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IJPIs Journal of Pharmaceutics and Cosmetology

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Formulation Development of Ketoprofen Liposomal Gel

Mansoori M.A.*, Jawade S., Agrawal S., Khan M.I.
Swami Vivekanand College of Pharmacy,
Indore-452020, State- Madhya Pradesh, INDIA

Corresponding author: Mansoori M.A.

Email address: mohammadazam86@gmail.com

ABSTRACT:
Liposomes are acceptable and superior carriers and have ability to encapsulate hydrophilic and
lipophilic drugs and protect them from degradation. Applied to the skin, liposomes act as a solubilizing
matrix, penetration enhancer and local depot for various poorly water-soluble drugs. The objective of this
study was to develop a ketoprofen (NSAID) liposomal gel for better anti-inflammatory activity and reduced
adverse effects. Liposome carriers, well known for their potential and topical drug delivery was chosen to
help transport ketoprofen molecule in skin layers. Ketoprofen was encapsulated in liposomes for topical
application. Ketoprofen liposomes were prepared by thin film hydration technique using soya lecithin,
cholesterol and drug in different weight ratios. Carbopol 934 was used as a vehicle for topical drug delivery
in the concentration range 1%. In evaluation study, the effect of varying concentrations of lipid on the
properties of liposomes such as encapsulation efficiency, particle size, invitro drug release and physical
stability were studied. Phase transition study was carried out to confirm the complete interaction of
ketoprofen with bilayer structure of liposome. Results of analysis revealed that the size of liposomes and
entrapment efficiency were dependent on the lipid concentration. Moreover, the release of the drug was also
modified and extended over a period of 8 h in all formulations. Drug lipid interaction study showed no
interaction between drug and lipid. Ketoprofen liposomal gel together, the result indicates that the
diclofenac liposomal gel is better than the regular gel without liposome.
Keywords: Liposome; liposomal gel; ketoprofen; topical delivery

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IJPIs Journal of Pharmaceutics and Cosmetology

1. INTRODUCTION
Skin has been considered as an alternative route for local and systemic treatment. Topical dosage forms
provide relatively consistent drug levels for prolonged periods and avoid gastric irritation, as well as the other typical
side effects of oral NSAID administration. Ketoprofen belongs to the group of substituted 2phenylproprionic acids
which has analgesic, antiinflammatory and antipyretic effects.
Ketoprofen is a non steroidal anti-inflammatory drug (NSAID) with analgesic and antipyretic properties.
Ketoprofen has pharmacologic actions similar to those of other prototypical NSAIDs, which inhibit prostaglandin
synthesis.
Presently ketoprofen is available in the market in the form of tablet, injection, and normal gel. All these
formulation having their own drawback in respect of their route of administration such as various side effect,
maintenance of plasma drug level and skin penetration problem respectively.
Like other NSAIDS, Following ketoprofen oral administration have several side effects such as abdominal
pain, peptic ulcer, nephrotoxicity, pencreatitis, G.I.T. bleeding, alveolitis, nausea, haematological disorder etc. on
continuous use.
Ketoprofen is drug with analgesic and antipyretic properties indicated for the treatment of rheumatoid arthritis,
osteoarthritis, muscular skeletal disorder, soft tissue injuries, tooth extraction, post partum, post operatively and acute
gout. Therapy of these diseases takes longer time for proper treatment; possibilities of side effects mentioned above are
high with continuous administration.
Parenteral route offer rapid onset of action with decline of systemic drug level. For effective treatment, it is
often desirable to maintain systemic drug level with therapeutic effective concentration range as long as treatment calls.
In case of chronic condition it requires frequent injection which ultimately leads to patient discomfort especially in old
persons.
Topical liposome formulation could be more effective and can penetrate deeper into skin layers and hence
gives better release of drug than conventional topical formulation.
For this reason research is focused on topical delivery of ketoprofen by encapsulation into liposomes in the
form of gel in order to avoid the side effects following oral administration, to maintain maximum therapeutic effect for
longer time, to provide sustain release of drug through enhancing circulation life time.
2. MATERIAL AND METHOD
2.1 Materials:
Ketoprofen was obtained as a gift sample from Ranbaxy Pvt. Ltd. Dewas. Soya lecithin and cholesterol were
purchased from Himedeia and carbapol from SDFCL. Carbopol 934 NF was purchased from Himedeia. All other
chemicals and reagents were of analytical grade.
2.2 Preparation of liposome(1):
Aqueous liposomal dispersions were prepared by conventional lipid film hydration method. Different weight
ratios of phospholipid, cholesterol and drug were weight and dissolved in chloroform in 250 ml round bottom flask. A
thin film was formed on evaporating organic solvent under vacuum in rotator evaporator at 35-40C. Subsequently the
flask was kept overnight under vacuum to ensure the complete removal of residual solvent. The dry lipid film was
hydrated with 15 ml of phosphate buffer solution (pH 7.4) at a temperature of 402 C. The dispersion was left
undistributed at room temperature for 2-3 hour to allow complete swelling of the lipid film and hence to obtain
vesicular dispersion.

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2.3 Gel Preparation:

Gel was made as a vehicle for incorporation of ketoprofen liposome for topical delivery. Carbopol 934 (1 g)
was dispersed in demineralised water (88 ml) by continuous stirring for 45 minutes. Then propylene glycol 10 ml was
added and the mixture was neutralized by drop wise addition of 10% sodium hydroxide. Mixing was continued until a
transparent gel appeared, while the amount of base was adjusted to achieve a gel with pH 6.5.
2.4 Incorporation of ketoprofen Liposome into Gel:
Liposomes containing ketoprofen were mixed into the 1% (w/w) carbopol hydrogel by a mechanical stirring at
25 rpm for 5 minute to get ketoprofen liposome gel.
Table 1: Composition of lipid for liposome preparation
Formulation code
F1
F2
F3

Drug
100
100
100

Ingredient (mg)
Lipid
Cholesterol
500
500
550
500
600
500

2.5 Drug entrapment efficiency (2):

The liposome suspension was centrifuged at 5000 rpm for 15 minute at 4C temperature by using semi cooling
centrifuge to separate the free drug. A supernatant containing liposome in suspended stage and free drug in the wall of
centrifuge tube. The supernatant was collected and again centrifuged at 1500 rpm at 4 C temperature for 30 minute. A
clear solution of supernatant and pellet was obtained. The pellet containing only liposomse was resuspended in distilled
water until further processing. The liposomes free from unentraped drug were soaked in 10 ml of methanol and then
sonicated for 10 minute. The vesicles were broken to release the drug, which were then estimated for the drug content.
The absorbance of drug was noted at 260 nm. The entrapment efficiency was then calculated using following equation.
% Drug Entrapped (PDE) = (Amount of drug in sediment / Total amount of drug) 100
2.6 In- vitro release study (3):
In vitro release studies were performed using modified Franz diffusion cell. Dialysis membrane (Hi Media
molecular weight 5000) was placed between receptor and donor compartments. ketoprofen liposomal was placed in the
donor compartment and the receptor compartment was filled with phosphate buffer, pH 7.4. The diffusion cell was
maintained at 370.5C with stirring at 500 rpm throughout the experiment. At fixed time intervals, 5ml of aliquots
were withdrawn from receiver compartment through side tube and analyzed by UV-Visible Spectrophotometer at
260nm. Percentage of drug release of different formulations are given in table no. 13.
2.7 pH of the liposomal gel( 4):
The pH of various gel formulations was determined by using digital pH meter. The measurement of pH of each
formulation was done in triplicates and average values were calculated. Same procedure results from pH measurement
are given in table no.4.
2.8 Storage- stability study(5):
The ability of vesicles to retain the drug (i.e., drug retentive behavior) was assessed by keeping the liposomal
suspension at three different temperature conditions, i.e., 4-8C (Refrigerator; RF), 252C (Room temperature; RT),
and 452C for a period of 4 weeks. The liposomal suspension was kept in sealed ampoules (10ml capacity) after
flushing with nitrogen. Sample was withdrawn periodically and analyzed for the drug content, in the manner described
under drug entrapment studies. Same procedure followed for all other formulations. Data from storage stability study
are given in table no.5.

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2.9 Differential Scanning Calorimetry (DSC) study( 6):

Differential scanning calorimetry (DSC) experiments were performed with differential scanning calorimeter
(model TA-60, Shimadzu, Japan). Samples of pure ketoprofen, mixture of soya lecithin and cholesterol and drugloaded multilamellar liposomes were subjected to DSC analysis. The analyses were performed on 5 mg samples sealed
in standard aluminum pans. Thermograms were obtained at a scanning rate of 10C/min. Each sample was scanned
between 0C to 250C. The temperature of maximal excess heat capacity was defined as the phase transition
temperature.
2.10 Scanning electron microscopy(7):
Scanning electron microscopy (XL 30 scanning microscope, Philip the Netherland) was employed to determine
the shape and surface morphology of produced liposome. A small amount of liposome was stuck on double sided
tapped metallic coated under vaccumm with a thin layer of gold before scanning. Structure of optimized liposome (F6 )
shown in fig 11.
3. RESULT AND DISCUSSION
Characterization of ketoprofen liposomal gel
Physicochemical properties
The liposome suspensions were white in colour, odourless and fluid in nature. Gel loaded with liposome
suspension were colourless, odourless with smooth appearance.
Drug entrapment efficiency
The drug entrapment efficiency of liposomal preparations were determined by centrifugation method. The
result of drug entrapment efficiency of liposomes table no.2 indicated that increase in with increase in concentration of
lipid, Drug entrapment efficiency of liposomes were increases which might be attributed to increase in number of
bilayers formation lipid, which having the drug molecule. The encapsulation efficiency of liposome is governed by the
ability of formulation to retain the drug molecule in aqueous core or in the bilayer membrane of the vesicle. The
entrapment efficiency data clearly suggested that the ratio of lipid to cholesterol in liposome entrapment is crucial
because enhanced cholesterol level disturbs entrapment due to increased separation between choline head group. From
the drug entrapment efficiency study, maximum drug encapsulation was found in F1, in which lipid and cholesterol in
equal ratio (1:2). In liposome preparation cholesterol was found to acts as fluidity buffer and provided stability and
rigidity to liposome.
Table 2: Drug entrapment of different formulations
Formulation Code

% Entrapment Efficiency

F1
F2
F3

97.51
95.36
93.69

In- vitro release study

The percentage drug releases of different formulations were determined by using Franz diffusion cell up to 24
hour. Drug releases of different formulations were compared with that of marketed product. The comparative in vitro
drug release profile summarized in table no.3, for marketed ketoprofen gel and for each formulation. It was observed
that marketed gel released approximately 92% of drug within 24 hour, while liposomal formulations F1, F2, F2, showed
81%, 87% and 84% drug release respectively in 24 hour. Liposomal formulations showed sustained drug release
compared to normal gel. All the liposome enriched gel formulation showed better drug release and also an increase in
release rate was observed after 12 hour.
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Table 3: Percentage Drug release of different formulations

Formulation
Code
Normal gel
F1
F2
F3

2
07
09
13
11

4
21
15
24
19

% Drug Release (hr)

6
8
10
35
48
62
29
42
57
38
49
62
31
46
62

12
78
73
78
74

24
92
81
87
84

Figure 1: Percentage drug release of different formulations

pH of the liposomal gel

The pH of various gel formulations was determined by using digital pH meter. pH of all the formulations was
found to be in the range of 5.86 to 6.06, which is around to the pH of skin. This showed that formulations are suitable
for topical use.
Table 4: pH of the liposomal gel
Formulation Code
F1
F2
F3

pH
6.06
6.03
5.86

Storage- stability study

One month stability study of liposomal suspension was conducted with respect to the liposomes ability to
retain an entrapped drug during a defined time period. All liposomal formulations were stored at different temperature
condition i.e. 2-8C and 25C and 45C for 4 week. All formulations showed same drug content as before their storage
table no.5. Result showed that liposome formulations of ketoprofen are expected to be stable for one month.
Table 5: Storage stability of the liposomal gel
Formulation
code
F1
F2
F3

Duration in weeks
25 C

4-8 C
1
96.86
94.86
92.60

Mansoori M.A. et al

2
96.75
94.51
92.45

3
96.54
94.33
92.31

4
96.31
94.09
92.10

1
92.90
91.65
89.69

2
92.78
91.41
89.50

3
92.55
91.39
82.99

4
92.31
91.11
89.13

45C
1
89.78
89.55
85.80

2
89.51
89.89
85.67

3
89.39
89.78
85.43

4
89.11
89.20
85.28
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Differential Scanning Calorimetry (DSC) study

DSC thermogram of ketoprofen showed endotherm at 940C. DSC thermogram of mixture of soya lecithin and
cholesterol showed endotherm at 1280. DSC thermogram of ketoprofen loaded liposome batches F1, composed of Soya
lecithin and cholesterol showed endotherm at 1050C, disappearance of melting endotherm of ketoprofen suggested that
all the lipid components interacted with each other to a grater extent while forming the lipid bilayers. The incorporated
ketoprofen associated with lipid bilayers and interact to a large extent with them. Absence of the melting endotherm of
ketoprofen and shifting of the lipid bilayers component endotherm suggested significant interaction of ketoprofen with
bilayers.
Figure 2: DSC thermograms A-(ketoprofen) B- Mixture (soya lecithin and cholesterol)
C- Liposome formulation
Pure drug (ketoprofen) - A

Mixture of samples - B (soya lecithin and cholesterol)

Liposome formulation

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Scanning electron microscopy

Morphology of liposome was studied under scanning electron microscope. The photomicrograph of optimized
batches revealed the presence of well identified spheres of multilamellar vesicles that consisted of many concentric
phospholipid bilayers.
Figure 3: Scanning electron microscopy picture of liposome

4. CONCLUSION
Liposomal product of ketoprofen was found to have reasonable drug loading, controlled release rate, particle size, and
stability and phase transition behaviour. The formulated ketoprofen liposomes have shown an appreciably enhanced
retention of drug molecules in the skin. Thus, the liposomal formulation, with desired characteristics for topical
administration, could be successfully prepared.
5. ACKNOWLEDGE
Authors are thankful to Ranbaxy Pvt. Ltd. Dewas for providing gift sample of ketoprofen. The author would
like thank to principle and management of Swami Vivekanand College of Pharmacy for providing necessary facilities
useful in conduction of this work.
6. REFERENCES
(1) A. V. Jithan and M. Swathi. Development of topical diclofenac sodium liposomal gel for better
antiinflamatory activity. International Journal of Pharmaceutical Science and Nanotechnology. 2010; 3(2);
986-993.
(2) Agarwal R. and O. P. Katare. Preparation and in vitro evaluation of miconazole nitrate loaded topical
liposome. Pharmaceutical Technology. 2002; 41(9); 48-60.
(3) Roopa Pai and Kusum Devi V. Lamivudine liposomes for transdermal delivery- Formulation,
characterization, stability and in vitro evaluation. International Journal of Pharmaceutical Science and
Nanotechnology. 2009; 1(4); 317-326.
(4) Gita Rao and R. S. R.Murthy. Preparation and evaluation of liposomal flucinolone acetonide gel for
intradermal delivery. Pharmacy and Pharmacology communication. 2000; 6(11); 447-483.
(5) Debnant Kumar Sujit. Formulation and evaluation of aceclofenac gel. International Journal of Chemtech
Research. 2009; 1(2); 204-207.
(6) Dr. Patel P. Rakesh. Formulation and evaluation of liposome of ketoconazole. International Journal of Drug
Delivery Technology. 2009; 1(1); 16-23.
(7) Omaina N. Preparation and evaluation of acetazolamide liposome as an ocular delivery system. APS
Pharmasci Tech. 2007; 8(1); 45-56.
Mansoori M.A. et al

Page 28

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(8) Amit Bhatia. Tamoxifen in topical liposomes development, characterization and in vitro evaluation. Journal of
Pharmaceutical Science. 2004; 7(2); 252-259.
(9) Natasha Skalko. Liposome with clindamicin hydrochloride in the treatment of Acne vulgaris. J Drug Target;
2002; 17(4); 223-230.
(10) B. V. Mikari. Formulation and evaluation of topical liposome gel for fluconazole. Indian Journal of
Pharmaceutical Education and Research. 2010; 44 (4); 324-333.
(11) S. Rathod. Design and evaluation of liposomal formulation of pilocarpine nitrate. Indian Journal of
Pharmaceutical Education and Research. 2010; 72(2); 155-160.
(12) Roopa Karki. Formulation and evaluation of coencapsulated rifrmpin and isoniazid liposome using different
lipid. Acta pharmaceutica sciencia. 2009; 51(6); 177-188.
(13) V. D. Shivhare. Formulation of pentoxifyllin liposome formulation. Digest Journal of Nanomaterials and Bio
structure. 2009; 4(4); 857-862.
(14) Nagasenker M. S. Preparation and evaluation of tropicamide for ocular delivery. International journal of
pharmaceutics. 1998; 1(3); 63-71.
(15) Reeta T. Mehta. Liposome encapsulation of Clofazimine reduced toxicity in vitro and in vivo and improved
therapeutic efficacy. Antimicrobial agent and chemotherapy. 1996; 40(8) 189-190.
(16) Afrouz Yousefi. Preparation and in vitro evaluation of pegylated nano liposomal formulation containing
docetaxel. Scientia pharmceutia. 2009; 77(14); 453-462.
(17) Gita Rao and R. S. R.Murthy. Preparation and evaluation of liposomal flucinolone acetonide gel for
intradermal delivery. Pharmacy and Pharmacology communication. 2000; 6(11); 447-483
(18) A. R. Mohammad. Liposomal formulation of poorly water soluble drug: optimization of drug loading and
ESEM analysis of stability. Indian journal of Pharmaceutic. 2008; 2 (1); 45-52.
(19) M. S. Nagarsenker. Preparation and evaluation of liposomal formulation of sodium cromoglicate.
International Journal of Pharmaceutics. 2003; 251(2); 49-56.
(20) Carol Cordeiro. Antibacterial efficacy of gentamicin encapsulated in pH sensitive liposome. International
Journal of Pharmaceutic. 1999; 195(2); 24- 36.
(21) Tang J. Pharmacokinetic and biodistribution of itraconazole in rats and mice following intravenous
administration in a novel liposome formulation. Drug Delivery. 2010; 17 (4); 223-30.
(22) Prabhakara Prabhu. Preparation and evaluation of liposome of brimonidine tartrate as an ocular drug delivery
system. International Journal of Pharmaceutical Science. 2010; 1(4); 502-508.
(23) Jain N.K. Controller and Novel Drug Delivery. CBS Publisher and distributors, New Delhi. 2009; 1; 278-283.
(24) Vyas S.P., Khar K.R. Trageted and Controlled drug delivery. CBS Publisher and distributors, New Delhi.
2002; 1; 181-187.
(25) Martin Alfred. Physical pharmacy. Lippincott Williams and Wilkins. Fourth edition. 2001; 32-33

Mansoori M.A. et al

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