Colloids and Surfaces B: Biointerfaces 102 (2013) 915–922

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Colloids and Surfaces B: Biointerfaces

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Preformulation studies of mucoadhesive tablets for carbamazepine

sublingual administration
Luana Perioli ∗ , Cinzia Pagano
Dipartimento di Chimica e Tecnologia del Farmaco, Università degli Studi di Perugia, Via del Liceo 1, Perugia 06123, Italy

a r t i c l e i n f o a b s t r a c t

Article history: The purpose of this research work was the realization of a bi-layered mucoadhesive dosage form intended
Received 3 February 2012 for carbamazepine sublingual administration and planned in order to obtain a unidirectional drug release
Received in revised form 1 October 2012 and diffusion only across buccal mucosa avoiding the liberation in the buccal environment.
Accepted 3 October 2012
Bi-layered tablets were constituted by an impermeable ethyl cellulose backing layer and a mucoad-
Available online 10 October 2012
hesive layer. The latter was composed by a blend of a semisynthetic polymer, as hydroxyethyl cellulose
or hydroxypropyl methylcellulose, and a synthetic polymer as, Carbopol® , physically mixed in different
Keywords:
ratios. The active ingredient carbamazepine was homogeneously dispersed in the mucoadhesive layer.
Carbamazepine
Mucoadhesion
The prepared formulations were carefully characterized by thickness, friability, swelling index, matrix
Polymers erosion, ex vivo and in vivo mucoadhesive force and time, moreover patient acceptability was evaluated
Buccal tablets as well.
Release Tablets constituted by Carbopol® :hydroxypropyl methylcellulose (25%:75%) and Carbopol® :
hydroxyethyl cellulose (75%:25%) showed the best properties and for this reason were submitted to
in vitro release studies.
Both tablet groups gave good results in terms of ex vivo and in vivo bioadhesive force and time giving
a sustained release.
© 2012 Elsevier B.V. All rights reserved.

1. Introduction this aspect represents a problem for the maintenance of therapeu-

Carbamazepine (CARBA) is an antiepileptic and psychotropic
drug mainly used in the treatment of secondarily generalized
tonic-clonic and partial seizures, trigeminal neuralgia and in the
prophylaxis of unresponsive lithium maniac depression (bipolar
disorder) [1]. According to Essential Medicines WHO Model List,
Core List, CARBA is labeled as class II drug of the Biopharmaceutics
Classification System (BCS) [2,3] because of its poor solubility. This
aspect is important as it affects drug absorption that results slow
and irregular through the GI tract reaching a lag time of 4–8 h [4,5];
the main consequence of this is the variable CARBA bioavailability.
CARBA is currently administered by oral route and because its
low water solubility (170 ␮g/ml at 25 ◦ C) high and frequent doses
are required in order to obtain the therapeutic effect. For exam-
ple, the suggested initial dose for epilepsy therapy is 100–200 mg
once or twice daily. It is gradually increased by increments of
100–200 mg every 2 weeks to an usual maintenance dose of
0.8–1.2 g daily, up to 2 g daily may be occasionally necessary. Usu-
ally, CARBA must be administered 2–4 times daily [1] however,
∗ Corresponding author. Tel.: +39 075 5855133; fax: +39 075 5855163.
E-mail address: luanaper@unipg.it (L. Perioli).
tic plasmatic levels as well as for patient compliance, in particular
for pediatric people.
The design and development of formulations for buccal admin-
istration could be an interesting approach in CARBA therapeutic
regimen improvement. In fact, CARBA administration by buccal
route was estimated by many researchers to be a suitable approach
to obtain a more efficient systemic absorption on such drugs [6,7].
Oral mucosa represents an interesting drug administration site
as it offers several advantages over both injectable and oral meth-
ods. Because of the high vascularization of this site, drugs absorbed
through the oral mucosa can enter directly into the systemic circu-
lation bypassing the GI tract and first-pass metabolism in the liver.
Moreover buccal devices can be easily administered and could be
rapidly removed if required. However, a series of factors could limit
the drug absorption across the oral mucosa, this include: exposure
to salivary flow, low absorption area, production of shearing forces
due to tongue movements, swallowing as well as short residence
time [8,9].
Since 1980s a great interest was oriented on buccal formula-
tions realized by using mucoadhesive polymers [10,11]; in fact,
these systems can offer numerous advantages in comparison to
non-adhesive solid oral dosage forms as: (i) a close contact between
the mucosa and the dosage form, (ii) a high drug concentration

0927-7765/$ – see front matter © 2012 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.colsurfb.2012.10.001
916 L. Perioli, C. Pagano / Colloids and Surfaces B: Biointerfaces 102 (2013) 915–922
maintenance at the absorption surface for a prolonged period of
time, (iii) the dosage form immobilization in a specific part of the
oral mucosa, (iv) drug protection from the environment [8,12].
The oral mucosa can be distinguished in four potential regions
suitable for drug delivery: buccal, sublingual, palatal, and gingival
[13]. Among them, the sublingual region can represent an inter-
esting administration site as it has some characteristics useful
to improve drug bioavailability in the case of active ingredients
presenting unsatisfactory biopharmaceutical properties. Thin and
highly permeable membrane of sublingual tissue is a perfect tar-
get if a prompt onset is desired. Considerable surface area and high
blood flow at this region provide a means for rapid access to the sys-
temic circulation. Moreover, sublingual region is easily accessible,
and generally well accepted from the patient [14].
The present research work deals with the realization and char-
acterization of simple bi-layered mucoadhesive tablets intended
for CARBA sublingual administration.
2. Experimental
2.1. Materials
Hydroxyethyl cellulose HEC (Natrosol® 250 HX PHARM) viscos-
ity 3400–5000 mPa sec in water solution 1% (w/v), hydroxypropyl
methylcellulose HPMC (Benecel® MP 844 PHARM) viscosity
29,000–43,000 mPa sec in water solution 2% (w/v) and ethylcel-
lulose EC (Aqualon® -T10) were purchased from Aqualon Hercules
Inc., (Wilmington, Delaware, U.S.A.). Carbopol® (Carbomer® 940)
viscosity 68,500 mPa sec in water dispersion 0.5% (w/v) was pur-
chased from Galeno (Firenze, Italy). Carbamazepine was purchased
from Sigma–Aldrich GmbH (Steinheim, Germany). Deionized water
was obtained by reverse osmosis process with a MilliQ system (Mil-
lipore, Roma, Italy). Other chemicals were of reagent grade and
were used without further purification.
Simulated salivary fluid (SSF) at pH 6.4 was prepared according
to Farmacopea Ufficiale Italiana (F.U. XII Ed.). One liter of SSF had
the following composition: Na2 HPO4 (1.79 g) KH2 PO4 (1.36 g), NaCl
(7.02 g) deionized water (1000 ml).
2.2. Tablet manufacturing
Bilayered tablets were prepared in 2 stages using a 12-mm-
diameter die, by a manual tableting press (PerkinElmer, Cambridge,
UK) using a compression force of 1.5 × 103 kg for a total time of 30 s.
Each tablet layer was prepared starting from a single powder (in the
case of backing layer) or a blend, gently mixed by mortar and pestle
in the case of the mucoadhesive layer.
Firstly the backing layer was prepared by EC (100 mg) compres-
sion for 15 s, then the punch was lifted and 100 mg of the second
mucoadhesive layer blend, mixed to 50 mg of CARBA, was added
to the first preformed layer and compressed for 15 s, obtaining
the bi-layered tablet. Tablet thickness was measured (n = 10) by a
micrometer (Borletti, Cremona, Italy).
Drug free tablets for in vivo mucoadhesion studies have been
also prepared following the same procedure described above (with-
out CARBA addition in the mucoadhesive layer).
2.3. Physical characterization: friability
Friability was determined according to F.U. (XII Ed.) by sub-
mitting 20 previously weighed tablets to falling shocks for 4 min
in a friabilator (Erweka TA 200), set at 25 rpm/min. After 4 min,
the tablets were reweighed and the percentage friability was
calculated.
2.4. Swelling studies
Tablet swelling properties (hydration %) and matrix erosion or
dissolution (DS), were measured following a procedure previously
reported [15,16]. Each tablet was weighed (W1) and put into a
petri plate (9 cm diameter) containing SSF (F.U. XII Ed.) then ther-
mostated at 37.0 ± 0.1 ◦ C in a ventilate heater (Orbital Incubator,
Sanyo Gallenkamp, Japan). The experiments were performed in
triplicate and data were calculated using Eqs. (1) and (2):
(W 2 − W 1)
% of Hydration = × 100 (1)
W2
W1 − W3
DS = × 100 (2)
W1
where W1 is the dry tablet weight, W2 is the tablet weight after
immersion in SSF for predetermined time intervals (0.5, 1, 2, 4, 8
and 12 h). W2 was measured after that SSF excess removing from
the tablets using filter paper. W3 is the weight of the swollen tablet
measured after its drying at 60 ◦ C for 24 h and then placed for 48 h
in a desiccator containing CaCl2 . This experiment was performed in
triplicate and expressed as mean ± SD (error bars are not reported
in the graphs in order to allow a clear analysis of each profile).
2.5. Ex vivo mucoadhesion time
Ex vivo mucoadhesion time studies were performed (in tripli-
cate) after tablet application on freshly cut porcine buccal mucosa.
Pig buccal mucosa, obtained from Large White pigs, weighing
∼165–175 Kg, was furnished by Veterinary Service of ASL N.1 Città
di Castello (Umbria, Italy) and used within 12 h from pig sacrifice.
The porcine buccal tissues (2 cm × 2 cm) were fixed in the internal
side of a beaker by cyanoacrylate glue. A side of each tablet was
wetted with 50 ␮L of SSF and then attached to the porcine buccal
tissue by applying a light force (∼0.5 N), by a fingertip, for 20 s. Then,
the beaker was filled with 1800 mL of SSF and kept at 37.0 ± 0.1 ◦ C;
after 2 min a stirring rate of 150 rpm was applied to simulate the
exposure to salivary flow in the buccal cavity. Tablet behavior and
mucoadhesive time were monitored until that complete detach-
ment or matrix erosion occurred.
2.6. Ex vivo mucoadhesion force
The ex vivo adhesion force was assessed by a dynamome-
ter (Didatronic, Italy) using the abovementioned porcine buccal
mucosa. The tablets (baking layer) were attached on a support, con-
nected to the dynamometer, using cyanoacrylate glue. A piece of
porcine buccal mucosa (2 cm × 2 cm) was glued onto a support and
kept in a vessel placed in a thermostatic bath at 37.0 ± 0.1 ◦ C. The
free side of the tablet was wetted with 50 ␮L of SSF and attached
to porcine buccal tissues by applying a light force with a fingertip
for 20 s. The vessel was filled with SSF and kept at 37.0 ± 0.1 ◦ C. The
measurements started after 2 min. The maximum adhesive forces
were the average of three measurements (n = 3) and the confidence
interval was determined at a 0.05 significance level.
2.7. In vivo evaluation of organoleptic properties and
acceptability
The followed procedures were in accordance with the ethical
standards of the responsible Committee on human experimenta-
tion (regional) and with the Helsinki Declaration. The studies were
performed (in triplicate) by applying drug free tablets on sublin-
gual mucosa of 3 healthy volunteers in order to assess the residence
time, the organoleptic characteristics, the fragment loss, the sali-
vary level variation, and the possible production of irritation or
 L. Perioli, C. Pagano / Colloids and Surfaces B: Biointerfaces 102 (2013) 915–922
pain. Each tablet was attached to the sublingual mucosa by applying
a light force with a fingertip for 20 s.
2.8. Stability studies
CARBA bi-layered tablets were put into a glass vial, then placed
at 40 ± 2 ◦ C and 75% RH ± 5% and then examined after 1, 3 and 6
months.
2.9. In vitro release studies
F.U. (XII Ed.) standard paddle apparatus, properly modified, was
used to evaluate drug release by using SSF added by 1% of sodium
lauryl sulfate (SLS) as dissolution medium [17–19] thermostated at
37.0 ± 0.1 ◦ C. The dissolution test was performed by two different
methods: A and B.
2.9.1. Method A [20]
The tablet backing layer was attached by cyanoacrylate glue
at a height of 7.0 cm to the inside of the glass dissolution vessel
then filled with 1000 ml the dissolution medium maintained at
37.0 ± 0.1 ◦ C. Paddle rotation was set to 150 rpm.
2.9.2. Method B [21]
The tablet backing layer was fixed by cyanoacrylate glue to a
support and positioned on the bottom of the vessel then filled with
1000 ml of the dissolution medium maintained at 37.0 ± 0.1 ◦ C. Pad-
dle rotation was set to 50 rpm.
In both cases four-milliliter samples were collected at pre-
determined time intervals and replaced with an equal volume of
fresh SSF. The samples, collected at established times, were fil-
tered through a cellulose membrane (Filter paper Whatman 41,
Whatman GmbH, Dassel, Germany) and drug concentration was
determined by a UV-vis spectrophotometer (Agilent mod. 8453).
Drug calibration curve in SSF added by SLS 1% was previously pre-
pared (max = 285.0 nm, r = 0.9998). Tests were made in triplicate
and results were registered as an average ± SD.
3. Results and discussions
3.1. Tablet preparation and characterization
The design of mucoadhesive buccal tablets must take into
consideration some important points in order to meet patient
compliance. The main requirements are: low size, flexibility and
compatibility with mucosal surface, pain free, good taste, easy
application and easy removal (if necessary).
On the basis of these characteristics, bi-layered mucoadhesive
buccal tablets intended for sublingual mucosa application have
been designed and developed. These formulations were planned
in order to promote drug release only from the side in contact
to the sublingual mucosal surface avoiding the release in sali-
vary fluids. For this reason tablets were provided for two layers: a
mucoadhesive layer, constituted by a simple blend of two mucoad-
hesive polymers, in which the drug is dispersed, and a backing
layer (Fig. 1). In particular, the mucoadhesive layer (100 mg) was
prepared by mixing a semisynthetic polymer (cellulose deriva-
tive) with a synthetic polymer (polyacrylic derivative) combined
to CARBA (50 mg), whereas the backing layer was realized by EC
(100 mg). The cellulose derivative combination to a polyacrylic
derivative comes from the necessity to produce a formulation
showing good mucoadhesion performances and able to guarantee,
at the same time, a high patient acceptability. It is noteworthy in
fact, that polyacrylic derivatives alone possess better bioadhesive
properties than cellulose derivatives as the high carboxylic groups
917
Fig. 1. Schematic representation of swellable by-layered tablet.
concentration makes them able to adhere strongly to mucosal sur-
face provoking sometimes pain. The employment of a mixture
of polyacrylic polymers and polymers with different characteris-
tics, as cellulose derivatives, can mitigate the strong interaction
with mucosal surface obtaining a good mucoadhesion and avoid-
ing pain for the patient. Among the synthetic polymers available,
Carbomer® 940 (Carbopol® ) was the polyacrylic derivative chosen
for tablet preparation while HEC and HPMC were selected among
the cellulose derivative polymers; nine kinds of tablets of different
compositions have been prepared (Table 1).
The bi-layered tablets were prepared by direct compression
(DC) in two different steps. DC represents an efficient and easier
process, if compared to others, because it involves only dry blend-
ing and compaction of drug and polymers. Additionally, DC is a
cheap process, requires low processing times, low labor costs, few
manufacturing steps, low number of equipments, low validation
and reduced power consumption, moreover, heat and moisture
are avoided. Although DC simplicity and cost-effectiveness, this
process is highly influenced by powder characteristics thus, the
selection of suitable excipients is one of the crucial steps that could
make a success in DC processing. In fact, powder properties can
influence greatly blend compaction behavior and tablet perform-
ances thus, the excipients chosen for tablets preparation by DC
must be manageable and machinable.
In regard to the prepared tablets (Table 1), tablets 1–6 were the
formulations showing high manageability and machinability while
in the case of tablets 8 and 9 (Table 1) some problems were encoun-
tered. In particular, tablet 8 was characterized by sticking while 9
underwent to capping during its ejection from the die, for these
reasons, these two tablets were discarder from the study. Pow-
ders behavior during compression can be explained analyzing the
compositions reported in Table 1. Polymers blend displaying high
manageability and machinability were those prepared by HPMC
alone (tablet 1) or mixed to Carbopol® (tablets 2, 3 and 4). In fact, t is
noteworthy, that HPMC is an excipient largely used in DC due to its
high compactability and good flow properties while the bad tablet-
ing properties in the case of polymeric mixtures used for tablets 8
and 9 could be attributable to HEC poor flow properties.
The polymeric composition must be chosen in order to obtain
final tablets with possessing mechanical properties able to confer
resistance to capping, abrasion or breakage under storage condi-
tions, transport and handling before usage. Thus, this aspect was
evaluated by submitting tablets 1–7 to friability test. The weight
loss resulted lower than 1% (0–0.17%) for all tablets meaning that
all the polymeric blends used confer good compactness, mechani-
cal resistance and low friability to final tablets. This is an important
aspect as tablets must keep their original shape and characteris-
tics during their packaging, shelf-life until their removal from the
blister and the application in the buccal cavity.
The adhesion of a mucoadhesive formulation to mucosal surface
is the results of a series of events involving the mucoadhesive poly-
mers the mucin, the main component of the gelled layer (mucus)
covering the mucosa. Once in contact with salivary fluids, the
mucoadhesive layer swells generating a gelled layer (Fig. 2). When
a polymer is immersed into a fluid, water penetrates into the
918 L. Perioli, C. Pagano / Colloids and Surfaces B: Biointerfaces 102 (2013) 915–922
Table 1
Tablet compositions.
Tableta Mucoadhesive layer
Carbopol® 940 (mg) HEC (mg) HPMC (mg)
1 0 0 100
2 25 0 75
3 50 0 50
4 75 0 25
5 100 0 0
6 75 25 0 100
7 50 50 0 100
8 25 75 0 100
9 0 100 0 100
a
For each tablet the mucoadhesive layer (100 mg) was added by 50 mg of CARBA
free spaces of the surface among the macromolecular chains. As
water content increases, the polymer becomes hydrated forming a
tridimensional network in which water molecules are entrapped,
generating a gel-like structure. The swelling capacity is very impor-
tant as it increases the polymer chain conformational mobility
enhancing the mucoadhesive force due to a concomitant increase in
chain diffusivity and degree of entanglement with the possibility
to expose a great number of hydrophilic groups to mucosal sur-
face. Moreover, polymer swelling degree plays an important role
in conditioning drug release kinetic. For these reasons the swelling
capacity, expressed as hydration percentage, was determined for
each tablet.
The highest swelling was obtained for tablets 1 and 7 that
hydrated rapidly reaching 48% and 53% respectively after 1 h and
86% and 91% respectively after 8 h (Fig. 3). In the case of tablets
2, 3, 4 and 6 similar hydration profiles were registered with an
hydration % of ∼30% after 1 h and ∼87% after 8 h. Tablet 5 showed
a reduced swelling capacity reaching 72% after 8 h. These results
could be explained considering that polymer swelling is influenced
by (a) the initial wetting processes involving tablet surface, (b) the
amount and type of polymer in tablet composition.
The wetting process is strictly connected to tablet excipient
nature. As hydrophilic polymers show high water affinity, thus
polymer chains enter in close contact with water molecules. This
means that the prevalence of a cellulose derivative (HPMC or HEC)
in tablet composition will make the formulation more hydrophilic
with enhanced wetting properties.
In regard to point b, the behavior of tablets 1 and 7 could be
explained considering that cellulose derivatives HPMC (tablet 1)
and HEC (tablet 7), are able to establish numerous bonds with
water molecules generating rapidly a gelled structure because of
the large number of hydrophilic groups in their structure. Tablet 7
(containing 50% of HEC) swelling capacity prevails against tablet
1 (containing 100% of HPMC), despite HEC shows the higher
Fig. 2. Picture of the by-layered tablet after swelling in SSF. The backing layer is
unaltered while the mucoadhesive layer hydrated generating a gelled structure. In
order to distinguish the mucoadhesive layer it was colored in methylene blue to the
mucoadhesive polymers mixture.
Backing layer EC (mg) Thickness (mm ± SD) (n = 10, ˛ = 0.05)
100 1.503 ± 0.51
100 1.393 ± 0.51
100 1.380 ± 0.88
100 1.353 ± 0.51
100 1.333 ± 0.33
1.373 ± 0.22
1.380 ± 0.00
Not measured
Not measured
hydrophilicity compared to HPMC [22]. The low hydration capac-
ity observed for tablet 5 is ascribable to Carbopol® (100%) low
water affinity that makes it unable to bind a high amount of
water molecules. Regarding tablets 2, 3 and 4 the obtained results
reveal that the increase of Carbopol® content in their composi-
tion (Carbopol® :HPMC ratios of 1:3, 1:1 and 3:1 tablets 2, 3 and
4 respectively) does not affect significantly the swelling capacity.
In the case of tablet 6 the prevalence of Carbopol® on HEC (75%
vs 25%) is responsible for the low hydration percentage. During the
first hour HEC chains relaxation takes place, with consequent tablet
swelling, as water penetrates into the matrix.
From the analysis of hydration curves obtained for each tablet,
two different phases could be distinguished (Fig. 3). Error bars have
not been indicated for graph clarity as these may be confusing since
many profiles overlap. The first phase is represented by 0–8 h range,
in which the gradual water content increase is evident for all the
formulations. Then, in the range 8–12 h (second phase), a reduced
hydration percentage is registered, justified by tablet fragment lost.
The latter was assessed by the erosion matrix assays (DS) evaluated
along 12 h. During the first 8 h all the tablets show similar behav-
iors, then DS% values increase as fragment loss occurred (Fig. 4).
Also in this case error bars have not been indicated for graph clarity
as these may be confusing since many profiles overlap. The high-
est value was measured for tablet 5 which, after 12 h, reached a
DS percentage of 8.77%. These observations are in according to the
low hydration % registered for tablet 5. The low Carbopol® water
affinity in fact, induces a slow and limited swelling combined to low
water penetration with high tendency to give fragments. Moreover,
Fig. 3. Hydration % vs time determined for all tablets in SSF at 37.0 ± 0.1 ◦ C at estab-
lished times (0.5, 1, 2, 4, 8, 12 and 24 h).(n = 3, error bars are not reported for a clear
graph analysis.)
 L. Perioli, C. Pagano / Colloids and Surfaces B: Biointerfaces 102 (2013) 915–922 919

attributable to HPMC presence. In fact, HPMC hydrophilic character

Fig. 4. Tablet DS% vs time determined for all tablets in SSF at 37.0 ± 0.1 ◦ C at estab-
lished times (0.5, 1, 2, 4, 8, 12 and 24 h). (n = 3, error bars are not reported for a clear
graph analysis.)
Carbopol® hydration generates a thick and viscous gel that eas-
ily undergoes to fragmentation because of hydrodynamic pressure
effect, exerted on the polymer network by water. Polymer swelling
is an important process involved in tablet adhesion to mucosal
surface. In fact, this water-activated process produces polymer
swelling and improves the consolidation step that increases the
polymer chains mobility facilitating the interpenetration with the
mucin chains of the mucus layer [23].
The bioadhesion capacity of the employed polymers is
attributable to the presence of amino and carboxylic groups in
their structure that make them able to form hydrogen bonds
with the mucosal components [23]. Tablet ability to interact with
buccal mucosa was firstly evaluated by ex vivo mucoadhesion
assays with the aim to measure the adhesion force and time
of tablets 1–7 (Table 2). Obtained data revealed that all tablets
require a short time to bind mucosal surface and that are able to
adhere for a prolonged time (Table 2). Tablet 6 (Carbopol® :HEC
75%:25%) and 7 (Carbopol® :HEC 50%:50%) showed the lower
mucoadhesion time in comparison to other tablets and this can be
explained by HEC presence in both tablets 6 and 7 compositions. As
mentioned before, tablet swelling behavior influences the mucoad-
hesion performances. Tablet 1 (HPMC 100%) required less swelling
time (15 minutes) in comparison to tablets 2 (Carbopol® :HPMC
25%:75%) and 3 (Carbopol® :HPMC 50%:50%, about 3 h), while tablet
4 (Carbopol® :HPMC 75%:25%) and 5 (Carbopol® 100%) didn’t swell
during all the period of contact with mucosa. Tablets 3 and 7
displayed the highest time (after 71 h and >10 h respectively) in
comparison to the remaining formulations that were discarded
from the successive studies. All the tablets maintained the bi-
layered structure for the entire experiment.
From ex vivo mucoadhesion force data analysis some consider-
ations can be drawn. For tablet 1 a low force value was registered,
favors tablet swelling in a short time after the contact with the SSF
because of the high water affinity, HPMC chains bind a high number
of water molecules thus, a small amount of chemical groups remain
available to interact with mucin. In regard to Carbopol® :HPMC
blend, the 1:3 ratio (tablet 2) confers high mucoadhesion capacity
as HPMC hydrophilicity allows an easy water penetration into the
matrix promoting the relaxation of Carbopol® chains then avail-
able to come in a very close contact with the mucus layer. Using
the same ratio of Carbopol® blended to HPMC or HEC (tablets 3
and 7 respectively) the mucoadhesion force decreases as tablet
swelling capacity is reduced because only a limited number of
groups are available for binding. The same observation could be
done for tablet 6 for which the low amount of HEC is responsible
for a limited swelling. In the case tablet 4, in which Carbopol® (75%)
content prevails, the presence of HPMC (25%) is important because
water molecules are able to penetrate and to promote a gradual
chain enlargement. In this way numerous carboxylic groups are
available to interact with mucin chains. This promotes a close and
strong contact with the mucosal surface, as demonstrated from
ex vivo mucoadhesion force data (Table 2) that could be respon-
sible for pain and irritation. In the case of tablet 5 the lack of a
cellulose derivative reduces the water access in the inner space
of the mucoadhesive layer bringing to a reduced interaction with
the mucosal surface. The characterization of tablet mucoadhesion
capacity was completed by in vivo assays having the objective to
evaluate patient acceptability and compliance. In particular, the
investigated parameters were: (1) bioadhesion capacity, (2) adhe-
sion time, (3) swelling capacity and patient acceptability, (4) pain
or irritation, (5) taste, (6) fragment lost. These studies are very
important in the choice of the best formulations to submit to the fol-
lowing in vitro release studies. With this aim drug free tablets were
prepared and applied on the sublingual mucosa of three healthy
volunteers. Formulation behavior was evaluated in the elapsed
time from tablet administration to the total detachment. During
this period the volunteers were able to eat and drink. Tablet 1 did
not show adhesion to mucosal surface (Table 2) and this can be
due to the high and quick water absorption due to HPMC pres-
ence (as observed in the hydration studies, Table 2) forming a
slippery non-adhesive mucilage which limits tablet bioadhesion.
Tablet hydration in fact, generates a weak gel which could be eas-
ily removed from the mucosal surface. Tablets 4 and 5 showed a
high adherence to mucosa, however it was so strong to provoke
pain and irritation after few minutes from the application. In both
cases the high percentage of Carbopol® in the mucoadhesive layer
(75% and 100% for tablets 4 and 5 respectively) is responsible for
the high binding to mucosal surface. In fact, Carbopol® shows a high
water absorption capacity that, associated to carboxylic groups ion-
ization, is responsible for the gradual relaxation of the polymeric
chains. In these conditions the carboxylic groups are able to enter
in close contact with the mucosal surface establishing hydrogen

Table 2
Ex vivo mucoadhesion times and forces and in vivo performances.

Ex vivo In vivo

Tablet Mucoadhesion Swelling Mucoadesion Bilayer integrity Mucoadhesion Pain after Fragment loss Taste
time (h) force (N ± SD) time (h) application
(n = 3; ˛ = 0.05)

1 >73 After 15 min 0.70 ± 0.12 Yes No adhesion – — —

2 >73 After 4 h 2.20 ± 0.27 Yes 17 No No Good
3 >73 After 71 h 1.66 ± 0.11 Yes No adhesion – – –
4 >73 no 5 Yes –a Yes – –
5 >73 no 1.03 ± 0.45 Yes –a Yes – –
6 44–46 After 3 h 1.66 ± 0.26 Yes 12 No – –
7 44–46 >10 h 1.73 ± 0.56 Yes No adhesion – – –
a
The mucoadhesion time, fragment loss and taste have not been estimated as the table have been removed after application because painful.
920 L. Perioli, C. Pagano / Colloids and Surfaces B: Biointerfaces 102 (2013) 915–922

bonds with the mucus layer. In the case of tablet 4, HPMC (25%)
improves the mucoadhesion capacity in comparison to Carbopol®
alone as, because of its lower water absorption capacity, it is able to
promote a gradual swelling encouraging the interaction with mucin
chains. Good adhesion for long time was observed for tablets 2 and
6 that were kept in the mouth for all the tests without pain for the
volunteers.
During the manufacturing process, as well as the storage con-
ditions, the dosage form is exposed to physical agents, such as
humidity, that could provoke changes of the shape and layers
adherence that compromise the efficiency of the dosage form.
This aspect is also important as tablet constituents are mucoad-
hesive polymers that, in presence of water, become sticky. Thus,
In order to evaluate tablet susceptibility to humidity, stability tests
were executed following the indications proposed from the Euro-
pean Agency for the Evaluation of Medicinal Products [24]. For
solid dosage forms, the guidelines prescribe to keep the samples
at 40 ± 2 ◦ C and 75 ± 5% RH for 6 months (accelerated tests). Each
sample (in triplicate) was put into a glass vial and then placed
at constant temperature and humidity (40 ± 2 ◦ C and 75% RH ± 5%
respectively). After 1 month each tablet was accurately examined;
tablets 1, 3 and 7 showed a partial layer separation (7 > 3 > 1), tablets
4 and 5 became sticky, while tablets 2 and 6 resulted unmodi-
fied and for this reason were kept at 40 ± 2 ◦ C and 75% RH ± 5% for
further 5 months. At the end of this period these tablets resulted
unchanged.
From the characterization studies performed, resulted that
tablets 2 and 6 were the best candidate for CARBA sublingual
administration as they adhere very well on sublingual mucosa, for
a long time without pain or irritation for the patient. Thus, these Fig. 5. In vitro release profiles of tablets 2 and 6 in SSF added by SLS 1% at 37.0 ± 0.1 ◦ C
two formulations have been submitted to further studies and the (n = 3, mean ± SD). (A) Dissolution performed positioning the tablet on internal
in vitro drug release was investigated. vessel surface at 7 cm from the bottom (Method A), (B) dissolution performed posi-
tioning the tablet on vessel bottom (Method B).

3.2. In vitro release studies

As official assays for mucoadhesive sublingual tablets are not
reported, this study was carried out by using the paddle apparatus
(F.U. XII Ed.), properly modified. In particular, in order to make the
assay bio-relevant, two different methods were followed in dissolu-
tion tests considering two possible tablet positions in the vessel as
reported from Miyazaki et al. (Method A) and Remuñán-López et al.
(Method B) [20,21]. The dissolution test was performed (in tripli-
cate) following the methods A and B described in method section. In
fact, the swelling capacity plays an important role in drug release,
connected to the formed gel properties as well as the dissolution
method used.
Regarding the release profiles of tablets 2 and 6 submitted to
method A (Fig. 5A) it results that the amount of CARBA released
from tablet 2 is higher in respect to tablet 6 for the entire test.
Specifically, after 8 h tablet 2 releases 35% of drug vs 17%, reaching
75% vs 68% after 24 h. The different release is attributable to the
different Carbopol® amount in the two tested formulations. The
high amount of Carbopol® in tablet 6 (75%), in fact, once in contact
with the dissolution medium, creates a viscous gel that delays and
reduces drug diffusion, moreover, the low HEC amount is respon-
sible for a limited formulation swelling. In the case of tablet 2,
HPMC (75%) predominance confers high water affinity to the for-
mulation allowing an easy water penetration into the matrix; the
polymer network results enough enlarged to promote a rapid and
high drug diffusion through the gelled layer. Also in the case of
the release profiles obtained following method B, CARBA release
from tablet 2 results better than tablet 6 (Fig. 5B), however the total
amount of released drug from both formulations is lower if com-
pared to the results coming from method A. These differences can
be ascribed to (i) the different hydrodynamics in dissolution test-
ing, mainly attributed to the influence exercised changing paddle
rotation speeds and (ii) location of the formulation in the dissolu-
tion vessel [25,26].
In regard to the first aspect, many studies performed in this field
revealed that the round bottom of a dissolution vessel, in combi-
nation to stirring devices, results in a varied flow dynamics and
surface area within the vessel inducing a high test variability [27].
Moreover, it was assessed that different fluid velocities exist at dif-
ferent points in the dissolution vessel meaning that the amount

Table 3
Ritger and Peppas’s kinetic mathematical model and first order kinetics model fitting.

Tablet Mt /M∞ = Ktn

n = 1 (zero n = 0.9 n = 0.8 n = 0.7 n = 0.6 n = 0.5 n = 0.5

order) release 0–60%
Higuchi

2 y = 3.096x + 3.451 y = 4.218x + 1.896 y = 5.812x − 0.101 y = 8.025x − 2.522 y = 11.120x − 5.538 y = 15.502x − 9.422 y = 14.626x − 8.156
r = 0.983 r = 0.991 r = 0.994 r = 0.995 r = 0.993 r = 0.985 r = 0.981
6 y = 2.443x − 0.883 y = 3.333x − 2.058 y = 4.550x − 3.446 y = 6.220x − 5.122 y = 8.528x − 7.202 y = 11.755x − 9.874 y = 10.592x − 8.147
r = 0.988 r = 0.985 r = 0.979 r = 0.970 r = 0.957 r = 0.940 r = 0.955
 L. Perioli, C. Pagano / Colloids and Surfaces B: Biointerfaces 102 (2013) 915–922
of released drug from the same formulation changes in relation
to its position in the vessel. Some studies revealed that a ‘dead
zone’ forms at the bottom of the vessel where the agitation rate
is minimum leading to low dissolution rates [28].
Concerning data coming from the dissolution tests (methods
A and B), it is possible to assert that in the case of the latter the
low paddle speed (50 rpm) and the formulation location at the bot-
tom of the vessel are responsible for the low amount of CARBA
released from both tablets 2 and 6. When the tablet is placed at
7.0 cm from the bottom of the vessel (method A) and the paddle
speed is increased to 150 rpm, the tablet is localized in a region in
which the dissolution fluid velocity is high resulting more solicited.
In these conditions tablet swelling results faster as well as drug
release.
Taking into account that this kind of tablet must be applied on
subligual mucosa and that in this region the formulation can receive
different solicitations (as salivary flow and shearing forces due to
tongue movements) method A appears to be more predictable of
physiological conditions than method B.
3.3. In vitro release mathematical model
In vitro release data were submitted to statistical investigation
in order to know the kinetic mechanism involved in CARBA release
from both tablets 2 and 6. The projected tablets are classified as
swellable matrix drug delivery systems as they are constituted
from a polymeric mixture able to hydrate in presence of water.
In these systems the drug is released from the matrix by diffusion,
which is controlled from the polymer network. In order to inves-
tigate what mechanism is involved in CARBA release, data of drug
dissolution coming from tablets 2 and 6 were fitted to Ritger and
Peppas kinetics mathematical model Mt /M∞ = Ktn [29], applied to
swellable matrices. The release mechanism can be controlled by
water penetration rate, responsible for hydration and drug diffu-
sion, and polymeric chain relaxation time. Diffusional exponent
value of one (n = 1) means that drug release occurs as an appar-
ent zero-order mechanism when is time dependent while value of
n = 0.5 means that release is controlled by a pure Fickian diffusion
mechanism. A n value between 0.5 and 1 indicates an anomalous
mechanism (not Fickian) meaning that both liquid penetration rate
and polymeric chain relaxation rate control the drug release.
Analyzing the results (Table 3) it is recognized that the best
fitting for tablet 2 was obtained for n = 0.7, meaning that drug is
released by an anomalous mechanism. For tablet 6 the best fitting
was obtained for n = 1 thus, in this case drug release follows a zero
order kinetic.
As in tablet 2 the main constituent is HPMC (75%), a polymer
with high water affinity, drug release is mainly due to diffusion
mechanisms. In regard to tablet 6 the main factor governing CARBA
release is the relaxation rate of polymeric chains. In fact, the high
Carbopol® (75%) amount reduces tablet affinity to water thus, in
the case of tablet 6 the relaxation rate is the main factor governing
CARBA release.
4. Conclusions
Mucoadhesive bi-layered tablets, constituted by an imperme-
able backing layer and a mucoadhesive layer, intended for CARBA
sublingual administration have been prepared and characterized.
The best formulations in terms of swelling capacity, mucoadhe-
sion performances and patient acceptability were those having
Carbopol® :HPMC 25%:75% or Carbopol® :HEC 75%:25% as mucoad-
hesive layer. The in vitro release studies, performed by two
different methods, showed that the highest drug release in 24 h
921
was obtained from tablet which mucoadhesive layer compositions
Carbopol® :HPMC 25%:75%.
This buccal formulation could represent an interesting approach
in the treatment of chronic disease as it is able to promote a sus-
tained CARBA release useful to reduce daily administrations in
comparison to conventional formulations. Moreover, the devel-
oped tablet is very manageable and easy to apply, favoring patient
compliance.
Conflict of interest
The authors declare that they have no personal financial or non-
financial conflicts of interest in this research.
Acknowledgments
Authors are very much grateful to Dr. Renato Innamorati from
ASL N. 1 (Umbria, Italy), for providing porcine mucosa and to Mr.
Marco Marani for technical assistance.
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