtain a thin uniform film on the surface of RBF. It
was observed that in the absence of glass beads,
the organic solvents evaporated rapidly even
under slow application of vacuum which lead to
poor film deposition on the walls of RBF, whereas
excessive bead quantity resulted in attrition and
inability of the solvents to evaporate. Glass beads
to solvent ratio 3:1 was found to be optimum as it
gave thin uniform film with no visible aggregates.
22 to 25 g glass beads were found to be
optimum for homogeneous film formation.
d. Vesicle sizing technique
Thin film hydration method gives large,
heterogeneous multilamellar vesicles. Vesicle size
analysis revealed Z-average (d-nm) size of 1360
nm with high PDI of 0.98, but good entrapment
efficiency was achieved. Techniques such as Ultra
Turrax and high pressure homogenization (HPH)
Aparajita Varshneya et al /Formulation and Characterization of Rutin Liposomes
International Journal of Pharmaceutical Research | July – Sept 2015 | Vol 7 | Issue 3|35
were tried to obtain size reduction and narrow
size distribution i.e. lower PDI. The results
indicated satisfactory size reduction and low PDI
but affected the entrapment efficiency adversely
which could be due to vesicle rupturing at high
pressure and speed. Hence, bath sonication was
used for further batches which gave vesicles in the
size range of 100-500 nm with PDI of 0.6 without
loss in drug entrapment.
e. pH of the hydrating media
According to Handerson Hasselbalch equation,
pKa is defined as the extent to which a drug is
available in the ionized form at a given pH.
pH = pKa + log (ionized/unionised)
Hence, the extent of ionization is influenced by
the pKa of any drug. Thus, entrapment of any
drug depends on its pKa to a considerable extent.
The pKa of Rutin trihydrate is cited as 6.4 in the
literature. Maximum ionization (~90%) would
hence occur at pH values ± one unit of the pKa
value. Phosphate buffer pH 5.4 was selected as it
yielded uniform suspension with minimal
sedimentation and maximum entrapment
efficiency. Sedimentation was observed when pH
of the buffer was raised beyond 7 whereas phase
separation was observed towards acidic pH.
Hence, further batches were prepared at pH value
of 5.4.
Formula optimization
a. Lipid concentration was gradually increased
from 0.5 to 2%. Increase in concentration of lipid
phase leads to increase in viscosity and thus the
settling rate was decreased. Liposomal
suspension with 0.5% lipid was a thin suspension
which exhibited faster settling rate. After 24 h, the
sediment formed took time to redisperse.
Formulation containing 2% lipid was much thicker
and showed less settling even after 24 h. But the
microscopic evaluation showed presence of larger
vesicles and aggregates. In comparison 1% lipid
suspension was found to give satisfactory results
in terms of settling, redispersion and microscopic
observation (Table 1).
b. Drug to lipid ratio was changed from 1:10,
2:10 and 3:10. The molecular weight of Rutin
trihydrate is high (664.56). This factor is of
importance as the amount of drug that enters the
lipid bilayer is also governed by its molecular
weight. The bilayer can only incorporate certain
amount of drug after which the drug starts
sedimenting immediately upon formation.
Microscopic observation also confirms this fact as
the unentrapped drug is clearly visible outside
vesicles. The threshold for Rutin trihydrate was
found to be 1 part of drug per 10 parts of lipid
employed. Hence, 1:10 drug to lipid ratio (10 mg
drug in 100 mg lipid) was found to be optimum
as maximum entrapment efficiency was obtained
by this ratio and optical microscopy showed
minimal unentrapped drug.
c. Phospahatidylcholine: Cholesterol ratio: In
principle, liposomes can be prepared using only
phosphatidylcholine. However, cholesterol is
added to improve stability and other structural
properties. Cholesterol has the effect of making
the membrane less permeable by filling up holes
or disruptions. Liposomes formed only with
phosphatidylcholine gives porous membranes
resulting in drug leakage whereas cholesterol acts
a cementing material thus improving drug
loading capacity of vesicles. Highest entrapment
was obtained with the batches having
Phosphatidylcholine: Cholesterol ratio as 1:0.2.
This could be because, the addition of cholesterol
in this ratio provides optimum rigidity to the
bilayer. Increase in the quantity of cholesterol
causes decrease in entrapment. This can be
explained by the fact that cholesterol might be
replacing Rutin trihydrate in the bilayer (Table 2).
Vesicle Size and Zeta Potential Measurement
MLV’s of size 0.5 to 1 μm were visualized easily
for all the batches using optical microscope (Motic
microscope) at magnification of 100x. TEM
imaging of optimized formulation showed drug
loaded liposomes of somewhat spherical shape.
Vesicles showed inner dark spherical core,
surrounded by comparatively faint background.
Vesicle size of optimized batch was approximately
600 nm (Figure 2).
The zeta potential was used as an indicator of the
stability of vesicles formed by thin film hydration
method. The zeta potential of the optimized batch
was -30.7 which indicates sufficient stability.
Liposomal gels
Liposomal suspensions were converted to gel
formulations since simple mixing of liposomal
dispersions with polymers gives the corresponding
gels. This is advantageous because the fusion of
vesicles can be effectively minimized or avoided
as the polymer molecules serve as a spacer
between the liposomes. Moreover, the size of the
vesicles is not affected because it is a controlled
liposome/polymer interaction process. Liposomal
creams were not chosen as stability problems are
reported in emulsion-based products due to
presence of excess oil and surfactant which may
interact with the vesicles. Various properties like
viscosity can be easily controlled by varying the
amount and type of polymer. Thus liposomal gels
were formulated as they circumvent the stability
issue, provide controlled release, have aesthetic
appeal as skin care cosmetics and are easy to
prepare [15].
Lipogels were pale yellow (colour of Rutin
trihydrate) and opaque. 1% w/w Carbopol gel
Aparajita Varshneya et al /Formulation and Characterization of Rutin Liposomes
36 | International Journal of Pharmaceutical Research | July – Sept 2015 | Vol 7 | Issue 3
was found to be of good consistency and
acceptable feel with smooth appearance devoid
of any aggregation. Drug content for the gels
was found to be in the range of 97% to 101%.
Viscosity of gels was in the range of 8500 cps to
9500 cps. Spreadability of gels was around 5-7
g.cm/s.
in vitro / ex vivo drug release
The stratum corneum is built like a wall with
protein bricks and lipid mortar. The intercellular
lipids are important in controlling the
percutaneous absorption. In the case of
liposomes, the phospholipids may mix with the
intercellular lipids and thereby cause the swelling
of lipids without altering the multiple bilayer
structure of the stratum corneum. These swollen
lipids cause accumulation of the drug and thereby
form an intracutaneous depot. Although the
mechanism of enhancement using topically
applied liposomes is not fully understood, drug
disposition is primarily dependent on lipid
composition, liposome lamellarity and surface
charge. Studies have indicated topical drug
delivery is also influenced by the size of liposomes
[17,18].
in vitro and ex vivo application of the liposomal
gel formulation showed sustained release effect.
The drug release was sustained upto 12 hrs in
comparison to 5-6 hrs of conventional gel (Figure
3, Figure 4). This could be due to depot effect of
large multilamellar vesicles. ex vivo drug release
profile of lipogel showed that it follows zero order
kinetics as its R2
value was closest to unity (Figure
5).
Anti-elastase assay
The 0.1 % Rutin trihydrate lipogel showed a mean
anti-elastase activity of 32.01 ± 1.40 % at a
concentration of 3000 μg/ml. 32% inhibition at
0.1% concentration of Rutin trihydrate indicates
good activity of the formulation (Figure 6).
Skin Irritation Study
Skin irritation study conducted on albino Wistar
rats when observed for sensitivity and reaction at
the end of 24, 48 and 72 h showed absence of
erythema or edema for both the formulations
(control as well as test). This indicates that the gels
do not cause any skin irritation as no sensitivity
reaction (redness/ erythema) was observed.
CONCLUSION
Liposomes have been reported to be efficient
colloidal carriers for the delivery of therapeutics
into skin. This work too confirms the promising
role of liposomal topical formulation.
Multilamellar vesicles with good entrapment
efficiency were successfully prepared by thin film
hydration method and the drug containing
liposomes were formulated into a gel. The
liposomal gel showed prolonged drug release
upto 12 h and also exhibited anti-elastase activity.
Due to the striking similarity between liposome
components and skin lipids, they are safe and
effective. In addition to delivering the drug in
higher concentration into skin layers, they
enhance skin hydration making them an ideal
vehicle for anti-aging remedies. Thus liposomes
can be considered as effective carriers for better
topical delivery.
ACKNOWLEDGEMENTS
The authors are thankful to VAV Life Sciences Pvt.
Ltd., Mumbai for gift sample of Leciva-S70
(soya lecithin). They also sincerely thank NIRRH,
Mumbai for TEM imaging, Dr. Kshitij Vasant,
Kelkar College, Mumbai for anti-elastase assay
and NMIMS’s SPTM College, Mumbai for helping
in carrying out skin irritation studies.
REFERENCES
1. Moody ML, Topical Medications in the
Treatment of Pain, Pain Medicine News, 2010:
15–21.
2. Morrow DIJ, McCarron PA, Woolfson AD, and
Donnelly RF, Innovative Strategies for Enhancing
Topical and Transdermal Drug Delivery, The
Open Drug Delivery Journal, 2007; 1: 36–59.
3. Rahimpour Y and Hamishehkar H, Niosomes as
Carrier in Dermal Drug Delivery, Recent
Advances in Novel Drug Carrier Systems, 2012;
1: 141–163.
4. Padamwar MN and Pokharkar VB, Development
of Vitamin Loaded Topical Liposomal
Formulation Using Factorial Design Approach:
Drug Deposition and Stability, International
Journal of Pharmaceutics, 2006; 320: 37–44.
5. Dua JS., Review Article Liposome : Methods of
Preparation And Applications, International
Journal of Pharmaceutical Studies and Research,
2012; 3 (2): 14–20.
6. Gaspar M, Bakowsky U, and Ehrhardt C, Inhaled
Liposomes – Current Strategies and Future
Challenges, Journal of Biomedical
Nanotechnology, 2008; 4 (3): 1–13.
7. Alinaghi A, Rouini MR, Johari Daha F, and
Moghimi HR, Hydrogel-embeded vesicles, as a
novel approach for prolonged release and
delivery of liposome, in vitro and in vivo., Journal
of Liposome Research, 2013; 23 (3): 235–43.
8. 8. Egbaria K and Weiner N, Liposomes as a
topical drug delivery system, Advanced Drug
Delivery Reviews, 1990; 5: 287–300.
9. 9. Shahi S and Athawale R, Development and
Evaluation of Cosmeceutical Nanolipogel,
Aparajita Varshneya et al /Formulation and Characterization of Rutin Liposomes
International Journal of Pharmaceutical Research | July – Sept 2015 | Vol 7 | Issue 3|37
Research J. Topical Cosmetic Sci., 2010; 1 (1):
18–24.
10. 10. Shaji J and Iyer S, Preparation, Optimization
and In-Vivo Hepatoprotective Evaluation of
Quercetin Liposomes, International Journal of
Current Pharmaceutical Research, 2012; 4 (2):
24–32.
11. 11. Yamamoto, Role of active oxygen species and
antioxidants in photoaging, PubMed Commons,
Journal of Dermatological Science, 2001: S1–4.
12. 12. Rahimpour Y and Hamishehkar H, Liposomes
in cosmeceutics, Expert Opinion on Drug
Delivery, 2012; 9 (4): 443–55.
13. 13. Laouini, Jaafar-Maalej, Limayem-Blouza, et al.,
Preparation, Characterization and Applications of
Liposomes: State of the Art, Journal of Colloid
Science and Biotechnology, 2012; 1 (2): 147–168.
14. Dash S, Review Kinetic Modeling on Drug
Release from Controlled Drug Delivery Systems,
Acta Poloniae Pharmaceutica ñ Drug Research,
2010; 67 (3): 217–223.
15. 1Sharma R., Walker RB and Pathak K, Evaluation
of the Kinetics and Mechanism of Drug Release
from Econazole Nitrate Nanosponge Loaded
Carbapol Hydrogel, Indian Journal of
Pharmaceutical Education and Research, 2010; 45
(1): 25–31.
16. Moon JY, Yim EY, Song G et al., Screening of
Elastase and Tyrosinase Inhibitory Activity from
Jeju Island Plants, EurAsian Journal of
BioSciences, 2010; 53: 41–53.
17. Verma D, Particle Size of Liposomes Influences
Dermal Delivery of Substances into Skin,
International Journal of Pharmaceutics, 2003;
258: 141–151.
18. Diec and Sokolowski, New Liposome Gels by
Self Organization of Vesicles and Intelligent
Polymers, Cosmetics and Toiletries, 2002; 117
(8): 55–62.

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Formulation of Rutin trihydrate Liposomes for Topical Delivery

  • 1. International Journal of Pharmaceutical Research | July – Sept 2015 | Vol 7 | Issue 3|29 Research Article Formulation and Characterization of Rutin trihydrate Liposomes for Topical Delivery APARAJITA VARSHNEYA AND PADMINI RAVIKUMAR Department of Pharmaceutics, SVKM’s Dr. Bhanuben Nanavati College of Pharmacy, Gate No: 1, Mithibai College Campus, V.M. Road, Vile Parle (W), Mumbai-400 056, India ABSTRACT In the present study, liposomes were formulated as drug carriers for enhancing the delivery of an antioxidant Rutin trihydrate for topical administration. Formulation of liposomes was done by thin film hydration method. Optimization was achieved by varying the lipid to cholesterol ratio and gradually increasing the amount of drug until maximum entrapment efficiency was achieved. The formulation and process parameters were optimized to attain multilamellar liposomes with homogeneous size and good entrapment. The optimized batch gave entrapment efficiency of 88%. Gels were prepared by using Carbopol 940 as the gelling agent. The in vitro and ex vivo release profile of liposomal gels was compared with conventional gel formulations. The liposomal gel showed prolonged drug release upto 12 h and also exhibited anti-elastase activity. The safety of liposomal gels was ensured by conducting skin irritation studies on albino Wistar rats. Keywords: Liposomes, Rutin trihydrate, topical delivery, gels.  Address for correspondence  Aparajita Varshneya, SVKM’s Dr. Bhanuben Nanavati College of Pharmacy, Gate No.1, Mithibai, College Campus, V.M.  Road, Vile Parle (West), Mumbai ‐ 400 056, India.  Phone no: +91‐9870687261 e‐mail: aparajita.varshney@yahoo.com  Received:18/02/15, Revised:02/04/15, Accepted:18/04/15  INTRODUCTION Topical drug delivery has long been sought after, as it is coupled with well-established advantages like localised drug delivery, avoidance of first pass metabolism, improved patient compliance owing to its non-invasive nature and reduced systemic side effects [1]. Despite these merits, the brick and mortar structure of stratum corneum i.e. the horny layer serves as the principal barrier to the percutaneous absorption of drugs [2,3]. Owing to these barrier properties of the skin, the delivery of active substances from conventional formulations is generally compromised. Thus, there arises the need for a suitable carrier to enhance drug delivery [4]. Liposomes Liposomes were first described by British haematologist Dr. Alec D Bangham in early 1960’s [5]. Liposomes are highly organized structures, composed of concentric bi-layered vesicles in which an aqueous volume is enclosed by a membranous lipid bilayer [6]. The main advantages that have made these vesicular systems favourable for drug delivery are their tissue compatibility, biodegradability, safety and their ability to entrap almost any drug in the bilayer core or in the outer domain. Additionally, liposomal drug formulations can be used to overcome a drug’s non-ideal properties and unfavourable pharmacokinetic profiles [7]. Regarding topical application, liposomes offer a wide range of advantages including moisturization and prolonged dermal release [8]. Topical use of antioxidants has been adopted as an important strategy in cosmeceutical industry to avoid wrinkling of the skin and protect it from degenerative effects such as photoaging, sunburn, photocarcinogenesis etc. [9]. Antioxidants help in preventing and repairing damage caused due to oxidative stress. They scavenge free radicals which are postulated to be the mediators in skin aging [10]. They are found to be useful in delaying the signs of aging such as fine lines, wrinkles and age spots. Antioxidants are promising in photoprotection with negligible side effects at physiological concentrations [11]. Of the various antioxidants, bioflavonoids are one of the most powerful antioxidants. They are a class of plant secondary metabolites possessing a broad range of pharmacological activities. Rutin trihydrate was chosen as the active molecule as it is a powerful antioxidant which inhibits free-radical mediated cytotoxicity and lipid peroxidation. It is sparingly soluble in water (0.125 g/L). This limits its use in topical delivery. Thus liposomes were chosen as drug carriers as they are amphiphilic in nature and lipophilic molecules get embedded in their concentric bilayers. Liposomes being physiologically similar to cell membrane, are nontoxic in nature and are easily re-absorbed from the epidermis into the deepest layers [12]. Thus, the objective of the study was to enhance the accumulation of drug at the site of administration and achieve prolonged release by means of liposomal formulation.
  • 2. Aparajita Varshneya et al /Formulation and Characterization of Rutin Liposomes 30 | International Journal of Pharmaceutical Research | July – Sept 2015 | Vol 7 | Issue 3 MATERIALS AND METHODS Phosphatidylcholine (Leciva S-70) was a generous gift sample from VAV Life Sciences, Mumbai, India. Rutin trihydrate was purchased from Loba Chemie, Mumbai and Cholesterol was procured from Qualigens, Mumbai. Carbopol 940, Glass beads (2.5 mm-3.5 mm) and all other AR grade reagents were purchased from S D Fine-Chem Ltd., Mumbai. Dialysis membrane 150 was purchased from HiMedia Laboratories Pvt. Ltd., Mumbai and porcine skin was procured from local market. Healthy male albino Wistar rats (Rattus norvegicus sp.) were purchased from Bharat Serum, Mumbai, India. Liposomes were prepared using Rotary Evaporator, Roteva, Equitron and UV-visible spectrophotometer, Shimadzu 1800 was used for evaluation. I. Preparation of Rutin Trihydrate Liposomes: Multilamellar vesicles were prepared by thin film hydration technique as described by Bangham et al. [13]. Phosphatidylcholine, Cholesterol and Rutin trihydrate constituted the lipid mixture. This lipid mixture was dissolved in chloroform and methanol (2:1) in a round bottom flask (RBF) and 25 g glass beads were added for homogeneous film formation. The RBF was attached to the rotary flash evaporator (Roteva, Equitron) and allowed to rotate at 80 r/min in a thermostated water bath at 40°. The organic solvents were then removed by slow application of vacuum leading to lipid film formation on the walls of the flask. The film was allowed to dry for 60 min to ensure complete removal of solvent. The dried lipid film was hydrated with phosphate buffer (pH: 5.4) at 150 r/min and 46° that is above the transition temperature of lipid. Rotation was continued for 30 min. The Rutin trihydrate liposomal suspension was kept at room temperature for 2 h for complete hydration and for annealing structural defects. The liposomes produced by this method are large and heterogeneous in size, thus they were downsized by bath sonication for 20 min. The suspension was also vortexed for few minutes. The liposomal suspension was characterized for physical appearance, settling time, re-dispersion time, entrapment efficiency and vesicle size by optical microscopy. It was then filled in amber colored glass bottles and stored at 4° until further use. II. Characterization of Liposomes: Rutin trihydrate liposomes were characterized for following parameters: Optical Microscopy A drop of the liposomal suspension was placed on a clean glass slide and observed under high power 45x and 100x of the optical microscope (Motic microscope). Multilamellar vesicles (MLV) were clearly identified (Figure 1). Microscopic observations were used to study presence of aggregates, precipitation or leakage of drug. Size and shape of the vesicles was noted for all liposomal batches. Figure 1: (a) Large multilamellar vesicles as visible under 100x magnification Determination of Vesicle Entrapment Efficiency Entrapment efficiency for the batches prepared during the optimization process was estimated by the following procedure: The liposomal pellet i.e. the drug loaded liposomes were separated from the aqueous phase by centrifugation at 15,000 r/min using Microspin centrifuge for 30-45 min until a clear supernatant was obtained. The amount of drug loaded was estimated by both direct and indirect method. Direct method: After centrifugation, the supernatant was decanted. The formed pellet was washed with phosphate buffer and vortexed, in order to remove the free drug adsorbed on the surface of liposomes. This step was repeated thrice to completely separate the unentrapped drug. The resulting pellet was then dissolved in methanol and sonicated for 20 min. (This causes rupturing of pellet and release of entrapped drug in the solution). The resulting methanolic solution was then analyzed by UV spectrophotometry using the developed analytical method. % Entrapment Efficiency (E.E.) =   (Amount of drug quantified in the pellet) x 100 Total amount of drug added Indirect method: The supernatant of the liposomal dispersion decanted after centrifugation and the solutions collected after washing of pellet were combined and analyzed by developed UV spectrophotometric method for the amount of drug present. % Entrapment efficiency was estimated as follows: % Entrapment Efficiency (E.E) = (Total amount of drug added - Amount of drug detected in the supernatant) x 100_ Total amount of drug added Determination of vesicle size and size distribution Vesicle size was determined by dynamic light scattering using a computerized inspection system (Malvern particle size analyzer). Freshly prepared batches of vesicles were diluted ten-fold with filtered
  • 3. Aparajita Varshneya et al /Formulation and Characterization of Rutin Liposomes International Journal of Pharmaceutical Research | July – Sept 2015 | Vol 7 | Issue 3|31 distilled water and then analyzed at 25° using ten runs at each scan. Optimization time was fixed at 120 s. Vesicle size and polydispersity index were measured for the liposomal formulation. The measurements were done in triplicate. Zeta potential measurement Liposomal suspension was diluted with filtered distilled water and zeta potential was measured in triplicate with the help of Malvern zetasizer. Transmission Electron Microscopy (TEM) TEM analysis was carried out for optimized drug loaded liposomal suspension. A drop of the diluted formulation was placed with the aid of a micropipette on a coated copper grid 3 mm in diameter. Negative staining was carried out by uranyl acetate. The grid was allowed to dry under an IR lamp for about 20 min. The grid was then loaded on a probe which was inserted in the slot designed for it in the electron microscope. The grid containing the sample was then bombarded with electrons accelerated at 200 kV and the sample visualized over a fluorescent stage. Vesicle size of the sample was measured and marked and its morphology observed on the computer (Figure 2). Figure 2: Negative staining TEM imaging of Rutin trihydrate liposomesMagnification 100,000x (size approx. 600 nm) III. Optimization of Process and Formulation Parameters: Many factors influence the formation of liposomes and its characteristics. Therefore, the following process parameters were optimized to obtain liposomes with maximum entrapment efficiency, homogeneous size and stability. a. Process Parameters Speed of the rotary evaporator It is desirable to obtain a thin and uniform film as it governs the final output of liposomal preparation. The speed of rotation was varied from 60 r/min to 150 r/min during film formation as well as during hydration. The ratio and volume of solvent system The solvent system was optimized by taking various combinations of organic solvents i.e. chloroform and methanol. The ratios 1:1, 2:1 and 3:1 were tried and the film was evaluated in terms of its uniformity. Quantity of glass beads Thin film hydration technique is readily employed for its feasibility on a laboratory scale, but problems related to limitations of surface area exist. Glass beads are commonly used for this purpose which helps to increase the surface area to provide a thin film. Liposomal dispersions were prepared using different ratios of glass beads (diameter: 2.5 mm-3.5 mm) and solvent volume such as 0:1, 2:1, 3:1 and 4:1. The formulations were evaluated in terms of uniformity of film and separation of vesicle aggregates (microscopic observation). Vesicle sizing technique Size reduction of the vesicles is done to obtain a homogeneous population of vesicles with low polydispersity index. The various techniques studied for size reduction were bath sonication using ultrasonic bath, high pressure homogenization and by using ultra turrax. Bath sonication was carried out at room temperature for 20-30 min. Size reduction using Ultra turrax was done at 6000 r/min for 15 min. High pressure homogenization was also tried by carrying out approximately 10 cycles at a pressure of 5000 psi for 10 min. The effect of these techniques was studied by estimating entrapment efficiency, vesicle size and polydispersity index. pH of the hydrating media The effect of pH of the phosphate buffer was studied on the formulation. The effect of pH on the entrapment of the drug is related to its pKa. Hence the pH of the hydrating buffer was adjusted at values closer to the pKa of the drug and entrapment efficiency was estimated. Distilled water, phosphate buffer pH 5.4, 6.8 and 7.4 were used as hydrating media and formulations were studied in terms of colour, sedimentation, redispersibility and entrapment efficiency. Table 1: Formula optimization based on lipid concentration Lipid Conc. (%) Consistency Settling rate Redispersibility Vesicle size (μ) 0.5 Thin suspension +++ + 1-4 1 Moderate, smooth suspension ++ +++ 0.25-3 2 Thick suspension + ++ 2-6
  • 4. Aparajita Varshneya et al /Formulation and Characterization of Rutin Liposomes 32 | International Journal of Pharmaceutical Research | July – Sept 2015 | Vol 7 | Issue 3 Settling rate: + Slow ++ Moderate +++Fast Redispersibility + Poor ++ Good +++Excellent Table‐2: Formula optimization based on Drug: Lipid and PC: CH ratio Batch No. PC:CH (molar ratio) Total lipid: 100 mg Globule Size (μ) EE (%)Amount of PC (mg) Amount of CH (mg) B1 1:0 100 0 0.5-5 56 B2 1:0.2 90.75 9.25 0.5-1.5 88 B3 1:0.5 79.68 20.32 0.5-3 79 B4 1:1 66.21 33.79 0.25-3.5 74 B5 1:0.1 95.15 4.85 1-4 49 B6 1:0.2 90.75 9.25 1-3 53 B7 1:0.5 79.68 20.32 2-4.5 58 B8 1:1 66.21 33.79 1-5 51 B9 1:0.1 95.14 4.85 1-5 33 B10 1:0.2 90.75 9.25 2-5 41 B11 1:0.5 79.68 20.32 2-4 35 B12 1:1 66.21 33.79 1-6 39 PC: Phosphatidylcholine, CH: Cholesterol, EE: Entrapment Efficiency Formulation parameters The basic components of formulation such as lipid concentration, drug to lipid ratio and phosphatidylcholine to cholesterol ratio were optimized to obtain reproducibility in the quality of liposomes. The key parameters considered for optimization of formula were entrapment efficiency and vesicle size. Percent drug entrapment i.e. the amount of drug inside the vesicles is the most important parameter for liposomal formulations. Lipid Concentration Liposomal formulations were prepared with increasing concentration of phosphatidylcholine 0.5%, 1% and 2% i.e. the quantity of lipid taken was 50 mg, 100 mg and 200 mg for 10 ml liposomal batch. Rutin trihydrate at low, fixed concentration was incorporated in the formulations. The formulations were studied on the basis of physical observations such as consistency, settling rate, redispersibility and vesicle size. Drug to Lipid ratio This is an important factor that determines the amount of drug entrapped within the liposomes. It varies as per the drug, its molecular size, charge and physical properties. For lipid soluble drugs, larger quantities of drug can be encapsulated and hence drug to lipid ratio can be large. Water soluble drugs have limitations of encapsulation efficiency, as comparatively smaller volume of aqueous portion is available for entrapment. Phosphatidylcholine to cholesterol ratio Liposomes were formulated in different molar ratios of phosphatidylcholine and cholesterol, with increasing concentration of drug and parameters such as % entrapment efficiency and globule size were estimated. IV. Preparation of Liposomal Gels Rutin trihydrate lipogel was prepared by using Carbopol 940 gel base. Carbopol is a pH sensitive polymer and gel formation occurs after inducing neutralization. Definite amount of Carbopol 940 was kept for hydration in double distilled water containing preservatives overnight. Liposomal pellet (obtained after centrifugation and separation of unentrapped drug) was dispersed in distilled water by sonication and added to hydrated Carbopol solution with stirring. Gelling was induced by neutralization using triethanolamine. The gels were evaluated for their visual appearance, colour, texture, feel upon application (like greasiness, smoothness), pH, viscosity, spreadability, drug content and in vitro/ex vivo drug release.
  • 5. Aparajita Varshneya et al /Formulation and Characterization of Rutin Liposomes International Journal of Pharmaceutical Research | July – Sept 2015 | Vol 7 | Issue 3|33 Figure 3: Comparative in vitro drug release through dialysis membrane Figure 4: Comparative ex vivo drug release through porcine ear skin in vitro / ex vivo drug release from lipogel and conventional gel in vitro / ex vivo drug release studies were performed using Franz diffusion cells having surface area of 3.91 cm2 and receptor compartment having a capacity of 22 ml. Phosphate buffer pH 5.4 was used as receptor fluid. The pH of the buffer approaches the natural pH value of human skin. in vitro: Dialysis membrane filters (molecular weight cut off 12 000 to 14 000) which were pre- hydrated by soaking in buffer overnight were mounted on the cells. Receptor compartment was filled with buffer as diffusion medium (37±0.5°). Reservoir solution was stirred at 300 r/min using magnetic bead. 0.5 g gel was placed in the donor compartment on the membrane. Aliquots were collected at 1, 2, 3, 4, 5, 6, 7, 8 & 24 h intervals and analyzed by UV- spectrophotometry (Figure 3). ex vivo: Porcine ear skin was shaved and carefully separated. Subcutaneous fat was removed using a scalpel. Skin sections were soaked in buffer overnight. Skin sections thus obtained were mounted on Franz Diffusion cells. Epidermal side of the skin was exposed to ambient condition while dermal side was kept facing the receptor solution. Receptor compartment was filled with buffer as diffusion medium (37±0.5°). Reservoir solution was stirred at 300 r/min using magnetic bead. 0.5 g gel was placed in the donor compartment on the skin. All bubbles were carefully removed between the underside of the skin and solution in the receiver compartment. Aliquots were collected at 1, 2, 3, 4, 5, 6, 7, 8 & 24 h intervals and analyzed by UV- spectrophotometry (Figure 4). Kinetic modeling The release data was analysed using zero order, first order, Higuchi, Hixson-Crowell and Korsmeyer-Peppas models, to evaluate the mechanism of drug release from the liposomal gel. Mathematical models aid in predicting the drug release rate and diffusion behaviour, thus reducing the number of experiments needed. It also helps in understanding the physics of a particular drug transport phenomenon, thus enabling the development of new formulations [14,15] . Figure 5: Pharmacokinetic release profile of lipogel (ex vivo) Anti-elastase assay Elastase is the only enzyme that is capable of breaking down elastin, an insoluble elastic fibrous protein that, together with collagen, determines the mechanical properties of connective tissue. Several studies have demonstrated that both skin- aging and anti-wrinkle effects are significantly correlated with decreased elastase activity [16]. Therefore anti-elastase assay was carried out of Rutin trihydrate liposomal gel to confirm its utility as an anti-wrinkle product. Porcine pancreatic elastase enzyme was incubated with the sample in the cuvette. The amount of enzyme left uninhibited was detected by reacting it with substrate N-Succ-Ala-Nitroanilide that gives p- nitroaniline as the final product. This was read spectrophotometrically at 405 nm. Higher the amount of product formed, lower is the inhibitory effect of the sample on the enzyme.
  • 6. Aparajita Varshneya et al /Formulation and Characterization of Rutin Liposomes 34 | International Journal of Pharmaceutical Research | July – Sept 2015 | Vol 7 | Issue 3 % Elastase inhibition = (Absorbance of control –Absorbance of test) x 100 Absorbance of control Figure 6: Anti‐elastase inhibition by Rutin trihydrate V. Skin Irritation Studies Healthy male albino Wistar rats (Rattus norvegicus sp.), 180-220 g, 3 months old were selected as test models. Six animals were used for the study, each animal served as its own control. The research project protocol was approved by Local Institutional Animal Ethics Committee (Approval No. CPCSEA/IAEC/BNCP/P-10/2014). All the animals were allowed to acclimatize to laboratory conditions prior to study. Hair was removed from the back side of rats and an area of 1 cm2 was marked on both the sides. One side served as control while the other as test. Control and test formulations (200 mg/rat) were applied on each side and the site was covered with cotton bandage. Observations for sensitivity and reaction were made and graded as follows Grade 1: No reaction, Grade 2: Slight, patchy erythema, Grade 3: Slight but confluent or moderate but patchy erythema, Grade 4: Moderate erythema, Grade 5: Severe erythema. RESULTS AND DISCUSSION Method of preparation of liposomes Thin film hydration method was selected for preparation of multilamellar liposomes. The reason behind the selection of thin film hydration method was that Rutin trihydrate is lipophilic in nature and has low aqueous solubility, so multilamellar vesicles are more capable of loading a higher mass of a hydrophobic drug than are unilamellar vesicles. Thin film hydration method was found to give excellent drug loading with efficiency over 80%. Hence, this method was used for preparation of liposomes. Optimization of process parameters a. Speed of rotation Uneven film was observed at higher speed of rotation (150 r/min) during film formation. This may be due to improper distribution of heat at higher speed. During film formation, lower speed increases the time of contact of the film with the hot water in the water bath. At higher speed, since sufficient time is not available for the lipid to form film, liquid to gel transitions of the lipid does not occur. Based upon these observations 80 r/min was considered optimum for formation of uniform film. But, during hydration of lipid film sufficient energy and vigorous shaking is required for the dried film to get hydrated and form liposomes, thus 100-150 r/min was found adequate to form uniform liposomal dispersion. b. The ratio and volume of solvent blend The choice of solvent is usually chloroform owing to its low boiling point (61.15°) and easy removal under vacuum leading to thin film deposition on the walls of the flask. Rutin trihydrate is insoluble in chloroform but soluble in methanol (Boiling point 64.7°) and thus methanol was used in combination with chloroform. Chloroform: methanol in a 2:1 ratio was selected. This ratio gave an even, uniform film. When the volume of solvents is low (5 ml or less), solvents evaporate rapidly under vacuum leading to improper film formation. On the other hand when the volume of solvent is high (10 ml or more), inadequate film drying was observed. The presence of organic solvent often leads to emulsion formation. Volume of the solvent system used was 7.5 ml (5 ml Chloroform and 2.5 ml methanol). c. Quantity of glass beads Optimum quantity of glass beads is required to obtain a thin uniform film on the surface of RBF. It was observed that in the absence of glass beads, the organic solvents evaporated rapidly even under slow application of vacuum which lead to poor film deposition on the walls of RBF, whereas excessive bead quantity resulted in attrition and inability of the solvents to evaporate. Glass beads to solvent ratio 3:1 was found to be optimum as it gave thin uniform film with no visible aggregates. 22 to 25 g glass beads were found to be optimum for homogeneous film formation. d. Vesicle sizing technique Thin film hydration method gives large, heterogeneous multilamellar vesicles. Vesicle size analysis revealed Z-average (d-nm) size of 1360 nm with high PDI of 0.98, but good entrapment efficiency was achieved. Techniques such as Ultra Turrax and high pressure homogenization (HPH)
  • 7. Aparajita Varshneya et al /Formulation and Characterization of Rutin Liposomes International Journal of Pharmaceutical Research | July – Sept 2015 | Vol 7 | Issue 3|35 were tried to obtain size reduction and narrow size distribution i.e. lower PDI. The results indicated satisfactory size reduction and low PDI but affected the entrapment efficiency adversely which could be due to vesicle rupturing at high pressure and speed. Hence, bath sonication was used for further batches which gave vesicles in the size range of 100-500 nm with PDI of 0.6 without loss in drug entrapment. e. pH of the hydrating media According to Handerson Hasselbalch equation, pKa is defined as the extent to which a drug is available in the ionized form at a given pH. pH = pKa + log (ionized/unionised) Hence, the extent of ionization is influenced by the pKa of any drug. Thus, entrapment of any drug depends on its pKa to a considerable extent. The pKa of Rutin trihydrate is cited as 6.4 in the literature. Maximum ionization (~90%) would hence occur at pH values ± one unit of the pKa value. Phosphate buffer pH 5.4 was selected as it yielded uniform suspension with minimal sedimentation and maximum entrapment efficiency. Sedimentation was observed when pH of the buffer was raised beyond 7 whereas phase separation was observed towards acidic pH. Hence, further batches were prepared at pH value of 5.4. Formula optimization a. Lipid concentration was gradually increased from 0.5 to 2%. Increase in concentration of lipid phase leads to increase in viscosity and thus the settling rate was decreased. Liposomal suspension with 0.5% lipid was a thin suspension which exhibited faster settling rate. After 24 h, the sediment formed took time to redisperse. Formulation containing 2% lipid was much thicker and showed less settling even after 24 h. But the microscopic evaluation showed presence of larger vesicles and aggregates. In comparison 1% lipid suspension was found to give satisfactory results in terms of settling, redispersion and microscopic observation (Table 1). b. Drug to lipid ratio was changed from 1:10, 2:10 and 3:10. The molecular weight of Rutin trihydrate is high (664.56). This factor is of importance as the amount of drug that enters the lipid bilayer is also governed by its molecular weight. The bilayer can only incorporate certain amount of drug after which the drug starts sedimenting immediately upon formation. Microscopic observation also confirms this fact as the unentrapped drug is clearly visible outside vesicles. The threshold for Rutin trihydrate was found to be 1 part of drug per 10 parts of lipid employed. Hence, 1:10 drug to lipid ratio (10 mg drug in 100 mg lipid) was found to be optimum as maximum entrapment efficiency was obtained by this ratio and optical microscopy showed minimal unentrapped drug. c. Phospahatidylcholine: Cholesterol ratio: In principle, liposomes can be prepared using only phosphatidylcholine. However, cholesterol is added to improve stability and other structural properties. Cholesterol has the effect of making the membrane less permeable by filling up holes or disruptions. Liposomes formed only with phosphatidylcholine gives porous membranes resulting in drug leakage whereas cholesterol acts a cementing material thus improving drug loading capacity of vesicles. Highest entrapment was obtained with the batches having Phosphatidylcholine: Cholesterol ratio as 1:0.2. This could be because, the addition of cholesterol in this ratio provides optimum rigidity to the bilayer. Increase in the quantity of cholesterol causes decrease in entrapment. This can be explained by the fact that cholesterol might be replacing Rutin trihydrate in the bilayer (Table 2). Vesicle Size and Zeta Potential Measurement MLV’s of size 0.5 to 1 μm were visualized easily for all the batches using optical microscope (Motic microscope) at magnification of 100x. TEM imaging of optimized formulation showed drug loaded liposomes of somewhat spherical shape. Vesicles showed inner dark spherical core, surrounded by comparatively faint background. Vesicle size of optimized batch was approximately 600 nm (Figure 2). The zeta potential was used as an indicator of the stability of vesicles formed by thin film hydration method. The zeta potential of the optimized batch was -30.7 which indicates sufficient stability. Liposomal gels Liposomal suspensions were converted to gel formulations since simple mixing of liposomal dispersions with polymers gives the corresponding gels. This is advantageous because the fusion of vesicles can be effectively minimized or avoided as the polymer molecules serve as a spacer between the liposomes. Moreover, the size of the vesicles is not affected because it is a controlled liposome/polymer interaction process. Liposomal creams were not chosen as stability problems are reported in emulsion-based products due to presence of excess oil and surfactant which may interact with the vesicles. Various properties like viscosity can be easily controlled by varying the amount and type of polymer. Thus liposomal gels were formulated as they circumvent the stability issue, provide controlled release, have aesthetic appeal as skin care cosmetics and are easy to prepare [15]. Lipogels were pale yellow (colour of Rutin trihydrate) and opaque. 1% w/w Carbopol gel
  • 8. Aparajita Varshneya et al /Formulation and Characterization of Rutin Liposomes 36 | International Journal of Pharmaceutical Research | July – Sept 2015 | Vol 7 | Issue 3 was found to be of good consistency and acceptable feel with smooth appearance devoid of any aggregation. Drug content for the gels was found to be in the range of 97% to 101%. Viscosity of gels was in the range of 8500 cps to 9500 cps. Spreadability of gels was around 5-7 g.cm/s. in vitro / ex vivo drug release The stratum corneum is built like a wall with protein bricks and lipid mortar. The intercellular lipids are important in controlling the percutaneous absorption. In the case of liposomes, the phospholipids may mix with the intercellular lipids and thereby cause the swelling of lipids without altering the multiple bilayer structure of the stratum corneum. These swollen lipids cause accumulation of the drug and thereby form an intracutaneous depot. Although the mechanism of enhancement using topically applied liposomes is not fully understood, drug disposition is primarily dependent on lipid composition, liposome lamellarity and surface charge. Studies have indicated topical drug delivery is also influenced by the size of liposomes [17,18]. in vitro and ex vivo application of the liposomal gel formulation showed sustained release effect. The drug release was sustained upto 12 hrs in comparison to 5-6 hrs of conventional gel (Figure 3, Figure 4). This could be due to depot effect of large multilamellar vesicles. ex vivo drug release profile of lipogel showed that it follows zero order kinetics as its R2 value was closest to unity (Figure 5). Anti-elastase assay The 0.1 % Rutin trihydrate lipogel showed a mean anti-elastase activity of 32.01 ± 1.40 % at a concentration of 3000 μg/ml. 32% inhibition at 0.1% concentration of Rutin trihydrate indicates good activity of the formulation (Figure 6). Skin Irritation Study Skin irritation study conducted on albino Wistar rats when observed for sensitivity and reaction at the end of 24, 48 and 72 h showed absence of erythema or edema for both the formulations (control as well as test). This indicates that the gels do not cause any skin irritation as no sensitivity reaction (redness/ erythema) was observed. CONCLUSION Liposomes have been reported to be efficient colloidal carriers for the delivery of therapeutics into skin. This work too confirms the promising role of liposomal topical formulation. Multilamellar vesicles with good entrapment efficiency were successfully prepared by thin film hydration method and the drug containing liposomes were formulated into a gel. The liposomal gel showed prolonged drug release upto 12 h and also exhibited anti-elastase activity. Due to the striking similarity between liposome components and skin lipids, they are safe and effective. In addition to delivering the drug in higher concentration into skin layers, they enhance skin hydration making them an ideal vehicle for anti-aging remedies. Thus liposomes can be considered as effective carriers for better topical delivery. ACKNOWLEDGEMENTS The authors are thankful to VAV Life Sciences Pvt. Ltd., Mumbai for gift sample of Leciva-S70 (soya lecithin). They also sincerely thank NIRRH, Mumbai for TEM imaging, Dr. Kshitij Vasant, Kelkar College, Mumbai for anti-elastase assay and NMIMS’s SPTM College, Mumbai for helping in carrying out skin irritation studies. REFERENCES 1. Moody ML, Topical Medications in the Treatment of Pain, Pain Medicine News, 2010: 15–21. 2. Morrow DIJ, McCarron PA, Woolfson AD, and Donnelly RF, Innovative Strategies for Enhancing Topical and Transdermal Drug Delivery, The Open Drug Delivery Journal, 2007; 1: 36–59. 3. Rahimpour Y and Hamishehkar H, Niosomes as Carrier in Dermal Drug Delivery, Recent Advances in Novel Drug Carrier Systems, 2012; 1: 141–163. 4. Padamwar MN and Pokharkar VB, Development of Vitamin Loaded Topical Liposomal Formulation Using Factorial Design Approach: Drug Deposition and Stability, International Journal of Pharmaceutics, 2006; 320: 37–44. 5. Dua JS., Review Article Liposome : Methods of Preparation And Applications, International Journal of Pharmaceutical Studies and Research, 2012; 3 (2): 14–20. 6. Gaspar M, Bakowsky U, and Ehrhardt C, Inhaled Liposomes – Current Strategies and Future Challenges, Journal of Biomedical Nanotechnology, 2008; 4 (3): 1–13. 7. Alinaghi A, Rouini MR, Johari Daha F, and Moghimi HR, Hydrogel-embeded vesicles, as a novel approach for prolonged release and delivery of liposome, in vitro and in vivo., Journal of Liposome Research, 2013; 23 (3): 235–43. 8. 8. Egbaria K and Weiner N, Liposomes as a topical drug delivery system, Advanced Drug Delivery Reviews, 1990; 5: 287–300. 9. 9. Shahi S and Athawale R, Development and Evaluation of Cosmeceutical Nanolipogel,
  • 9. Aparajita Varshneya et al /Formulation and Characterization of Rutin Liposomes International Journal of Pharmaceutical Research | July – Sept 2015 | Vol 7 | Issue 3|37 Research J. Topical Cosmetic Sci., 2010; 1 (1): 18–24. 10. 10. Shaji J and Iyer S, Preparation, Optimization and In-Vivo Hepatoprotective Evaluation of Quercetin Liposomes, International Journal of Current Pharmaceutical Research, 2012; 4 (2): 24–32. 11. 11. Yamamoto, Role of active oxygen species and antioxidants in photoaging, PubMed Commons, Journal of Dermatological Science, 2001: S1–4. 12. 12. Rahimpour Y and Hamishehkar H, Liposomes in cosmeceutics, Expert Opinion on Drug Delivery, 2012; 9 (4): 443–55. 13. 13. Laouini, Jaafar-Maalej, Limayem-Blouza, et al., Preparation, Characterization and Applications of Liposomes: State of the Art, Journal of Colloid Science and Biotechnology, 2012; 1 (2): 147–168. 14. Dash S, Review Kinetic Modeling on Drug Release from Controlled Drug Delivery Systems, Acta Poloniae Pharmaceutica ñ Drug Research, 2010; 67 (3): 217–223. 15. 1Sharma R., Walker RB and Pathak K, Evaluation of the Kinetics and Mechanism of Drug Release from Econazole Nitrate Nanosponge Loaded Carbapol Hydrogel, Indian Journal of Pharmaceutical Education and Research, 2010; 45 (1): 25–31. 16. Moon JY, Yim EY, Song G et al., Screening of Elastase and Tyrosinase Inhibitory Activity from Jeju Island Plants, EurAsian Journal of BioSciences, 2010; 53: 41–53. 17. Verma D, Particle Size of Liposomes Influences Dermal Delivery of Substances into Skin, International Journal of Pharmaceutics, 2003; 258: 141–151. 18. Diec and Sokolowski, New Liposome Gels by Self Organization of Vesicles and Intelligent Polymers, Cosmetics and Toiletries, 2002; 117 (8): 55–62.