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Indian Journal of Pharmaceutical Education and Research

Association of Pharmaceutical Teachers of India

Development and Evaluation of Sorbitan Monostearate Organogels


as a Topical Delivery System for Aceclofenac
1*

Kamble S.R. , Prachi Udapurkar , Nakhat P.D. , Yeole P.G. and Biyani K.R.
1

Anuradha College of Pharmcy, Anuradha Nagar, Sakegaon Road Chikhli, Buldana-443 201

Wockhardt Research Centre, MIDC, Chikhalthana, Aurangabad - 431 201

Institute of Pharmaceutical Education & Research, Borgaon (Meghe), Wardha-442 001

ABSTRACT

Submitted: 15/4/2010

Revised: 30/6/2010

Accepted: 28/8/2010

The studies were conducted with an object to develop stable safe and efficient delivery system for aceclofenac. During the course of studies
different organogel formulations of aceclofenac for topical application were prepared by using sorbitan monostearate (span 60), isopropyl
myristate, purified water, sorbic acid, potassium sorbate, tween 20, vitamin E, methyl salicylate and menthol. The formulated organogels were
evaluated for psychorheological characteristic, drug content, pH, spreadability. The viscosities of different formulations were determined by
using Brookfield Viscometer at 25C, the viscosity of formulations increases as the surfactant concentration increases. The developed
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formulation then subjected to in vitro diffusion study. The two formulations showed best release having flux > 0.2 mg/cm /hr were selected and
modified with incorporation of 10% methyl salicylate and 5% menthol. Further efficacy of these formulations was evaluated and compared with
standard marketed formulation by anti-inflammatory activity on albino rats using carageenan-induced rat paw edema model, promissing
edema inhibition withen three hr was observed. Safety was determine through skin irritancy testing on Guinea pigs for seven days showing no
signs of skin irritation. Finally stability studies were carried out for three months showed no separation of gel indicating overall stability. The
result indicates that Sorbitan Monostearate organogels serves as a better vehicle for topical delivery of aceclofenac.
KEYWORDS: Aceclofenac, Sorbiton monostearate, Organogel

INTRODUCTION
Aceclofenac is a phenylacetic acid derivative which has an
established use in inflammation caused by soft tissue injury
and rheumatic diseases including osteoarthritis.1 The main
concern with this drug is that relatively high quantities of drug
must first be systemically administered to treat inflammation
which may be the cause of many adverse effects. Hence in the
present study attempt has been made to improve its
therapeutic performance by developing topical gel
formulation which will improve penetration across skin,
increase availability at desired site and improve patient
compliance with avoidance of undesirable effects. Gels are
intermediate states of matter containing both solid and liquid
components. The solid component comprises a three
dimensional network of interconnected molecule or
Address for Correspondence:
Kamble S.R, Anuradha College of Pharmcy, Anuradha Nagar, Sakegaon Road Chikhli,
Buldana-443 201
E- mail: ksantosh_49@rediffmail.com

Ind J Pharm Edu Res, Jan-Mar, 2011/ Vol 45/ Issue 1

aggregates which immobilize the liquid continuous phase


when the liquid component is aqueous, the gel is termed a
hydrogel and when the liquid is an organic medium, the gel is
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called an organogel. Organogel formation is based on the
spontaneous self-association of individual gelator molecules
into three-dimensional networks of randomly entangled
fiber-like structures, thereby confining the solvent at the
3
microscopic level. Murdan, S. et al. (2004) studied sorbitan
monostearate and sorbitan mono palmitate hydrophobic nonionic surfactant (10% w/v) organogels using solvents such as
hexadecane, isopropyl myristate etc. And non-ionic
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surfactant based organogel incorporating niosomes.
MATERIALS AND METHODS
Aceclofenac was kindly supplied as gift sample by Suyash
Laboratories, Mumbai (India). Sorbitan monostearate and
isopropyl myristate were purchased from Loba chemie
Mumbai. All the other materials used were of analytical
grades.

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Kamble et al.: Development and Evaluation of Sorbitan Monostearate Organogels as a Topical Delivery System for Aceclofenac

Preperation of Organogel:
Organogels (F1-F7) were formulated by preparing oil phase
and aqueous phase at 60. The composition of oil phase was
sorbitan monostearate (7-13%), sorbic acid (0.2%), tween 20
(2%) and isopropyl merystate while that of aqueous phase
was potassium sorbate (0.2%) in purified water finally
aceclofenac (1.5%) was incorporated in oil phase and
aqueous phase was slowly added to it with stirring. Organogel
(F8 and F9) were formulated by modifying F1 and F2
respectively with incorporation of 10% methyl salicylate and
5% menthol respectively. The composition of different
formulation is summarized in Table No 1.
Evaluation of Organogel:
Psychorheological Characterization:
The formulated gels were inspected visually for colour,
presence of any clog and sudden viscosity changes. To
evaluate the feel, the formulations were applied on the skin
and the feel was experienced psychorheologically. 5

using Brookfield Viscometer (Model: RV-DV-E 230,


Brookfield Engineering Lab., Inc., Middleboro. USA).
Spindle no. 6 was used for organogels and the spindle speed of
2 rpm at 25 C.
Spreadability:
Spreadability of formulations was determined by an
apparatus suggested by Multimer et al. which was fabricated
in laboratory and used for study (Fig.1). An excess of gel
sample 2.5 g was placed between two glass slides and a 1000g
weight was placed on slides for 5 minutes to compress the
sample to a uniform thickness. Weight (60g) was added to the
pan. The time (seconds) required to separate the two slides
was taken as a measure of spreadability. It was calculated
using the formula, S = m.l / t, where, S - Spreadability in g.cm
/sec, m - Weight tied to upper slide, l - Length of glass slide
and t - Time in seconds. Length of glass slide was 11.3 cm and
weight tied to upper slide was (60g) throughout the
experiment.8
In vitro Skin Permeation Studies:

Drug content:
Each formulation (0.5g) was dissolved in 50 ml ethanol. The
solution was filtered through 42 membrane; 1ml of the
above filtrate was pipetted out and diluted to 10ml with
ethanol. The absorbance was measured at 276.5 nm, using
double beam UV visible spectrophotometer (Model: UV2401 PC, Shimadzu Corporation, Singapore).6
pH Determination:
The pH of formulated organogels was determined using pH
meter. The electrode was immersed in organogels and
readings were recorded on pH meter.7
Viscosity:

The developed formulations were subjected to in vitro


diffusion study through dialysis membrane (HIMEDIA);
with molecular weight cut off 12000-14000 KD, using
modified Keshry Chien diffusion cell. Pieces of dialysis
membrane were soaked in medium used for diffusion study
for 24 hr before mounting on a diffusion cell. The receptor
compartment was filled with saline phosphate buffer (pH 7.4)
and methanol (90:10). Methanol was added in medium to
maintained sink condition. The whole assembly was
maintained at 37 1 C and receptor solution was stirred
with a magnetic stirrer at 600 rpm throughout the experiment.
Care was taken that no air bubbles were trapped under the
membrane. Aliquots (1ml) were withdrawn at regular interval

Viscosities of the formulated organogels were determined


Table 1: Formulation of sorbiton monostearate Organogels

Contents

F1

F2

F3

F4

F5

F6

F7

Aceclofenac (% wt)

1.5

1.5

1.5

1.5

1.5

1.5

1.5

Span 60 (% wt)

10

11

12

13

Tween 20 (% wt)

Sorbic acid (% wt)

0.2

0.2

0.2

0.2

0.2

0.2

0.2

Vitamin E (% wt)

0.1

0.1

0.1

0.1

0.1

0.1

0.1

Isopropyl myristate up to (% wt )

100

100

100

100

100

100

100

Potassium sorbate (% wt)

0.2

0.2

0.2

0.2

0.2

0.2

0.2

30

30

30

30

30

30

30

Purified water (ml)

*Values are Mean SEM. (n=3)

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Kamble et al.: Development and Evaluation of Sorbitan Monostearate Organogels as a Topical Delivery System for Aceclofenac

of 1hr for a period of 8 hr and replaced with equal volume of


fresh medium equilibrated at 37 1 C. All the samples were
suitably diluted with medium and analyzed
spectrophotometrically at 274 nm for aceclofenac content.
The cumulative quantity of aceclofenac diffused per unit area
of the membrane was calculated for each time interval.9
The mean cumulative amount of drug permeated per unit
surface area was plotted versus time. The slope of the linear
2
10
portion of the plot was calculated as flux J (mg/cm /hr) and
the permeability coefficient was calculated using equation,
Kp = J/Cd, where, Kp is the permeability coefficient and Cd is
11
the initial drug concentration in drug compartment.

confluent or moderate but patchy erythema, 2-Moderate


13
erythema, 3-Severe erythema with or without edema.
Stability study:
Freeze-thaw and thermal cycling test:
The selected organogels were exposed to different
temperature conditions in a cyclic pattern that simulate the
changes likely to be encountered during its use or in
distribution process. During the course of study the selected
formulations were subjected to refrigerated temperature
(2-8C) for two days, followed by 40 C for two days, one
cycle. Such types of three cycles in twelve days were
completed and major changes were observed.

In vivo Anti-inflammatory study:


In vivo anti-inflammatory study was conducted using 30
albino rats of either sex weighing (100-150g) and divided into
5 groups. The experimental protocol was approved by
Institutional Animal Ethical Committee under the guidelines
of CPCSEA, New Delhi. In all groups, acute inflammation
was induced by sub-planter injection of 0.1ml of freshly
prepared 1% w/v suspension of carrageenan in normal saline
in left hind paw of the rats. The medicated formulation (0.25g)
or base was applied topically with gentle rubbing to the paw of
each rat of respective group one hour before and one hour
after the carrageenan challenge. The paw edema volume was
measured using plethysmometer (model: 520 (Almemo
2290-4), IITC Life Sciences, Woodland Hills, USA) at 1, 2, 3
and 4 hr after injection of carrageenan. The average paw
edema volume of all the groups were calculated and
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compared with that of control. The percent inhibition of
edema was calculated by using following formula.
% Edema inhibition = (1- Vt / Vc) 100, where, Vt - mean
edema volume of test; Vc - mean edema volume of control.
Data were analysed statistically using one way analysis of
variance (ANOVA) followed by Dunnet test.
Skin irritation study:
In the present study 10 guinea pigs of either sex weighing
between (400-500g) were used. Animals were divided in to 2
groups of 5 animals each. Hairs were depleted from the back
of guinea pig with the help of depilatories and area 4 cm2 was
marked on both the sides. One side served as control while the
other as test and animals were used after 24 hrs after hair
depletion. Gel was applied (500mg / guinea pig) once a day
for 7 days and site was covered with cotton bandage and
observed for any sensitivity and the reaction if any was graded
as 0-No reaction, 0.5-Slight; patchy erythema, 1-Slight but
Ind J Pharm Edu Res, Jan-Mar, 2011/ Vol 45/ Issue 1

The selected organogels were also subjected to 25C 2C at


75 5% RH and 40C 2C at 75 5% RH of temperature
and relative humidity during stability studies and were
evaluated for various parameters after every month for three
months. The parameters of the organogels studied were Drug
content, viscosity, pH and in vitro diffusion study.
Statistical analysis: All values were expressed as mean
SEM. Data were analysed statistically using one way
analysis of variance (ANOVA) followed by Dunnet test. The
level of significance was considered at P<0.01.
RESULT AND DISCUSSION
Various organogels of Nonionic surfactants (Span 60, Tween
20) incorporating water were formulated. The prepared
formulation were evaluated for their psychorheological
characteristic, drug content, pH, viscosity, spreadability,
in vitro diffusion study. It was found that organogels were
smooth and showed no clogging which indicate good texture
of formulation. The drug content of organogels were in the
acceptable range, indicating no interaction of drug with
component of base. The pH of all the formulations was in the
range of 5.1 to 5.6 that suits the skin pH, indicating skin
compatability. The organogels showed viscosities in the range
of 27500-47200 cps and spreadability in the range of 32.19 52.70 g.cm/sec (Table No. 2).
Effectiveness of topical applications mainly depends upon its
rate and extent of drug release from base. All formulations
were evaluated for their release patterns from bases and
compared with the marketed formulation. Cumulative
2
amount of Aceclofenac permeated (mg/cm ) in 8 hr from F1, F2
and marketed formulations were 1.897 0.023, 1.7534
0.011 and 1.737 0.020 respectively when saline phosphate
buffer pH 7.4 and methanol (90:10) was used as medium for
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Kamble et al.: Development and Evaluation of Sorbitan Monostearate Organogels as a Topical Delivery System for Aceclofenac

Table 2: Evaluation of sorbiton monostearate Organogels

Formulation

Drug
content* (%)

pH

Viscosity
(centipoise)

Spreadability*
(gcm/sec)

F1
F2
F3
F4
F5
F6
F7
F8
F9

99.89 0.41
98.66 0.40
99.61 1.02
100.2 0.43
99.07 0.82
99.61 1.438
98.07 0.40
99.89 0.41
98.66 0.40

5.16
5.25
5.40
5.53
5.30
5.58
5.31
5.11
5.20

27500
30400
33000
36800
38500
42600
47200
27500
30400

52.70 1.03
50.18 1.05
45.56 0.46
43.07 0.79
39.11 0.52
37.23 0.37
32.19 0.20
52.70 1.03
50.18 1.05

In vitro parameters
Flux
Permeability
2
(mg/cm /hr)
coefficient

0.216
0.201
0.201
0.192
0.174
0.164
0.184
0.216
0.201

0.029
0.027
0.028
0.026
0.023
0.022
0.024
0.029
0.027

*Values are Mean SEM. (n=3)

diffusion study. Further increase in concentration of


organogelators decreases cumulative amount permeated,
these percentages of gelator could be consider as optimum.
The decrease in amount of drug permeated could be attributed
to formation of micelles. At higher concentrations there is
more extensive entanglement of long reverse cylindrical
polymer like micelles with each other, forming a network like
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structure with very high viscosity. The entrapment of the
drug within this network lowers the amount of free drug
available for release. The result revealed that maximum
in vitro permeation of aceclofenac was observed in case of
F1 and F2 formulations (Fig. 2).
The percent inhibition of paw edema of the albino rats was
taken as measure of the anti-inflammatory activity. In vivo
studies in albino rats indicated that all the formulations
showed a significant reduction in paw volume when
compared with control. Formulation F8, F9 and marketed had
showed 86.53 %, 76.42 % and 71.14% edema inhibition after
rd
(Fig 1a)
3 hour of carrageenan challenge respectively when Nonionic
surfactant gel base was used as a control (Table No. 3). The
formulations showed no signs of skin irritations after
continuous application of formulation to guinea pigs for 7
days. Stability study of F8 showed no precipitation, no

CONCLUSION
Findings of this study suggest that sorbiton monostearate
based organogels is able to provide desired anti-inflammatory
action. In vitro permeation study demonstrated that organogel
was effective in providing faster drug release. In vivo study
confirmed the findings of in vitro study. The result indicates
that organogel exhibits useful pharmaceutical properties and
serves as a better vehicle for topical delivery of aceclofenac.
Hence, sorbiton monostearate based organogels seems to be a
promising novel topical delivery system for aceclofenac.
However, the role of the formulation developed in this study
can only be settled with clinical investigations on humans
with emphasis on therapeutic index and side effects followed
by pilot scale studies of manufacturing the product for
commercial use.
ACKNOWLEDGMENT
The Authors are grateful to Suyash Laboratories, Mumbai
(India) for gift sample of aceclofenac.

aggregation, no phase separation and no significant loss of


viscosity in freezethaw and thermal cycling test. The
formulations was also found to be stable at 25C 2C at 75
5% RH and 40C 2C at 75 5% RH with respect to drug
content, pH, release rate. However, slight changes in
rheological characteristics were observed, indicating overall
stability of the formulation (Table No. 4).

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Kamble et al.: Development and Evaluation of Sorbitan Monostearate Organogels as a Topical Delivery System for Aceclofenac

Kamble et al.: Development and Evaluation of Sorbitan Monostearate Organogels as a Topical Delivery System for Aceclofenac

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