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ISSN: 1071-7544 (print), 1521-0464 (electronic)

Drug Deliv, Early Online: 1–10

! 2014 Informa Healthcare USA, Inc. DOI: 10.3109/10717544.2014.936987

RESEARCH ARTICLE

Formulation and characterization of acetaminophen nanoparticles in

orally disintegrating films
Nusaiba K. Al-Nemrawi and Rutesh H. Dave

Division of Pharmaceutical Sciences, Long Island University, Brooklyn, NY, USA

Abstract Keywords
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The purpose of this study was to prepare orally disintegrating films containing nanoparticles Hydroxypropyl methyl cellulose, in vitro,
loaded with acetaminophen. Nanoparticles were prepared by the emulsion-solvent evaporation poly(lactide-co-glycolide acid), solvent
method where acetone phase containing acetaminophen and poly(lactide-co-glycolide acid) evaporation method, strips
(PLGA) was added to water phase containing hydroxypropyl methyl cellulose, poly ethylene
glycol, polyvinyl alcohol (PVA) and aspartame in a rate of 1.5 drop s1 and agitated at 1200 rpm. History
The size, polydispersity index (PI) and drug entrapment (DE) were measured. The emulsions
were cast to form films, which were evaluated physico-mechanically. The effect of different Received 3 April 2014
degrees of hydrolization of PVA and polymerization of PLGA and the effect of different ratios of Revised 16 June 2014
PVA to PLGA was studied. Films with acceptable physico-mechanical properties were further Accepted 17 June 2014
studied. The size and PI of the nanoparticles was dependent on PVA hydrolization, PLGA
polymerization and the ratio of PVA to PLGA. All films disintegrated in less than one minute, but
acetaminophen was not free in the dissolution media even after six days. These results may
For personal use only.

indicate that although the nanoparticles released from the films immediately when impressed
in solution the drug is sustained in the nanoparticles for longer time, which is to be clarified in
future work.

Introduction using PLGA nanoparticles in drug delivery (e.g. Lupron

DepotÕ ) (Farokhzad & Langer, 2006), there are still few
In the past few decades, poly(lactide-co-glycolide acid)
formulas in the market, which may be due to a number of
(PLGA) has been extensively used for developing both
different reasons.
microparticulate and nanoparticulate drug delivery systems.
First, low physical stability of nanoparticles is a key
PLGA has several advantages, such as good mechanical
problem that has not yet been solved. Nanoparticles in
properties, low immunogenicity, low toxicity, excellent bio-
solutions have high surface free energy, which may be related
compatibility and predictable biodegradation kinetics
to their large surface to volume ratio. Therefore, nanoparticles
(Soppimath et al., 2001). Furthermore, PLGA is believed to
tend to precipitate or aggregate into larger particles within a
increase the amount of drug absorbed through mucosal
short time in order to overcome the high energy level. In order
membranes (Astete & Sabliov, 2006). Because of that
to solve these instability issues, nanoparticles are usually
enhancement effect, the polymer was used for the delivery
lyophilized and stored in a solid form, then redispersed
of a variety of drug classes ranging from classical drugs to
directly before being used (Pomogailo & Kestelman, 2005).
peptides and even for proteins (Vandervoort & Ludwig,
Second, nanoparticles properties are highly sensitive to the
2002). A large number of publications employed PLGA for
method of preparation. For instance, the drug is either
the preparation of sustained release preparations using nano
dissolved, entrapped, encapsulated or attached to a nanopar-
and microparticulate systems (Anderson & Shive, 1997;
ticle matrix, and depending upon the method of preparation,
Fonseca et al., 2002). The administration routes vary from
nanoparticles, nanospheres or nanocapsules can be obtained
parenteral (Mu & Feng, 2003), oral (Coombes et al., 1997),
(Gupta & Kompella, 2006; Schmid, 2010). It has also been
dermatological (de Jalon et al., 2001), pulmonary (O’Hara &
shown that changing the rate of adding the organic phase to
Hickney, 2000) and nasal (Tobio et al., 1998) to ocular(Veloso
the aqueous phase or the viscosity of the aqueous layer affects
et al., 1997). Unfortunately, after years of FDA approval for
the particle size, and hence, the drug release. These variations
lead to variations in size, which might range from 10 to
1000 nm or sometimes lead to high polydispersity of the
prepared patches.
Address for correspondence: Rutesh H. Dave, Associate Professor, Long Furthermore, drug solubility is one of the challenges; it is
Island University, Division of Pharmaceutical Sciences, 75 DeKalb
Avenue, Health Sciences Center, 6th floor 614A, Brooklyn, NY 11201, considered easier to entrap hydrophobic drugs into PLGA
USA. Tel: (718) 488-1660. E-mail: rutesh.dave@liu.edu nanoparticles compared to hydrophilic drugs due to the
 2 N. K. Al-Nemrawi & R. H. Dave
hydrophobic nature of PLGA molecule. This hydrophobic
nature of the polymer drives away hydrophilic drugs while the
nanoparticles are formed, leading to low entrapment effi-
ciencies. As a consequence, a large number of studies have
used hydrophobic but not hydrophilic drugs (Teixeira et al.,
2005; Budhian et al., 2007).
Fast-dissolving films are rapidly gaining interest in the
pharmaceutical industry. These thin films are designed to
dissolve within a few seconds without the need for water or
chewing. The introduction of fast-dissolving dosage forms has
solved some problems encountered in the administration of
drugs to pediatric and elderly patients. This convenience
provides both marketing advantages and higher patient
compliance (Mashru et al., 2005; Kulkarni et al., 2010).
Generally, fast-dissolving films are made of plasticized
hydrocolloids or blends. Therefore, formulation of these
systems is usually simple. This can be accomplished either by
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the solvent-casting method or the hot melt extrusion method.
In the solvent casting method, the polymer and drug are
dissolved or dispersed in the solvent, which is often ethanol or
water, and a film is cast by solvent evaporation. In the
extrusion process, the drug and other ingredients are mixed in
a dry state, subjected to the heating process and then extruded
out in a molten state. In this process, solvents are completely
eliminated. The strips are further cooled and cut to the desired
size. The high temperature used in the latter method may
degrade thermolabile active pharmaceutical ingredients. For
For personal use only.
this reason, the solvent-casting method is mostly employed in
manufacturing strips (Sharma et al., 2005).
Different polymers had been used to formulate these films,
such as polyvinyl alcohol (PVA), polyvinylpyrrolidone, malto-
dextrin, microcrystalline cellulose, hydroxypropyl methyl
cellulose (HPMC), modified starch, chitosan, gums or blends
of these polymers (Dixit & Puthli, 2009). One of the drawbacks
of using orally disintegrating film (ODF) is the drug solubility,
where the drugs used should have sufficient water solubility
and high intraoral absorption. If poorly soluble drugs are to be
used, then the solubility should be enhanced. This was
achieved previously using the salt form or a complex of the
drug with other polymers (Gupta et al., 2011).
Absorption of the drug from the sublingual mucosa is
another challenge facing ODF formulation. The sublingual
mucosa is considered relatively permeable due to its thin
membrane and large veins beneath (Ah, 1998). However,
most of the studies carried out concluded that there is a need
to co-administer a penetration enhancer in order to improve
drug absorption. Penetration enhancers can range from
nonionic to ionic surfactants, and even to steroidal detergents
(Aungst et al., 1996); unfortunately, most of these enhancers
are considered to have irritating effect on the oral mucosa,
which in turn limits their clinical application. Furthermore
both cationic and anionic surfactants are considered toxic
since they can damage the mucosa even at relatively low
concentrations (Lindenberg et al., 2004). Therefore, new safe
and effective approaches are needed.
Acetaminophen was studied thoroughly in the past few
decades and is considered to be safe. It is widely used as
an analgesic/antipyretic that is routinely sold as an over-
the-counter product world-wide. Acetaminophen is classified
as a BCS III drug (high solubility and low permeability).
Drug Deliv, Early Online: 1–10
Acetaminophen is highly soluble with 12.78 mg dissolved in
1 mL of water at 20  C. Its low permeability was referred to
the first pass effect and degradation of the drug while passing
through the GI tract. The absolute bioavailability of 325 mg
tablet was found to be 79% (Ameer et al., 1983).
In this study, we tried to prepare acetaminophen-loaded
nanoparticles to be cast directly, while still in the emulsion
form, into ODF. By casting the nanoparticles in the films, we
expected to keep the particles in a stable form where the
nanoparticles would be away from each other to prevent their
aggregation. Once the films were applied on the buccal mucosa,
they were supposed to dissolve within seconds, releasing the
nanoparticles. Then the nanoparticles could be directly
absorbed through the mucosa to the blood stream and deliver
acetaminophen there. Since nanoparticles are small enough in
size, they are expected to pass through the buccal mucosa
easily. This is why we decided not to add any penetration
enhancers, which in turn may have added another advantage to
these films of protecting the mucosal layers from harmful
chemicals. Acetaminophen was chosen to be the model drug
due to its safety as well as its challenging physic-chemical
properties (high water solubility and low permeability).
Experimental
Materials
Acetaminophen was purchased from MP Biomedicals: Santa
Ana, CA, while Methocel A3, Methocel E5 and Methocel E15
were supplied from Dow Chemical Corp.: Center Midland,
MI. Different grades of PVA (80% molecular weight (Mw) of
9000–10 000, 87–89% Mw of 85 000–124 000 and 98–99%
Mw 146 000–186 000) were purchased from Sigma-Aldrich,
St. Louis, MO. PLGA (75:25) Mw 66 000–107 000, (65:35)
Mw 40 000–75 000 and (50:50) Mw 30 000–60 000 were also
purchased from Sigma. Poly ethylene glycol (PEG) 400 was
provided by J.T. Baker: Boston, MA. All other materials and
liquids are of chemical grade.
Screening of the components for formulation of
blank ODFs
HPMC is known for its good film-forming properties and has
an excellent acceptability. Various grades of HPMC, namely
Methocel A3, Methocel E5 and Methocel E15, were
evaluated as primary film formers. Other ingredients, such
as PEG, were used as a plasticizer, and aspartame was used as
a sweetener. PVA was used for nanoparticle formation
purposes. Initially, films containing different concentrations
of HPMC, PEG and PVA were studied to scan their effect on
the film properties. The composition of various films is listed
in Table 1. All films were prepared using the solvent-casting
method (Ghorwade et al., 2011).
Briefly, HPMC was dispersed uniformly in heated water
(70  C) and left to cool down to room temperature with slow
agitation. Meanwhile, PVA, PEG and aspartame were
dissolved in water. After the addition of HPMC, the volume
was adjusted to 10 ml with distilled water; 10 ml of this
dispersion was dried in a tray drier at 40  C. The films
were carefully removed from the Petri dish and cut into strips
with the dimensions 2.5  2.5 cm and stored in an airtight
 DOI: 10.3109/10717544.2014.936987 ODFs containing acetaminophen nanoparticles 3
Table 1. Different formulation of unloaded ODF (amounts are in g ml1).

Formulae
Component F1 F2 F3 F4 F5 F6 F7 F8 F9 F10 F11 F12 F13 F14 F15 F16
HPMC A5 6% – – – – – – – – – – – – – – –
HPMC E5 – 6% – – – – – – – – – – – – – –
HPMC E15 – – – 2% 4% 6% 10% 12% 6% 6% 6% 6% 6% 6% 6% 6%
PEG 400 5% 5% 5% 5% 5% 5% 5% 5% 5% 5% 5% 5% – 10% 15% 20%
Aspartam 0.1% 0.1% 0.1% 0.1% 0.1% 0.1% 0.1% 0.1% 0.1% 0.1% 0.1% 0.1% 0.1% 0.1% 0.1% 0.1%
PVA 80% 2% 2% 2% 2% 2% 2% 2% 2% – 1% 4% 6% 1% 1% 1% 1%
water 10 ml 10 ml 10 ml 10 ml 10 ml 10 ml 10 ml 10 ml 10 ml 10 ml 10 ml 10 ml 10 ml 10 ml 10 ml 10 ml
PLGA 1% 1% 1% 1% 1% 1% 1% 1% 1% 1% 1% 1% 1% 1% 1% 1%
Acetone 5 ml 5 ml 5 ml 5 ml 5 ml 5 ml 5 ml 5 ml 5 ml 5 ml 5 ml 5 ml 5 ml 5 ml 5 ml 5 ml

glass bottle to be evaluated and compared to ListerineÕ was pulled out at the rate of 0.5 mm per second and the force
(Dinge & Nagarsenker, 2008). required to pull the films was measured.
Characterization Effect of different variables on the formulation of
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Thickness measurements nanoparticles

The thickness of each film was measured at five different
locations (center and four corners) using Vernier caliper
micrometer. Data are represented as a mean ± SD of five
replicate determinations (El-Setouhy & El-Malak, 2010).
Tensile strength measurement
The tensile strength of the ODF was measured using a texture
analyzer (TA; XT.plus, Texture Technology Corp, Hamilton,
For personal use only.
All of the nanoparticles in this work were prepared by the
emulsion-solvent evaporation method (Md et al., 2013). Five
milliliter of acetone phase was added to 10 ml of water phase
at a rate of 1.5 drops s1. The emulsions were stirred at
1200 rpm during the addition of acetone phase to the water
phase and for additional 10 min after that, at the same stirring
rate. These emulsions were studied before and after being cast
into films. In order to prepare the films, the emulsions were
poured in Petri dishes and oven-dried at 40  C for 1 h.

MA). The samples of ODF with dimension of 2.5  2.5 cm

were held vertically between two clamps of 1 cm apart at room
temperature. The ODF was pulled by the clamp at a rate of
1 mm/s and contact force of 0.05 N (Liew et al., 2012). The
tensile strength was defined as the maximum load force used to
break the ODF and calculated by dividing the applied load at
rupture with the cross-sectional area of the film. For each
formulation, six samples were measured (Dixit & Puthli,
2009).
force at break  100
Tensile strength ¼
strip thickness  strip width
Film burst
The pressure that will burst the films was measured using a
TA-108S probe TA. With a trigger force of 5 g, the
penetration distance was measured as the film was
compressed.
Disintegration time
The tensile strength of the ODF was measured using a TA.
The samples of ODF with dimensions of 2.5  2.5 cm, were
attached to a platform and tightened with screws. The probe
(TA-108S5) moved at a speed of 0.5 mm/s until the probe
touched the films. Once the probe touched the film, 1 ml of
artificial saliva was added and a trigger force of 5 g was
maintained.
Stickiness test
This test was performed using a TA-10 probe TA. The test
was run at a pre-speed of 0.5 mms1 and a trigger force of
500 g. The films were placed on a flat surface and the probe
Effect of PVA grade on nanoparticles formation
In order to determine the effect of PVA grade on
nanoparticles, different grades of PVA were used. Typically,
a solution of 1% of PLGA (75:25) and 1% acetaminophen in
5 mL acetone was added to 10 mL water phase at a rate of
1.5 drops/s and under 1200 rpm. The water phase contained
2% PVA-80%, 89% or 99%. The emulsions were studied for
particle size distribution and PI. In later steps, 6% HPMC E5
and 5% PEG 400 were added to the water phase before
dropping the acetone phase in order to clarify the effect of
HPMC and PEG on the nanoparticles. Table 2 lists the
different component of different nanoemulsions.
Effect of PLGA polymerization on nanoparticles formation
Different polymers of 1% PLGA and 1% of acetaminophen
were dissolved in 5 ml of acetone. This solution was dropped
into 10 ml water phase in a rate of 1.5 drops s1 and stirred
under 1200 rpm. The water phase contained 2% PVA-80% at
the beginning and later on contained 6% HPMC E5 and 5%
PEG 400. The emulsions were studied for particle size
distribution and PI. Table 2 lists the different components of
the different nanoemulsions.
Effect of PLGA to PVA ratio on nanoparticles formation
Finally, the effect of the ratio of PLGA (75:25) to PVA-80%
was considered by preparing different ratios (1:2, 1:1 and 2:1)
to be tested. The emulsions were prepared either with HPMC
E5 and PEG 400 or without them. Table 2 lists the different
component of different nanoemulsions.
 4 N. K. Al-Nemrawi & R. H. Dave Drug Deliv, Early Online: 1–10

Table 2. Different component of nanoparticles. the same method described previously except that the
reference films do not contain PLGA. For that reason, the
Formulae
reference films will have free-drug, that is, not inside
Component N1 N2 N3 N4 N5 N6 N7 nanoparticles. Films containing the drug equivalent of 5 mg
HPMC E15 6% 6% 6% 6% 6% 6% 6% were submerged into 20 ml glass tubes; then 10 ml of
PEG 400 5% 5% 5% 5% 5% 5% 5% phosphate buffer, pH 7.4, was added to the test tubes. These
PVA 80% 2% – – 2% 2% 1% 1% tubes were kept in a shaker (environmental incubator shaker,
PVA 89% – 2% – – – – – Edison, USA) that shook the samples vertically at a rate of
PVA 99% – – 2% – – – –
Water 10 ml 10 ml 10 ml 10 ml 10 ml 10 ml 10 ml 120 rpm at 37  C (Dinge & Nagarsenker, 2008). Samples
Acetaminophen 1% 1% 1% 1% 1% 1% 1% (200 ml) were withdrawn at 0.5, 1, 2, 4, 6 and 12 h and 1, 1.5,
PLGA 50:50 – – – 1% – – – 2, 4 and 6 d time intervals. An equal volume of fresh
PLGA 65:35 – – – – 1% – – dissolution medium maintained at the same temperature was
PLGA 75:25 1% 1% 1% – – 1% 2%
Acetone 5 ml 5 ml 5 ml 5 ml 5 ml 5 ml 5 ml added after withdrawing sample to maintain the volume.
Samples were centrifuged at 12 000 rpm at 4  C for 15 min,
and the supernatant was diluted and analyzed using
HPLC-UV. The area under the curve values were transformed
Film preparation
to concentration by reference to a standard calibration curve
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The emulsions were dried in a tray dryer at 40  C for 1 h. Then that was obtained experimentally.
the films were carefully removed from Petri dish and cut into
strips of dimensions 2.5  2.5 cm and stored in an airtight Results and discussion
glass bottle to be assayed and evaluated in vitro.
Screening of the components for formulation of
Nanoparticles characterization blank ODFs

Particle size distribution and PI
The mean particle size and PI of the empty and loaded
nanoparticles and for the films after dissolution was assessed
For personal use only.
Films that were found to be translucent, colorless and spotless
were evaluated in terms of their physico-mechanical proper-
ties, and the results are presented in Figure 1(A–C) and
summarized in Table 3. Various grades of water-soluble

by using light scattering method (Malvern Instruments,
Malvern, UK). The analysis was performed at a scattering
angle of 90 at a temperature of 25  C using samples appropri-
ately diluted with ultra-purified water. For each sample, the
mean diameter of three determinations was calculated.
Instrumentation and HPLC parameters
HPLC system (Shimadzu), equipped with an automatic
injection system (model SIL-10AF), dual solvent delivery
pumps (model LC10AS), a system controller (model SCL-
10A) and a visible/ultraviolet wavelength detector (model
SPD-10AV). Data were collected and analyzed using com-
puter software (TotalChrome, Shimadzu Scientific
Instrument: Columbia, MD).
Samples of 50 ml were injected into HPLC-UV at 207 nm.
The mobile phase consisted of acetonitrile and water (1:3), and
the pH was adjusted using phosphate buffer to 3. The flow rate
was set at 0.7 ml min1 at room temperature. Separation was
achieved using 150 mm  4 mm C18 column with a pore size
of 5 mm Spectrum Chemicals, New Brunswick, NJ).
Scanning electron microscopy
Field emission scanning electron microscope (SEM)
LEO1530VP GEMINI (Carl Zeiss, Inc., Peabody, MA) was
used to observe the distribution of the nanoparticles in the
films. A 1 mm 1 mm films were placed on top of carbon
tape. Samples were carbon-coated using a sputter coater (Bal-
Tec MED 020 HR) before analysis.
In vitro release profiles
Both reference and test films were prepared and studied
in vitro. The reference and the test films were prepared with
polymers, such as HPMC and polyethylene alcohols were
selected as primary film formers in order to obtain ODF with
rapid disintegration, good mouth feel and mechanical proper-
ties (Gupta et al., 2011). In order to consider any film for
future work, it should have the highest TS and FB properties,
which make it easier to be handled; but at the same time, the
films should still disintegrate within one minute. HPMC was
used as a primary film former due to its excellent film-
forming properties. Primary studies indicated that among
various grades of HPMC, Methocel E15 gave films the most
desired properties; highest TS and FB compared to E5 with
DT less than 1 min. Methocel A3 films were found to be so
soft that they could not be removed from the Petri dishes and
were excluded from study (F1). On the other hand, films made
with Methocel E5 and E15 were removable from the Petri
dishes and easy to be handled; but when they were compared
E15, films were 10 times more tensile than E5. For that
reason, Methocel E15 was used for future work. The initial
letter following the Methocel identifies the type of cellulose
chemistry, while the number that follows the chemistry
designation identifies the viscosity of that product in mPas,
measured at 2% concentration in water at 20  C. This might
explain why that the films prepared with Methocel E15 had
higher TS, FB and DT compared to E5. Methocel E15 has
higher viscosity, which is three times higher than that of E5,
which give it harder properties (Mahesh et al., 2010). Same
results were found when Methocel E3 was compared to E5 in
other studies were E5 had higher TS and DT compared to E3
but E15 was not studied in that reference (Dinge &
Nagarsenker, 2008).
Increasing the amount of Methocel E15 from 0 to 12% in
the films shows an increase in the TS, FB and DT. For
instance, films made with 0% and 2% Methocel were too
 DOI: 10.3109/10717544.2014.936987 ODFs containing acetaminophen nanoparticles 5
Figure 1. (A) Tensile strength (N m2) of
different formulations; (B) Film burst (N) of
different formulations; and (C) disintegration
time(s) of different formulation.
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For personal use only.

Table 3. Physico–mechanical properties of different unloaded ODFs. brittle to be removed from Petri dishes and were excluded (F3
and F4). Six percent Methocel E15 was found to provide good
Formula TS (N m2) FB (N) DT (sec) TS and FB properties with DT less than one minute (around
F2 5050 ± 2.00 822.3 ± 2.0 5 ± 0.05 30 s) and was used in the formulation of the ODF. The same
F3 780 ± 0.03 20 ± 1.0 5 ± 0.06 results were found previously in the literature (Kulkarni et al.,
F5 2650 ± 3.30 54.7 ± 1.7 30 ± 0.07 2010).
F6 13812.5 ± 3.50 457.1 ± 1.4 70 ± 0.08
F7 15297.5 ± 2.95 957.7 ± 2.0 140 ± 0.02 PVA is one of the well-known polymers that is used as a
F8 27437.5 ± 1.50 1006.4 ± 2.5 213 ± 0.1 film forming polymer (El-Setouhy & El-Malak, 2010;
F9 9915 ± 1.30 1425.6 ± 1.2 160 ± 0.9 Kulkarni et al., 2010). Another advantage of PVA is its
F10 12110 ± 1.50 880.7 ± 1.1 233 ± 0.09 stabilization effect on nanoparticles, which will be beneficial
F11 7585 ± 1.62 1776.8 ± 1.3 77 ± 0.08
F12 16940 ± 1.30 1344.4 ± 1.0 165 ± 0.05 when nanoparticles are incorporated in the films at a later
F14 10185 ± 0.98 750 ± 1.3 37 ± 0.04 stage. Unlike HPMC, PVA had no direct relation between its
F15 672.5 ± 1.14 270.4 ± 1.2 180 ± 0.09 concentration and film properties as can be seen from
F16 1005 ± 2.10 290.2 ± 1.1 105 ± 0.08
Figure 1 (F6 and F9–F12). For instance, the TS showed a
reflection point at 4%. Below this concentration, the TS was
decreased as PVA amount increases, but after 4%, the TS
 6 N. K. Al-Nemrawi & R. H. Dave Drug Deliv, Early Online: 1–10

starts to increase. In contrast, both DT and FB showed
minimal at 2%. From the results, 2% PVA was considered to
be the optimum amount to be used for preparing the ODF that
gives balanced film properties. The incorporation of PVA was
found to decrease the DT of the films compared to the films
prepared without PVA (F9). In Kulkarni’s work, a mixture of
HPMC E15 and PVA was used to prepare ODF and gave films
with a DT of 78 s. This unclear role for PVA on the films’
physical properties may be explained by the fact that PVA is
classified as an average film-forming polymer. When mixed
with a good film-forming polymer, such as HPMC, the latter’s
properties dominate and determine the final film properties
(Kulkarni et al., 2010).
As we can see from Figure 1 (F14–F16 and F6), as
PEG400 concentration increases, both FB and TS decreases.
On the other hand, the DT increases as PEG amount
increases. PEG was used as a plasticizer in this work in
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properties; 1018.5 N m2, 750 N and 37 s for TS, FB and DT,
respectively. Finally, when these films were compared to the
commercially available strips (ListerineÕ ), they were found to
have 1007.2 N m2, 136 N and 20 s for TS, FB and DT,
respectively. Therefore, these film ingredients were used in
later steps to prepare nanoparticles in films. All films
thickness was to find in the range of 0.22–0.24 mm.
Screening the components for formulation of empty
nanoparticles
From Table 4, it can be seen that the degree of hydrolyzation
of PVA plays a significant role in the nanoparticles mean size,
where the size changes from 228 nm when 80% PVA is used,
compared to 645 nm when 99% PVA is used. This might be
explained by the nature of PVA and PLGA interaction, where
the hydrocarbon chains of PVA are adsorbed on the surface of
nanoparticles via hydrophobic bonding and a large number of

order to increase the elasticity of the films (Ghorwade et al.,
2011). Five percent PEG was considered to provide optimum
TS, FB and DT; the films had the highest TS and still
disintegrated in less than one minute. The average thickness
of all films prepared in this work was 0.71 ± 0.02 mm.
All films studied recorded zero stickiness when measured
using a TA.
The primary screening showed that films with 6% of
HPMC E15, 2% PVA 80% and 5% PEG400 to have good
For personal use only.
hydroxyl groups of PVA are hydrated at the surface to
stabilize nanoparticles. As the degree of hydrolyzation
increases, stronger hydrogen bonds are formed via hydroxyl
groups between PVA molecules. This may produce
aggregated nanoparticles, leading to aggregation, and as a
consequence, an increase of the nanoparticles size.
Figure 2 also shows that as the ratio of PLGA to PVA
increases, the particle size increases from 228 nm up to
273 nm for ratios of 1:2 and 2:1, respectively. At the same
time, the PI increases as the ratio changed from 1:2 to 2:1 of

Table 4. Effect of different hydrolyzation of PVA, PLGA polymerization PLGA to PVA. The particles prepared with 2% PVA were
and PLGA to PVA ratio on the particle size and PDI. found to be stable over a sufficient period of time (data not
shown) and with acceptable size around 228 nm; hence, 2% of
Formula Mean particle size (nm) PDI DEa DEb
PVA was used in this work. PLGA polymerization also
N1 228 ± 2.0 0.0848 96.0% 34.2% affected the particle size. PLGA consists of two monomers:
N2 488 ± 0.3 0.0991 95.2% 28.9% lactic acid and glycolic acid. The data show that as the ratio of
N3 645 ± 1.0 0.1021 93.4% 40.0%
N4 303 ± 0.7 0.1770 96.1% 28.7% lactide to glycolide increases, the particle size decreases.
N5 339 ± 1.0 0.1300 96.3% 33.5% In general, adding HPMC and PEG to the water phase
N6 243 ± 3.2 0.1540 92.8% 30.0% showed an increase in the particle size compared to the
N7 273 ± 0.7 0.1879 95.0% 36.1% particles prepared in pure water as shown in Figure 2.
a
DE when acetaminophen was dissolved in water phase. This can be explained by the fact that HPMC and PEG
b
DE when acetaminophen was dissolved in acetone phase. have a major role in increasing the viscosity of the water.

Figure 2. Mean particle size of nanoparticles

before and after adding HPMC and PEG and
after loading with acetaminophen.
 DOI: 10.3109/10717544.2014.936987
The higher the viscosity of the water phase during the
preparation of nanoparticles was found to increase the particle
size. SEM images for the surface of different films are shown
in Figure 3(A–F). Images shown represent typical cases
selected from all observations. The visual observation
provides an indication of particle dispersion in the
films. Nanoparticles are well-dispersed through the film
with few aggregation of particles were observed. The
aggregation of particles might have occurred during casting
or film drying.
The size of the nanoparticles was not affected by the
method of preparation; the size was almost the same whether
acetaminophen dissolved in water or in acetone. In prelim-
inary studies, the DE was highly affected by the phase in
which we dissolved the acetaminophen. Dissolving acet-
aminophen in acetone gave above 90% DE in all formulas,
while dissolving the drug in water gave amount that is less
Drug Delivery Downloaded from informahealthcare.com by University of Newcastle on 09/14/14
than 40% entrapment.
In vitro release profile
Figure 4 shows the release profile of different films. All films
were completely disintegrated after one minute, and in less
than 5 min, they were completely dissolved. The reference
films, which were prepared in the same way as the films
with nanoparticles but without PLGA in the acetone
phase, released almost 90% of the drug after 10 min, while
For personal use only.
the test films released the drug in a sustained manner over
six days.
According to the fact that the films were dissolved within
10 min but the drug was still not free in the dissolution
medium was explained by the effect of nanoparticles on the
drug release. Although the nanoparticles were released from
the films, they retained the drug inside them and released it
very slowly. In order to test this hypothesis, a portion of the
samples withdrawn at 5, 10 and 15 min were extracted and
analyzed by HPLC-UV. These samples were not centrifuged
and instead were extracted with 2 ml of ethyl acetate and
stirred for 2 min. The supernatant was separated and
evaporated under nitrogen gas at room temperature. The
remnant was reconstituted with 150 ml mobile phase and
injected in the HPLC (West, 1981). The samples showed 90,
Figure 3. Different SEM magnifications of nanoparticles distributed in films.
ODFs containing acetaminophen nanoparticles 7
93 and 92% release of the drug, respectively. This indicates
that the film releases the nanoparticles in less than 10 min,
while the nanoparticles release the drug over a much longer
time.
Most of nanoparticle formulations show a biphasic release
with initial burst release of drug followed by sustained
release, which we also noticed in all formulas. All films
showed a burst effect release at the beginning, almost 6 h with
less than 20% of drug released (Mu & Feng, 2003). The high
burst release at the beginning may be related to free drug or
weakly bonded drug in the nanoparticles (Mittal et al., 2007).
Figure 4(A) shows that the degradation of the nanoparticles
depends on the lactic acid to glycolic acid ratio. As the lactic
acid increased from PLGA (50:50) to PLGA (75:25), the drug
release becomes slower; more sustained. This can be
explained by the fact that PLGA degrades by hydrolysis of
its ester linkages between the lactic and glycolic acids
oligomeres in an aqueous medium. And as the lactide
increases, which is more hydrophobic in nature, the degrad-
ation of the polymer decreases.
It can be seen from Figure 4(B) that the dissolution of the
nanoparticles has a relation to the PLGA to PVA ratio; the
higher the ratio of PLGA to PVA, the slower the release of the
drug. This can be explained by the hydrophobic and
hydrophilic nature of PLGA and PVA, respectively. PVA is
believed to surround PLGA nanoparticles (Sahoo et al., 2002;
Menon et al., 2012); and as its amount increases, the
hydrophilic characteristics of the surrounding environment
increase, and therefore, more water molecules are attracted to
the nanoparticles, which increase the dissolution. Other
hydrophilic surfactants such as d-a-tocopheryl polyethylene
glycol were found to have the same effect when used to
stabilize PLGA nanoparticles (Mu & Feng, 2003).
PVA is classified according to its degree of hydrolyzation
of the acetyloxyl groups, which might be completely or
partially hydrolyzed. As the degree of hydrolyzation
increases, the polymer makes more inter and intra hydrogen
bonds that makes the polymer more hydrophobic in an
aqueous solution (Nagano et al., 1970). From this fact, we
may expect that the release of the drug from nanoparticles
shall decrease as the degree of PVA hydrolyzation increases.
This relation was clear when the hydrolyzation of PVA
 8 N. K. Al-Nemrawi & R. H. Dave Drug Deliv, Early Online: 1–10

Figure 4. (A) Effect of PLGA polymerization

on acetaminophen release from nanoparti-
cles; (B) effect of PLGA to PVA ratio on
acetaminophen release from nanoparticles;
and (C) effect of PVA polymerization on
acetaminophen release from nanoparticles.
Drug Delivery Downloaded from informahealthcare.com by University of Newcastle on 09/14/14
For personal use only.
 DOI: 10.3109/10717544.2014.936987
increased from 80% to 89% as shown in Figure 4(C).
Surprisingly, nanoparticles prepared with PVA 99% showed
the fastest release rather than being the slowest. In order to
explain this, the particle size of nanoparticles was measured
during the dissolution experiment. All formulations showed
stable nanoparticle size and low PI during the six dissolution
days, except the one that was prepared with PVA 99%.
The latter had a high PI (above 0.5), which might
indicate instability, and hence explains the strange fast release
of the drug. It was suggested in other works that when
the degree of hydrolyzation of PVA is higher, the hydration
of hydroxyl groups would be hindered due to increasing
the hydrogen bonds between inter and intramolecules, which
may produce aggregated nanoparticles (Murakami et al.,
1997).
Conclusion
Drug Delivery Downloaded from informahealthcare.com by University of Newcastle on 09/14/14
This work indicates that the formulation of films with
acceptable physical-mechanical properties containing nano-
particles in one formulation step is possible. Both the film and
the nanoparticle properties were dependent on the ingredients
used in the formulation of the films. The size of the
nanoparticles increased as PVA and HPMC concentration
increase. The DE was much higher when the drug dissolved in
acetone rather than water. The drug release profile implies
that although the nanoparticles released from the films within
For personal use only.
less than one minute, the drug release from those nanopar-
ticles was sustained over 12 h.
Declaration of interest
The authors report no conflicts of interest. The authors alone
are responsible for the content and writing of this article.
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