FORMULATION AND EVALUATION OF BIFONAZOLE ORGANOGEL AS A NOVEL TOPICAL DRUG DELIVERY SYSTEM

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Shubhrajit Mantry. / International Journal of Pharmacy, 3(1), 2013, 1-8.
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FORMULATION AND EVALUATION OF BIFONAZOLE
ORGANOGEL AS A NOVEL TOPICAL DRUG DELIVERY SYSTEM
1Shubhrajit Mantry, 2Arabinda Patnaik, 3N.Sriram, 4V.Bharath Raju
1,4Kottam Institute of Pharmacy, Mahabub nagar, Andhrapradesh, India- 509125
2Sanskrit College of pharmacy, Rangareddy, Andhrapradesh, India.-500084
3Smt.Sarojini Ramulamma College of Pharmacy, Mahabub nagar, Andhrapradesh, India-509001
INTRODUCTION
Topical delivery can be defined as the application
of a drug containing formulation to the skin to directly treat
cutaneous disorders (e.g. acne) or the cutaneous
manifestations of a general disease (e.g. psoriasis) with the
intent of containing the pharmacological or other effect of
the drug to the surface of the skin or within the skin. Semi-
solid formulation in all their diversity dominate the system
for topical delivery, but foams, spray, medicated powders,
solution, and even medicated adhesive systems are in use.
Topical delivery includes two basic types of
product: i. External topical that are spread, sprayed, or
otherwise dispersed on to cutaneous tissues to cover the
affected area. ii. Internal topical that are applied to the
mucous membrane orally, vaginally or anorectal tissues for
local activity.
Organogels have an organic solvent as the liquid
continuous medium. Organogels are formed by specific
kind of small organic molecules, which in many solvents
very effectively self-assemble into a three dimensional
network of nanoscale dimension, thereby turning a liquid
into a gel. 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 microscopic level. Organogels can be
distinguished from hydrogels by their predominantly
organic continuous phase and can then be further
subdivided based on the nature of the gelling molecule:
polymeric or low molecular weight (LMW)
organogelators. Polymers immobilize the organic solvent
Corresponding Author:- Shubhrajit Mantry Email:- manu28pharmacy@gmail.com
International Journal of Pharmacy
www.ijpjournal.org e-ISSN: 2249 – 7684
Print ISSN: 2249 – 7692
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ABSTRACT
Bifonazole is a novel imidazole antifungal drug which has been used in many skin and nail infections caused by fungi.
The aim of the present study is to improve availability of drug
at the desired site by use of organogels such as pluronic lecithin
and to check the suitability of pluronic lecithin organogels containing Bifonazole for topical application. Eight formulations
were developed using Bifonazole, lecithin, Pluronic F127, isopropyl myristate, water, sorbic acid and potassium sorbate were
coded as FL1 to FL8.
For the preparation of FL1, FL2, FL3 and FL4, 20 % oil phase and 80 % aqueous phase were used while
for FL5, FL6, FL7 and FL8, 30 % oil phase and 70 % aqueous phase were used.
The formulated organogels were evaluated for
appearance by psychorheologically, in vitro diffusion study, drug content, viscosity and pH.
It was found that the pH of all the
formulations is in the range of to 5.29 -6.58 that suits the skin pH, indicating skin compatability. This is the primary
requirement for a good topical formulation.
This fact reflects higher spreadability of pluronic organogels as compared to
sorbitan Monostearate organogels. All formulation showed spreadability in the range of 12.79- 52.15 g.cm/sec.
Key words: Pluronic lecithin organogel, Bifonazole, Topical delivery.
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by forming a network of either cross-linked or entangled
chains for chemical and physical gels, respectively. The
latter is possibly further stabilized by weak inter-chain
interactions such as hydrogen bonding, van der Waals
forces, and π-stacking [1,2].
Advantages of organogels: i. Organogels are
thermoreversible and offer improved microbial resistance
as compared to aqueous solutions of hydrogels. ii. Also the
network structure formed in organogels is highly ordered at
the supramolecular level iii. Improved drug permeability
across the skin iv. Ease of preparation and administration
[3].
Organogels in drug delivery
Despite the large abundance and variety of
organogel systems, relatively few have current applications
in drug delivery, owing mostly to the lack of information
on the biocompatibility and toxicity of organogelator
molecules and their degradation products. Bifonazole is a
novel imidazole antifungal drug which has been used in
many skin and nail infections caused by fungi. Although,
topical formulation of the drug is available but, to
investigate the potential of organogel as a novel topical
drug delivery system, the above drug candidate was
selected. The objective of the present study is Development
of a novel topical vehicle for the delivery of an antifungal
agent in terms of effectiveness and elegance as compared
to conventional ointment and creams, improved
availability at the desired site by use of organogels and
Improved patient compliance [4,5].
Lecithin Organogels
The most investigated Organogels for topical
delivery of active agents are Lecithin organogels. Lecithin
organogels in the biocompatible organic solvents such as
high purity hydrocarbon oils may be used in dermatology
and cosmetic as a base of topical compositions. These gels
have the following advantages:
 Their micellar structure can contain both water-soluble
and oil-soluble ingredients. It is known that lecithin gels in
vitro accelerate transdermal delivery of drugs.
 The lecithin gels in vaseline oil soften a human skin,
they are quickly absorbed by the skin without greasy shine.
It was shown the lecithin organogels don't irritate the skin.
 The lecithin gels are thermodynamically stable
systems, so they can be kept in a closed vessel for a very
long time
 It is easy to produce a large amount of the lecithin
gels. The technological process doesn't require a
complicated device and lecithin is an available commercial
product [6-8].
METHOD OF PREPARATION OF ORGANOGELS
Pluronic Lecithin Organogels
For the preparation of FL1, FL2, FL3 and FL4, 20
% oil phase and 80 % aqueous phase were used while for
FL5, FL6, FL7 and FL8, 30 % oil phase and 70 % aqueous
phase were used.
Oil phase: Weighed amount of lecithin soya, sorbic acid,
vitamin E were placed in a beaker and isopropyl myristate
was added upto the mark. The mixture was kept overnight
in order to dissolve lecithin.
Aqueous phase: Weighed amount of Pluronic F-127 and
potassium sorbate were taken and cold water was added
upto the mark and shaken. The mixture was stored in
refrigerator overnight for dissolution of Pluronic F-127.
Drug was incorporated with stirring in oil phase.
Finally, aqueous phase was slowly added in oil phase .This
process was carried out in tissue homogenizer [10,11].
The compositions of different formulations are
summarized in Table 1
EXPERIMENTAL AND RESULTS
Melting Point
The melting point of bifonazole was determined
by capillary method. The melting point was found to be in
the range of 142-145°C. The reported melting point is in
the range of 142-143°C.
Identification tests by
a) Ultra violet spectrophotometry
Bifonazole (200 mg) was taken in a 100 ml volumetric
flask and volume was made up with methanol. One ml of
the solution was pipetted out and diluted to 100 ml with
methanol. Examined between 200-400nm.The solution
showed maximum absorption at 254.4nm [12].
b)
c) Infrared absorption spectrophotometry:
A FT-IR spectrum of drug sample was taken
using kBr pellet between 4000 to 500 cm-1. Spectra are as
shown in Fig 4 and interpretations are shown in Table 2.
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
[13]. Results are shown in Table 5.
Drug content
a) Standard curve of Bifonazole in methanol
Bifonazole (200 mg) was taken in a 100 ml volumetric
flask and volume was made up with methanol. One ml of
the solution was pipetted out and diluted to 100 ml with
methanol. Then aliquots were further diluted with
methanol to get concentration of 2, 4, 6, 8, 10, 12, 14, 16,
18 and 20 μg/ml. Absorbance were recorded
spectrophotometrically at λmax 254.4nm. Absorbance

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values are shown in Table 4 and standard curve is shown in
Fig.5.
Table 4: Absorbance values of Bifonazole in Methanol
at 254.4 nm
b) Procedure
Each formulation (0.5g) was taken in a 50 ml volumetric
flask, diluted with methanol and shaken to dissolve the
drug in methanol. The solution was filtered through
Whatman filter paper no. 42, one ml of the above filtrate
was pipetted out and diluted to 10 ml with ethanol. The
content of the drug was estimated spectrophotometrically
by using standard curve plotted at λmax 254.4 nm. Results
are shown in Table 5.
pH Determination
The pH of formulated organogels was determined
using pH meter. 2 g of gel was stirred in distilled water to
get an uniform suspension.Volume was made upto 40 ml.
The electrode was immersed in organogel - distilled water
suspensions and readings were recorded on pH meter.
Results are shown in Table 6.
Viscosity
Viscosities of the formulated organogels were
determined using Brookfield Viscometer. Spindle no. 6
and Spindle no. 7 were used for nonionic surfactant
organogels and Pluronic Lecithin organogels respectively
and the spindle speed of 2 rpm at 25° C was used. Results
are shown in Table 7.
Spreadability
Spreadability of formulations was determined by
an apparatus suggested by Multimer et al. which was
fabricated in laboratory and used for study. The apparatus
consists of a wooden block, with a fixed glass slide and
movable glass slide with one end tied to weight pan rolled
on the pulley, which was in horizontal level with fixed
slide. Apparatus is shown in Fig.8.
Procedure
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 [14].
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
t - Time in seconds
Length of glass slide was 11.3 cm and weight tied to upper
slide was (60g) throughout the experiment. Results are
shown in Table 8.
In vitro antifungal activity
The in vitro antifungal activity of bifonazole from
the selected formulations FL2 was carried out using
Candida albicans as a representative fungi, adopting the
cup – plate method. Commercial clotrimazole ointment
was taken as a reference standard [15]. Clotrimazole is a
well-known effective antifungal drug and it is available as
a topical formulation. Suspension of Candida albicans was
inoculated in sabouraud dextrose agar medium and then
poured into the sterile petridish, and allowed to solidify.
Wells were done in plate using borer and the formulations
were poured into wells. These plates were incubated at 37º
C for 24 hours. The standard and test were tested at a
concentration of 0.1mg/ml. The mean zone of inhibition
was calculated using Hiantibiotic Zone scale for each plate,
and this value was taken as an indicator for the antifungal
activity.The zones of inhibition are shown in the figures 6.
FTIR Study of Organogels
FT-IR spectrum of prepared organogels were
taken using kBr pellet between 4000 to 500 cm-1. Spectra
is as shown in Fig 7.
Fig 1. Classification of Organogels

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Preparation of organogels
Fig 2. Schematic representation of formation of Lecithin organogel
Fig 3. Spectrum showing absorbance maxima of Bifonazole in methanol
Fig 4. FT-IR spectrum of bifonazole sample
5007501000125015001750200022502500275030003250350037504000 1/cm
20
40
60
80
100
120
140
160
%T
bifonazole

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Fig 5. Standard curve of Bifonazole in Methanol at 254.4 nm
Fig 6. The zones of inhibition shown by the gels and standard
Control
FL2
Clotrimazole
Fig 7. FTIR study of organogels
Fig 8. Spreadability Test Apparatus
y = 0.029x
R² = 0.996
Concentration (µg/ml)
Absor
bance
5007501000125015001750200022502500275030003250350037504000 1/cm
0
5
10
15
20
25
30
35
%T
FS4

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Table 1. Pluronic Lecithin Organogels
Components
Content
Formulations
FL1
FL2
FL3
FL4
FL5
FL6
FL7
FL8
Drug
Bifonazole (%)
1
1
1
1
1
1
1
1
Oil phase
Lecithin soya (%)
3
5
7
10
3
5
7
10
Sorbic acid (%)
0.2
0.2
0.2
0.2
0.2
0.2
0.2
0.2
Vitamin E (%)
0.1
0.1
0.1
0.1
0.1
0.1
0.1
0.1
Isopropyl myristate upto(%)
100
100
100
100
100
100
100
100
Aqueous phase
Pluronic F-127 (%)
20
20
20
20
20
20
20
20
Potassium sorbate (%)
0.2
0.2
0.2
0.2
0.2
0.2
0.2
0.2
Purified water (ml)
100
100
100
100
100
100
100
100
Table 2. Details of FT-IR spectrum of Bifonazole
Table 3. Psychorheological characterization of Organogels
Sr. No.
Parameters
ORGNOGELS
Pluronic Lecithin Organogels
1.
Colour
Slight yellowish
2.
Clogging
-
3.
Sudden Viscosity change
No change
4.
Feel
Smooth
Table 4. Absorbance values of Bifonazole in Methanol at
254.4 nm
Sl. No.
Concentration (μg/ml)
Absorbance*
1.
0
0.000
2.
2
0.053±0.002
3.
4
0.101±0.006
4.
6
0.163±0.008
5.
8
0.228±0.012
6.
10
0.276±0.023
7.
12
0.360±0.007
8.
14
0.409±0.009
9.
16
0.479±0.014
10.
18
0.508±0.017
11.
20
0.588±0.011
Table 5. Drug content of Organogels
Sl.no
Formulation
% Drug
Content
1
FL1
98.96±0.014
2
FL2
99.65±0.020
3
FL3
98.27±0.017
4
FL4
99.31±0.023
5
FL5
97.93±0.032
6
FL6
98.62±0.019
7
FL7
97.58±0.013
8
FL8
100.34±0.025
Table 6. pH of Organogels
Sl. No.
Formulation
pH*
1.
FL1
6.30
2.
FL2
6.12
3.
FL3
5.81
4.
FL4
5.98
5.
FL5
5.30
6.
FL6
5.29
7.
FL7
5.74
8.
FL8
5.48
9.
Marketed
5.78
Table 7. Viscosities of Organogels at 25°C
Sl. No.
Formulations
Viscosity (centipoise)
1.
FL1
96500
2.
FL2
99400
3.
FL3
103400
4.
FL4
107500
5.
FL5
138300
6.
FL6
159200
7.
FL7
165000
8.
FL8
188000
9.
Marketed
105000
SR. NO
FREQUENCY (CM-1)
ASSIGNMENTS
1
3025
Aromatic C-H stretching
2
2950 & 2830
Aliphatic C-H stretching
3
1487.01
Aromatic C-C stretching
4
1452.30
C-H deformation
5
1220.86
Possibility of ethereal linkage

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Table 8. Spreadability of Organogels
Sl. No.
Formulations
Spreadability
1.
FL1
25.11±0.043
2.
FL2
23.37±0.051
3.
FL3
22.59±0.024
4.
FL4
19.94±0.028
5.
FL5
16.95±0.019
6.
FL6
15.54±0.026
7.
FL7
13.83±0.006
8.
FL8
12.79±0.062
16.
Marketed
21.18±0.005
Table 9. Antimicrobial activity of gels in comparison to reference standard using Candida albicans
Zone of inhibition (mm)
Formulation
1
2
3
4
Mean
FL2
17
22
23
24
21.5
Clotrimazole
15
18
19
20
18
DISCUSSION
The procured sample of Bifonazole was tested for
its identification. The quality of Bifonazole was confirmed
by melting point and UV-absorption maxima in alcohol.
The results of these entire tests were in compliance with
official specification. The IR- spectrum obtained with
Bifonazole confirmed its quality. Thus, the Bifonazole
used in the entire study was of pure quality. The
Organogelators used in study were first confirmed for their
purity by their identification tests official in
pharmacopoeia [16]. Organogelators showed compliance
with official specifications. The different formulations
using various Organogelators of Lecithin were developed.
These organogelators were selected based on their use in
topical formulation for other drugs by various scientists
across the world. Lecithin has become recognized as a
possible permeation enhancer in the transdermal delivery
of drugs but to produce proper viscosity Pluronic F-127
was used in combination. Sorbitan monostearate, a
hydrophobic nonionic surfactant form gel with isopropyl
myristate and the gel stability was improved with the
addition of Tween 20.
Various organogels of Pluronic Lecithin
combination organogel incorporating water were
formulated. All the formulations were evaluated for their
Psycho-rheological characteristic, Drug content, pH,
Viscosity, Spreadability. It was found that, Pluronic
Lecithin organogels were smooth and showed no clogging
which indicate good texture of formulation. All the
formulations were evaluated for the drug content, the
results were found in the acceptable range, indicating no
interaction of drug with component of base. The data is
shown in It was found that the pH of all the formulations is
in the range of to 5.29 -6.58 that suits the skin pH,
indicating skin compatibility [17]. This is the primary
requirement for a good topical formulation. This fact
reflects higher spreadability of Sorbitan Monosterate
organogels as compared to Pluronic Lecithin organogels.
All formulation showed spreadability in the range of
12.79- 52.15 g.cm/sec. Standard calibration curves for
Bifonazole were prepared in methanol and in combination
of saline phosphate buffer pH 7.4 and sodium lauryl sulfate
(2%) as Bifonazole is practically insoluble in water. The
equation of line was found to be y = 0.029x (R2 = 0.996)
for calibration curve in methanol and y = 0.024x (R2 =
0.998) for calibration curve in saline phosphate buffer pH
7.4 and SLS. Correlation coefficient (R2) values indicate
the linear correlation between concentration and
absorbance.
CONCLUSION
From the entire discussion it was concluded that
FL2 is the best formulation. It showed higher cumulative
amount of drug permeation, higher anti-fungal activity as
compared to the marketed. Therefore, FL2 could be used
as an effective formulation for topical delivery of
Bifonazole as compared to marketed.
ACKNOWLEDGMENT
The authors are thankful to COMPRIME Lab Pvt.
Ltd. Hydernagar, Hyderabad, A.P. for completion of this
work.
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