Journal of Scientific & Industrial Research

Vol. 74, February 2015, pp. 88-92

Nanoemulsion based Hydrogels of Itraconazole for Transdermal Drug Delivery

S Sampathi1*, S K Mankala2, J Wankar1 and S Dodoala1
*1

Department of Pharmaceutics, National Institute of Pharmaceutical Education and Research (NIPER-HYD),

Balanagar, Hyderabad, A.P. - 500037, India
2
Department of Pharmaceutics, Sri Krupa Institute of Pharmaceutical Sciences, Vill: Velkatta, Siddipet. District: Medak,
A.P. - 502103, India
Received 29 January 2013; revised 20 January 2014; accepted 31 October 2014
The present study aimed to formulate a nanoemulsion and nanoemulsion based hydrogel of itraconazole for transdermal
delivery in the treatment of on chomycosis. The nanoemulsions were prepared using lecithin and sodium cholate as
surfactant and co-surfactant. The prepared nanoemulsions were characterized for particle size and zeta potential.
The optimized nanoemulsion was incorporated into 3% carbopol-934 solution to get a gel for improving convenience in
superficial application. In vitro and ex vivo drug penetration studies of nanoemulsions and gels were determined using
dialysis membrane and rat skin. The particle size was found around 223.9 nm to 154.3 nm. The viscosity of the
nanoemulsions and nanoemulsion gel were found around 1964.89 mPa.S to 1644.82 mPa.S and 28.3 mPa.S to 8.58 mPa.S
respectively. The polydispersibility value was found very low indicating uniformity of droplet size of the formulations.
The drug content in gels was found in between 86.2% to 98.26%. The drug release was found to be 44.33 % to 73.6% after
24 h with permeation flux around 296.3 to 203.1 (g/cm2/hr1). The results indicated that nanoemulsion based hydrogels as
promising vehicle for transdermal delivery of itraconazole. Further in vivo studies are to be performed to access
its suitability for topical application.
Keywords: Itraconazole, nanoemulsion, nanoemulsion based hydrogel, lecithin, sodium cholate, carbopol.

Introduction
Recently research has been focused on the colloidal
drug delivery systems such as micro emulsions,
solid lipid nano-particles and liposomes for topical
delivery of drugs because of low side effects
high bioavailability, good patient compliances1,2.
Nanoemulsions are non-equilibrium heterogeneous
systems consisting of two immiscible liquids in which
one liquid is dispersed as droplets in another liquid
with droplet diameter in the range of 10-100 nm. Due
to their unique physicochemical properties NE offer
many advantages over traditional topical and
transdermal drug delivery formulations3-7. Previously
attempts have been made in the development of ITZ
loaded nano-particles for parenteral application8. This
article is intended to demonstrate the feasibility of
new nanoemulsion system for transdermal delivery of
anti-fungal drug itraconazole (ITZ).
Methods
Materials

Itraconazole was obtained as gift sample from Dr.

Reddys Labs., Pvt. Ltd. (Hyderabad, India). Lecithin

*Author for correspondence

E-mail: sunithasampathiphd@gmail.com

and sodium cholate were purchased from Sigma

Aldrich, eugenol was purchased from Loba
chemicals. Pvt. Ltd. (Mumbai, India) and all other
reagents and solvents used were of analytical grades.
Analytical method development for itraconazole

A spectrophotometric method was developed for

analysis of itraconazole. Briefly, the stock solution
was prepared by dissolving 100 mg of itraconazole in
100 mL of saline phosphate buffer with 2% SLS and
subsequent dilutions were made to get concentrations
of 5 - 25 g/mL. The absorbance was measured using
double beam Jasco UV spectrophotometer at 263 nm.
Preparation of Nanoemulsion (Ultrasonication method)

The aqueous phase was prepared by dissolving

weighed amount of lecithin and sodium cholate
in 4.5 mL of deionized distilled water (Table 1).
Itraconazole (50 mg) was accurately weighed and
dissolved in 0.5 mL of eugenol, heated to 75-80C for
2-3 min. The aqueous phase was added slowly
to the oil phase and mixed using electronic stirrer at
15000 rpm for a period of 5 min. The droplet size
in the course emulsion was further reduced by
ultrasonication at 30% and 50% amplitude duty cycle
using ultrasound instrument (Vibra-Cell VC 505,

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SAMPATHI et al.: NANOEMULSION BASED HYDROGELS FOR DRUG DELIVERY

Sonic Instruments, Newtown, CT) for 15 min and

stored at 4C for further studies.
Preparation of nanoemulsion based hydrogel (NBH)

The best nanoemulsion formulation was

incorporated into 3% of carbopol 934 to get a gel of
the nanoemulsion. Weighed quantity of the carbopol
934 was dissolved in 15 mL of distilled water and
stirred thoroughly to get homogenous slurry. The best
and stable nanoemulsion was incorporated and mixed
thoroughly and the pH was adjusted to neutral with
triethanolamine. Limonene 0.1% was added as the
permeation enhancer and as odor masking agent in the
formulations.

for the measurement of droplet size. The average

diameter and polydispersity index of samples were
measured by photon correlation spectroscopy
(Nano ZS90, Malvern Instruments, U.K) at 633 nm.
The measurement was performed at 25 C using a
He-Ne laser.
Drug content

The drug content of the nanoemulsion based gel

was determined by dissolving an amount of gel
containing 5mg of drug. The drug content was
determined after appropriate dilutions at 263 nm by
UV Spectrophotometer.
Viscosity determination

The viscosity of the formulations was determined

by using a Brookfield viscometer using spindle #S16
at 25 0.3C.

Evaluation
Characterization of nanoemulsions
Photon correlation spectroscopy

The mean particle size and the size distribution

were determined by photon correlation spectroscopy.
The prepared nanoemulsions were 40 times diluted

Zeta potential

The surface charge was analyzed using a Zeta-sizer

at 25C by measuring the zeta potential of the
prepared formulation. The formulations were diluted
with distilled water and then the zeta potential was
measured.
Scanning electron microscopy (SEM)

Morphological characterization of the nanoemulsion

was carried using scanning electron microscopy under
the reduced pressure (0.001torr). The nanoemulsion
were viewed at an accelerating voltage of 15-20kv.
Thermodynamic stability study

Fig. 1Globule size distribution curve of ITZ nanoemulsion F4

The optimized NE was centrifuged at 4000 rpm

for 30 min and observed for phase separation,
creaming, and cracking. Then it was subjected to
heating and cooling cycle. Six cycles between the
refrigerator temperature (4C) and 45C temperature
were performed with storage at each temperature

Table 1Composition, Particle size, Polydispersity index and Zeta potential of the ITZ nanoemulsion formulations
Formulation

Lecithin
(mg)

Sodium cholate
(mg)

Particle size
(nm)

Polydispersity index
(PDI)

Zeta potential

Viscosity

F1
F2
F3
F4
F5
F6
F7
F8
F9

21.03
40.98
60.11
80.56
101.05
121.59
140.53
161.04
180.92

20.55
21.08
21.53
20.44
21.23
21.83
21.03
20.40
20.64

199.2
193.3
181.6
178.7
164.3
223.9
159.7
154.3
163.6

0.0091
0.082
0.079
0.134
0.136
0.252
0.145
0.203
0.192

-20.6
-19.9
-19.4
-18.4
-17.5
-14.9
-18.0
-18.0
-17.7

73.5
34
46.8
35.1
102
113.2
109.5
122.7
117.6

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J SCI IND RES VOL 74 FEBRUARY 2015

for not less than 48 h. Finally it was subjected

to freeze thaw cycle. Formulations were exposed for
three freeze thaw cycles between -21C and +25C
with storage at each temperature for not less than
48 h to check the thermodynamic stability of the
formulations.
In vitro diffusion studies using dialysis membrane

The in vitro study was performed by using Franz

diffusion cells with an effective diffusion area of
2cm2. The dialysis membrane (12,000 MW) was
soaked for 12 h and clamped between the donor and
receptor compartment of the cell. The nanoemulsion
and the NBH containing itraconazole were placed in
the donor compartment. The receptor compartment
was filled with saline phosphate buffer with 2% SLS
as the medium and maintained at 37 0.5 C and
stirred at 600 rpm and samples were withdrawn at
regular intervals for 24 h and analyzed by UVSpectrophotometer. The experiment was conducted in
triplicates.
Ex vivo studies

The best nanoemulsion was taken and studied for

the ex vivo release studies by using albino rat skin as
the membrane. Ex vivo skin permeation study was
performed by using Franz diffusion cells as
mentioned earlier. Adult Wistar rats of 180 -200 g
were depilated with trimmer and skin samples were
excised and were clamped between the donor and the
receptor chamber of Franz diffusion cells with the
stratum corneum facing the donor chamber. The
nanoemulsion and the NBH containing itraconazole
were placed in the donor compartment. The study was
performed in the similar way as invitro diffusion
study for 24h and analyzed the samples by UV
spectrophotometer.
Steady state flux (Jss/Cm/h) and lag time (T-lag/h)
were calculated from the slope and intercept of
the straight line obtained by plotting the cumulative
amount of itraconazole permeated versus time in
steady condition. Permeability Coefficient (Kp.Cm/h)
was calculated by dividing the flux obtained
by the initial concentration of drug in the donor
compartment9,10.
Kinetic analysis of drug release

To analyze the mechanism of drug release from

itraconazole nanoemulsion and nanoemulsion based
gel the in vitro dissolution data were fitted to zero
order, first order, Higuchi release model, and

Korsemeyer and Peppas model and the model with

higher correlation coefficient was considered to be the
best model.
Skin irritation test

The test was performed on male albino rats

weighing around 180 - 200 g and all the procedures
were approved by institutional animal ethical
committee
[Ethical
Committee
Reg.
No:
1548/PO/a/11/CPCSEA]. The animals were divided
into 4 groups each group consists of 4 rats:
Group A served as control, Group B received 0.5ml of
0.8% V/V aqueous formalin solution as a standard
irritant. Group C and D received ITZ-NE and
ITZ-NBH prepared in plain carbopol gel. Standard
irritant was applied on the left dorsal surface of
each rat and formulations were applied on the right
dorsal surface of the albino rat. The formulation
was removed after period of 24 h with the help
of an alcohol swab. The skin was examined for
erythema or edema and primary irritancy index (PII)
was scored as per Draize et al.,11.The results
obtained were statistically compared using
students T- test and statistical significance was set at
p < 0.05.
Results and discussion
Itraconazole
nanoemulsions
with
different
concentrations of lecithin and sodium cholate as
surfactant and co-surfactant were formulated. The
mean particle size of the nanoemulsion formulation
of ITZ was found to be 154.3 - 223.9 nm.
The polydispersity index was found to range from
0.0091 to 0.203 which indicates uniformity of droplet
size within the formulations with zeta potential values
between -14.9 to -20.6 respectively (Table 1).
The small mean particle size and PdI values
below 0.2 (Table 1), indicated a narrow droplet size
distribution and thus assured better stability of the
formulation. The viscosity of all nanoemulsion was
ranging from 34 to 122.7 mPa.S. The optimized
nanoemulsion was incorporated 3 % carbapol 934 as
the gelling agent to obtain the desired NBH and
evaluated for drug content, which showed uniformity
of 5 mg in all the prepared NBH formulations, hence
the data has not been included.In vitro drug
release studies from all the 9 formulations of
nanoemulsions were determined and higher release
was observed with the formulation F5 73.6% and
lowest with formulation F4 i.e. around 44.33%.

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The significant difference in the release between

formulations was probably due to the mean size of
internal phase droplets. Hence, the maximum drug
release found with F5 and F8 formulations having
least size of internal phase droplets (Figure 2).
Fitting of the release data into first order, Higuchis,
and Peppas equations were done for formulations
to known the mechanism of drug release and
almost all formulations were following first order
release kinetics by fickian transport (Table 2).
In order to evaluate the skin targeting potential of the

NE and NBH, the skin permeation and penetration

ability of ITZ cross the rat skin were examined
ex vivo. Flux and permeability coefficients for
optimized formulations are given in Table 3 and
cumulative drug permeated in Figure 3. Compared
to ITZ solution, the skin permeation and
penetration ability were more pronounced with ITZ
NE and ITZ NBH. The nanoemulsions can interact
with the lipid bilayers of the stratum corneum
and thereby contribute significantly to the penetration
enhancing effect which was the reason for enhanced
flux and permeation coefficient with nanoemulsions
as compared with the gels as the consistency of
the gels inhibit the drug permeation. The skin
irritation studies shows that the ITZ-NE and
ITZ-NBH to be non-irritant with the PII of
ITZ-NE 1.25 0.4 and1.8 0.38 for ITZ-NBH
(p > 0.05) when compared to the control group
treated with formalin solution with a PII of 6.86
0.89 (p < 0.05). The innovated formulations were
safe to be applied to the skin for the intended period
of time.
Table 3Flux values and permeability coefficient values of the
formulations
Sl. No Formulation code

Fig. 2In vitro drug release profiles of itraconazole nanoemulsion

formulations F1-F9

1
2
3
4

F8 NE
F8 NE Gel
F5 NE
Flux of F5 Gel

Flux
(g/cm2/hr1).

KP
(Cm.hr-1)

296.3
203.1
296.5
211.1

29.63
20.31
29.65
21.11

Table 2Curve fitting data for all formulations of itraconazole nanoemulsions using lecithin as surfactant
Formulation

ZERO ORDER
2

0.959
0.923
0.913
0.965
0.959
0.943
0.855
0.960
0.974
0.982
0.974
0.919
0.835

r
F1
F2
F3
F4
F5
F6
F7
F8
F9
F5 (ex vivo)
F8 (ex vivo)
F5 Gel (ex vivo)
F8 Gel(ex vivo)

FIRST ORDER

9.050
6.770
8.586
4.617
9.050
7.550
7.505
6.018
3.734
4.776
10.51
13.95
16.25

0.976
0.981
0.986
0.991
0.995
0.995
0.917
0.996
0.998
0.987
0.996
0.880
0.955

HIGUCHI

2.002
1.973
1.970
1.988
1.975
1.984
1.973
1.993
1.990
2.040
1.995
1.881
1.911

0.985
0.994
0.994
0.981
0.972
0.992
0.944
0.981
0.984
0.928
0.902
0.93
0.954

PEPPAS
K

-10.63
-6.780
-7.655
-7.548
-10.08
-7.923
-9.689
-12.76
-9.510
-7.532
3.604
10.24
8.869

0.611
0.840
0.823
0.776
0.682
0.787
0.834
0.735
0.772
0.688
0.732
0.888
0.727

2.029
2.017
2.026
2.018
2.054
2.025
2.034
2.053
2.027
2.240
2.222
1.971
1.991

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frequency of administration and improves the patient

compliance.
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Fig. 3Ex vivo drug release of ITZ nanoemulsions and hydrogel

based nanoemulsions of F8 and F5 formulations

Conclusion
The results obtained in the present work show
that NE based hydrogel containing eugenol,
lecithin and sodium chlolate as a suitable carrier
system for incorporation of itraconazole and
satisfies the best attributes for transdermal application
with good particle size in the nano range with
negative zeta potential, viscosity, good spread
ability, and with suitable release profile. Prepared
nanoemulsion based gel formulations are highly
stable and safe for the transdermal delivery.
The developed system could able to release the drug
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