Assessment of solubilization characteristics of different surfactants

for carvedilol phosphate as a function of pH

Subhashis Chakraborty, Dali Shukla, Achint Jain, Brahmeshwar Mishra, Sanjay Singh
*
Department of Pharmaceutics, Institute of Technology, Banaras Hindu University, Varanasi 221 005, India
a r t i c l e i n f o
Article history:
Received 14 January 2009
Accepted 4 March 2009
Available online 5 April 2009
Keywords:
Carvedilol phosphate
Surfactant
Colloidal drug delivery system
pH dependent solubility
Solubilization characteristics
a b s t r a c t
The effect of surfactants on the solubility of a new phosphate salt of carvedilol was investigated at differ-
ent biorelevent pH to evaluate their solubilization capacity. Solutions of different classes of surfactants
viz., anionicsodium dodecyl sulfate (SDS) and sodium taurocholate (STC), cationiccetyltrimethylam-
monium bromide (CTAB) and non-ionicTween 80 (T80) were prepared in the concentration range of
535 mmol dm
3
in buffer solutions of pH 1.2, 3.0, 4.5, 5.8, 6.8 and 7.2. The solubility data were used
to calculate the solubilization characteristics viz. molar solubilization capacity, water micelle partition
coefcient, free energy of solubilization and binding constant. Solubility enhancement in basic pH was
in following order: CTAB > T80 > SDS > STC. CTAB and T80 showed remarkable solubility enhancement
in acidic pH as well. Among the anionic surfactants, solubility in acidic medium was retarded except
at pH 1.2 in case of SDS. Cationic and non-ionic surfactants were found to be suitable for enhancing
the solubility of CP which can be employed for maintaining the in vitro sink condition in the basic dis-
solution medium. While anionic surfactants showed solubility retardant behavior which may be
exploited in increasing the drug entrapment efciency of a colloidal drug delivery system formulated
by emulsication technique.
2009 Elsevier Inc. All rights reserved.
1. Introduction
The key biopharmaceutical parameters responsible for effective
bioavailability and good in vitro in vivo correlation are the drugs
solubility and permeability [1]. Especially in case of ionizable
drugs, their solubility, dissolution and level of sink condition
(when the saturation solubility is at least three times more than
the drug concentration in the dissolution medium as outlined in
USP) will depend upon the degree of ionization in the dissolution
medium at different pH [24]. Several approaches to achieve the
sink condition in the dissolution medium include increasing the
volume of the aqueous medium or removing the dissolved drug,
solubilization of the drug by co-solvents up to 40%, addition of cat-
ionic, anionic or non-ionic surfactants to the dissolution medium
above critical micelle concentration (cmc) and use of a hydroalco-
holic surfactant solution [5,6]. Of the above mentioned methods,
the use of media containing surfactants is the most popular meth-
od because of its simulation with various surfactants present in the
gastrointestinal uid, e.g., bile salts, lecithin, cholesterol and its es-
ters [1,7]. Surfactant monomers above their cmc, aggregate to form
colloidal moieties known as micelles which are capable of encap-
sulating the drug molecules resulting in reduction in the interfacial
tension and improved solubility of the drug in the medium. Ionic
surfactant may preferably solubilize oppositely charged species
than uncharged or similarly charged species. However, there may
be cases where oppositely charged species and surfactant may
interact to form an insoluble salt and result in desolubilization as
reported in case of erythromycin and PG-300995 (an anti-HIV
agent) after addition of sodium dodecyl sulfate due to the forma-
tion of an estolate salt [8,9].
Carvedilol (()-1-(Carbazol-4-yloxy)-3-[[2-(o-methoxyphen-
oxy) ethyl] amino]-2-propanol) is an alpha and beta blocker used
to treat high blood pressure and heart failure. Carvedilol is practi-
cally insoluble in water and exhibits pH dependent solubility. Its
solubility is <1 lg/ml above pH 9.0, 23 lg/ml at pH 7 and about
100 lg/ml at pH 5 at room temperature. In between pH 1 and 4
in various buffer systems, solubility of carvedilol is limited due
to its protonation, resulting in in situ hydrochloride salt formation
which exhibits less solubility in acidic medium [10]. Its extremely
low solubility in the alkaline pH may prevent the availability of the
drug for absorption in the small intestine and colon, thus making it
a poor candidate for an extended release dosage form. Moreover,
the secondary amine attached to the drug core is susceptible to
degradation. In view of the above limitations, a phosphate salt of
carvedilol was recently developed to improve the aqueous solubil-
ity and chemical stability by protonation of the secondary amine
group as a salt [10]. Carvedilol phosphate (CP) is available for
once-a-day administration as controlled-release oral capsules con-
taining 10, 20, 40, or 80 mg of the active moiety.
0021-9797/$ - see front matter 2009 Elsevier Inc. All rights reserved.
doi:10.1016/j.jcis.2009.03.047
* Corresponding author. Fax: +91 542 2368428/2369162.
E-mail address: sanjays@bhu.ac.in (S. Singh).
Journal of Colloid and Interface Science 335 (2009) 242249
Contents lists available at ScienceDirect
Journal of Colloid and Interface Science
www. el sevi er . com/ l ocat e/ j ci s
Our interest in CP arose from the nding in our laboratory that
though the salt had a signicantly improved aqueous solubility
over the free base, its solubility in the intestinal pH range was
<140 lg/ml. Though, this may not be a problem for the lower
strengths (10, 20 and 40 mg), but it would be difcult to maintain
in vitro dissolution sink condition for the highest strength of
80 mg. To maintain the sink condition in 1000 ml of dissolution
medium for the highest strength, the saturation solubility should
be greater than 240 lg/ml. This indicates that there is a need to im-
prove the solubility of the drug in the dissolution medium, which
otherwise would pose problems in the early stages of drug product
development. In the present study, different classes of surfactants
viz. anionicSDS and STC, cationicCTAB and non-ionicT80 have
been employed for solubility enhancement. Furthermore, analysis
of the solubilization characteristics viz., molar solubilization ratio,
micellewater partition coefcient, free energy of solubilization
and binding constant would be valuable in evaluation of the solu-
bilization capacity of the surfactants for CP.
Molar solubilization ratio (v) is the number of moles of the sol-
ute (drug) that can be solubilized by one mole of micellar surfac-
tant and it characterizes the ability of the surfactant to solubilize
the drug. It can be calculated using the following equation [11].
v S
t
S
cmc
=C
t
C
cmc
: 1
where S
t
and S
cmc
are total drug solubility in aqueous surfactant
solution and drug solubility at cmc of surfactant, respectively; C
t
and C
cmc
are total surfactant concentration and surfactant concen-
tration at cmc, respectively. It can also be determined from the
slope of the solubility of drug in presence of surfactant vs. concen-
tration of surfactant (above cmc) plot.
Micellewater partition coefcient (P
m
) is the ratio of mole fac-
tion of solubilizate in the micellar phase to that in aqueous phase
and it is the measure of the effectiveness of solubilization of the
solubilizate between the micelle and aqueous phases and can be
calculated using the following equation [12].
P
m
v=fS
cmc
V
m
1 vg: 2
where V
m
is molar volume of water at 37 C and is equal to
0.01815 dm
3
mol
1
, v is molar solubilization capacity and S
cmc
is
drug solubility at cmc.
The standard free energy of solubilization (DG
s

) is indicative of
the enthalpy changes during solubilization and can be represented
by the following equation [13,14].
DG
s

kj mol
1
RT lnP
m
: 3
where R is universal gas constant, T is absolute temperature and P
m
is water micellar partition coefcient.
Binding constant (K) of drug surfactant system is the measure of
interaction between surfactant and solubilizate and is related to
the total surfactant concentration (C
t
), critical micelle concentra-
tion (C
cmc
) and aggregation number (N) of micelles through the fol-
lowing equation [14,15].
S
t
S
cmc
=S
cmc
K=NC
t
C
cmc
: 4
The value of K/N is obtained from the slope of (S
t
S
cmc
)/S
cmc
vs.
(C
t
C
cmc
) plot.
N is an important parameter that characterizes the micelles and
corresponds to the average number of surfactant monomers in
each micelle of a micellar solution. In a micellar solution, N is
approximately constant for a broad total concentration range (up
to about 100 times the cmc), while the number of micelles vary
[15]. To simplify the interpretation of results, the N values of the
surfactants have been assumed to be constant and were used as
obtained from the literature as 4, 62, 170 and 58, for STC, SDS,
CTAB and T80, respectively [16,17].
To our knowledge, the solubility issues related to CP has not
been addressed and reported earlier. Therefore, this investigation
would be helpful in the design of a suitable in vitro dissolution
medium for this drug. This study would also provide signicant
preformulation information about the drugs behavior in the pres-
ence of different type and concentration of surfactants which are
essential components used in the fabrication of colloidal drug
delivery systems, e.g., polymer/lipid based nanoparticles and
emulsions. We have also tried to correlate the ndings of this
investigation with the colloidal behavior of the drug and physio-
logical surfactants in the process of dissolution which play a signif-
icant role in improving the bioavailability of the drug.
2. Materials and methods
2.1. Materials
The drug CP was provided by Lupin Ltd. (Pune, India). SDS and
T80 were purchased from Sigma Chemical Co. (St. Louis, MO) and
Merck (Germany), respectively. CTAB and STC were kind gifts of
Tatva Chintan Pharma Chem Pvt Ltd. (Ankeleshwar, India) and
New Zealand Pharmaceuticals (NZ), respectively. All other reagents
were of analytical grade. The ingredients were used as received.
2.2. Methods
2.2.1. Analytical method
In order to determine the solubility of the drug, standard curves
were prepared in distilled water and all buffer solutions. The
absorbance of the standard solutions prepared in the concentration
range of 428 lM/dm
3
was measured at 240.2 nm(k
max
) using Shi-
madzu UV-1205 UVvis spectrophotometer. In all cases, standard
curves prepared for the determination of drug concentration in
samples, were linear with R
2
> 0.999. Absorbance of samples was
measured using blank as distilled water or buffer solutions with/
without the appropriate molar concentration of surfactants as in
the sample. Samples containing surfactants at all concentration
were scanned everytime to detect any shift in k
max
.
2.2.2. Solubility study in water
Drug was added in incremental amount (1060 mg) in six con-
ical asks each containing 20 ml of distilled water. The asks were
tightly corked and placed in a thermostated water bath at
37.0 0.5 C agitated at 70 rpm for at least 24 h. Samples were ta-
ken at 8, 12, 16 and 24 h to ensure that the equilibrium solubility
had been reached. After this period, aliquots were withdrawn, l-
tered through 0.45-lm lter, diluted with the medium and the
drug concentration in the nal sample solutions was determined
spectrophotometrically. Each solubility value was determined in
triplicate and the results reported are the mean of the three. All
the glassware and lters used were maintained at 37 2.0 C prior
to the experiment to avoid precipitation of drug during analysis.
2.2.3. Solubility study in buffer
pH 1.2 hydrochloric acid buffer solution, pH 3.0 acid phthalate
buffer, pH4.5 acetate buffer and pH5.8, 6.8 and 7.2 potassiumphos-
phate buffers were prepared as per USP. The buffers were prepared
using freshly prepared double distilled water. The ionic strength
(IS) of the media was adjustedto 0.1 M(representative of physiolog-
ical IS) withsodiumchloride[18]. Thesolubilityof druginthebuffers
was measured in the similar manner as in water.
2.2.4. Solubility study in buffer with surfactants
Solutions containing different concentration of surfactants at
their respective cmc and in the range of 535 mmol dm
3
were
prepared in above mentioned freshly prepared buffer solutions.
S. Chakraborty et al. / Journal of Colloid and Interface Science 335 (2009) 242249 243
The solubility study of drug in these solutions was carried out in
similar manner as in buffer solutions using excess amount of the
drug. The solubility data were used to calculate the solubilization
characteristics using their respective equations.
3. Results
3.1. Solubility in distilled water and buffer solutions
Solubility of the drug in distilled water and buffer solutions was
checked to study the impact of excess solids on the apparent solu-
bility [19]. The results indicated a signicant and gradual increase
in solubility of drug (0.22.8 mg/ml) in water with increase in the
excess drug quantity (supporting information). At the end of
equilibration (after 24 h), it was found that the pH of water
(7.0) reduced with the increase in amount of excess drug
(approximately by 22.5 U). Higher the amount of drug added,
more was the shift towards lower pH.
In case of buffer solutions, the variation in solubility upon equil-
ibration with similar increase in the drug amount was insignicant
and the reduction in pHwas found to be within 0.10.2 U (support-
ing information). Solubility at different pH was in following order:
pH 3.0 > pH 4.5 > pH 1.2 > pH 5.8 > pH 6.8 > pH 7.2, as shown in
Fig. 1.
3.2. Solubility in buffer solutions with surfactant
In order to judge the effect of surfactants on solubility, the buf-
fer solutions used have been divided in two broad groups; acidic
solutions (pH 1.2, 3.0 and 4.5) and basic solutions (pH 5.8, 6.8
and 7.2).
3.2.1. Effect of SDS
The effect of SDS concentration on the solubility of CP at differ-
ent pH is presented in Fig. 2. With 5 mmol dm
3
concentration of
SDS, there was a drastic reduction in solubility in acidic solutions
while an increase in solubility was observed in basic solutions. At
this concentration, the solubility was almost same at all pH, indi-
cating no effect of variation in pH on drug solubility. Throughout
the pH range studied, the solubility increased gradually with fur-
ther increase in the SDS concentration up to 35 mmol dm
3
. In
comparison to the drugs solubility at different pH without
surfactant, the solubility with the highest concentration of SDS
was found to be reduced in acidic solutions except at pH 1.2 and
signicantly increased in basic solutions. The solubility at
8.1 mmol dm
3
(cmc of SDS) [20] concentration was found to lie
between the solubility range of 5 and 10 mmol dm
3
solutions.
The difference in solubility due to change in pH was minimal at
all SDS concentrations studied than in absence of SDS.
3.2.2. Effect of STC
The effect of STC concentration on the solubility of CP at differ-
ent pH is presented in Fig. 3. The solubility was found to reduce at
all pH on addition of STC at 3 mmol dm
3
(cmc of STC is 3
11 mmol dm
3
) [21] and 5 mmol dm
3
concentration. However,
at 10 mmol dm
3
concentration, the solubility reduced signi-
cantly in acidic solutions and no marked change in solubility was
observed in basic solutions. In this case, similar solubility values
at all pH were attained at 10 mmol dm
3
concentration. As the
concentration of STC was increased from 10 to 35 mmol dm
3
,
the increase in solubility was observed throughout the pH range
studied. Similar pattern of reduced pH dependence of drug solubil-
ity, as in case of SDS, was observed at and above 10 mmol dm
3
concentration of STC.
3.2.3. Effect of T80
The effect of T80 concentration on the solubility of CP at differ-
ent pH is presented in Fig. 4. The solubility in presence of T80 was
found to increase continuously from 5 to 35 mmol dm
3
concen-
Fig. 1. Solubility of CP in buffers of different pH.
Fig. 2. Effect of concentration of SDS on solubility of CP in buffers of different pH.
Fig. 3. Effect of concentration of STC on solubility of CP in buffers of different pH.
244 S. Chakraborty et al. / Journal of Colloid and Interface Science 335 (2009) 242249
tration at all pH solutions. The increase in solubility at the highest
concentration of T80 was approximately 2.53 times at pH 3.0 and
4.5, 10 times at pH 1.2 and 5.8, 25 times at pH 6.8 and 40 times at
pH 7.2 than the solubility of the drug in the buffer solutions with-
out surfactant. The increase in solubility at cmc level
(0.012 mmol dm
3
) [17] of T80 was not signicant than the solu-
bility without surfactant at all pH.
3.2.4. Effect of CTAB
The effect of CTAB concentration on the solubility of CP at dif-
ferent pH is presented in Fig. 5. Results of solubility study of CP
in presence of CTAB were quiet similar to that of T80. Solubility
was found to increase linearly with increase in surfactant concen-
tration at all pH which was comparatively higher than that
achieved in presence of T80. The increase in solubility at the high-
est surfactant concentration was approximately 34 times at pH
3.0 and 4.5, 1319 times at pH 1.2 and 5.8 and 56 times at pH
6.8 and 75 times at pH 7.2. Unlike anionic surfactants, the solubil-
ity of CP at cmc level of CTAB (0.81 mmol dm
3
) [12] was higher
than that of its solubility in absence of surfactant and lower than
that observed with higher concentrations of CTAB.
The solubility data were used to calculate the molar solubiliza-
tion ratio (Fig. 6), micellewater partition coefcient (Fig. 7), free
energy of solubilization (Fig. 8) and binding constant (Fig. 9).
Fig. 4. Effect of concentration of T80 on solubility of CP in buffers of different pH.
Fig. 5. Effect of concentration of CTAB on solubility of CP in buffers of different pH.
Fig. 6. Comparison of v values of the surfactants for CP at different pH.
Fig. 7. Comparison of P
m
values of all four surfactants for CP at different pH.
Fig. 8. Comparison of DG
s

values of all four surfactants for CP at different pH.

S. Chakraborty et al. / Journal of Colloid and Interface Science 335 (2009) 242249 245
4. Discussion
4.1. Solubility in water and in buffer without surfactant
It was reported recently that the amount of excess drug added
for solubility studies may have a signicant effect on the apparent
solubility, likely to be caused by a competition between the crys-
tallization and dissolution rates [19]. To conrm the same phe-
nomenon in the present work, preliminary solubility studies
were carried out in water and buffer solutions with addition of
incremental quantities of the drug. The increase in solubility of
the drug in water can be attributed to its lack of buffering capac-
ity. As the drug comes in contact with water, it undergoes ioniza-
tion to release phosphate ions. The amount of phosphate ions
released depends on the amount of drug in contact with water
which results in increase in acidity. This in turn increases the sol-
ubility of the drug as it exhibits pH dependent solubility and is
more soluble at lower pH (higher solubility at pH 34.5 than
pH 57) as shown in Fig. 1. Thus, the increase in solubility of
the drug as a function of its quantity is a self induced phenome-
non which indicates that the solubility of the phosphate salt of
carvedilol in water cannot be measured exactly. The implication
of the above observation to the salt form of other drugs is yet
to be ascertained.
On the other hand, the buffering capacities of the buffered solu-
tions are sufcient enough to resist the pH change due to release of
phosphate ions, thus preventing signicant variations in solubility.
Fig. 1 indicates that CP exhibits pH dependent solubility. The
solubility increases with the decrease in pH up to 3.0 due to in-
crease in the ionization of the drug. This may be attributed to the
presence of an alpha-hydroxyl secondary amine functional group,
which has a pK
a
of 7.8. However, CP undergoes in situ protonation
in the gastric medium of pH 12 to generate its corresponding HCl
salt which has limited solubility in such medium [10].
4.2. Solubility in buffer solutions with surfactant
Anionic surfactants were selected based on the fact that the
electrostatic interactions between the oppositely charged surfac-
tant and drug molecules would cause a decrease in the repulsive
forces between the head groups of the surfactant molecules, con-
tributing to enhanced solubilization [14]. However, the results ob-
tained in both the anionic surfactants (SDS and STC) were
contradictory to the above hypothesis in acidic solutions where
the drug exists predominantly in cationic state except at pH 1.2.
4.2.1. Effect of SDS
The effect of SDS on the solubility prole of CP at different pH is
shown in Fig. 2. The prole reects an increasing trend in solubility
except a drop at 5 mmol dm
3
(below cmc) concentration in acidic
solutions. In acidic solutions of pH 3 and 4.5, the overall solubility,
even with the highest concentration of SDS used, remains drasti-
cally below the baseline level obtained without surfactant. This
phenomenon can be explained on the basis of the ionic state of
the molecules (Fig. 10). As the drug is basic in nature (pK
a
7.8),
its ionization, hence solubility increases with the decrease in pH.
SDS being an anionic surfactant (pK
a
1.9) shows its surface active
properties only in ionized state. It is approximately 10% ionized
in 0.1 N HCl and its ionization and hence wetting property in-
creases with pH [22]. Therefore, in acidic solutions, the drug exhib-
its its ionic (cationic) state while SDS is predominantly in the non-
ionic state which is less capable of increasing solubility. The
remaining molecules of SDS in the ionic state (anions) interact with
the cationic species of drug to form an insoluble salt which precip-
itates out resulting in desolubilization of the drug. This phenome-
non, in acidic solutions at low SDS concentration, is responsible for
the overall reduction in solubility. Similar interactions of SDS have
been reported with erythromycin and an anti-HIV agent [8,9]. In
basic solutions, the drug is predominantly in the non-ionic state
which is unsuitable for ionic interaction while the surfactant is in
ionic (anionic) state which is highly suitable for exhibiting surface
active properties. This results in a favorable situation for solubility
enhancement. Increase in basicity results in higher solubilization
due to less ionic interaction and higher surface activity of SDS.
The solubilization capacity of surfactants below cmc is due to their
wetting property which results in increased effective surface area
for solubilization and dissolution by deaggregation of drug parti-
cles while above cmc it is due to micellar solubilization [23]. On
increasing the concentration of SDS above cmc (10 mmol dm
3
),
a rise in solubility is observed at all pH due to solubilization of
insoluble salt formed, by entrapment in the micelles. Increase in
SDS concentration (1535 mmol dm
3
) results in increase in the
number of micelles which further enhances the solubility.
4.2.2. Effect of STC
The cmc and pK
a
of STC has been reported to be 3-
11 mmol dm
3
and 1.4, respectively [21,24]. At 3 mmol dm
3
con-
centration, the solubility of the drug is reduced at all pH solutions
with further reduction at 5 mmol dm
3
concentration which is
more pronounced in acidic than in basic solutions (Fig. 3). The rea-
sons may be similar to that as explained with SDS as it is also an
Fig. 9. Comparison of K values of all four surfactants for CP at different pH.
Fig. 10. Effect of pH on ionization of CP and SDS.
246 S. Chakraborty et al. / Journal of Colloid and Interface Science 335 (2009) 242249
anionic surfactant with comparable pK
a
value. Up to
10 mmol dm
3
STC concentration, the trend of solubility reduction
continues in the acidic solutions while there was no marked
change in solubility in the basic solutions. With further increase
in the surfactant concentration above the cmc (15 mmol dm
3
), a
rise in solubility is observed throughout the pH range indicating
micellar solubilization of the insoluble salt formed, which contin-
ues further with increase in the surfactant concentration (25 and
35 mmol dm
3
) due to increased number of micelle available for
solubilization. Like SDS, even with the highest concentration of
surfactant at pH 3.0 and 4.5, the solubility remained well below
that obtained in pure buffer solutions.
The above observation can be extended to explain the pharma-
cokinetic and pharmacodynamic behavior of the drug in the hu-
man body. The bile salt concentration in the duodenum and
upper jejunum are approximately 23 times higher in fed state
(1015 mmol dm
3
; range 335 mmol dm
3
), as compared to fast-
ing conditions (5 mmol dm
3
, range 014 mmol dm
3
) [2527].
Thus, the absorption of BCS Class II drugs like CP [28] showing dis-
solution rate limited uptake should enhance, when administered in
the fed state, due to the surfactant effects of bile components. This
is in agreement with the result of clinical trials reported in litera-
ture [29]. Administration of extended release capsule of CP with
a high-fat meal resulted in increase in AUC (20%) and C
max
when
compared with administration with a standard meal. This increase
in bioavailability could be explained on the basis of higher STC (a
major bile salt component) concentration in fed state which fur-
ther increases in the presence of high-fat meal [30,31]. Increase
in the bile salt concentration helps in effective colloidal solubiliza-
tion in the intestine which may be the major factor responsible for
faster absorption and enhanced bioavailability of the drug [32]. A
decrease in AUC (27%) and C
max
(43%) were observed when the
capsules were administered in the fasted state compared to admin-
istration after a standard meal. The reason for reduced bioavailabil-
ity could be probably due to interaction between the drug and the
STC molecules belowcmc. Therefore, the extended release capsules
of CP have been recommended to be taken with food.
One of the less prevalent side effects of CP is increased levels of
serum glutamic pyruvic transaminase and serum glutamic oxaloa-
cetic transaminase enzymes, which are indicative of liver damage
[29]. Such incidences of liver damage may be correlated with the
present ndings. CP undergoes extensive rst pass metabolism
and is excreted primarily through bile into the feces. Liver releases
the un-metabolized and metabolized drug in the bile duct where
the former could interact with bile salts to cause its precipitation
and block the duct. Owing to bile secretory failure, bile salts and
other biliary constituents are retained in the hepatocyte and this
leads to progressive liver damage [33]. Such cases may occur
where the release of STC is below cmc where micellar solubiliza-
tion of the complex formed is not possible. Similar incidence of bile
secretory failure because of interaction of drug and STC has been
reported for chlorpromazine hydrochloride [34].
4.2.3. Effect of T80
Non-ionic surfactants have the potential to provide a combina-
tion of good molar solubilization capacity and high micellar con-
centration due to their extremely low cmc [13]. The solubility of
CP increases gradually at all concentrations (535 mmol dm
3
) of
T80, owing to micellar solubilization of the drug (Fig. 4). Being
non-ionic in nature, it does not exhibit any kind of ionic interac-
tion. It was observed that the solubilization effect is greater at high
pH than at low pH indicating better surface activity in the basic pH.
Due to the extremely low cmc level, a higher molar fraction of T80
is available in the micellar form which is responsible for exhibiting
comparatively higher solubility than other surfactants. From the
pharmacological perspective, the lower cmc value of T80 is bene-
cial in improving the stability of the drug incorporated micelles. As
on intravenous administration, large volume of the blood causes
dilution of the administered solution and only micelles of surfac-
tants with very low cmc value can exist, while micelles of surfac-
tants with high cmc value may dissociate into monomers and
their content may precipitate in the blood [35].
4.2.4. Effect of CTAB
CTAB is a cationic surfactant and it also has low cmc of
0.81 mmol dm
3
, although it is higher than T80. Being cationic, it
could exert repulsive electrostatic forces on coming in contact with
drug ions and reduce the chances of micellar entrapment of drug. It
was expected that it would show higher solubility enhancement
than anionic surfactant because of lack of ionic interaction, but
lower than non-ionic T80 owing to common ion repulsion between
drug and CTAB. However, to our surprise, the results were contrary
to above mentioned presumption. It yielded highest solubility
enhancement than all the surfactants in all the media (Fig. 5).
The reason can be related to its relatively higher N value (170) than
T80 (60) which is responsible for greater micellar size of CTAB
which helps to accommodate more drug molecules per micelle.
4.3. UVvis spectrum analysis
The possible interaction between CP and the ionic surfactants
was also observed in the UV spectrum (supporting information). A
red shift in the absorption maxima of CP (240.2 nm) in the presence
of both SDS and STC (approximately 56 nm and 23 nm shift was
observed with SDS and STC, respectively) is indicative of change in
themicroenvironmental polarityof CPprobablydue toionic interac-
tion between the head group of SDS and STC and positively charged
alpha-hydroxyl secondaryaminemoietyof thedrug. Maximumshift
was observedat the cmc level of the surfactants. Withthe increase in
the surfactant concentration above the cmc, the shift reduced which
maybe due todecrease inpolarityas a result of incorporationof drug
molecules or complexes intothemicelles. Similar shift intheabsorp-
tionmaxima due toionic interactionwas alsoreportedbyShahet al.,
[36]. The shift in k
max
was found to be independent of pH. No signif-
icant shift was observed in the UV spectrumof CP in the presence of
CTAB and T80 indicating absence of any kind of ionic interaction be-
tween the drug and surfactants.
4.4. Solubility characterization
The molar solubilization capacities (v) at the cmc values of the
different surfactants followed the following trend: CTAB > T80 >
SDS > STC (Fig. 6). The maximum v value of the ionic surfactants
is at pH 4.5, probably due to best balance between the ionic and
non-ionic species of the drug and surfactant molecules. The low
v values of STC and SDS in comparison to T80 and CTAB is probably
because of interaction between the cationic species of the drug and
the anionic species of the surfactants resulting in formation of an
insoluble salt. Thus, a part of the molar fraction is utilized in the
interaction while on further addition of these anionic surfactants
above cmc results in micellar solubilization. The involvement of
the surfactant molecules in the interaction causes an overall shift
in its effective cmc towards higher side resulting in reduced solu-
bilization. The relatively low v value of STC in comparison to SDS
can be discussed on the basis of their cmc values. It is evident from
Fig. 4 that the solubility of CP in presence of STC increases above
10 mmol dm
3
concentration, which indicates that this concentra-
tion of STC is insufcient for micelle formation. Therefore, as the
cmc level of STC is higher than SDS, more amount of the former
is required to achieve the same level of solubilization as in the
presence of the latter. Further, the v value of CTAB was lesser than
T80 at pH 3 which can be attributed to the electrostatic repulsion
S. Chakraborty et al. / Journal of Colloid and Interface Science 335 (2009) 242249 247
between the cations of surfactant and the drug. The increase in v
value of CTAB in comparison to T80 above pH 3 is due to the reduc-
tion in the electrostatic repulsion because of the increase in non-
ionic state of the drug. Another factor contributing to the increase
in v value of CTAB than T80 at higher pH is its N value. Higher the N
value, larger will be the size of micelle and more will be number of
drug molecules per micelle. Therefore, CTAB with high N value
incorporates the maximum number of drug molecules within it
exhibiting high solubility. On the other hand, STC with a very
low N value shows very low solubilizing ability. The least solubiliz-
ing capacity of STC can also be explained on the basis of the molec-
ular weights of CP and STC (513.5 and 537.7, respectively). Due to
comparable molecular weight and extremely low N value, the mi-
celle formed will not have adequate space to incorporate the drug
and/or the complex of either equal or larger size, respectively. Sim-
ilar case also exists with SDS of lower molecular weight (288.38)
but due to its larger N value, it dominates over STC in terms of sol-
ubilization capacity.
As can be seen fromFig. 7, the water micelle partition coefcient
(P
m
) of CTAB and T80 is higher than SDS and STC in basic pH range.
Further, SDS shows higher P
m
than STC for the same reason as dis-
cussedfor v. T80exhibits higher P
m
thanCTABwhichcouldbeattrib-
uted to its extremely low cmc value for which it initiates micelle
formation at very low concentration. The higher micellar partition-
ingof T80thanCTABcanbeexplainedonthebasis of their hydropho-
bic chain length: T80 (oleic acid-C18) and CTAB (hexadecyl group-
C16). Being lipophilic in nature, the drug would have higher afnity
for a longer chain with higher hydrophobicity and hence higher
micellar partitioning. For all the surfactants, the P
m
increases from
pH 3.07.2. This is because of increasing non-ionic nature of drug
at higher pH which causes better partitioning in the micelle, thus
reducing chances of interaction or electrostatic repulsion. In all
cases, the P
m
value is higher at pH 1.2 than at pH 3.0. At pH 1.2, the
drug tends to form insoluble hydrochloride salt (non-ionic state)
whichexhibits higher partitioningthanat pH3.0where drugis max-
imum ionized and has minimum P
m
value.
DG
s

is a measure of the interfacial thermodynamic interactionof

thesolubilizateandthemicelles, i.e. theenergyutilizedintheforma-
tion of micelles and incorporation of the solute molecules within
them. A greater negative DG
s

indicates an energetically favorable

condition for the uptake of the solute within the micelles. DG
s

val-
ues obtained(Fig. 8) are negative all throughout the systems indicat-
ing spontaneous solubilization. The values of DG
s

follow exactly
similar trend as that of P
m
but on the negative side. Comparatively
larger negative DG
s

values with high solubility data (i.e. v and P

m
)
were observed for CTAB and T80 micellar systems in basic solutions,
showing effective solubilizationof CP by cationic andnon-ionic than
anionic micellar system. Therefore, CTAB and T80 are the suitable
surfactants to be used to improve the sink condition during dissolu-
tion study in the intestinal pH range.
Analyses of the binding constant values (Fig. 9) indicate that
irrespective of pH, STC micelles have the minimum afnity for
the drug molecules than the other surfactants as is also evident
from its solubility prole. Thus, STC can be selected as the best sur-
factant, to be used as solubility retardant throughout the pH range
studied. Its application becomes more specic in the development
of colloidal formulations for this drug. In colloidal formulations like
solid lipid nanoparticles, surfactants are essentially added in the
external dispersion medium to reduce the interfacial tension be-
tween the two immiscible phases which results in a thermody-
namically stabilized system [37]. However, the presence of
surfactant increases the solubility of the drug (usually with poor
aqueous solubility) in the external medium, resulting in reduced
drug entrapment into the colloidal particles during the emulsica-
tion process. Therefore, surfactants capable of reducing the solubil-
ity of drug in the external medium may play a signicant role in
improving the drug entrapment efciency of the colloidal particles,
along with stabilization of the system. On the other hand, the bind-
ing constant of SDS is highest among all the surfactants throughout
the pH range indicating the formation of a very stable insoluble
estolate salt [8]. If incorporated within the colloidal particles along
with the drug, its high afnity for the drug would be effective in
controlling the release of the drug.
5. Summary
The solubility of CP in the presence of different types and con-
centrations of surfactants, at different pH has been reported for
the rst time in this study. Interestingly, our ndings revealed that
the use of anionic surfactants signicantly minimized the pH
dependent variation of the drugs solubility. While the micellar sol-
ubilization of the drug in presence of non-ionic and cationic surfac-
tants can be applied for the dissolution method development, the
solubility retarding property of the anionic surfactants may be uti-
lized for the stabilization and enhancing the drug entrapment ef-
ciency of colloidal formulations. Although the data presented in
this work is applicable to CP, we hope that the present investiga-
tion will provide pharmaceutical researchers, adequate insight
concerning the conduct of preformulation studies to select suitable
surfactants for formulation and dissolution method development
of colloidal drug delivery system of other drugs as well which
would save a great deal of valuable time and energy.
Appendix A. Supplementary data
The supporting information contains additional supplementary
material. Fig. S1 shows the chemical structure of CP. Table S1
shows the effect of excess amount of CP on its solubility in water
and Table S2 shows the effect of excess amount of CP on its solu-
bility in buffer solutions. Table S3 presents the shifts in the wave-
length maxima of CP in various buffer solutions containing
different surfactants at cmc. Supplementary data associated with
this article can be found, in the online version, at doi:10.1016/
j.jcis.2009.03.047.
References
[1] G.L. Amidon, H. Lennernas, V.P. Shah, J.R. Crison, Pharm. Res. 12 (1995) 413
420.
[2] K.G. Mooney, M.A. Mintun, K.J. Hemmelstein, V.J. Stella, J. Pharm. Sci. 70 (1981)
2232.
[3] United States Pharmacopeial Convention, In vitro and in vivo evaluation of
dosage forms, USP 28, United States Pharmacopeial Convention Inc., Rockville,
MD, 2005.
[4] J.G. Aunins, M. Southard, R.A. Myers, K.J. Hemmelstein, V.J. Stella, J. Pharm. Sci.
74 (1985) 13051316.
[5] M.A. El-Massik, I.A. Darwish, E.E. Hassan, L.K. E1-Khordagui, Int. J. Pharm. 140
(1996) 6976.
[6] S.H. Park, H.K. Choi, Int. J. Pharm. 321 (2006) 3541.
[7] S.H. Yalkowsky, Solubility and Solubilization in Aqueous Media, Oxford
University Press, New York, 1999.
[8] C. Stubbs, I. Kanfer, Int. J. Pharm. 63 (1990) 113119.
[9] A. Jain, Y. Ran, S.H. Yalkowsky, AAPS PharmSciTech. 5 (3) (2004) 6567 (Article
45).
[10] C.S. Brook, W. Chen, P.G. Spoors, US Patent 7,268,156, 2007.
[11] F.A. Alvarez-Nez, S.H. Yalkowsky, Int. J. Pharm. 200 (2000) 217222.
[12] A.A. Dar, G.M. Rather, A.R. Das, J. Phys. Chem. B. 111 (2007) 31223132.
[13] C.O. Rangel-Yagui, P. Adalberto, C.T. Leoberto, J. Pharm. Pharm. Sci. 8 (2005)
147163.
[14] C.O. Rangel-Yagui, H.W.L. Hsu, J.A. Pessoa, L.C. Tavares, Braz. J. Pharm. Sci. 41
(2005) 237246.
[15] P.C. Hiemenz, R. Rajagopalan, Principles of Colloid and Surface Chemistry, third
ed., Marcel Decker, New York, 1997.
[16] Sigma Product Information, <http://www.sigmaaldrich.com/catalog/search/
ProductDetail/SIGMA/T4009/>, 2008 (accessed 29.08.08), part of Sigma
Aldrich Co. <http://www.sigmaaldrich.com/> (accessed 29.08.08).
[17] Amresco Detergents Selection Chart: Mandel Scientic Company, Inc., <http://
www.mandel.ca/products/microbiology/Buffers/Detergents_Selection_Chart/
248 S. Chakraborty et al. / Journal of Colloid and Interface Science 335 (2009) 242249
index.htm/>, 2008 (accessed 30.08.08), part of <http://www.mandel.ca/
index.htm/ (accessed 30.08.08).
[18] S.D. Mithani, V. Bakatselou, C.N. TenHoor, J.B. Dressman, Pharm. Res. 13 (1)
(1996) 163167.
[19] K. Kawakami, K. Miyoshi, Y. Ida, Pharm. Res. 22 (9) (2005) 15371543.
[20] H. Akbas, M. Taliha, Turk. J. Chem. 27 (2003) 357364.
[21] Sigma Product Information, <http://www.sigmaaldrich.com/sigma/product%
20information%20sheet/t4009pis.pdf/> (accessed 29.08.08), part of Sigma
Aldrich Co. <http://www.sigmaaldrich.com/> (accessed 29.08.08).
[22] J.W. McGinity, Drugs and the pharmaceutical sciences, Aqueous Polymeric
coatings for Pharmaceutical Dosage Forms, second ed., vol. 79, Marcel Dekker,
Inc., NY, 1996.
[23] J. Swarbrick, J.C. Boylan, Encyclopedia of Pharmaceutical Technology, Marcel
Dekker, New York, 2001.
[24] Entry# 9242, The Merck Index, 12th ed., Merck Research Laboratories, division
of Merck and Co., Inc., Whitehouse Station, NJ, USA, 1996.
[25] O. Fausa, Scand. J. Gastroenterol. 9 (1974) 567570.
[26] A. Tangerman, A. VanSchaik, E.W. Vander Hook, Clin. Chem. Acta 159 (1986)
123132.
[27] T.L. Peeters, G. Vantrappen, J. Janssens, Gastroenterology. 79 (1980) 678681.
[28] T. Loftsson, S.B. Vogensen, C. Desbos, P. Jansook, AAPS PharmSciTech 9 (2)
(2008) 425430.
[29] Rxlist website, <http://www.rxlist.com/cgi/generic/coreg_cr.htm/>, 2008
(accessed 29.08.08), part of http://www.rxlist.com/ (accessed 29.08.08).
[30] D. Fleisher, C. Li, Y. Zhou, L. Pao, A. Karim, Clin. Pharmacokinet. 36 (3) (1999)
233254.
[31] L.X. Yu, A.B. Straughn, P.J. Faustino, Y. Yang, A. Parekh, A.B. Ciavarella, E. Asafu-
Adjaye, M.U. Mehta, D.P. Conner, L.J. Lesko, A.S. Hussain, Mol. Pharm. 1 (5)
(2004) 357362.
[32] C.J.H. Porter, C.W. Pouton, J.F. Cuine, W.N. Charman, Adv. Drug. Del. Rev. 60
(2008) 673691.
[33] J.S. Frederick, J.S. Ronald, F.B. William, Liver Disease in Children, third ed.,
Cambridge University Press, 2007.
[34] M.C. Carey, P.C. Hirom, D.M. Small, Biochem. J. 153 (1976) 519531.
[35] M. Yokoyama, Crit. Rev. Ther. Drug Carrier Syst. 9 (1992) 213248.
[36] S.S. Shah, R. Ahmad, S.W.H. Shah, K.M. Asif, K. Naeem, Colloids Surf. A:
Physicochem. Eng. Aspects 137 (1998) 301305.
[37] W. Mehnert, K. Mader, Adv. Drug. Del. Rev. 47 (2001) 165196.
S. Chakraborty et al. / Journal of Colloid and Interface Science 335 (2009) 242249 249