Latin American Journal of Pharmacy Regular article

(formerly Acta Farmacéutica Bonaerense) Received: September 18, 2014

Revised version: March 30, 2015
Lat. Am. J. Pharm. 34 (5): 1022-9 (2015) Accepted: March 31, 2015

Transdermal Diclofenac Potassium Gels:

Natural Penetration Enhancers, Can Be Effective?
Abid HUSSAIN 1, Gul M. KHAN 2, Nauman R. KHAN 4 *, Amjad KHAN 3, Saif ur REHMAN 1,
Hafiz M. ASIF 5, Muhammad AKRAM 5, & Yusuf ALI 6 6
1Department of Pharmacy, Poonch University Rawalakot, AJ&K, Pakistan.
2 Department of Pharmaceutics & 3 Department of Biotechnology,

Quaid-e-Azam University Islamabad, Pakistan.

4 Department of Pharmaceutics, Faculty of Pharmacy Gomal University D.I.Khan, Pakistan.
5 Department of Eastern Medicine and Surgery, Faculty of medical and health sciences,

Poonch University Rawalakot, AJ&K, Pakistan.

6 Faculty Medical and Health Sciences, Poonch University, Rawalakot, AJ&K, Pakistan.

SUMMARY. The aim of the current study was to develop the novel Diclofenac Potassium (DP) gel formu-
lations to show sustained release of the model drug used. Turpentine oil and olive oil were used as penetra-
tion enhancers to investigate the permeation enhancing capability of DP. 1.5% w/v diclofenac potassium
gels were developed with carboxypolymethylene with and without penetration enhancer. Enhancers were
incorporated in increasing concentration, i.e. 1, 2, 3, 4, 5, and 10% of the total gel formulation. Gels were
evaluated on physical basis for pH, viscosity, spreadability, extrudability, smoothness and appearance. To
study the in vitro and ex vivo permeation potential of formulated DP gels, Franz diffusion cell was used, us-
ing silicon membrane and excised rabbit abdominal skin. Flux, permeability coefficient (Kp), lag time and
enhancement ratios of DP were measured over 24 h and compared with control formulation. Diclofenac
Potassium gel with 10% turpentine oil showed maximum flux 159.65 μg/cm2/h while 10% olive oil showed
flux of 147.33 μg/cm2/h through rabbit skin.
RESUMEN. El objetivo del presente estudio fue desarrollar formulaciones novedosas de gel de diclofenac potási-
co (DP) para lograr una liberación sostenida del fármaco modelo utilizado. Se utilizaron aceite de trementina y
aceite de oliva como potenciadores de la penetración para investigar la capacidad de mejorar la permeación de ge-
les de DP. Los geles de DP al 1.5% w/v se desarrollaron con carboxipolimetileno con y sin potenciador de la pe-
netración. Los potenciadores se incorporaron en concentración creciente,es decir, 1, 2, 3, 4, 5 y 10% de la formu-
lación total de gel. Los geles fueron evaluados para pH, viscosidad, extensibilidad, capacidad de extrusión, suavi-
dad y apariencia. Para estudiar el potencial de permeación de geles DP in vitro y ex vivo, se utilizó la célula de di-
fusión de Franz, usando membrana siliconada y piel abdominal de conejo. El flujo, el coeficiente de permeabili-
dad (Kp) y las relaciones de tiempo de DP se midieron durante 24 h y se compararon con la formulación control.
El gel de diclofenac de potasio con un 10% de aceite de trementina mostró un flujo máximo 159,65 g/cm2/h,
mientras que el que contiene 10% de aceite de oliva mostró un flujo de 147,33 g/cm2/h en piel de conejo.

INTRODUCTION a low molecular weight (334.24 daltons) with a

Diclofenac is well known non-steroidal anti-
inflammatory drug (NSAID). NSAIDs produce
their therapeutic activities by inhibition of cy-
clooxygenase (COX); this enzyme is responsible
for the synthesis of prostaglandins (PGs) 1 .
However, they have some undesirable systemic
side effect and gastrointestinal irritation at the
usual dose of oral administration 2. Since the
NSAIDs are used for prolonged period, it is nec-
essary to overcome these problems by any pos-
sible mean. Thus bypass gastric irritation and al-
so reduces adverse systemic effects. DP having
short half-life makes it a suitable candidate for
transdermal drug delivery system (TDDS).
TDDS bears number of advantages over tra-
ditional drug delivery systems. All these advan-
tages have been quoted by number of authors
and included avoidance of first pass
metabolism, decrease in side effects (e.g. gastric
irritation), improved patient compliance, and
enhanced therapeutic efficacy 3. Recently, devel-
opment of TDDS has been focused on the for-
mulation that can achieve the desirable constant
rate of drug penetration into systemic circula-

KEY WORDS: Diclofenac potassium, Turpentine oil, Olive oil and Transdermal.
* Author to whom correspondence should be addressed. E-mail: naumanpharma@gmail.com

ISSN 0326 2383 (printed ed.)

1022 ISSN 2362-3853 (on line ed.)

tion. Many approaches have been suggested us-
ing these polymers and enhancers to overcome
to deliver the drug transdermally and overcome
the main barrier stratum corneum 4.
Chemical penetration enhancers modify bar-
rier properties of the stratum corneum and
hence increase drug permeability across skin.
Ideally, the effects of the penetration enhancer
on the skin should be reversible, nontoxic, non-
allergic, compatible with drugs and excipients
and nonirritating. Many enhancers possess these
properties e.g. azone and its analogues 5 fatty
acids, alcohol and pyrrolidones 6. Due to the
systemic and localised toxicity of these penetra-
tion enhancers, many of them have not been
explored so far. Hence, natural products have
increasingly been used as enhancers due to
their better safety profile. The use of the pene-
tration enhancer in the formulation is to im-
prove the permeability of the drugs and drug
flux through the epithelium. Various classes of
the penetration enhancers have been studied to
check their effectiveness such as surfactants,
bile salts, the glycols and there derivatives, natu-
ral oils terpenes, fatty acids and chelators 7. Ter-
penes are popular permeation enhancer in
transdermal drug delivery 8. This category in-
cludes a heterogeneous range of members and
the effect of a specific terpene on skin depends
upon its exact physiochemical properties, in
particular its lipophilicity. They have been
found effective penetration enhancers for a
number of hydrophilic and lipophilic drugs 9.
Terpenes are considered as “Generally Recog-
nized as Safe” (GRAS) and have been extensive-
ly used to enhance the permeation of several
drugs including ketoprofen 10, hydrocortisone 11,
imipramine hydrochloride 12, estradiol 13 and
haloperidol 14.
Carbopol is best gelling agent having desir-
able properties with high viscosity at low con-
centration is quite stable to heat with negligible
batch to batch variability. It increases the stabili-
ty of the formulation and does not support bac-
terial and fungal growth. Formulations obtained
by carbopol have a pleasant texture and is not
irritating 15.
MATERIALS AND METHODS
Diclofenac potassium (Shanghai Mepherkang
International China), carboxypolymethylene
(Zhong Tang Dalian Materials Co. Ltd, China)
and triethanolamine (Siga Aldrich) were re-
ceived as gift samples from Global Pharma Is-
Latin American Journal of Pharmacy - 34 (5) - 2015
lamabad Pakistan, the enhancer used were tur-
pentine oil and olive oil (Siga Aldrich Germany),
ethanol (96%), potassium dihydrogen phosphate
and sodium hydroxide (Merk Germany). All
chemicals were used without further purifica-
tion.
DP (1.5% w/v) gel formulations
The required amount (1 g) of gelling poly-
mer (CPM) was accurately weighed and dis-
persed slowly in 50 mL of distilled water on
magnetic stirrer at 400-600 rpm for 1 h. Weighed
amount of DP (1 g) was dissolved in 20 mL of
ethanol. The ethanolic drug solution was slowly
added into previously prepared CPM suspension
with continues stirring at 400-600 rpm till ho-
mogenous dispersion is achieved. Turpentine oil
and Olive oil was incorporated in increasing
concentration. One mL of Triethanolamine was
added to adjust the pH of carbopol gels. The fi-
nal quantity was made up to 100 g with distilled
water and 1 mL Triethanolamine. The formula-
tion is given in Table 1.
Evaluation of formulated gels
The formulated DP gels were evaluated
physically for pH, viscosity, spreadability, ex-
trudability, smoothness and for appearance. The
pH of the various hydrogel formulations was
determined, using digital pH meter (Denver,
USA). It was calibrated before each use with
buffered solutions at pH 4, and 7) by dipping
the pH meter electrode in gel samples. The de-
termination was carried out in triplicate and av-
erage of the three readings was recorded.
Spreadability was determined by wooden block
and glass slide apparatus. This apparatus con-
sists of a wooden block with fixed glass slide
and a pulley. About 20 g of hydrogel was
placed in the fixed glass slide, another glass
slide (movable) with a pan attached to it, was
placed over the fixed glass slide. Hydrogel was
sandwiched between the two slides for 5 min.
The time was noted for upper slide (movable)
to separate completely from the fixed slide 16.
Spreadability was determined using following
formula: S = M.L/T, where S = spreadability in
g.cm/s, M = weight tide to upper slide, L =
length of glass slide, and T = time taken to sep-
arate the slide completely from each other.
All the developed hydrogels were tested for
homogeneity by visual inspection. They were
tested for their appearance and feel on applica-
tion. The viscosity of formulated hydrogels was
measured by Brookfield digital viscometer (DV-
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HUSSAIN A., KHAN G.M., KHAN N.R., KHAN A. & ASIF M.

Enhancer
Drug Weight CPM Ethanol D. Water
Batch No
(DP) (g) (mL) QS to 100 mL
Turpentine oil Olive oil

DP Control 1 gram 1 10 mL – – do
DPT-oil 1 1 gram 1 10 mL 1% do
DPT-oil 2 1 gram 1 10 mL 2% do
DPT-oil 3 1 gram 1 10 mL 3% do
DPT-oil 4 1 gram 1 10 mL 4% do
DPT-oil 5 1 gram 1 10 mL 5% do
DPT-oil 10 1 gram 1 10 mL 10 % do
DPO-oil 1 1 gram 1 10 mL 1% do
DPO-oil 2 1 gram 1 10 mL 2% do
DPO-oil 3 1 gram 1 10 mL 3% do
DPO-oil 4 1 gram 1 10 mL 4% do
DPO-oil 5 1 gram 1 10 mL 5% do
DPO-oil 10 1 gram 1 10 mL 10 % do
Table 1. Composition of DP gel with Turpentine oil and Olive oil as enhancers. Each formulation contain 1 mL
of TEA.

1 + Viscometer, USA) using spindle S04 for 20
min. The spindle of viscometer was rotated at
2.5, 4, 5, and 10 rpm. Samples were measured
at 30 °C. Each sample was recorded three times
and averaged. For extrudability, closed collapsi-
ble tube containing hydrogel was pressed firmly
at the crimped end, the hydrogel extruded until
pressure dissipated. Weight in g was determined
to extrude 0.5 cm ribbon of hydrogel in 10 s.
Preparation of Rabbit skin
The animal skin was prepared according to
procedure described elsewhere 17 . Briefly,
young and healthy rabbits, each weighing 2-2.5
kg was obtained from the animal house of the
F/Pharmacy, Gomal University, D.I. Khan. Ani-
mals were housed and handled in the standard-
ized conditions at animal facility of the Faculty
of Pharmacy, Gomal University, D.I. Khan. All
animal procedures were performed in accor-
dance with and to the prior approval of the Eth-
ical Committee, Faculty of Pharmacy, G.U.D.I.
Khan, and later followed the recommended
guidelines to keep and maintain the in vivo test
laboratory properly. The animals were free to
move and to take standard food easily at their
own will. The water was also allowed ad libi-
tum. The rabbits were sacrificed after giving ex-
cess ether anaesthesia. Before testing the ani-
mals, hair from the abdominal region was
shaved properly by electric and hand razors.
The skin from the abdominal region was then
carefully removed surgically. The dermis (adher-
ing fats) removed by heating the skin in water
at 60 °C for 2 min. Epidermis wrapped in alu-
minium foil and stored at -80 °C till further use.
Before each use, skin was thawed at room tem-
perature for 2 h.
Gels in vitro and ex vivo permeation study
The permeation studies were performed in
Franz cell apparatus (Perme Gear USA) with
available diffusion area of 1 cm2 across cellulose
as well as hairless rabbit skin. Phosphate buffer
pH 7.4 used as receptor medium. The artificial
membrane was mounted on the receptor com-
partment of the Franz cell and donor compart-
ment was covered with parafilm. The tempera-
ture of the cell was maintained at 37 ± 0.5 °C 18
by continues circulation of water through cell
jackets. The donor compartment was maintained
at ambient temperature of 25 °C. The donor
compartment was loaded with 1 g gel samples
contact with cellulose membrane. The receiving
medium as spun with magnet bar which stir at
600 rpm. The effect of drug concentration on its
release from the gels was studies according to
drug concentration of 0.5, 1, and 1.5%, and ef-
fect of temperature on drug release was per-
formed at 25, 27, and 37 oC by water bath. Sam-
ple volume with drawn was 2 mL at different
time intervals for 24 h. Each time, the fresh re-
ceiving medium was replaced in volume equal
to the volume withdrawn. The samples were
analysed at 276 nm on UV/Visible spectropho-
tometer (UV-1601, Shimadzu, Japan) taking
phosphate buffer pH 7.4 as blank. The amount
of the drug permeated per square centimetre at
each time interval was calculated and plotted
against time.

1024
 Latin American Journal of Pharmacy - 34 (5) - 2015

Analysis of permeation data
The cumulative amount of the permeated
drug was plotted versus time, and the flux was
calculated from the steady-state part of the
curve. The lag time (Tlag) was determined by ex-
trapolating the linear portion of cumulative
amount permeated versus time curve. Perme-
ability values were calculated by the following
equation: Permeability= J/Cd, where J is steady
state flux and Cd is concentration in donor com-
partment.
The effectiveness of penetration enhancer
was determined by comparing the flux of DP in
the presence and absence of enhancer. It was
defined as the enhancement ratio (ER) that was
calculated using the following equation: E.R =
Kp with treatment / Kp without treatment. The
values reported are mean ratios from a mini-
mum of three replicates.
Statistical analysis
All the data were statistically analyzed by
analysis of variance (ANOVA). Results were
quoted as significant where p < 0.05.
ritation upon application to the skin 19. All the
gel formulations were found to have the desired
viscosities and thereby ensuring an optimum/
reasonable spreadability and ease of application.
Gel spreadability is considered one of the im-
portant parameters which ultimately enhance
the patient compliance by provide the uniform
applicability of the gel on the desired site. A
good gel is one which takes less time to spread
with simple application 20. The calculated values
of spreadability shows the test gel formulation
was well spreadable with simple application
with being rubbed. And it is also noted the gel
spread well with very small amount of shear
and also showed very good adhesion of site of
application. Content uniformity test also showed
the drug contents were in good acceptable limit
of ± 10% thus proving the content uniformity.
All the preparations were clear. The extrudabili-
ty reflects the capacity of the hydrogel, to get
ejected in uniform and desired quantity when
the tube is squeezed. Extrudability in terms of
weight in grams varies from 183 to 191 g as
shown in Table 2.

RESULTS AND DISCUSSION In vitro DP gels permeation

Physical evaluation of gels
The results of different physicochemical pa-
rameters for the evaluation of the formulated
gels of diclofenac potassium are shown in the
Tables 2 and 3, respectively. All the gel formula-
tions were clear and transparent in appearance
and were found to have a reasonable homo-
geneity without the presence of any lumps. The
pH of all gel formulations lies in the range of
6.2-7.0 which is the physiologicaly acceptable
pH range and in principle devoid of any skin ir-
The effects of different enhancers on the per-
meation of DP across the synthetic membrane
were investigated. Tables 3 and 4 show the per-
meation data of DP gel with/without enhancers.
The permeation of gels formulations containing
enhancer having good results as compared to
those which containing no enhancer. In our
findings, DP has good solubility in ethanol, as it
used in the formulation which favors the drug
partitioning towards the skin, Ethanol also pre-
sent in the formulation which causes the extrac-

pH Viscosity Spreadability Extrudability

F . Code Homogeneity
(10 rpm) (g.cm/sec) (g)

DPT-oil 1 6.5 15734 5.82 191 +++

DPT-oil 2 6.6 16703 6.56 187 ++
DPT-oil 3 6.8 16410 6.65 188 ++
DPT-oil 4 6.6 15645 6.76 188 +++
DPT-oil 5 6.3 16621 5.88 183 +++
DPT-oil 10 6.4 16601 5.83 186 +++
DPO-oil 1 6.2 16170 6.37 186 +++
DPO-oil 2 6.8 15234 5.22 187 ++
DPO-oil 3 6.1 16083 5.43 181 +
DPO-oil 4 6.4 16190 6.86 187 +++
DPO-oil 5 6.8 15645 5.62 185 ++
DPO-oil 10 6.4 15829 5.64 188 +++
Table 2. Physiochemical evaluation of different hydrogels of diclofenac potassium containing turpentine and
olive oil as enhancer. +++ Excellent, ++ Good, + Satisfactory.

1025
HUSSAIN A., KHAN G.M., KHAN N.R., KHAN A. & ASIF M.

Synthetic membrane Rabbit skin

Flux Kp Tlag Flux Kp Tlag

F. Code ER ER
(µg /cm2.h) ± SD (cm/h) ± SD (h) ± SD (µg /cm2.h) ± SD cm/h) ± SD (h) ± SD

Control 8.25 ± 1.44 0.161 ± 0.008 1.00 3.68 ± 0.001 10.01 ± 1.04 0.229 ± 0.001 1.00 4.25 ± 0.005
DPT-oil 1 15.66 ± 3.82 0.109 ± 0.0011 0.67 5.12 ± 0.021 11.92 ± 2.32 0.136 ± 0.0011 0.59 2.72 ± 0.029
DPT-oil 2 29.71 ± 2.71 0.191 ± 0.0032 1.19 5.78 ± 0.029 33.91 ± 3.41 0.401 ± 0.0012 1.75 1.58 ± 0.001
DPT-oil 3 42.29 ± 1.05 0.337 ± 0.0035 2.09 3.17 ± 0.014 63.85 ± 1.45 0.427 ± 0.0065 1.86 4.11 ± 0.044
DPT-oil 4 75.043 ± 4.14 0.788 ± 0.007* 4.89 4.12 ± 0.071 97.643 ± 0.34 1.083 ± 0.027* 4.73 2.79 ± 0.033
DPT-oil 5 136.18 ± 2.43 1.737 ± 0.006* 10.79 3.27 ± 0.022 151.78 ± 2.43 2.506 ± 0.004* 10.9 3.12 ± 0.062
DPT-oil 10 141.52 ± 3.86 1.796 ± 0.007* 11.2 3.66 ± 0.043 159.65 ± 4.26 2.608 ± 0.018* 11.4 3.35 ± 0.013
Table 3. Diclofenac potassium skin permeation parameters at the presence of T-oil as enhancer through syn-
thetic membrane and rabbit skin, the values are mean and standard deviations of 3 determinations. * Kp is sig-
nificantly different from control formulation, p < 0.05.

Synthetic membrane Rabbit skin

Flux Kp Tlag Flux Kp Tlag

F. Code ER ER
(µg /cm2.h) ± SD (cm/h) ± SD (h) ± SD (µg /cm2.h) ± SD cm/h) ± SD (h) ± SD

Control 8.25 ± 1.44 0.161 ± 0.008 1.00 3.68 ± 0.001 10.01 ± 1.04 0.229 ± 0.001 1.00 4.25 ± 0.005
DPO-oil 1 16.64 ± 2.37 0.367 ± 0.01 2.28 3.34 ± 0.031 15.043 ± 5.31 0.409 ± 0.076 1.79 3.34 ± 0.031
DPO-oil 2 31.45 ± 3.73 0.69 ± 0.051* 4.30 2.41 ± 0.03 51.753 ± 3.33 1.182 ± 0.059* 5.16 2.41 ± 0.01
DPO-oil 3 64.59 ± 5.85 1.46 ± 0.025* 9.08 3.79 ± 0.04 77.29 ± 2.85 1.754 ± 0.045* 7.66 3.79 ± 0.004
DPO-oil 4 80.56 ± 2.84 1.78 ± 0.027* 11.07 3.52 ± 0.033 87.26 ± 4.24 2.017 ± 0.02* 8.81 3.52 ± 0.033
DPO-oil 5 113.3 ± 3.13 2.10 ± 0.034* 13.07 2.32 ± 0.045 138.46 ± 3.63 3.165 ± 0.012* 13.82 2.56 ± 0.014
DPO-oil 10 121.4 ± 2.83 2.28 ± 0.077* 14.1 2.62 ± 0.045 147.33 ± 4.13 3.316 ± 0.042* 14.48 2.32 ± 0.014
Table 4. Diclofenac potassium skin permeation parameters at the presence of olive oil as enhancer through syn-
thetic membrane and rabbit skin, the values are mean and standard deviations of 3 determinations. * Kp is sig-
nificantly different from control formulation, p < 0.05.

tion of lipids from SC and enhanced percuta-

Cumulative drug permeated ug/cm2

neous absorption of the DP 21. It have been in-

vestigated that alcohol affect skin permeability 22
and when its used in combination in water in-
creases the permeability of drugs through the
skin 23. Due to this permeability of DP increased
when increased solute diffusivity through the
partially delipidized SC.

Effect of turpentine oil

Turpentine oil is one of the naturally occur-
ring volatile oil. Research studies have proved
that it might be used as acceptable enhancers
Time (h)
effectively. They were reported to have good
toxicological profiles, high percutaneous en- Figure 1. Cumulative amount of Diclofenac potassium
hancement abilities, and low cutaneous irritancy hydrogel permeation containing different concentra-
at low concentrations (1-5%) 24. Percutaneous tion of Turpentine oil as enhancer across synthetic
absorption of various hydrophilic and lipophilic membrane. Mean ± SD (n = 3).
drugs has been increased using turpentine oils
25. In this study the effect of different concentra-

tions of turpentine oil on the permeation of di- potassium through the synthetic membrane and
clofenac potassium is investigated. Six different rabbit skin from the gels in the presence of dif-
concentrations (1, 2, 3, 4, 5, and 10%) were ferent concentrations of the enhancers are
studied. The permeation profiles of diclofenac shown in the Figs. 1 and 2, respectively.

1026

Cumulative drug permeated ug/cm2
Cumulative drug permeated ug/cm2
Time (h)
Figure 2. Cumulative amount of Diclofenac potassium
hydrogel permeation containing different concentra-
tion of Turpentine oil as permeation enhancer across
rabbit skin. Mean ± S.D (n = 3)
Cumulative drug permeated ug/cm2
Time (h)
Figure 3: Cumulative amount of diclofenac potassium
hydrogel permeation containing different concentra-
tion of olive oil as permeation enhancer across syn-
thetic membrane. Mean ± SD (n = 3).
Table 3 illustrates the parameters of the
drug penetration measurments. Reported results
showed that turpentine oil used in the current
study has promoted the permeation of di-
clofenac potassium efficiently through skin port.
Highest concentration (10%) of turpentine oil
showed the increased permeation of drugs. The
flux of the formulation containing the highest
concentration of the turpentine oil is significant-
ly different (p < 0.05) from the control formula-
tion (without enhancer). The maximum flux of
the drug through the synthetic membrane is
141.523 ± 3.86 µg/cm2.h and permeability coeffi-
cient is 1.796 ± 0.007 cm/h which is significantly
different (p < 0.05) than the flux through the
control formulation which is 8.25 ± 1.44
µg/cm2.h and permeability coefficient is 0.161 ±
0.008 cm/h. There is no significance difference
among the flux of the formulations containing
the 1, 2, and 3% of the enhancer but significant
difference was obtained from 4, 5 to 10%. The
Latin American Journal of Pharmacy - 34 (5) - 2015
Time (h)
Figure 4. Cumulative amount of diclofenac potassium
hydrogel permeation containing different concentra-
tion of olive oil as permeation enhancer across rabbit
skin. Mean ± SD (n = 3).
flux obtained from the rabbit skin is little higher
than the synthetic membrane which is 159.65 ±
4.26 µg /cm 2 .h while using 10% of the en-
hancer. There is no significant difference be-
tween the flux of the formulation containing 5
and 10% enhancer which is 136.187 ± 2.34 and
141.523 ± 3.86 µg /cm2.h (synthetic membrane)
and 151.78 ± 2.43 and 159.65 ± 4.26 µg /cm2.h
(rabbit skin), respectively. To determine the ef-
fect of concentration of turpentine oil on di-
clofenac potassium permeation enhancement ra-
tio was calculated and it was found that al-
though increasing concentration of turpentine
oil (1 to 10%) resulted in increase the flux, there
is no significant difference between 5% and 10%
turpentine oil (p > 0.05). Thus, addition of 5%
turpentine oil seems to be optimum for enhanc-
ing skin permeation rate. It is also possible that
great enhancing effect of turpentine oil may be
due to the presence of ethanol in the formula-
tion. It also enhances the permeability by modi-
fying the intercellular packing, disrupting highly
ordered structure of lipids 25.
The synergic effect between ethanol and
monoterpenes has been reported earlier by
Obata 23. Since the gels also contain ethanol,
which is also reported by Manabe 26 have an in-
creasing effect on drug permeation, as it dis-
solves away the skin fats. The permeation kinet-
ic parameters indicate that with increase in the
enhancer concentration, the permeability coeffi-
cient, flux, and enhancement ratio increases. It
has been reported by Barry and Chow that the
enhancer concentration in the formulation af-
fects the promotion of transdermal drug delivery
27,28. Among many chemical enhancers exam-
ined, terpenes have been extensively investigat-
1027
HUSSAIN A., KHAN G.M., KHAN N.R., KHAN A. & ASIF M.
ed for their clinical use as penetration enhancer
and suggested to increase drug diffusivity in the
skin by disrupting the intercellular lipid packing
in the horny layer 4,14. Considering the balance
between efficiency and toxicity, many terpenes
clinical used as chemical enhancer 29.
Effect of olive oil
DP permeation measured across the synthet-
ic membrane and rabbit skin gave following re-
sults (Figs. 3 and 4).
Table 4 shows the flux, permeability coeffi-
cient and lag time of formulations containing
different concentrations of olive oil as enhancer
(1, 2, 3, 4, 5, and 10% w/v).
The results from this study show that addi-
tion of concentrations of the enhancer from 1 to
5 % significantly increases (p < 0.05) the perme-
ability coefficient, flux and ER of the diclofenac
potassium through synthetic membrane and rab-
bit skin as compared to control gel. When the
concentration increases from 5 to 10% there is
no significant increase in the permeation rate.
Aggarwal et al. 30 also found that olive oil en-
hance the permeation of resperidone up to 5%
concentration and no significance difference be-
tween 5 and 10% olive oil. The enhancement ra-
tios were found to be 14.1 and 3.63 for 1 and
5% olive oil, respectively. At the lowest concen-
tration of enhancer (1%) the flux and permeabil-
ity coefficient are very low (16.643 ± 2.37
µg/cm2.h and 0.367 ± 0.008 cm/h through syn-
thetic membrane and 15.043 ± 5.31 µg/cm2.h
and 0.409 ± 0.076 cm/h through rabbit skin)
which are near to the control gel (p > 0.05). As
the concentration of olive oil increases, it may
cause delay of partitioning of the drug out of
the base gel to the stratum corneum. There is
significance increase (p < 0.05) in the flux of the
drug from 3 to 5% increase in the enhancer con-
centration. Many studies have shown that sever-
al fatty acid esters increased the fluidity of the
lipid portions of stratum corneum by disrupting
lipid packing and thereby enhancing the perme-
ability of drugs through the skin 31. A proposed
mechanism of the oleic acid is the enhancement
of the drug permeation by increasing the diffu-
sivity and partitioning of the drug in the stratum
corneum 32. Fatty acids incorporated into the
skin lipids, disrupt molecular packaging and
change the level of hydration, thus allowing
drug penetration 33. Results also shown that the
lag time also decreases by increasing the en-
hancer concentration in case of turpentine oil
1028
while there is no significant effect of enhancer
on lag time in case of olive oil results shown in
Tables 3 and 4.
It also clear from the results that cumulative
drug penetrated through the rabbit skin is high-
er than the synthetic membrane. The possible
reasons could be; that rabbit skin has a large
number of hair follicles. It has been suggested
that permeation effect may be higher due to
these hair follicles, more hair follicles translates
good permeation 34. Furthermore, the anatomy
of skin may also allow more drug diffusion via
fluidization of skin lipidic domain in intercellu-
lar path, an action exerted by oleic acid (the
major component in olive oil). Lastly but not the
least, barrier thickness might also be a differenti-
ating factor which was 88 ± 1.9 mm for rabbit
skin and 129 ± 1.2 mm for synthetic membrane,
which means more resistance to drug diffusion
may have been offered by more thicker skin.
CONCLUSION
On the basis of above results and discussion,
it is well justified to conclude that the turpentine
oil causes an increase in permeation of DP from
gels . The peremation increases as the concen-
tration of turpentine oil and olive oil increases.
The primary mechansim of enhancing drug per-
meation might be the disruption of stratum
corneum. Turpentine oil holds great potential to
be used as natural penetartion enhancer for for-
mulation of hydrogels as compared to olive oil.
From the permeation result, the gel formulations
prepared with 1.5% carbopol 940 containing
10% turpentine oil as enhancer, was found to be
most satisfactory, compared to other formula-
tions with respect to physico-chemical, and me-
chanical property. The permeation kinetic pro-
file indicates that, with increase in the enhancer
concentration, permeability coefficient, and en-
hancement factor increases. Thus the release
rate of DP can be achieved by altering the en-
hancer concentration in the formulation.
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