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WORLD JOURNAL OF PHARMACY AND PHARMACEUTICAL SCIENCES
Drais.
World Journal of Pharmacy and Pharmaceutical Sciences
SJIF Impact Factor 6.041
Volume 5, Issue 6, 308-318
Research Article
ISSN 2278 – 4357
DEVELOPMENT, CHARACTERIZATION AND EVALUATION OF
THE PIROXICAM NANOEMULSION GEL AS TOPICAL
DOSAGE FORM
Hayder Kadhim Drais*
Ministry of Health and Environment, Babylon Health Directorate, Babylon, Iraq.
Article Received on
13 April 2016,
ABSTRACT
Piroxicam is a non-steroidal anti-inflammatory drug. It is practically
Revised on 01 May 2016,
Accepted on 22 May 2016
insoluble in water. It is associated with many gastrointestinal unwanted
DOI: 10.20959/wjpps20166-7031
effect. The objective of this investigation to prepare an
evaluate
nanoemulsion gel of piroxicam as a topical dosage form to increase
*Corresponding Author
solvability of piroxicam and decrease the gastrointestinal side effect
Dr. Hayder Kadhim
that lead to enhance piroxicam bioavailability and therapeutic activity.
Drais
The pseudoternary phase diagrams were constructed from the double
Ministry of Health and
distilled water, surfactant mixture (Smix) was tween 80 and ethanol at
Environment, Babylon
ratios of (1:1, 2:1, 3:1, and 4:1) and oil mixture. The pseudoternary
Health Directorate,
Babylon, Iraq.
phase plot that has larger region of nanoemulsion was Smix ratio (3:1)
from which take six of nanoemulsion formulas (NE1-NE6) for
characterization of nanoemulsions and to formulate nanoemulsion gel formulas (NF1-NF6).
The six of nanoemulsion gel formulas were evaluated. NF6 was the optimized formula and
tested for stability and morphology. Atomic force microscopy (AFM) shows that the selected
formula (NF6) is highly stable. It can be concluded that the selected (NF1) was an effective
alternative for the topical delivery of piroxicam.
KEYWORDS: Nanoemulsion, Piroxicam, Nanoemulsion gel.
INTRODUCTION
Piroxicam a non-steroidal anti-inflammatory drug utilized to relieve the symptoms of
inflammatory arthritis such as (rheumatoid arthritis, psoriatic arthritis, ankylosing spondylitis,
systemic lupus erythematosus), degenerative arthritis such as osteoarthritis, in acute gout and
in post-operative pain. It is practically insoluble in water, however, its use has been
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associated with a number of undesirable side-effects on the stomach and kidneys in addition
to gastric mucosal damage.[1]
Nanoemulsion (NE) which is nano scale delivery systems in has great interest as a part of
nanotechnology. It is transparent with the range of particle sizes between 100 to 500 nm. The
aqueous phase, oil phase, surface active agent, co-surfactant represent the composition of
nanoemulsion.[2] However, the rheological properties of nanoemulsion make it unsuitable
for application to the skin. When combine gelling agent with nanoemulsion, this make more
desirable for transdermal delivery.[3] When gelling agent and nanoemulsion are utilized in
combination form the dosage form is named nanoemulgel. This provide double release
control system i.e. Emulsion and gel.[4] The solubility problem of most of the hydrophobic
drugs make these drugs cannot be incorporated directly into the gel base.The nanoemulsion
gel help that the hydrophobic drugs solubilized into the oily phase and then oil droplets are
dispersed in aqueous phase leading to oil in water (o/w) nanoemulsion then
this
nanoemulsion can be incorporate into gelling agent to get nanoemulsion gel.[5]
Thus, the aim of this research to formulate and evaluate nanoemulsion gel of piroxicam
employed topically to increase solubility of piroxicam and decrease oral unwanted effect of
piroxicam that lead to improve piroxicam therapeutic activity and patient compliance.
MATERIALS AND METHODS
MATERIALS
Piroxicam supplied by Wadi Al-Rafidian factory for pharmaceutical products, Baghdad, Iraq.
peppermint oil and ginger from BAR-SUR-loop Grasse A. M Franc. Tween 80 from SD fine
Chemlimited (SDFCL) Mumbai, India. Methanol and ethanol from grin land chemical comp,
United. Carbomer 934 from Hengshui Taocheng Chemicals Auxiliary Co., Ltd. China.
KH2Po4 and Na2Po4 from Merck & Co., Inc. Germany.
METHODS
1. FORMULATION OF PIROXICAM NANOEMULSION AND DRAWING OF
PSEUDOTERNARY PHASE DIAGRAM
The oil mixture was produced by mixing of ginger oil and peppermint oil in a ratio (1:2) to
obtain oil mixture. The aqueous phase, oil mixture and surfactant mixture (Smix) are
ingredients of the pseudo ternary phase diagram that is get by use the aqueous phase titration
method. The surface active agent and co-surfactant is combined in various weight ratios (1:1,
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2:1, 3:1, 4:1) to obtain Smix. The piroxicam loaded in oil mixture mix with Smix in different
weight ratios for each phase diagram, so that all parts of the pseudoternary phase diagram
will be covered. The slow titration of double distilled water to admixture of the oil mixture
and Smix was occurred to get the borderline of phases and ocular investigation were made for
transparency. When there is a clear liquid to the eyes, stop further adding double distilled
water and different o/w nanoemulsions are formalised. The pseudoternary phase plot was
designed. Take seven formulas from pseudoternary phase diagram that has wider
nanoemulsion area to prepare nanoemulsion gel of piroxicam.[6]
2. FORMULATION OF PIROXICAM NANOEMULSION GEL
The aqueous phase was made by combining double distilled water and tween80. The oil
mixture that is previously formulated constitute the oily phase. The oil mixture was sonicated
for 15 minutes. The aqueous phase heated to 80 and oily phase to 70°C on hot plate; the oily
phase gradually was poured to the aqueous phase with continuous stirring with magnetic
stirrer at 2000 RPM and stirring was continued for 5-10 min until becoming cool at room
temperature. The gel bases were formulate by dispersing carbopol 934 with specified
concentration in double distilled water with constant stirring at a moderate speed using
mechanical stirrer. Few drops of triethanolamine were added to obtain a pH of about ( 5.5).
The nanoemulsion was then combine with the gel in (1:1) ratio to obtain homogenous
nanoemulsion gel.[7]
Table (1): The piroxicam nanoemulsion gel formulations.
Formulation
components
(%w/w)
Piroxicam
Carbopol 934
Oil mix(1:2)
Smix(3:1)
Propylene glycol
Ethanol
Methylparaben
Propylparaben
Sodium sulphite
Double distilled
water up to
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Formulation code
F1
F2
F3
F4
F5
F6
Piroxicam gel
1
1
5
35
0
0
0.02
0.02
0.1
1
1
5
32.5
0
0
0.02
0.02
0.1
1
1
5
30
0
0
0.02
0.02
0.1
1
1
5
27.5
0
0
0.02
0.02
0.1
1
1
5
25
0
0
0.02
0.02
0.1
1
1
5
22.5
0
0
0.02
0.02
0.1
1
1
5
0
5
5
0.02
0.02
0.1
100
100
100
100
100
100
100
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CHARACTERIZATION OF THE FORMULATED PIROXICAM NANOEMULSION
MEASUREMENT OF THE GLOBULE SIZE
The globule size was determined throug ABT-9000 nanolaser particle size analyzer. The
average of globule size and globule size distribution plot was obtained. All samples were
taken in three trails.[6]
POLY DISPERSITY INDEX (PDI) MEASUREMENT
It can obtained by ABT-9000 nanolaser particle size tester. It determine the uniformity of
the globule size in nanoemulsion preparation.. The measurements were occuring in triplicate.
PDI range from 0.0 to 1.0. As the polydispersity index value is closer to zero, the globules
with high uniformity.[8]
EVALUATION PIROXICAM NANOEMULSION GEL
PHYSICAL APPEARANCE
The prepared nanoemulsion gel formulas were visually analyzed for color, appearance and
homogenicity. The tested samples were occurred in triplicate.[9]
THE pH DETERMINATION
The pH of all the nanoemulsion gel preparations was tested by a pH meter at room
temperature. The human skin is naturally acidic with a normal pH of 4-6. The women have a
pH more acidic skin than men.One of the high important factors in the evaluation of
nanoemulsion gels is the pH. The pH values affect on ionization of the drug i.e. ionized and
unionized forms of the drug. The tested samples were occurred in triplicate.[10]
THE DRUG CONTENT MEASUREMENT
By UV-visible spectrophotometer, the drug content was measured(6). The samples were
diluted with methanol then read in the UV analyzer. The tested samples were occurred in
triplicate. The absorbance was observed at 200 nm and the drug content was determined by
the following equation.
Drug content = (Analyzed content/Theoretical content) x 100 ….. (Eq2)
SWELLING INDEX ESTIMATION
Swelling index is the separation of molecules of formulas by water molecules cause an
expansion in volume of the mass. It associated to application area of nanoemulsion gel. One
gram of piroxicam nanoemulsion gel formulations was draped with aluminium foil that is
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punched to get holes and put in phosphate buffer pH 6.8 for 6 hours, then the swollen
samples wiped with the filter paper to eliminate access of absorbed distilled water on the
surface and then it was rapidly weighed on an electronic balance. All samples were taken in
three trails.
The swelling index was measured by utilizing the following equation(Eq).
Sw = [(Wt – Wo) / Wo] × 100….. (Eq1)
Where, SW = percentage of swelling index of piroxicam nanoemulsion gel
Wt = the weight (g) of the nanoemulsion gel at time t
Wo = initial weight (g) of the piroxicam nanoemulsion gel.[11]
IN VITRO RELEASE MEASUREMENT
A United States Pharmacopeia (USP) dissolution tester apparatus II is the device that is used
to measure the in vitro release of piroxicam from formulas. One gram of piroxicam
nanoemulsion gel formulations (NF1-NF6) and piroxicam gel was put in class tube with
1.5cm diameter, covered with a cellulose acetate membrane which was formerly soaked in
phosphate buffer of pH (5.5) for about 24 hr. The membrane adquately sealed and inverted
under the surface of 900 mL of phosphate buffer of pH (5.5) containing 1% polysorbate 80, at
37 ± 0.5 °C with stirring speed of 100 RPM. The sample of 5 mL was taken at 0, 15, 30, 60,
90,120, 150, 180, 210, 240 min and replaced with an equivalent amount of dissolution
medium. The drug content of the samples was determined by a UV spectrophotometer
at
200 nm. The tested samples were occurred in triplicate.[12,13]
ATOMIC FORCE MICROSCOY (AFM) ANALYSIS
By AFM angstrom advanced inc. AA3000 USA, the morphology of selected piroxicam
nanoemulsion gel formula was determined. AFM analysis was occurred by placing drops of
the sample onto a glass slide and then measure.[14]
STATISTICAL ANALYSIS
The tested samples were occurred in triplicate and use analysis of variance (ANOVA) at level
(P<0.05) to explain the results.[6]
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RESULTS AND DISCUSSION
FORMULATION
OF
PIROXICAM
NANOEMULSION,
DRAWING
OF
PSEUDOTERNARY PHASE DIAGRAM AND FORMULATION OF PIROXICAM
NANOEMULSION GEL
The pseudoternary phase plots were made by drawing double distilled water, oil mixture (1:2)
and variable Smix ratios as 1:1, 2:1, 3:1, and 4:1 and as shown in figure (1). The shaded area
of pseudoternary phase plot give the region of nanoemulsion, whereas the nonshade area
offer the emulsion region. The Smix ratio (3:1) of pseudoternary phase plot has a greater
nanoemulsifying region and from which a six formulas of nanoemulsion (NE1-NE7) were
taken for characterization of nanoemulsions and to formulate nanoemulsion gel formulas
(NF1-NF6).
Figure (1): The pseudoternary phase diagram of double distilled water, oil mixture(1:2)
and Smix (1:1, 2:1, 3:1, and 4:1).
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CHARACTERIZATION OF THE FORMULATED PIROXICAM NANOEMULSION
MEASUREMENT OF THE GLOBULE SIZE
The results globule size range was NE1 (5-6.1nm), NE2 (5-7.29 nm), NE3 (7.2-9.2 nm), NE4
(13.1-15 nm), NE5 (18-23 nm) and NE6 (5-7 nm). The results indicate that all the formulas
had droplets in the nanoscale size. Analysis of variance show significant differences between
globule size and the concentration (%w/w) of Smix where (p<0.05).
POLY DISPERSITY INDEX (PDI) MEASUREMENT
PDI was from (0.01 to 0.019). The results of PDI indicate to that nanoemulsion formulations
had a homogenous composition.
EVALUATION PIROXICAM NANOEMULSION GEL
PHYSICAL APPEARANCE
The physical appearance of nanoemulsion gel formulas were yellowish in color,transparent
and excellent homogenicity as shown in table (2).
THE pH DETERMINATION
The pH of all the nanoemulsion gels formulas was between 5.3 to 5.89 as in table (2) which
are within the normal pH value of human skin.
Table (2): The physical appearance, pH, percent of swelling index and percent of drug
content in piroxicam nanoemulsion gel formulations.
Formulation
code
NF1
NF2
NF3
NF4
NF5
NF6
color
clarity
uniformity
yellow
yellow
yellow
yellow
yellow
yellow
clear
clear
clear
clear
clear
clear
excellent
excellent
excellent
excellent
excellent
excellent
Phase
separation
none
none
none
none
none
none
pH
5.3
5.55
5.89
5.66
5.44
5.51
% Swelling
index
17
20
23
24
28
32
% Drug
content
98.1
96.4
96.2
97.6
96.4
95.21
THE DRUG CONTENT MEASUREMENT
The drug content was in a range of (95.21 – 98.1%) as shown in table (2).
SWELLING INDEX ESTIMATION
The outcome described in table (2). The result shows the effect of Smix on swelling index at
constant gelling agent concentration. As the Smix concentration increase that lead to create
more hydrophobic media that retard water molecule from enterance to formula therefore it
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was found NF6 significantly higher swelling index due to it contain least Smix quantity and
NF1 with higher Smix concentration has significantly lower swelling index (p<0.05).
IN VITRO RELEASE MEASUREMENT
The drug release profile in figure (2) indicates that the NF6 was significantly highest in
dissolution rate while piroxicam gel formula has lower in dissolusion rate which was
significantly lowest (P<0.05).
The comparability drug release diagram of piroxicam nanoemulsion gel (NF1-NF6) and
piroxicam gel show that the drug release was followed the descending order:
NF6>NF5>NF4> NF3>NF2>NF1> piroxicam gel.
The drug release profile from piroxicam nanoemulsion gel preparations (NF1-NF6) show the
effect of Smix concentration at constant concentration of oil mixture and gelling agent,as the
concentration Smix increase this lead decrease drug release this may be due to the high
concentration of Smix increase hydrophobicity and retarding forces of formulations and
resist dissolution media molecules passage through hydrophobic matrix and release the drug
also the piroxicam molecules will have greater diffusional pathway to reach the dissolution
media therefore it was found NF6 has greater release profile compare to other of piroxicam
nanoemulsion gel formulations due to it has the least concentration of Smix that make less
retarding effect for dissolution molecules and lower diffusional pathway to drug molecules to
reach the dissolution media.
also, the piroxicam gel release profile has lowest release rate compare to other piroxicam
nanoemulsion gels this due to nanoemulsion found in nanoemulsion gels
give many
advantages in compare to gel such as nanosize droplets and improve solubility of piroxicam
this will enhance passage of dissolution media to hydrophobic matrix and lower diffusional
pathway for piroxicam molecules to reach the release media that make piroxicam
nanoemulsion gels superior and highly attractive in compare to piroxicam gels.[6, 15] From in
vitro release analysis, it was found that NF6 is a selected formula where it has lower globule
size (5-7 nm ) as shown in figure (3) , low PDI (0.019), excellent physical appearance, the
normal value of pH (5.51 ), a higher percent of swelling index (32), accepted percent of drug
content (95.21%) and higher release rate makes it faster in relieving inflammatory conditions
and enhance therapeutic efficacy. The selected formula (NF6) subject for further analysis of
atomic force microscopy (AFM) study.
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Figure (2): Represent drug release profile for piroxicam nanoemulsion gel and
piroxicam gel formulations.
Figure (3): The particle size distribution for NF6.
ATOMIC FORCE MICROSCOY (AFM) ANALYSIS
The result of AFM shows that the selected formula(NF6) has nano scale particle nearly
spherical in shape and smooth surface globules, this mean the
stability of optimized
nanoemulsion gel (NF6) as shown in figure (4).
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Figure (4): The AFM image of NF6 where scanning area is 5 μm * 5 μm.
CONCLUSION
1. The aqueous phase titration method is a cheap and easy method that employed in the
preparation of piroxicam nanoemulsion.
2. The piroxicam nanoemulsion gel (NF6) with its attractive physical characterstics and
higher in vitro release rate will improve piroxicam solubility and bioavailability that lead
to increase piroxicam therapeutic efficacy and enhance patient compliance.
3. The AFM study indicate that the optimized formula (NF6) with nanometer scale has good
stability that makes it promising formula to local and systemic delivery of piroxicam.
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