European Journal of Pharmaceutical Sciences 39 (2010) 272–278

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European Journal of Pharmaceutical Sciences

journal homepage: www.elsevier.com/locate/ejps

Improving sublingual delivery of weak base compounds using pHmax concept:

Application to propranolol
Yanfeng Wang a , Zhong Zuo a , Xiao Chen b , Brian Tomlinson c , Moses S.S. Chow d,∗
a
School of Pharmacy, Faculty of Medicine, The Chinese University of Hong Kong, Sha Tin, Hong Kong SAR, PR China
b
Department of Pharmacy, The First Affiliated Hospital, Sun Yat-sen University, Guangzhou, PR China
c
Department of Medicine and Therapeutics, Faculty of Medicine, The Chinese University of Hong Kong, Sha Tin, Hong Kong SAR, PR China
d
Center for Advancement of Drug Research and Evaluation, College of Pharmacy, Western University of Health Sciences, 309 E. Second Street, Pomona, CA 91766-1854, USA

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

Article history: The purpose of the present work was to provide theoretical and experimental support in generating an
Received 10 June 2009 optimal pH (pHmax ) for a representative weak base compound (propranolol), that can lead to enhanced
Received in revised form 23 October 2009 sublingual absorption. Initially equations for pH-solubility and pH-permeability profiles were derived
Accepted 23 December 2009
and compared to the profiles obtained experimentally. Excellent correlation (R2 = 0.999) of solubility
Available online 8 January 2010
profiles was obtained using non-linear regression, and the permeability profiles further predicted that at
certain pH (pHmax ), optimal mucosal permeation could be achieved. Subsequently, in a pharmacokinetics
Keywords:
study, a buffered sublingual propranolol tablet, designed to achieve its pHmax (when dissolved in saliva),
Sublingual
Weak base
were compared to that from a marketed product (Inderal® which could not achieve pHmax ) in 8 healthy
Propranolol subjects. Each subject received the products sublingually for 15 min followed by swallowing the remaining
Permeability drug–saliva. The plasma propranolol concentrations of AUC during first 30 min from the buffered tablet
Pharmacokinetics were significantly higher than that from the Inderal® tablet (p < 0.05), and no significant differences in the
remaining AUC were observed. These in vitro and in vivo results on propranolol provided experimental
confirmation of the pHmax concept as well as its utility in sublingual drug delivery. Such an approach may
be applicable to other similar compounds to improve sublingual drug delivery.
© 2010 Elsevier B.V. All rights reserved.

1. Introduction For weak base compounds that involve passive diffusion as a

Sublingual mucosal delivery can offer attractive therapeutic
advantages. Not only its thin non-keratinized, stratified epithelium
which can provide drug permeation rate 4–4000 times that of the
skin (Shojaei et al., 1998; Galey et al., 1976), the enzymatic degra-
dation in the gastrointestinal tract and first-pass metabolism in the
liver can be also avoided (Squier and Hall, 1985). Thus, sublingual
drug delivery is an attractive route in achieving high bioavailability
and rapid onset of action.
Despite the above advantages, relatively small number of
sublingual products are currently successfully developed and mar-
keted. An important limitation is the low quantity of drug molecule
that can be permeated across the limited area of sublingual mucosa.
Thus, new methods to improve drug transport or permeation are
needed in order to enhance sublingual drug product development.
∗ Corresponding author. Tel.: +1 909 4695301; fax: +1 909 4695539.
E-mail addresses: yfwang2000@yahoo.com (Y. Wang), msschow@westernu.edu
(M.S.S. Chow).
mechanism of transport, one approach of improving their sub-
lingual permeation or diffusion is to increase the pH of the drug
solution (Beckett and Triggs, 1967; Hao and Heng, 2003). It is well
known that based on the Henderson–Hassebalch equation, increas-
ing the pH of the solution of a weak base can increase the ratio of
unionized to ionized species, which in turn can lead to improved
transmucosal permeation.
While maximizing the unionized fraction of a weak base com-
pound is an important factor for trans-membrane permeation,
recent data indicated that the ionized species of certain weak base
compounds should likewise be taken into consideration since such
species are also capable of diffusing across the mucosal membrane
(Chen et al., 1999; Chetty et al., 2001). Thus, maximizing the solu-
bility of both the ionized and unionized species of such compounds
instead of just the unionized species can be potentially an effective
approach in improving sublingual permeation.
Previous work on pH-solubility profiles showed that it is pos-
sible to saturate simultaneously the unionized and ionized species
at a particular pH called pHmax (Kramer and Flynn, 1972). Thus at
pHmax , maximal aqueous solubility of a weak base compound is
expected and this should lead to higher transmucosal permeabil-

0928-0987/$ – see front matter © 2010 Elsevier B.V. All rights reserved.
doi:10.1016/j.ejps.2009.12.011
 Y. Wang et al. / European Journal of Pharmaceutical Sciences 39 (2010) 272–278
ity than at any other pH. The aims of this paper are (1) to derive
the quantitative relationship of pHmax , solubility and permeabil-
ity, and (2) to demonstrate the application of its pHmax concept in
improving sublingual absorption or permeation of a representative
weak base compound, propranolol, using laboratory and human
experiments.
2. Theory
The theoretical equations describing pH-solubility and pH-
permeability (flux) profiles are shown bellow.
2.1. pH and solubility profile of weak base
When a weak base is dissolved in an aqueous solution, equilib-
rium exists between the unionized species (B) and ionized species
(BH+ ), and can be described by Eq. (1):
Ka
BH+ + H2 OB + H3 O+
where Ka is the dissociation constant of the weak base and can be
expressed as:
[B][H3 O+ ] Cu [H3 O+ ]
Ka = + or
BH Ci
where [B] or Cu is the concentration of the unionized species and
[BH+ ] or Ci is the concentration of the ionized species. Eq. (2) can
be also re-written as:
[BH+ ] C At pH > pHmax
pH = pKa − log or pKa − log i
[B] Cu
At a particular pH (pHmax ), when both the unionized and ionized
species can be simultaneously saturated (Kramer and Flynn, 1972;
Ledwidge and Corrigan, 1998), Eq. (3) can be re-expressed at the
saturated state as:
Si
pHmax = pKa − log
Su
where Su and Si are the intrinsic solubilities of unionized and
ionized species, respectively. Si can be experimentally estimated
at pH  pHmax since the fraction of unionized species is very
low at such pH. Conversely, Su can be experimental estimated at
pH  pHmax since the fraction of ionized species is very low at such
pH. From these Si and Su values, the maximum solubility or total
solubility, ST , of an ionizable compound can be obtained (i.e. Si + Su ).
At any pH lower than pHmax , the concentration of the soluble
ionized species, Ci or [BH+ ] should approach its intrinsic solubility
(Si ); and the concentration of the soluble unionized species Cu or
[B] at such pH, can be then estimated by the solution pH and Si
according to Eq. (5). Similarly, at any pH higher than pHmax , Cu
should approach its intrinsic solubility (Su ); thus, Ci or [BH+ ], can
be estimated by solution pH, Ka and Su according to Eq. (6).
At pH < pHmax , ST can be represented as
Si · Ka
ST = Si + Cu = Si +
[H3 O+ ]
At pH > pHmax , ST can be represented as:
Su · [H3 O+ ]
ST = Su + Ci = Su +
Ka
2.2. Relation of permeability to solubility at different pHs
For a highly lipid soluble weak base compound like propranolol,
permeation of the total weak base across the lipid membrane is
expected to include both the unionized and ionized species. The
273
steady-state flux (amount permeated across the membrane per
unit time and area) of the ionized (Jss(i) ), unionized (Jss(u) ), and total
species (Jss(T) ) at a given constant pH solution can be expressed
using Fick’s second law:
dQi
Jss(i) = = Pi Ci (7)
dt · A
dQu
Jss(u) = = Pu Cu (8)
dt · A
dQ
Jss(T ) = = Jss(u) + Jss(i) = Pi Ci + Pu Cu (9)
dt · A
where Qi , Qu and Q are the accumulated amount of the ion-
ized, unionized and total species permeated across the membrane,
respectively. A is the permeation area, Ci and Cu are the soluble
concentrations of ionized and unionized species, respectively, and
Pi and Pu are the individual permeability coefficients of ionized and
unionized species, respectively. (Pi and Pu should be independent
of solution pH provided that the structure and physicochemical
(1)
properties of each species are not influenced by pH).
Theoretically, at pH = pHmax , both Ci and Cu will be saturated or
maximized (i.e. equal to Si and Su , respectively), therefore, the high-
est Jss(T) can be expected. Jss(T) values at pH > pHmax and pH < pHmax
(2) can be obtained using Pi and Pu and Ci and Cu (or Si , Su ) from Eqs.
(10) and (11) below.At pH ≤ pHmax
Ka
Jss(T ) = Pi Si + Pu Si (10)
[H3 O+ ]
(3)
[H3 O+ ]
Jss(T ) = Pu Su + Pi Su (11)
Ka
To generate Jss(T) -pH profile using Eqs. (10) and (11), Pi and Pu
need to be first estimated by using Eq. (9) at 2 different pHs with
corresponding Ci and Cu obtained as Su ·[H3 O+ ]/Ka and Su ·Ka /[H3 O+ ],
respectively (see Eqs. (5) and (6)). For example, for propranolol, to
(4)
simultaneously solve Pi and Pu using Eq. (9), Jss(T) values were deter-
mined experimentally at pH 6.8 and pH 9.0 (see methods below)
and used with Ci and Cu at these pH values (as discussed above)
together with value of Si and Su and pKa = 9.23. Once Pi and Pu are
known, the theoretical Jss(T) -pH profile were generated by Eqs. (10)
and (11) and are shown in Fig. 3.
3. Materials and methods
3.1. Materials
(±)-Propranolol hydrochloride, verapamil, sodium dihydrogen
phosphate dihydrate, disodium hydrogen phosphate, ortho-
phosphoric acid, lactose monohydrate hydroxypropyl methylcel-
lulose and magnesium stearate were obtained from Sigma–Aldrich
Co. (St. Louis, MO, USA). Triethylamine was purchased from BDH
Laboratory Supplies, England. HPLC grade organic solvents, includ-
ing methanol, n-hexane, isopropyl alcohol and acetonitrile, were
(5)
obtained from Lab Asian Co. (Bangkok, Thailand). Hydrochloric acid
(37%) was supplied by Merk Co. (USA). Disposable syringe filters
(0.20 ␮m) were obtained from IWAKI GLASS (Japan).
(6)
3.2. Determination of propranolol solubilities at various pHs
To determine the solubility of propranolol at different pH values,
an excess amount of propranolol hydrochloride was first added to
tubes containing isotonic phosphate buffer and their pH adjusted
using 85% (w/w) phosphoric acid or 20% (w/v) sodium hydroxide
(to achieve a pH range of 5.4–10.0). The tubes were then immersed
274 Y. Wang et al. / European Journal of Pharmaceutical Sciences 39 (2010) 272–278
in a shaking water bath at 37 ◦ C for 24 h and the pH of the solu-
tions were verified. The final mixture was filtered through a 0.2 ␮m
syringe filter (Iwaki Glass, Japan), and the total concentration of
propranolol or ST (in the filtrate) was determined by HPLC using
the method described by Nielsen and Rassing (2000) with slight
modification.
3.3. Determination of propranolol permeability
The permeability experiment was performed using fresh
porcine sublingual mucosa obtained from local slaughter house
(excised from the floor of the mouth of male adult pigs with the
epithelium layer separated from the underlying connective tissue)
within 2 h of procurement. A side-by-side diffusion chamber sys-
tem (PermeGear Co., PA, USA) was used for the permeability study.
The temperature of the system was maintained at 37 ◦ C and the
solution (isotonic phosphate buffer) in each chamber was stirred
with magnetic bars. The sublingual mucosa was then mounted
between the donor and receiver chamber with the surface of
mucosa facing the donor chamber. After a 30 min equilibration, the
receiver chamber and donor chamber were freshly filled with 4 ml
pre-warmed phosphate buffer and saturated propranolol solution
at different specified pHs, respectively. A 200 ␮l sample was subse-
quently withdrawn from the receiver chamber every 15 min until
2 h and the propranolol concentration was determined (by HPLC,
described below). Jss(T) was then obtained according to Eq. (9).
3.4. Preparation of special buffered sublingual propranolol tablet
to enhance permeability
To verify the applicability of the pHmax concept for sublingual
delivery, a buffered sublingual propranolol tablet was prepared by
direct compression of 40 mg propranolol with excipients (disodium
hydrogen phosphate, lactose, hydroxypropyl methylcellulose and
magnesium stearate). This formulation was designed to achieve a
saliva pH of 7.4–7.6, i.e. close to pHmax when dissolved in saliva
under the tongue. In addition, a marketed propranolol formulation
(Inderal® ), which achieved pH 6.7–6.9 when mixed with human
saliva, was selected for comparison.
Our buffered tablets were prepared according to the fol-
lowing steps: (1) weigh and mix the propranolol powder with
magnesium stearate in a V-blender (ERWEKA Co., Germany) for
30 min; (2) add other excipients to the mixed powder prepared
in step (1), and then commingle the powder in the V-blender for
30 min; (3) directly compress the blended composition into plain
tablets (100 mg/tab) using a rotary tablet press machine (Mei-King
Machinery Engineering Co., Hong Kong), with 6 mm in diameter
and 2–3 mm in thickness. Each tablet should contain 40 mg pro-
pranolol hydrochloride and 40 mg disodium hydrogen phosphate.
The tablet hardness, friability, weight variation, content uniformity,
disintegration time, and dissolution profile were also measured
according to the USP monograph.
3.5. Human pharmacokinetic study
To verify the enhanced absorption (permeation) using the
pHmax concept, a pharmacokinetic study was carried out in 8
healthy male volunteers (aged 18–30 years with body weight
between 50 and 90 kg) who received the special buffered propra-
nolol tablet and Inderal® tablet in a random crossover manner.
The study followed the tenets of the Declaration of Helsinki (1964)
and the study protocol was approved by the Joint Chinese Univer-
sity of Hong Kong—New Territories East Cluster Clinical Research
and Ethics Committee. The approved Informed Consent Form from
the Committee was signed by all subjects. Prior to the studies,
all subjects underwent ECG and mouth examination and received
instruction not to take any medications 2 weeks prior to and during
the period of the study. The subject inclusion criteria were: (a) male,
18–55 years old, (b) body weight within 15% of ideal weight, (c)
accessible vein for blood sampling, (d) high probability for compli-
ance and completion of the study, (e) no significant abnormalities
in general physical examination, (f) ECG recording within normal
limits, and (g) biochemical and haematological parameters within
normal limits.
The subject exclusion criteria consisted of: (a) history of hepatic,
renal, biliary, cardiovascular, gastrointestinal, haematological and
other chronic and acute diseases within 3 months prior to the study,
(b) clinically relevant abnormality in physical examination, ECG
evaluation, urine test, blood chemistry or haematological test, (c)
tobacco use in any form, (d) regular consumer of alcohol, (e) blood
donation within 4 weeks prior to the start of the study, (f) treatment
with propranolol or similar class of drugs within 4 weeks before the
study, (g) participation in any clinical drug study within 2 months
prior to the study, and (h) hypersensitivity to propranolol or other
␤-blockers.
The subjects were randomly assigned to two equal size groups
with one group receiving a single 40 mg dose of buffered tablet and
the other group receiving a 40 mg Inderal® tablet. After a washout
period of at least 1 week, each subject was crossed over to receive
the other formulation.
Alcohol, caffeine-containing products and smoking were not
permitted for 24 h prior to, and for the duration of, the study. At
the time of the clinical study, each subject was required to place
the tablet under the tongue and not to swallow any tablet or saliva
until 15 min, at which time the saliva (containing the dissolved
propranolol) was swallowed. Venous blood samples (5 ml) were
collected at 0, 3, 6, 10, 15, 30, 45, 60, 90, 150, 300 and 420 min
after the start of sublingual administration. The samples were cen-
trifuged at 2000 rpm for 10 min at 4 ◦ C and then plasma obtained
and stored at −80 ◦ C until analysis.
3.6. Propranolol assay
An HPLC system consisted of a Waters 600 controller, a 717plus
auto-sampler, a Waters 2487 dual ␭ absorbance detector, Waters
464 fluorescence detector and a Thermo Hypersil-Keystone Col-
umn (250 mm × 4.6 mm, 5␮ Hypersil BDS C18 ) was used for the
assay.
Propranolol samples in both solubility and permeability studies
were determined using Waters 2487 dual ␭ absorbance detector,
the mobile phase consisted of 35% acetonitrile and 65% 25 mM PBS
(pH 4.0, containing 0.5% triethylamine) and the samples were run
at a flow rate of 1 ml/min. The chromatograms were recorded at
230 nm. Excellent linearity of the calibration curve (R2 = 0.9998)
within the concentration range studied was obtained. The assay
detection limit was 55 ng/ml and the inter-day and intra-day coef-
ficients of variation were less than 5%.
For plasma propranolol concentration determination, 25 ␮l
internal standard (verapamil 40 ␮g/ml), 50 ␮l 2 M NaOH and 5 ml
n-hexane plus isopropyl alcohol mixture were added to 1 ml
plasma sample. After shaking (10 min) and centrifugation (10 min
at 4000 rpm), the organic phase (4 ml) was transported to a clean
tube and 200 ␮l of 0.05 M H3 PO4 was added. After mixing (1 min)
and centrifugation (2 min at 10800 rpm), the upper organic phase
was removed and 100 ␮l of the aqueous layer was injected into an
HPLC system for analysis.
The plasma propranolol concentration was determined using
Waters 464 fluorescence detector. The mobile phase, composed of
0.05 M KH2 PO4 buffer (pH 3.0, adjusted by 2 M HCl) and methanol
(40:60), was run at a flow rate of 1 ml/min. The assay calibra-
 Y. Wang et al. / European Journal of Pharmaceutical Sciences 39 (2010) 272–278 275

Table 1
Unionized fraction of propranolol and its solubility at various pH levels (n = 4).

pH Unionized (%)a Solubility (mg/ml)

5.4 0.01 22.12 ± 0.02

6.8 0.37 22.30 ± 0.30
7.0 0.49 22.48 ± 0.12
7.2 0.99 23.21 ± 0.18
7.4 1.46 23.05 ± 0.08
7.6 2.29 22.73 ± 0.24
7.8 3.58 14.11 ± 0.08
9.0 37.06 1.28 ± 0.01
10.0 85.48 0.63 ± 0.00
a
Calculated based on Eq. (3) assuming propranolol pKa = 9.23.

tion curve had excellent linearity (R2 = 0.9996) at the concentration

range (1–100 ng/ml) with extraction recoveries ranging from 69%
to 71%.
Fig. 1. Comparison of the observed and predicted solubility of propranolol at various
3.7. Data analysis pH levels.

All data were expressed as mean ± SD. A non-parametric test

(Mann–Whitney U-test) was used to analyze the data between
two groups. A p < 0.05 was considered statistically significant for
all tests. All statistical analyses were performed with the SPSS 11.0
(SPSS Inc., Chicago, IL).
The pharmacokinetic parameters of propranolol were calcu-
lated using a non-compartmental model with the aid of WinNolin
2.0 (Pharsight, Mountain View, CA). Peak plasma concentration
(Cmax ) and time to reach Cmax (Tmax ) were directly determined from
the plasma concentration–time curve. The terminal elimination
rate constant (z ) was obtained by linear regression of the terminal
portion of the plasma concentration–time curve and the elimina-
tion half-life (t1/2 ) was calculated as 0.693/z . The area under the
plasma concentration–time curve (AUC) was estimated using the
trapezoidal rule and extrapolated to infinity.

4. Results
4.1. pH-solubility profile and pHmax
The solubilities of propranolol at pH 5.4–10.0 are shown in
Table 1. The pHmax was determined to be 7.62 (see below).
The fraction of ionized and unionized propranolol was cal-
culated according to Eq. (3). At pH 5.4, the total solubility of
propranolol was found to be 22.12 mg/ml, with 99.99% of propra-
nolol in the ionized form (Table 1). Therefore, the solubility of the
ionized form (Si ) was obtained from the overall solubility of pro-
pranolol at pH 5.4:
Fig. 2. Effect of pH on the steady-state flux of saturated propranolol across porcine
sublingual mucosa.
4.2. pH-permeability profile
The amount of propranolol permeated across the membrane per
unit time and area, i.e. Jss(T) , at different pH (determined experimen-
tally according to using Eq. (9)) were compared to that generated
theoretically (Eqs. (10) and (11)) are shown in Figs. 2 and 3,
respectively. The maximum Jss values corresponded to pH 7.40

Si = ST × Xi = 22.12 × (99.99%) = 22.12 (mg/ml)

At pH 10.0, the total solubility of propranolol was 0.63 mg/ml,

therefore the solubility of the unionized form (Su ) was estimated
as:

Su = ST (1 − Xi ) = 0.63 × (1 − 14.52%) = 0.54 (mg/ml)

With known Si , Su and pKa of 9.23 (Schoenwald and Huang,

1983), the pHmax of propranolol was calculated according to Eq.
(4).
The profile of total solubility of propranolol as a function of
pH was generated from Eqs. (5) and (6) by using Si = 22.12 mg/ml,
Su = 0.54 mg/ml from above and pKa = 9.23. The pH-solubility pro-
file so generated in comparison to that determined experimentally
is shown in Fig. 1. Excellent fit (R2 = 0.999) was observed when these
data were analyzed using non-linear regression (GraphPad Prism, Fig. 3. Predicted steady-state flux of propranolol across porcine sublingual mucosa
Version 4.0, GraphPad Software Inc., CA, USA). as a function of pH.
276 Y. Wang et al. / European Journal of Pharmaceutical Sciences 39 (2010) 272–278

Table 2
Pharmacokinetic parameters following sublingual administration of the special
buffered propranolol buffered and Inderal® tablet after in eight male healthy sub-
jects (mean ± SD, n = 8).

Parameter (unit) Formulations

Buffered tablet Inderal® tablet

Teff (min)a 8.5 ± 7.7 38.8 ± 20.4b

Fig. 4. Dissolution-time profiles of special buffered propranolol tablet () and
Inderal® propranolol tablet () using the USP II method at a stirring rate of 50 rpm
(n = 6).
from the experimental data and pH 7.62 from the theoretical
data.
4.3. Preparation of special buffered sublingual propranolol tablet
The mean hardness of the buffered tablet was 1.2 kg (n = 12),
with friability of 0.3% and mean disintegration time of 3.81 min
(n = 6). The dissolution profiles for the buffered tablet and Inderal®
obtained by the standard paddle method (USP II) were found to
be similar (Fig. 4), except for that a significant higher drug release
rate was found from the buffered tablets at 5 min (p < 0.001). Other
parameters, i.e. weight variation and content uniformity, for the
buffered tablet also meet USP specifications.
4.4. Pharmacokinetics following sublingual propranolol
The plasma propranolol concentration versus time curves fol-
lowing sublingual administration of our special buffered tablet and
Inderal® tablet are shown in Fig. 5. During the first 30 min, the
buffered tablet resulted in a significantly higher plasma concen-
tration or AUC0–30 min (p < 0.05) as compared to that from Inderal®
tablet. The mean time to reach minimal ␤-adrenoceptor blocking
concentration of 20 ng/ml (Johnsson and Regardh, 1976; Nies and
Shand, 1975) was about 8.5 min for the buffered tablet, as compared
to 38.8 min for Inderal® tablet (p < 0.01). Other mean pharmacoki-
Fig. 5. Plasma propranolol concentration versus time curves of the special buffered
propranolol tablet and Inderal® tablet in 8 male healthy subjects.
Tmax (min) 78.8 ± 50.6 99.4 ± 44.6
Cmax (ng/ml) 58.3 ± 21.3 51.7 ± 11.8
AUC0–30 min (ng min/ml) 909 ± 450 307 ± 209b
AUC30 min–last (ng min/ml) 15674 ± 6754 12998 ± 4567
AUC0–last (ng min/ml) 16582 ± 6634 13305 ± 4683
AUC0–∞ (ng min/ml) 22973 ± 10096 19864 ± 10142
z (h−1 ) 0.22 ± 0.07 0.22 ± 0.06
t1/2 (min) 203 ± 48 210 ± 68
MRT0–420 min (min) 179.6 ± 18.0 187.3 ± 24.4
a
Time to reach minimum plasma concentration to produce ␤-adrenoceptor
blockage (20 ng/ml).
b
Significantly different between the 2 formulations (p < 0.05).
netic parameters (Tmax , Cmax, z , AUC30 min–last , AUC0–last , and MRT)
derived from the plasma concentration–time curves are summa-
rized in Table 2 and were not significantly different between the 2
formulations.
Following propranolol administration of both tablets, mild
drowsiness and some irritation under the tongue were noted in the
majority of the subjects during the study. These effects were tran-
sient and none of the subjects objected to the repeated (crossover)
study during the second session.
5. Discussion
The present study provided for the first time, the equations
that can quantitatively relate permeability (Jss ) to pH and pHmax ,
in addition to confirming the theoretical pH-solubility and pHmax
concept of a representative weak base compound. Furthermore, our
study demonstrated the application of the pHmax concept in sublin-
gual drug development leading to rapid and enhanced propranolol
absorption.
The improved Jss(T) observed at pHmax (pH 7.62) is most likely
a reflection of optimal permeation from both the solubilized
ionized and unionized species. Although maximizing the union-
ized/ionized ratio of propranolol at high pH, e.g. higher than 7.62,
is expected to improve permeability based on the membrane par-
tition hypothesis and Henderson–Hasselbalch equation (Eq. (3)),
such an approach is unlikely to yield a higher permeability than that
at pHmax since the solubility of propranolol is dramatically lower at
such high pH (Table 1). The pHmax concept incorporates not only the
membrane partition hypothesis and Henderson–Hasselbalch equa-
tion but also maximal solubility of both the unionized and ionized
species. Thus, it should be a more powerful approach in enhancing
membrane permeation of certain weak base compounds, such as
propranolol.
Notable differences between the theoretically predicted (Fig. 3)
and experimental Jss(T) (Fig. 2) were observed: the increases in
the experimental Jss(T) around pHmax were more prominent than
that from the theoretical prediction. The reason for the difference
is uncertain. One possibility could be the alteration of the bar-
rier function of the mucous membrane by propranolol, which may
result from transient mucosal irritation. Such irritation has been
observed following its nasal and buccal administration (Johnsson
and Regardh, 1976; van de Donk and Merkus, 1982; Place et al.,
1998; Taylan et al., 1996) as well as following sublingual adminis-
tration in the present study.
Our in-vitro permeation study clearly revealed that at pH values
higher or lower than pHmax , Jss(T) is dramatically lower compared to
 Y. Wang et al. / European Journal of Pharmaceutical Sciences 39 (2010) 272–278
that at pHmax . In addition to this in-vivo observation, we found that
in a limited number of human subjects, the heart rate reduction fol-
lowing sublingual administration of propranolol was greater at a
saliva pH maintained around pHmax than that at higher or lower
pH values (unpublished data). Our significantly greater plasma
concentration data during initial 15 min following our buffered
tablet (achieving salvia pH 7.4–7.6) compared to Inderal® (achiev-
ing saliva pH 6.7–6.9) provide further support of the pHmax concept.
In the present study, the initial higher plasma concentrations
(up to 30 min) following the sublingual administration of our
buffered formulation compared to Inderal® tablet is most likely
a reflection of the differences in the absorption of the two for-
mulations during the first 15 min of sublingual administration.
There were no significant differences in the overall Tmax , Cmax ,
AUC30–420 min and AUC0–420 min values between the two formu-
lations, since these data largely reflected gut absorption after
swallowing of the saliva–drug solution at 15 min post-sublingual
administration. The two-peak observation was also found with vin-
camine when administered in a similar manner by others (Aiache
et al., 1990).
The faster dissolution from our buffered tablet may have
resulted in faster sublingual absorption in first 5–10 min. How-
ever, the mean propranolol concentration in plasma following
buffered tablets was abut 10-fold higher than that of Inderal®
tablets at 3 min, and about 5.9-fold at 6 min. These data were
disproportional to the 70% higher release from the buffered tablets
in comparison to the Inderal® tablets at 5 min. Moreover, the
propranolol plasma concentration following buffered tablets was
1–2 folds higher than Inderal® tablet at 10–30 min, when the drug
release profiles were similar between the two formulations. Since
both buffered and Inderal® tablets were completely dissolved
sublingually at about 15 min, it appeared that the major factor
contributing to the significantly faster sublingual absorption and
higher propranolol plasma concentration was the optimal saliva
pH generated by the buffered tablet.
The buffered tablet was designed for rapid disintegration
(<1 min). However, due to small volume of saliva generated per
minute, the amount/min that can be disintegrated is small. Nev-
ertheless, whatever the amount disintegrated, the buffering agent
will keep the saliva pH near pHmax thus allowing optimal absorp-
tion, leading to significantly earlier and higher propranolol concen-
trations in first 30 min as compared to the conventional formula-
tion without the appropriate buffering agent. To further enhance
and prolong mucosal absorption, bioadhesive technology may be
an attractive choice by utilizing various bioadhesive polymers, i.e.
Cabomer, HPMC, and chitosan, etc. in the future (Beyssac et al.,
1998; Yong et al., 2001; Hassan et al., 2009; Cardot et al., 2004).
The optimization of rapid sublingual absorption of propra-
nolol by utilizing the pHmax concept may offer a new therapeutic
approach to meet its clinical needs. Propranolol is widely used for
the long-term maintenance treatment and prophylaxis of many
cardiovascular conditions (Koch-Wesser and Frishman, 1981;
Regardh, 1982; Wood et al., 1978). For some of these conditions
which are acute but episodic, e.g. anxiety tachycardia, situational
tachycardia, paroxysmal atrial fibrillation and tachycardia that
can occur suddenly in the ambulatory setting, instead of current
approach of oral chronic maintenance or prophylaxis, administer-
ing buffered tablet at the onset of each episode or using “pill in
the pocket” approach, may conveniently provide rapid “targeted”
therapy for these episodes. Also, this can avoid unnecessary drug
consumption at the time between the episodes, especially if these
episodes are infrequent.
The present study has provided clearly a detailed example of
the application of the pHmax concept to permeation of propranolol,
a well-known representative weak base compound. We have now
277
investigated preliminarily the permeation profile of 2 additional
weak base compounds, nebivolol and verapamil and confirmed a
similar close relationship of the theoretical vs. experimental data,
using the pHmax concept illustrated by propranolol.
The present pHmax concept, however, is unlikely be applica-
ble to all weak base compounds. Those weak base compounds
with low molecular weight, high partition coefficient, and single
pKa value may be suitable. For most hydrophilic compounds, or
macromolecules (i.e. peptides and proteins), the transmucosal per-
meability of the ionized species are extreme low. Thus, the pHmax
concept cannot be applied for these compounds.
6. Conclusion
In summary, the present work demonstrated that the trans-
membrane flux for a weak base compound such as propranolol,
across a mucosal membrane, could be enhanced by applying the
pHmax concept, leading to the development of a potential sublin-
gual product with rapid enhanced absorption. Such a concept may
improve the development of certain other weak base compounds
suitable for sublingual or other mucosal administration to achieve
rapid onset of action for urgent medical conditions that require
such intervention.
Acknowledgements
This work was supported by a Direct Grant (No. CUHK 2041010)
from The Chinese University of Hong Kong and an ITF Grant (No.
ITS/174/00) from the Innovation & Technology Commission. The
authors are also thankful to Dr. Benny Fok, Mr. W.K. Fung and Ms.
Miranda W.S. Lai for their technical and logistic assistance.
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