A.Abdul Hasan Sathali, M.Pharm, Professor and Head, Department of pharmaceutics, College of pharmacy, Madurai Medical College, Madurai-625 020 CERTIFICATE This is to certify that the Dissertation entitled “FORMULATION AND EVALUATION OF DICLOFENAC POTASSIUM ETHOSOMES” submitted by Mr. M.R.VIJAYAKUMAR in partial fulfillment of the requirement for the degree of Master of Pharmacy in Pharmaceutics is a bonafide work carried out by him, under my guidance and supervision during the academic year 2009 – 2010 in the Department of Pharmaceutics, Madurai Medical College, Madurai-20. I wish him success in all his endeavors. Place: Madurai Date: (A.Abdul Hasan Sathali)
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A.Abdul Hasan Sathali, M.Pharm, Professor and Head, Department of pharmaceutics, College of pharmacy, Madurai Medical College, Madurai-625 020
CERTIFICATE
This is to certify that the Dissertation entitled “FORMULATION AND
EVALUATION OF DICLOFENAC POTASSIUM ETHOSOMES” submitted by
Mr. M.R.VIJAYAKUMAR in partial fulfillment of the requirement for the degree of
Master of Pharmacy in Pharmaceutics is a bonafide work carried out by him, under
my guidance and supervision during the academic year 2009 – 2010 in the Department
of Pharmaceutics, Madurai Medical College, Madurai-20.
I wish him success in all his endeavors.
Place: Madurai
Date: (A.Abdul Hasan Sathali)
FORMULATION AND EVALUATION OF
DICLOFENAC POTASSIUM ETHOSOMES
Dissertation submitted to THE TAMILNADU Dr. M.G.R. MEDICAL UNIVERSITY,
CHENNAI
in partial fulfillment of the requirement for the award of degree of
MASTER OF PHARMACY
IN
PHARMACEUTICS
March - 2010
DEPARTMENT OF PHARMACEUTICS
COLLEGE OF PHARMACY
MADURAI MEDICAL COLLEGE
MADURAI - 625020.
Chapter 1 Introduction
1
CHAPTER- I
INTRODUCTION
The skin covers a total surface area of approximately 1.8m2 and provides the
contact between the human body and the external environment. Dermal drug delivery is
the topical application of drugs to the skin in the treatment of skin diseases. This has the
advantage that high concentrations of drugs can be localized at the site of action, reducing
the systemic side effects. Transdermal drug delivery uses the skin as an alternative route
for the delivery of systemically acting drugs [1].
The alleviation of the psychological, physical suffering and the cure or at least
amelioration of disfiguring diseases such as eczema, psoriasis, icthyosis and the skin
cancers are noble aims of dermal and transdermal delivery of drugs. For example, in the
UK alone there are 44,000 skin carcinoma cases per year, of which 2000 are fatal (study
at 2001) [2]. The main reason for many problems with transdermal drug delivery is that
the impermeability of human skin limits the delivery of daily drug, for example from an
acceptable sized patch approximately 10mg. How to increase this low limit for topical
systems in general provides a major challenge to scientists and many university
laboratories worldwide.
Transdermal route of drug administration have several advantages like
circumvention of the variables that could influence gastro intestinal absorption such as
PH, food intake, gastrointestinal motility, circumvention of hepatic metabolism, constant
controlled drug input and targeting of the active ingredient for a local effect. Constant
drug input from the transdermal formulation decreases the variations in the drug plasma
levels, reducing the side effects particularly of drugs with narrow therapeutic window.
Chapter 1 Introduction
2
Although the skin as a route for drug delivery can offer many advantages, the
barrier nature of the skin makes it difficult for most drugs to penetrate into and permeate
through it[3]. Human skin effectively inhibits drug permeation mainly because of the
stratum corneum. Thus to maximize drug flux, formulations reduce the hindrance of this
barrier, although sometimes drug transport via the hair follicle might also be involved.
During past decades there has been wide interest in exploring new techniques to increase
drug absorption through the skin. But only few drugs are currently in market in
transdermal drug delivery system like clonidine, estradiol, nitroglycerine, fentanyl,
testosterone, scopolamine, nicotine and oxybutinin[4-8].
The structure of stratum corneum is often compared with a brick wall, with the
corneocytes as the bricks surrounded by the mortar of the intercellular lipid lamellae [9],
[figure-1]. Many techniques have been aimed to disrupt and weaken the highly organized
intercellular lipids in an attempt to enhance drug transport across the intact skin [10, 11].
One of the most controversial methods is the use of vehicle formulations as skin delivery
systems.
Chapter 2 Vesicles in transdermal drug delivery – A Review
3
CAHAPTER- II
VESICLES IN TRANSDERMAL DRUG DELIVERY- A REVIEW
The first paper to report the effectiveness of liposomes for skin delivery was
published by Mezei and Gulasekharam(1980) [12,13]. Conflicting the results continued to
be published concerning their effectiveness, enhancing the controversy of liposomes as
dermal and transdermal drug delivery vesicles [14]. The first therapeutic using lipid
vesicle on the skin was commercialized shortly before year 1990 and contained the
antimycotic agent econazole. A few other, relatively simple liposome based dermal
products followed [15-17]. Recently suggested, classic liposomes are of little or no value
as carriers for transdermal drug delivery as they do not deeply penetrate the skin.
Intensive research led to the introduction and development of new class of
lipid vesicles, the highly deformable (ultradeformable or elastic) liposomes that have been
termed transferosomes [18]. Several studies have been reported that deformable
liposomes were able to improve invitro and invivo skin delivery of various drugs [19-24]
with efficiency comparable with subcutaneous administration [25-28].
Ethosome is a novel lipid carrier, recently developed by Touitou et al showing
enhanced skin delivery.
Conventional liposomes [29,30]
Liposomes are lipid vesicles contain an aqueous volume enclosed by bilayer
lipid membrane . Lipid vesicles are usually phospholipids with or without some additives,
cholesterol may be included to improve bilayers characteristics of liposomes, increasing
micro viscosity of the bilayers, reducing permeability of the membrane to water soluble
molecules, stabilizing the membrane and increasing rigidity of the vesicles.
Chapter 2 Vesicles in transdermal drug delivery – A Review
4
From the study of Mezei and Gulasekharam(1980), four to five fold
triamcinolone acetonide concentrations in the epidermis and dermis with lower systemic
drug levels were observed when the drug was delivered from liposomal lotion in
comparison with conventional formulations. Similar findings were also observed for
triamcinolone acetonide liposomal gel formulations and for progesterone and
econozole[30]. Several invivo and invirto transport studies reported that conventional
liposomes only enhanced skin deposition with mostly reduction in percutaneous
permeation or systemic absorption of hydrocortisone [31], other corticosteroids [32],
lidocaine [33], tretinoin [34] and cyclosporine [35].
Although some authors suggested conventional liposomes as suitable carriers
for transdermal delivery of some drugs, it became recently evident that in most cases,
classic liposomes are of little or no values as carriers for transdermal drug delivery as
they do not deeply penetrate skin but rather remain confined to upper layers of the
stratum corneum. Confocal microscopy studies showed that intact liposomes were not
able to penetrate into granular layers of the epidermis.
An important role of the transappendageal route in improving skin delivery of
drugs by liposomes was also suggested [36, 37].
Highly deformable liposomes (transferosomes)
While conventional liposomes were reported to have mainly localizing or
rarely transdermal effects, deformable liposomes were reported to penetrate intact skin,
carrying therapeutic concentrations of drugs but only applied under non-occluded
conditions.
Chapter 2 Vesicles in transdermal drug delivery – A Review
5
Transferosomes are the first generation of elastic vesicles introduced by Cevic
and Blume (1992) [38]. They consist of phospholipids and an edge activator. An edge
activator is often a single chain surfactant, having a high radius of curvature that
destabilizes lipid bilayers of the vesicles and increases deformability of the bilayers.
The transition temperature (Tm) of vesicular lipids is measured using
modulated differential scanning colorimetry with programmed heating rate of 10oC/min
under constant nitrogen stream within a range of -50o to +50oC. The amount of sample
usually carried out this experiment is 20±5mg. Calorimetric studies demonstrate low Tm
values for ethosomal system as compared to liposomes suggesting a fluidizing effect of
ethanol on phospholipid bilayers. Thus ethosomes considered as a soft liquid state
vesicles with fluid bilayers. Further Tm of the drug loaded ethosomal system is similar to
that of ethosomal systems, suggesting presence of drug in ethosomal core. If this not
similar indicates presence of drug in bilayer.
(9) Storage and physical stability of ethosomes [51]
The vesicular suspensions are kept in sealed vials after flushing with nitrogen
and stored at different temperatures 4±2oC (actual storage temperature), 25±2oC (room
temperature). The stability of ethosomes was assessed quantitatively by monitoring size
and morphology of the vesicles overtime using dynamic light scattering technique and
TEM. For assessing the skin permeability of stored ethosomal system, confocal laser
microscopic studies were performed.
Chapter 4 Literature Review
22
CHAPTER- 4
LITERATURE REVIEW
1. E.Touitou et al., developed minoxidil and testosterone ethosomes. He proved
ethosomal systems were much more efficient at delivering a fluorescent probe to the skin
in terms of quality and depth than either liposomes of hydroethanolic solution. The
ethosomal system dramatically enhanced the skin permeation of minoxidil invitro
compared with either ethanolic of hydroethanolic solution or phospholipid ethanolic
micellar solution of minoxidil. In addition, the transdermal delivery of testosterone from
ethosomal patch was greater both invitro and invivo than from commercially available
patches. Skin permeation was demonstrated in diffusion cell experiments. Ethosomal
system composed of soy phosphatidyl choline 2%w/v, ethanol 30%v/v and water were
shown by electron microscopy to contain multilamellar vesicles. 31P-NMR studies
confirmed the bilayer configuration of the lipids. Calorimetry and fluorescence
measurements suggested that the vesicular bilyers are flexible, having a relatively low Tm
and fluorescence anisotropy compared with liposomes obtained in the absence of ethanol.
Dynamic light scattering was used in the average vesicle size measurement [50].
2. N.Dayan and E.Touitou characterized an ethosomal carrier containing
Trihexyphenidyl Hcl and investigated the delivery of drug from ethosomes versus classic
liposomes. As the drug concentration increased from 0 to 3% the size of the vesicles
decreased from 154nm to 94nm due to surface activity of trihexiphenidyl, measured in his
work. Zeta potential also increased from -4.5 to +10.4 as increasing the concentration
from 0 to 3% of drug. In contrast, trihexyphenidyl liposomes were much larger and their
charge was not affected by the drug. Ethosomes had a greater ability to deliver entrapped
fluorescent probe to the deeper layers of skin. The flux of drug through nude mouse skin
Chapter 4 Literature Review
23
from ethosomes was 87, 51 and 4.5 times higher than from liposomes, Phosphate buffer
and hydroethanolic solution respectively. The drug retention in the skin at the end of the
18-hour release experiment was statistically significantly greater than from liposomes or a
control hydroethanolic solution. The efficient drug delivery with the long term stability of
ethosomes made this system a promising candidate for transdermal delivery of
trihexyphenidyl [51].
3. E.Touitou et al., investigated the efficiency of transcellular delivery in to swiss
albino mice 3T3 fobroblasts of molecules with various physico-chemical characteristics
from ethosomes. The probes chosen were 4-(4-diethlyamino) styryl- N- methyl pyridinum
iodide (D-289), rhodamine red (RR) and fluorescent phosphatidyl choline (PC∗). The
penetration of these fluorescent probes into fibroblasts and nude mice skin was examined
by CLSM and flocytometry analysis. CLSM micrographs showed that ethosomes
facilitated the penetration of all probes into the cells. But from hydroethanolic solution or
classic liposomes, almost no fluorescence was detected. Enhanced delivery of molecules
from the ethosomal carrier was also observed in permeation experiments with the
hydrophilic calcein and lipophilic RR to whole nude mouse skin. Calcein penetrated the
skin to a depth of 160, 80 and 60µm from ethosomes, hydroethanolic solution and
liposomes were 150, 40, and 20 AU respectively. The highly efficient delivery exhibited
together with its non-toxicity, made this system a promising and chemical compounds to
both skin and cultured cells [52].
4. Elisabetta Esposito et al., produced ethosomes by the method described by Touitou
et al, addition of an aqueous phase to an ethanol solution of soy phosphatidylcholine
5%w/w and azelaic acid under mechanical stirring. Liposomes were obtained by the same
composition without ethanol with the reverse phase evaporation method. In order to
obtain homogenously sized vesicles, both ethosomal and liposomal dispersions were
Chapter 4 Literature Review
24
extruded through polycarbonate membrane with400nm and 200nm pore size, vesicle size
was measured by photon correlation spectroscopy and vesicles morphology was
characterized by freeze-fracture scanning electron microscopy. Free energy
measurements of the vesicle bilayers were conducted by differential scanning
calorimetry. Diffusion studies of ethosomal and liposomal incorporated gel was
investigated by a Franz cell assembled with synthetic membranes. The release rate was
more rapid from ethosomal systems than from liposomal systems. In particular,
ethosomes produced by the highest ethanol concentration released azelaic acid more
rapidly [64].
5. D.Ainbinder and E.Touitou designed and tested a testosterone non patch
formulation using ethosomes for enhanced transdermal absorption. The ethosomal
formulation was characterized by transmission electron microscopy and dynamic light
scattering for structure and size distribution and by ultracentrifugation for entrapment
capacity. The systemic absorption of drug from this formulation in rats was compared
with a currently used gel (Androgel®). Further, theoretical estimation of testosterone
blood concentration following ethosomal application in men was made. For this purpose,
invitro permeation experiments through human skin were performed to establish
testosterone skin permeation rules. This work showed that the ethosomal could enhance
testosterone systemic absorption and also be used for designing new products to solve the
weakness of the current testosterone replacement therapies [55].
6. Biana Godin and E.Touitou designed and characterized Erythromycin ethosomes
and their antibacterial efficiency was evaluated invitro and invivo. TEM, CLSM, DLS,
DSC studies performed and ultra centrifugation tests indicated erythromycin etrhosomes
are small unilamellar soft vesicles encapsulating 78.6% of erythromycin. Susceptibility
studies conducted on 3 bacterial strains. Ethosomal erythromycin applied to the skin of
Chapter 4 Literature Review
25
ICR mice inoculated with 107cfu S.aureus ATCC29213 resulted in complete inhibition of
infection. On the contrary, when hydroethanolic solution of erythromycin was applied,
deep dermal and subcutaneous abscesses developed within 5 days after challenge. For
these animals histological examination showed necrosis, destroyed skin structures and
dense infiltrates of neutrophils and macrophages. These findings showed that ethosomes
are efficient carriers for erythromycin delivery to bacteria localized within the deep skin
strata for eradication of staphylococcal infections [63].
7. J.Y.Fang et al., developed catechins encapsulated liposomes incorporating anionic
surfactants and ethanol. Liposomes were characterized for size, zeta potential and
entrapment efficiency. Both invitro and invivo skin permeation performed using nude
mouse skin as a model. Incorporation of anionic surfactants such as deoxy cholic acid and
dicetyl phosphate in the liposomes in the presence of 15% ethanol increased the (+)-
catechin permeation by five to seven fold as compared to the control. The stability and
invitro transepidermal water loss test indicated the safety of the practical use of liposomes
developed in this study [59].
8. M.M.A.Elsayed et al., investigated the possible mechanisms by which deformable
liposomes and ethosomes improve skin delivery of ketoprofen under non-occlusive
conditions. Invitro permeation and skin deposition behavior of deformable liposomes and
ethosomes , having ketotifen only inside the vesicles, only outside the vesicles and both
inside and outside the vesicles was studied using rabbit pinna skin. Results suggested that
both the penetration enhancing effect and the intact vesicle permeation into stratum
corneum might play a role in improving skin delivery of drugs by deformable liposomes
and that penetration enhancing effect was of greater importance in case of ketoprofen.
Regarding ethosomes, results indicated that ketoprofen should be incorporated in
Chapter 4 Literature Review
26
ethosomal vesicles for optimum skin delivery. Ethosomes were not able to improve skin
delivery of non entrapped ketoprofen [62].
9. V.Dubey et al., 2007 (Eur. J. Pharm. Sci) evaluated the transdermal potential of
novel ethanolic liposomes (ethosomes) bearing melatonin having poor skin permeation
and long lag time. TEM, SEM and dynamic light scattering defined ethosomes as
spherical, unilamellar structures; nanomeric size range. Entrapment efficiency was found
to be 70.71±1.4% stability profile assessed for 120 days revealed very low aggregation
and grouth in vesicular size. Ethosomal carriers showed an enhanced transdermal flux of
59.2±1.22µg/cm2/h and decreased lag time of 0.9 hours across human cadaver skin. FT-
IR data revealed a greater mobility of skin lipids on application of ethosomes as
compared to that of ethanol or plain liposomes. Confocal lasser scanning microscopy
revealed an enhanced permeation of rhodamine red loaded formulations to the deeper
layers of the skin (240nm). Further, a better skin tolerability of ethosomal suspension on
rabbit skin suggested that ethosomes may offer a suitable approach for transdermal
delivery of melatonin [60].
10. V.Dubey et al., 2007(J. Control. Release) investigated the transdermal potential of
novel vesicular carrier, ethosomes bearing methotrexate, an anti psoriatic, anti neoplastic,
highly hydrosoluble agent having limited transdermal permeation. Methotrexate loaded
ethosomes were prepared, optimized and characterized for vesicular shape and surface
morphology, stability, invitro human skin permeation and vesicle-skin interaction. The
formulation having 3% phospholipid content and 45% ethanol showing the greatest
entrapment about 68.71±1.4 and optimal nanomeric size range about 143±16nm was
selected for further transepidermal permeation studies. Stability profile for 120 days,
revealed very low aggregation and growth in vesicular size. Ethosomal carriers provided
an enhanced transdermal flux of 52.2±4.3422µg/cm2/h and decreased lag time of 0.9 hour
Chapter 4 Literature Review
27
across human cadaver skin. Confocal lasser scanning microscopy revealed an enhanced
permeation of rhodamine red [61].
11. Subheet Jain et al., investigated the mechanism for improved intercellular and
intracellular drug delivery from ethosomes using visualization techniques and cell line
study. Ethosomal formulations were prepared using lamivudine as model drug and
characterized invitro, exvivo and invivo. TEM, SEM and fluorescence microscopy were
employed to determine the effect of ethosome on ultra structure of skin. The optimized
ethosomal formulation showed 25 times higher transdermal flux (68.4±3.522µg/cm2/h)
across the rat skin as compared with that of lamivudine solution (2.8±0.222µg/cm2/h).
Results of cellular uptake of ethosomes (85.7±4.522µg/cm2/h) as compared with drug
solution (24.9±1.922µg/cm2/h) [57].
12. M.I.Tadros et al., compared the transdermal delivery of solbutamol sulphate, a
hydrophilic drug used as a bronchodilator, from ethosomes and classic liposomes
containing different cholesterol and dicetyl phosphate concentrations. All the ethosomal
systems were characterized for shape, particle size and entrapment efficiency percentage
by image analysis optical microscopy, laser diffraction and ultra centrifugation
respectively. Invitro drug permeation via a synthetic semipermeable membrane or skin
from newborn mice was studied in Franz diffusion cells. The selected formulations were
incorporated into pluronic F-127 gels and evaluated for both drug permeation and mice
skin deposition. The vesicle size was significantly decreased by decreasing cholesterol
concentration and increasing dicetyl phosphate and ethanol concentrations. The
entrapment efficiency percentage was significantly increased by increasing cholesterol,
dicetyl phosphate and ethanol concentrations. Invitro permeation studies of the prepared
gels containing the selected vesicles showed that ethosomal systems were much more
Chapter 4 Literature Review
28
efficient at delivering solbutamol sulphate into mice skin than were liposomes or aqueous
or hydroethanolic solutions [55].
13. Zhaowu et al., evaluated the preparation of matrine ethosomes and the percutaneous
permeation invitro and the ant-inflammatory activity invivo in the rat skin. The matrine
ethosomes were prepared by the ethanol injection-sonication method. The particle size,
entrapment efficiency was determined by laser particle size analyzer and
ultracentrifugation respectively. Anti-inflammatory activity invivo was determined by a
reflection spectrophotometer. Average particle size was in the range of 50-200nm with a
narrow size distribution and entrapment efficiency was in the range of 40-90%. Within 24
hours the cumulative permeation quantity is 60.5% and with no permeation quantity is
60.5% and with no permeation lag time. Matrine ethosomes able to disappear more
rapidly the induced erythema than non ethosomal formulations. Matrine ethosomes can
increases the percutaneous permeation of matrine in the experiment invitro and improve
the anti-inflammatory activity of matrine invivo in rat skin [56].
14. Massimo Fresta et al., evaluated the various ethosomal suspensions made up of
water, phospholipids and ethanol at various concentrations for their potential application
in dermal administration of ammonium glycyrrhizinate, a useful drug for the treatment of
various inflammatory-based skin diseases. Physicochemical characterization of
ethosomes was carried out by photon correlation spectroscopy and freeze fracture
electron microscopy. The percutaneous permeation of ammonium glycyrrhizinate
ethosomes was evaluated in vitro through human stratum corneum and epidermis
membranes by using Franz’s cells and compared with the permeation profiles of drug
solutions either in water or in a water–ethanol mixture. Ethosomal suspensions had mean
sizes ranging from 350 nm to 100 nm as a function of ethanol and lecithin quantities,
i.e.,high amounts of ethanol and a low lecithin concentration provided ethosome
Chapter 4 Literature Review
29
suspensions with a mean size of approximately 100 nm and a narrow size distribution. In
vitro and in vivo experiments were carried out by using an ethosome formulation made up
of ethanol 45% (v/v) and lecithin 2% (w/v). The ethosome suspension showed good skin
tolerability in human volunteers, also when applied for a long period (48 h). Ethosomes
elicited an increase of the in vitro percutaneous permeation of both methylnicotinate and
ammonium glycyrrhizinate. Ethosomes were able to significantly enhance the anti-
inflammatory activity of ammonium glycyrrhizinate compared to the ethanolic or aqueous
solutions of this drug [54].
Chapter 5 Scope of work
30
CHAPTER-V
SCOPE OF WORK
Nonsteroidal anti-inflammatory agents (NSAIDs) are class of drugs which
budded from the bark of willow in the mid-eighteenth century. Now a days, there has
been a rapid increase in the number of products that have been designed to deliver
NSAIDs. These include creams, gels, and more complex transdermal systems. A number
of approaches have been continuousely investigated so as to enhance dermal delivery by
use of prodrugs, ultrasound, ionotophorosis and microneedles. But the choice of the most
appropriate drug depends on a number of factors which includes its potency, its ability to
permeate the stratum corneum, its lack of local skin toxicity and stability towards
metabolizing enzymes present on the skin surface.
Diclofenac potassium is one of the NSAID widely used for musculoskeletal
complaints, especially arthritis, rheumatoid arthritis, polymyositis, dermatomyositis,
osteoarthritis, dental pain, spondylarthritis, ankylosing spondylitis, gout attacks, and pain
management in cases of kidney stones and gallstones. An additional indication is the
treatment of acute migraines. Diclofenac potassium is used commonly to treat mild to
moderate post-operative or post-traumatic pain, particularly when inflammation is also
present, and is effective against menstrual pain and endometriosis. It can also be used to
reduce menstrual pain and dysmenorrhea.
Oral dose of Diclofenac potassium causes an increased risk of serious
gastrointestinal adverse events including bleeding, ulceration and perforation of the
stomach or intestines which can be fatal. These events can occur at any time during use
Chapter 5 Scope of work
31
and without warning symptoms. This drug may also cause an increased risk of serious
cardiovascular thrombotic events, myocardial infarction and stroke.
Due to the presence of these oral adverse effects, necessitates the need for
investigating other routes of drug delivery fo Diclofenac Potassium. Transdermal delivery
of the drug can improve its bio activity and transdermal effect, reduce the side effects and
enhance therapeutic efficacy. This can be achieved only when the drug has entered the
lower layers of the skin, then only it can be absorbed by blood and transported to the site
of action, or penetrate deeper in to areas where inflammation occurs. Pure drug or
liposomal formulations not reach the lower layers of the skin.
Ethosome, a novel liposome, is especially suitable for topical and transdermal
administration carrier. Compared to other liposomes, the physical and chemical properties
of ethosomes make the delivery of the drug through the stratum corneum in to a deeper
layer efficiently or even into the blood circulation. Diclofenac Potassium is a water
soluble drug and generally the entrapment efficiency of the ethosomes of a water soluble
drug is higher than that of the other vesicle formulation. So the Diclofenac potassium
ethosomal formulation may be better than other transdermal or topical formulation of
Diclofenac potassium.
Chapter 6 Plan of work
32
CHAPTER- VI
PLAN OF WORK
PART –I
1. Determination of λmax of Diclofenac Potassium. 2. Calibration curve for the drug in phosphate buffer saline PH: 7.4.
PART-II
1. Formulation of Diclofenac Potassium ethosomes using different concentration of phosphatidylcholine and ethanol by classic mechanical dispersion method.
PART-III
1. Determination of drug entrapment efficiency by centrifugation method. 2. Invitro release characteristics of ethosomes in phosphate buffer saline PH: 7.4 using synthetic semipermeable membrane.
PART-IV
1. Formulation of Diclofenac Potassium liposomes and determination of drug entrapment efficiency.
PART-V
1. Invitro release characteristics of the selected ethosomal formulation, liposomal formulation, hydroethanolic drug solution and phosphate buffer saline PH: 7.4 drug
solution through rat membrane.
2. Estimation of drug retention in the rat skin by ethanolic extraction method after 12 hours of invitro skin permeation study.
PART-VI
1. Morphological studies of ethosomes using scanning electron microscopy.
PART-VII
1. Determination of vesicular size distribution of ethosomal and liposomal formulations using dynamic light scattering (DLS) technique.
PART-VIII
1. IR studies to determine the interaction between ethosomal membranes with drug.
Chapter 6 Plan of work
33
PART-IX
1. Stability studies of ethosomal formulations at refrigerated temperature and room temperature.
PART-X
1. Formulation of gel containing Diclofenac Potassium ethosomes.
PART-XI
1. Pharmacodynamic studies to compare the ethosomal gel formulation and marketed gel formulation.