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ABSTRACT: This paper describes the preparation and characterization of transdermal patches
impregnated with naproxen. A mixture of ethylene vinyl acetate and Eudragit R
E100 (80:20,
w/w) is used as a polymeric matrix to obtain a thin membrane to be impregnated. Drug impreg-
nation is carried out under pressurized CO2 as a processing medium according to a two-step
procedure. The patch is first soaked at 1000 psi and 22◦ C for 2 h, and then foamed as a result of
the rapid release of CO2 pressure in order to increase the porosity of the surface. Subsequently,
the naproxen solution is placed in contact with the membrane and then soaked in CO2 at 450
psi and 37◦ C for 2.5 h to enhance the mass transfer of drug into the polymer matrix. The char-
acterization of the resulting samples by liquid chromatography, microscopy, and calorimetry
provides information on naproxen content and distribution. Patches synthesized in this way are
loaded with about 1% naproxen. The drug release and diffusion process through a membrane
have been studied chromatographically using a Franz diffusion cell. Results have shown that
a sustained delivery for more than 24 h is obtained. © 2010 Wiley-Liss, Inc. and the American
Pharmacists Association J Pharm Sci 100:992–1000, 2011
Keywords: controlled delivery; transdermal drug delivery; processing; in vitro models; poly-
meric drug carrier; supercritical fluids
formation of a molecular dispersion of drug into the Chem Station for data acquisition and analysis (Rev.
matrix. A 10.12), all of them from Agilent Technologies
Although a wide variety of polymers have been (Waldbronn, Germany). The analytical column was a
used in the past 40 years as drug carriers, recent reverse phase C18 (Synergi Hydro-RP, Phenomenex,
trends rely on water-soluble matrices such as PVP 150 × 4.6 mm2 d.i., 4 µm particle size). Naproxen was
or its copolymer with vinylacetate (PVP-VA 64).16 eluted isocratically with 10 mM of formic acid/formate
Other polymers such as cellulose derivatives [e.g., aqueous solution (pH 3.2) + MeOH (20/80, v/v) as a
ethylcellulose (EC), methylcellulose] 17,18 ethylene mobile phase. The flow rate was maintained at 1 mL
vinyl acetate (EVA), 19,20 and pH-dependent polymers × min−1 and the injection volume was 20 µL. Ultra-
such as Eudragit R
E100 (Evonik Degussa, Essen, violet (UV) spectrophotometric detection was carried
Germany), (polymethacrylate copolymer)18 are being out at 270 nm. Fluorescence detection was carried out
increasingly used. at 270 and 357 nm as excitation and emission wave-
This study is focused on the preparation and char- lengths, respectively. A magnetic stirrer IKA R
RCT
acterization of a transdermal patch as a model sys- basic (Staufen, Germany) was used for controlling the
tem of sustained released using pressurized CO2 as release conditions.
a processing medium. Naproxen is the drug chosen Standard solutions for calibration were prepared in
here for this development. Naproxen is a member of methanol in the concentration range from 5.2 × 10−7
the 2-arylpropionic acid family of nonsteroidal anti- to 3.9 × 10−5 M. For UV spectrophotometric detection,
inflammatory drugs commonly used for the reduction a good linearity in the studied range was found with a
of mild-to-moderate pain, fever, inflammation, and regression coefficient r2 = 0.9989. Detection limit es-
stiffness.21 The US Food and Drug Administration timated for a signal-to-noise ratio of three was 1.8 ×
approved the use of naproxen sodium as an over the 10−7 M. Repeatability expressed as relative standard
counter drug in 1994. Analytical techniques, includ- deviation (RSD in %) for the peak area was calcu-
ing differential scanning calorimetry (DSC), confocal lated from eight replicates at a concentration of 2.5
fluorescence microscopy, and high-performance liq- × 10−6 M and was 2.1%. For fluorescence detection,
uid chromatography (HPLC), have been utilized for a the linearity was found with a regression coefficient
more rigorous characterization of naproxen samples. r2 = 0.9988. Detection limit was 1.1 × 10−7 M and
repeatability was 1.5%. The chromatographic method
was used in both the determination of the impreg-
MATERIALS AND METHODS nation percentage and in the monitoring of the drug
Materials release.
A differential scanning calorimeter (DSC-822e/400,
Sodium hydrogenphosphate, sodium dihydro- Mettler Toledo, Greifensee, Switzerland) was used to
genphosphate, formic acid, rhodamine (5,6- determine melting and glass transition temperatures.
carboxytetramethylrhodamine), and naproxen Thermograms were obtained at a heating rate of 10◦ C
(99%) were purchased from Sigma–Aldrich (St. × min−1 from 30◦ C to 250◦ C under a N2 purge of
Louis, Missouri). Methanol and methylene chloride 50 mL × min−1 .
(HPLC grade, Merck, Darmstadt, Germany) were A confocal microscope Leica TCS SPII (Leica Mi-
used as solvents. Carbon dioxide (CO2 , 99.99 mol% crosystems, Wetzlar, Germany) operating in both re-
purity) was supplied by Praxair (Columbus, Ohio). flectance and fluorescence modes was used. Excita-
Polymers used were EC 20 cps from Keyser & Mackay tion was at 351 and 364 nm using UV lasers and
(Brussels, Belgium), PVP-VA 64 (molecular weight = reflectance, and emission intensities were recorded
45,000–70,000 g mol−1 ) from BASF (Ludwigshafen, in the range of 400—800 nm. The objective used
Germany), Eudragit R
E100 (acrylic polymer, molecu- was a 10 × 0.3 N.A. HCPL FLUOTAR lens (Leica
lar weight = 150,000 g mol−1 ) from Evonik Degussa Microsystems). Images were processed using the
(Essen, Germany), and EVA (70 wt% of vinyl acetate) ImageJ (NIH Image; www.rsb.info.nih.gov/ij) and
from Sigma. Ultrapure water (Millipore, Milford, Photoshop 7.0 software (Adobe Corp., San Jose,
Massachusetts) was used for the preparation of California). Transversal sections were taken every
aqueous solutions. 2.4 µm.
Analytical Instrumentation
The chromatographic system consists of an HPLC Ag-
Preparation of Working Solutions and Naproxen
ilent 1100 Series instrument equipped with a G1311A
Impregnated Patches
quaternary pump, a G1379A degasser, a G1329B
standard autosampler (1200 Series), a G1315B diode- The working solution of rhodamine for preliminary
array detector furnished with a 13-mL flow-cell, impregnation studies consisted of 1.5 mg dissolved in
a G1321A fluorescence detector, and an Agilent 100 mL of sodium phosphate buffer solution (PBS)
DOI 10.1002/jps JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 100, NO. 3, MARCH 2011
994 ARGEMÍ ET AL.
at pH 6.2. Naproxen working solution consisted of allowed to ensure thermal equilibrium. Then,
100 mg in 25 mL of PBS at pH 7.4. the 500D syringe pump was used to pressurize
the vessel with CO2 until the working pressure
Patch Synthesis.
was reached. The temperature and pressure
Ten grams of EVA and 2.5 g of Eudragit R
E100 were held constant during this period. The sys-
were treated with 30 mL of CH2 Cl2 . The mixture tem was then depressurized over nearly 30 min
was left in an ultrasonic water bath (Unisonics ul- by slowly opening the purge valve. Finally, the
trasonic cleaner FXP8) for 90 min (until polymers patch was dried at room temperature for 8 days
were completely dissolved). This water bath sonica- at least and stored in a sealed plastic bag until
tor has an ultrasonic power of 50 W and provides a future characterizations. The experimental pro-
frequency of 40 kHz. The resulting solution was cast cessing window for impregnation was explored
on a microporous Teflon film placed on a glass plate. by experimental design. For this purpose, a full
The Teflon film was used as a release agent because factorial design with three factors (temperature,
polymer sticks on the glass. A thin layer of polymer pressure, and time) at two levels was utilized,
solution was obtained with the aid of an adjustable which corresponded to eight experiments.
thin film applicator (GARDCO, Paul N. Gardner Co.,
Pompano Beach, Florida). The solvent was allowed
to evaporate, first at ambient conditions, and then in Characterization Studies
the vacuum oven (P = 30 mmHg) at room temper-
Determination of the Percentage Impregnation.
ature overnight. The thin polymer layer was peeled
off and thickness was measured using an electronic The amount of naproxen entrapped in the polymeric
gauge Mitutoyo (Model 543-252B, Mitutoyo America matrix was determined by HPLC. For this purpose,
Corp., Aurora, Illinois) with an accuracy of ±0.5 µm. about 30–40 mg of sample were dissolved in 10 mL of
Membranes with a thickness ranging from 231.5 to CH2 Cl2 by ultrasonication for 1 h. Subsequently, the
242.4 µm were obtained. Finally, patches were cut in solvent was evaporated under nitrogen current and
square shape (4 × 4 cm2 size, approx.) and were stored the dry residue was redissolved in 25 mL of methanol.
at ambient conditions until further use. Twenty microliter of the resulting solution, previously
filtered through a 0.45 µm pore-size membrane, was
Patch Impregnation Using CO2 .
injected into the chromatograph.
The procedure for impregnation of patches with
naproxen consisted of two steps as follows:
Drug Release Studies.
(1) Patch foaming process: First, the synthesized The study of naproxen diffusion from the patches was
patches (see patch synthesis section) were carried out using a Franz glass cell with 3.14 cm2 of
treated with pressurized CO2 to induce the for- diffusion area and a receptor chamber of 12 mL vol-
mation of pores in the material. The polymeric ume. Naproxen is soluble in the receptor medium at
patch (placed on a piece of Teflon film) was lo- a concentration of 5.2 mg mL−1 . A sample amount of
cated inside a stainless steel high-pressure ves- 25–60 mg was distributed on a synthetic Nylon mem-
sel (Pressure Products Industries, Inc., Warmin- brane of 0.45 µm pore size (Whatman Int., Maidstone,
ster, Pennsylvania). Another piece of Teflon film Kent, UK) and placed in the donor chamber. Imme-
and a piece of stainless steel were placed on diately after that, the top of the cell cap was covered
top of the patch. The system was pressurized with Parafilm R
(Chicago, Illinois) to minimize evap-
to 1000 psi and held at constant pressure for oration during the test. The receptor chamber was
2 h with an ISCO Syringe pump 500D (ISCO, filled with 100 mM of PBS at pH 7.4 and the tem-
Lincoln, Nebraska). The temperature was main- perature was kept at 32 ± 0.5◦ C. The solution in the
tained at 22◦ C. The foaming process was initi- receptor chamber was stirred with the aid of a cylin-
ated with a rapid CO2 depressurization in 4 s drical magnetic stir bar at a constant rate of 70 rpm.
and the porous patch obtained was ready to be Aliquots of 300 µL withdrawn at preselected times of
impregnated with the drug. 1, 2, 4, 6, 8, and 24 h were analyzed by HPLC and,
(2) Impregnation process: 1000 µL of solution of immediately after, equal volumes of fresh temperate
model (rhodamine, 1.5 mg in 100 mL PBS at pH PBS solution were added to the receptor chamber.
6.2) and active drug (naproxen, 0.1 g in 25 mL Sink conditions were well ensured by correcting any
PBS at pH 7.4) were dipped onto the porous ma- volume losses, when necessary. At the end of the pro-
trix surface of the patch. The high-pressure ves- cess, the drug content remaining in the patch was
sel was then sealed airtight and heated with determined as indicated previously in section of the
a Peltier system up to the desired experimen- percentage impregnation determination. Kinetic re-
tal temperature. A minimum of 15 min was leases were performed in triplicate.
JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 100, NO. 3, MARCH 2011 DOI 10.1002/jps
NAPROXEN-POLYMERIC PATCH FOR TRANSDERMAL SUSTAINED RELEASE SYSTEM 995
DOI 10.1002/jps JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 100, NO. 3, MARCH 2011
996 ARGEMÍ ET AL.
Figure 1. Impregnation yields from the two-level three-variable experimental design. (a)
Processing time = 2.5 h; (b) Processing time = 6 h.
macroscopic drug distribution is shown in Figure 3. corners (from 0.1% to 0.6%, approx.). Sections around
As 1000 µL aliquot of sample solution was poured in the central square piece contained intermediate drug
the center of the patch, this area reasonably contained percentages (0.9%–1.4%, approx.).Differential scan-
a higher amount of naproxen. It was confirmed that ning calorimetry was used to study the thermal
the drug impregnation in the center corresponded to properties of the CO2 -treated samples. Thermo-
1.8% (w/w), whereas percentages were lower at the grams of raw drug, unloaded polymeric patch, and
JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 100, NO. 3, MARCH 2011 DOI 10.1002/jps
NAPROXEN-POLYMERIC PATCH FOR TRANSDERMAL SUSTAINED RELEASE SYSTEM 997
Figure 2. Pictures obtained from fluorescence confocal microscopy in two dimensions. Pictures
of samples 3a and 5b measured at a certain depth. Scale bars shown.
Figure 4. Result from differential scanning calorimetry (DSC) analysis. (1) Raw naproxen,
(2) unloaded patch, and (3) impregnated patch.
release in the first 6 h was obtained. After this pe- plots, being higher for experiments 7 and 8 (5.0 and
riod, a more prolonged release process was observed. 5.5 wt%h0.5 , respectively) than for experiment 2 (4.0
For all samples belonging to the experimental design, wt%h0.5 ). Experiments 7 and 8 corresponded to lower
the percentage of naproxen release over 24 h ranged impregnation yields and experiment 2 to a higher im-
from 15% to 25%. In addition, a more sustained drug pregnation yield. In conclusion, lower impregnation
delivery from the patches was attained in comparison of naproxen facilitated its diffusion and release.
with the kinetics of the raw naproxen dissolution, ob-
taining t1/2 > 24 h (Fig. 5a). No lag times were found
in the concentration delivery profiles. This fact was a
CONCLUSIONS
proof that the delivery was controlled by the pristine The novelty in this work consisted in the preparation
formulation, hence the polymeric matrix. Moreover, of a naproxen transdermal patch by using pressurized
there are no diffusion limitations due to the mem- CO2 . The device presented here seemed to be a promis-
brane. The release profile followed the well-known ing alternative approach to conventional formula-
Higuchi model27 for simple diffusion processes. Suc- tions. Although various procedures were assayed
cessful correlation with the experimental data was for the preparation of the membranes, including
achieved. Figure 5b plots the cumulative released melting processes in a press, the most successful
amount in function of the square root of time. The de- strategy relied on dissolution/evaporation of poly-
livery rates determined for all samples were from 3.8 mers. It has been evidenced that the patch pretreat-
to 5.5 wt%h0.5 . The relationship between delivery and ment with dense CO2 creates a higher porous mate-
the square root of time was associated with a mecha- rial, which was further impregnated with naproxen.
nism controlled by the polymeric matrix. In this way, After that, the membrane was efficiently loaded with
the longitudinal diffusion resistance was increased the drug at 450 psi and 37◦ C for 2.5 h. Significant
with residence time through the outer zone of the advantages were gained from the use of dense CO2 as
matrix (where a reduction of drug has happened) and a processing agent because it is a not toxic gas and is
the border of water–matrix layer. Naproxen concen- easily removable by depressurization. Characteriza-
tration in the donor chamber was gradually decreased tion in vitro assays proved excellent results, such as
and as a consequence the drug delivery. This trend ex- the naproxen delivery for a prolonged period of time,
plained the two different behaviors of the cumulative at least 24 h. Future perspectives should be focused
naproxen profiles: release stage and sustained deliv- on the processing of the patch, optimizing impregna-
ery to the skin. tion working parameters and paying attention in new
The respective rates of release were determined materials. Furthermore, drug capacity loading should
from the slopes of the regression lines in the Higuchi be appropriately studied and if possible, improved.
JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 100, NO. 3, MARCH 2011 DOI 10.1002/jps
NAPROXEN-POLYMERIC PATCH FOR TRANSDERMAL SUSTAINED RELEASE SYSTEM 999
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JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 100, NO. 3, MARCH 2011 DOI 10.1002/jps