Solubility Advantage of Amorphous Pharmaceuticals:

I. A Thermodynamic Analysis
SHARAD B. MURDANDE,
1,2
MICHAEL J. PIKAL,
1,3
RAVI M. SHANKER,
2
ROBIN H. BOGNER
1,3
1
Department of Pharmaceutical Sciences, University of Connecticut, Storrs, Connecticut 06269
2
Pzer Global R&D, Groton Labs, Groton, Connecticut 06340
3
Institute of Material Science, University of Connecticut, Storrs, Connecticut 06269
Received 29 December 2008; revised 6 July 2009; accepted 8 July 2009
Published online 20 August 2009 in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/jps.21903
ABSTRACT: In recent years there has been growing interest in advancing amorphous
pharmaceuticals as an approach for achieving adequate solubility. Due to difculties in
the experimental measurement of solubility, a reliable estimate of the solubility
enhancement ratio of an amorphous formof a drug relative to its crystalline counterpart
would be highly useful. We have developed a rigorous thermodynamic approach to
estimate enhancement in solubility that can be achieved by conversion of a crystalline
form to the amorphous form. We rigorously treat the three factors that contribute to
differences in solubility between amorphous and crystalline forms. First, we calculate
the free energy difference between amorphous and crystalline forms from thermal
properties measured by modulated differential scanning calorimetry (MDSC). Secondly,
since an amorphous solute can absorb signicant amounts of water, which reduces its
activity and solubility, a correction is made using water sorption isotherm data
and the GibbsDuhem equation. Next, a correction is made for differences in the
degree of ionization due to differences in solubilities of the two forms. Utilizing
this approach the theoretically estimated solubility enhancement ratio of 7.0 for
indomethacin (amorphous/g-crystal) was found to be in close agreement with the
experimentally determined ratio of 4.9. 2009 Wiley-Liss, Inc. and the American Pharmacists
Association J Pharm Sci 99:12541264, 2010
Keywords: solubility; thermodynamics; activity; amorphous; crystal
INTRODUCTION
Accurate estimate of the degree of improvement in
solubility provided by an amorphous drug would
enable the formulator to make an informed
decision of whether or not to pursue utilizing
the amorphous formfor enhancing bioavailability.
Experimental determination of the solubility of
an amorphous material has been found to be
extremely difcult because of the tendency of
amorphous materials to rapidly crystallize
upon contact with water. Hancock and Parks
1
reported a procedure for the estimation of the
solubility advantage of amorphous pharma-
ceuticals based on measured thermal properties
of amorphous and crystalline forms of drugs.
Their approach was based on the method of
Parks et al.
2,3
where the solubility ratio of the two
forms (Aamorphous and Ccrystalline) being
examined at any temperature (T) is expressed in
terms of the free energy difference (DG) between
those two forms:
DG
A;C
T
RTln
a
amorphous
a
crystal
(1)
Correspondence to: Michael J. Pikal (Telephone: 860-486-
3202; Fax: 860-486-2076; E-mail: michael.pikal@uconn.edu)
Journal of Pharmaceutical Sciences, Vol. 99, 12541264 (2010)
2009 Wiley-Liss, Inc. and the American Pharmacists Association
1254 JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 99, NO. 3, MARCH 2010
where a is activity of the solute in the saturated
solution, which in dilute solution where activity
coefcients are essentially unity, is the con-
centration at saturation or the solubility, R is
the gas constant, and T is the temperature. The
key issue with the application of Eq. (1) is how to
obtain the free energy difference from DSC data,
with the principle assumption involving the
estimation of the heat capacity difference between
amorphous and crystalline forms. There were
several procedures used by Hancock and Parks
to estimate the heat capacity difference. The
simplest procedure was to assume the heat
capacity difference between amorphous and crys-
talline forms (DCp
a,c
) is nearly zero and may be
neglected. However, the accuracy of this approach
is questionable. Another approach was to assume
that the difference in heat capacity between
amorphous and crystalline forms is the same as
the heat capacity gain upon going through the
glass transition, DCp
a,c
%DCp
Tg
. However, this
approximation is also likely to be inaccurate. A
third approach was to assume that DCp
a,c
%DS
f
(the entropy of fusion of the crystalline solute).
Again, this relationship may also be highly
inaccurate.
4
None of these approximations are
necessary if one obtains accurate heat capacity
data, which is relatively straightforward with
modulated DSC. That is, one can measure,
directly, the difference in heat capacity between
amorphous and crystalline forms, and this direct
measurement procedure is used in this research.
An implicit assumption in the Hancock and Parks
approach is that when equilibrium is established
between the neat amorphous solid and the
saturated solution, the amorphous solid remains
as single component system (i.e., free of solvent).
However, the amorphous form can absorb sig-
nicant amounts of water upon exposure to water
during the determination of its apparent aqueous
solubility. Thus, the amorphous solid is no longer
a one component system, and the free energy of
the amorphous solute changes due to solute-
solvent interactions and due to statistical con-
tributions from the entropy of mixing, thus
reducing the solubility enhancement ratio. In
the present work, the impact of water sorption on
the thermodynamic activity of the amorphous
solute is evaluated in order to obtain a more
accurate estimate of the solubility advantage.
Additionally, there is a reduction in the fraction
ionized in the un-buffered solution in equilibrium
with the amorphous solid relative to the crystal-
line solid, which will also lower the solubility
enhancement. The reduction is a result of the
higher concentration in the system containing
amorphous solute. The wide discrepancies bet-
ween observed and measured solubility enhance-
ment ratios in the Hancock and Parks
1
study can
be attributed to the three factors discussed above:
(a) inaccurate assumptions of the heat capacity
difference between the two forms; (b) changes in
the free energy of amorphous form due to sorption
of water; and (c) reduced fraction ionized at higher
concentration and its impact on entropy.
The objective of this work was to rene the
thermodynamic estimate of the ratio of the
solubility of amorphous and crystalline pharma-
ceuticals. In summary, the present approach was
based on the following: (a) making a correction for
the reduction of the degree of ionization in
saturated solutions of the amorphous form
relative to the crystalline form; (b) calculating
the activity of the solute in the hydrated
amorphous solid by determining the activity of
the water from water sorption isotherms and
applying the GibbsDuhemequation to determine
the corresponding solute activity; and (c) calculat-
ing free energy difference between dry amorphous
and crystalline forms by measuring melting point
and heat of fusion for the crystalline form,
determining the glass transition temperature
for the amorphous form, and measuring the heat
capacity difference between the two forms using
MDSC. This analysis renes the approach of
Hancock and Parks by eliminating many of their
assumptions. The solubility enhancement predic-
tion achieved by our approach is expected to be
close to the actual concentration one might
achieve in the absence of crystallization. Our
analysis does require measurement of some
experimental parameters (i.e., melting tempera-
ture, heat of fusion, ionization constant and
solubility for crystalline form; glass transition
temperature for amorphous form; heat capacity
difference between crystal and amorphous forms
and activity of hydrated amorphous solute).
DERIVATION OF THERMODYNAMIC
RELATIONSHIPS TO CALCULATE THE
SOLUBILITY ENHANCEMENT RATIO
(AMORPHOUS/CRYSTAL)
Solubility is a phase equilibrium in which the
chemical potential (m) of each component is equal
DOI 10.1002/jps JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 99, NO. 3, MARCH 2010
SOLUBILITY ADVANTAGE OF AMORPHOUS PHARMACEUTICALS 1255
in all phases in equilibrium. Using 2 to denote
the solute (pharmaceutical) component and 1 to
denote the solvent (water), at equilibrium we may
write
m
soln
2
m
solid
2
(2)
Form of the Chemical Potential in Solution
With Ionization
For an ionizable solute where a is the fraction
ionized, the chemical potential of the solute in
solution may be written as the sumof the chemical
potentials of all the species,
m
soln
2
am
0

RTlnc

m
0

RTlnc

1 a
m
0
u
RTlnc
u
g
u
(3)
The symbols, c and g, represent the concentration
and activity coefcient, respectively, of each
species (cation, anion, and uunionized);
R is the gas constant and T is the temperature in
degrees Kelvin.
5
The unionized and ionized
species are in equilibrium, so the free energy of
ionization is zero. Thus for a monoacidic or
monobasic compound, we may relate the compo-
nent chemical potentials as follows:
1 am
0
u
RTlnc
u
g
u

am
0

m
0

2RTlnc

(4)
where c

is the concentration of the negative or

positive ions formed, and g

is the mean ionic

activity coefcient. We choose the expression
written in terms of the concentration of unionized
species, c
u
, but we substitute as follows:
c
u
(1 a)c
T
, where c
T
is the total concentration
of solute species in solution, ionized and union-
ized, which is what would be measured in a
solubility experiment. Thus, with this choice of
components, combining Eqs. (3) and (4) yields the
chemical potential of the solute in solution as
m
soln
2
m
0
2
RTln1 ac
T
(5)
where we have assumed the solution is suf-
ciently dilute due to the lowsolubility of the solute
such that their activity coefcients are nearly
unity. The standard state chemical potential, m
0
2
,
refers to the Henrys lawstandard state where the
activity coefcient approaches unity as the solute
concentration approaches zero.
Similarly, we write the chemical potential of the
solute in the solid phase as
m
solid
2
m

2
RTlnX
s
2
g
s
2
(6)
where we choose m

2
as the hypothetical super-
cooled liquid for the amorphous system standard
state. If the solid is entirely crystalline, both the
mole fraction of 2 in the solid, X
s
2
, and the activity
coefcient of 2 in the solid, g
s
2
, are unity, and the
chemical potential is simply equal to the standard
state value, which is the chemical potential of the
pure crystalline material. If the solid is entirely
amorphous, neither the mole fraction of 2 in the
solid nor the activity coefcient is necessarily
unity. In general the activity will not be unity as
sorption of water by amorphous solid reduces the
chemical potential which results in lowering its
activity. Combining Eqs. (2), (5), and (6), we then
obtain
ln
1 ac
T
X
s
2
g
s
2
_ _

m
0
2
m

RT
constant (7)
Note that the bars above the symbols are used
to indicate that the properties refer to the
properties of solid and solution at equilibrium
between the two phases (i.e., saturated solution
and solid saturated with water). Eq. (7) is the
usual solubility equation, but since we are
accounting for ionization and for absorption of
solvent (water) in the solid amorphous phase, the
expression on the left hand side of the equation
is somewhat more complex than found in the
classical expression. The right hand side of Eq. (7)
is the negative of the free energy of solution,
divided by RT. The standard Gibbs free energy
of solution, denoted DG
0
soln
, is dened by the
hypothetical process where one mole of solute is
dissolved in an innite amount of solvent satu-
rated with solute, DG
0
soln
m
0
2
m

2
. Note that the
fraction ionized a, is obtained by solving the
ionization equilibrium equation
a
2
1 a

K
a
c
T
(8)
where K
a
is the ionization constant. For a
given compound, the fraction ionized decreases
with increasing concentration of ionizable
solute. The degree of ionization is different (i.e.,
lower) for a saturated solution in equilibrium
with an amorphous solid than for the correspond-
ing equilibrium with a crystalline solid. Thus,
the entropy gained on ionization is less for
the amorphous systemper mole solubilized, which
JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 99, NO. 3, MARCH 2010 DOI 10.1002/jps
1256 MURDANDE ET AL.
in turn moderates the solubility advantage of
the amorphous form. Therefore the theoretically
estimated solubility ratio needs to be corrected
for ionization differences due to the differences in
aqueous drug concentration between amorphous
and crystal forms of a drug. The amorphous
phase produces a higher concentration in a
saturated solution than does the crystalline
phase. Thus, the fraction of dissolved solute
ionized in an un-buffered system is less for the
amorphous solute, resulting in the solubility
advantage of the amorphous form being
dampened.
Impact of Moisture Sorption on the Activity of
Amorphous Solute
We now focus on the determination of the
impact of absorbed water on the chemical
potential of the solute. The value of X
s
2
g
s
2
in
Eq. (7) may be obtained from water sorption
isotherm data through the use of the Gibbs
Duhem equation. Application of the Gibbs
Duhem equation to calculate the activity of
drugs or other solids in the hydrated solid has
been described in the literature.
6,7
This approach
involves determination of moisture sorption
isotherm using a gravimetric moisture sorption
analyzer where the number of moles of water
absorbed per mole of solute are measured as a
function of relative humidity (i.e., water activity).
From the measured activity of water as function
of composition, the activity of the solute can be
calculated by integrating the GibbsDuhem
equation.
At constant pressure and temperature, the
GibbsDuhem equation may be written
X
2
dm
2
X
1
dm
1
0 (9)
Rearrangement of Eq. (9) and integration from
X
2
1 to the value of X
2
when the solid is in
equilibrium with solution yields
m
solid
2
m

2

_
X
s
2
X
s
2
1
X
s
1
X
s
2
dm
s
1
X
2
(10)
Recall that all symbols in Eq. (10) refer
to properties of the solid. When equilibrium
exists between water in the vapor phase
and water in the solid, the chemical potential
of water in the solid and in the vapor are equal,
yielding
dm
solid
1
dm
vapor
1
RTdlnP
1
(11)
where m
solid
1
is the chemical potential of solvent
(water) in the solid, m
vapor
1
is the chemical potential
of water in the vapor phase and P
1
is the partial
pressure of water vapor. Note that the activity of
water (a
1
), relative to a standard state of pure
water, is given by
a
1

P
1
P
0
1
(12)
where P
0
1
is the vapor pressure of pure water.
Thus, we may combine Eqs. (6) and (10)(12) and
write,
lnX
s
2
g
s
2

m
solid
2
m

2
RT

_
X
s
1
X
s
1
0
n
1
n
2
_ _
dlna
1
X
s
1

_
a
s
1
a
s
1
0
n
1
n
2
_ _
1
a
1
da
1
(13)
where n
1
and n
2
are moles of water and solute,
respectively. It is the ratio n
1
/n
2
that is directly
obtained from the water sorption experiment. For
convenience, we denote the integral on the far
right hand side of Eq. (13) as I(a
2
), which may be
written as
Ia
2

_
a
H
1
a
1
0
n
1
n
2
_ _
1
a
1
da
1
..
Henrys LawRegion
a
1
k
H
n
1
=n
2

_
a
s
1
a
H
1
n
1
n
2
_ _
1
a
1
da
1
..
HighWater Region

n
1
n
2
_ _
Henry

_
a
s
1
a
H
1
n
1
n
2
_ _
1
a
1
da
1
..
HighWater Region
(14)
In Eq. (14), we have divided the limits of
integration into the Henrys law region, which
ranges from zero water content to the limit of
linearity of water activity with n
1
/n
2
, denoted a
H
1
and a high water region. The value of the
integral over the Henrys law region is simply
the concentration value at the limit, (n
1
/n
2
)
Henry.
The integral in the high water region may be
DOI 10.1002/jps JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 99, NO. 3, MARCH 2010
SOLUBILITY ADVANTAGE OF AMORPHOUS PHARMACEUTICALS 1257
numerically evaluated
1
using water sorption
isotherm data where we measure the number
of moles of water per mole of solid solute,
n
1
/n
2
, as a function of relative humidity,
RH100a
1
. Since the water sorption isotherm
data normally would not be obtained at unit
water activity, a small extrapolation is required.
For this purpose, the term, exp(I(a
2
)), is
evaluated at each value of a
1
and the values
of exp(I(a
2
)) are plotted versus a
1
and extra-
polated a short distance to a
1
1 to obtain the
correction to solubility enhancement described in
Eq. (16).
Next, we combine Eqs. (7), (13), and (14) to
nd,
c
T

1
1 a
_ _
exp Ia
2
exp
DG
0
soln
RT
_ _
(15)
Eq. (15) represents the incorporation of both
ionization and water sorption effects into the
thermodynamic prediction of solubility.
Equations of the formof Eq. (15) may be written
for both amorphous and crystalline phases. For an
entirely crystalline solid, by denition I(a
2
) 0
(i.e., mole fraction and activity coefcient are
unity). Thus, the ratio of solubilities of amorphous
and crystalline solids, R
s
, can be written as
R
s

c
a
T
c
x
T

1 a
x

1 a
a

exp Ia
2
exp
m
a
2
m
x
2

RT
_ _
(16)
where c
a
T
is solubility of amorphous solute, c
x
T
is solubility of crystalline solute, and m
a
2
m
x
2
is the change in standard state chemical
potential on transforming from the crystal to
the dry amorphous state. These standard state
chemical potentials are also molar free energies,
and we may denote this difference in chemical
potentials as the standard state molar free
energy on transition from crystalline to amor-
phous states,
DG
xa
m
a
2
m
x
2
(17)
Free Energy Difference between Neat Amorphous
and Crystalline Forms
At the melting point (T
m
) of the crystalline phase,
DG
xa
DG
xa
=T 0, but at other temperatures,
the free energy of transformation is not zero. We
evaluate the effect of temperature on free energy
as usual,
@DG
xa
=T
@T

DH
xa
T
2
(18)
where the enthalpy change, DH
xa
is itself tem-
perature dependent through the corresponding
change in heat capacity. The change in heat
capacity in going from crystal to super-cooled
liquid at T
g
<T<T
m
denoted, DC
x;l
p
, is large, but
for T<T
g
is the heat capacity change in going
from crystal to glass, denoted DC
x;g
p
, which is
normally quite small. We relate the two heat
capacity changes through the quantity, g
DC
x;g
p
1 gDC
x;l
p
(19)
The irreversible enthalpy of melting at tempera-
ture, T, can be described as the sum of the
enthalpies of the three reversible processes:
heating of the crystalline solid to its melting
point, T
m
; melting the solid at its melting point;
and nally cooling the liquid back to temperature
T. Thus, the heat of melting at temperature, T,
can be related to its value at melting point, DH
f
by
DH
f
T DH
f
T
m
DC
x;l
p
T T
m
(20)
Substituting Eq. (20) into Eq. (18) and using the
relationship given in Eq. (19), we integrate
Eq. (18) from the melting point, T
m
, to the
temperature of interest, T, assuming that the
heat capacity changes are independent of tem-
perature. Thus, after considerable algebra, one
nds
DG
xa
T
DH
f
T
m
T
m
T TDC
x;l
p
ln
T
m
T
_ _ _
g ln
T
g
T
_ _

T
m
T
1 g g
T
m
T
g
1
_ __
(21)
Eq. (21) is utilized to calculate the free energy
difference between amorphous and crystal forms
to insert into Eq. (16). We acknowledge that the
glassy state is not an equilibrium state. The state
1
For example, one would plot (n
1
/n
2
)/a
1
versus a
1
, and the integral
would be the area under the curve froma
H
1
to the value of a
s
1
, the
latter being the activity of water in the solid in equilibrium
with the dilute solution. An effective procedure is to evaluate
the integral for the high water region numerically by summing
the products,
n1
n2
1
a1
_ _
i1;i
a
i1
1
a
i
1
from the end of the Henrys
law region (i H) to the a
1
value of interest. This gives the
partial area under the curve fromthe starting point (i.e., end of
the Henrys law region) to the water activity of interest, and
then the value of I(a
2
) is obtained from Eq. (14).
JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 99, NO. 3, MARCH 2010 DOI 10.1002/jps
1258 MURDANDE ET AL.
of the glass will depend on its thermal history and
physical properties may change with time. How-
ever, such variations and changes are small on the
time scales of interest. We treat the system as
an equilibrium state to evaluate relationships
between thermodynamic properties. Thermody-
namic calculations are commonly carried out on
systems undergoing slow changes; in fact, most
pharmaceutical systems are intrinsically unstable
chemically. In the present case, while changes in
enthalpy and heat capacity of the glassy state do
occur over time, such changes are very small.
8
The portion of Eq. (16) that remains to be
addressed is the evaluation of the term a
a
. This
evaluation may be facilitated by writing the
solubility enhancement ratio (i.e., Eq. 16) in the
form,
R
s

B
1 a
a

;
B 1 a
x
exp Ia
2
exp
DG
xa
T
RT
_ _
(22)
As noted in Eq. (22), the value of B may be
calculated from: (a) the known solubility of the
crystalline form and the ionization constant (to
give the degree of ionization), (b) the water
sorption isotherm data to give I(a
2
) according to
Eq. (14), and (c) the thermal data to give the free
energy term from Eq. (21). However, the degree of
ionization for the amorphous form is unknown
since the concentration of drug in solution at
equilibrium is unknown for the amorphous form.
However, the degree of ionization for the amor-
phous form is easily calculated utilizing the
ionization constant for crystalline solute, solubi-
lity of crystalline solute and DSC and water
sorption data. Using Eq. (8) and the denition of
R
s
(Eq. 22), one nds
a
a

K
a
=c
x
T
B
_
1

K
a
=c
x
T
B
_ (23)
where K
a
is the ionization constant, c
x
T
is the
solubility of the crystalline phase, and B is a
collection of properties obtainable from DSC and
water sorption data using Eq. (22). In summary,
the ratio of solubilities of amorphous and crystal-
line solids can be calculated using Eq. (16). This
equation has three terms. The rst term denes
the difference in degree of ionization between
crystalline and amorphous forms using experi-
mental data on ionization constant of crystalline
solid (K
a
) and solubility of the crystal form (c
x
T
)
(Eqs. 8 and 23). The second term denes activity
of the amorphous solute determined from moist-
ure sorption isotherm data using gravimetric
moisture sorption analyzer and application of
the GibbsDuhem equation (i.e., by numerically
integrating the moisture sorption data according
to Eq. 14). The third term determines the free
energy difference between amorphous and crystal
forms from thermal properties (melting point,
heats of fusion, glass transition temperature, and
heat capacity change at glass transition tempera-
ture), directly measured using modulated DSC,
according to Eq. (21).
EXPERIMENTAL METHODS
Crystalline indomethacin (1-(4-chlorobenzoyl)-5-
methoxy-2-methyl-1H-indole-3-acetic; MW 357.8
Da; thermodynamically stable g-form) of >99%
purity was purchased from Sigma Chemical
Company (St. Louis, MO). Amorphous indo-
methacin was prepared by melting the g-form of
crystalline indomethacin; the melt was kept at the
temperature of melting (i.e., 1628C) for 5min,
when it was obvious that 100% molten material
was obtained (visual observation). The molten
material was quench-cooled by immersion into
liquid nitrogen and ground in liquid nitrogen in a
previously cooled mortar and pestle in a dry box to
avoid condensation of moisture. The resulting
samples of indomethacin were conrmed to be
amorphous by the absence of birefringence under
cross polarizers by optical microscopy, absence of
a distinct diffraction pattern characteristic of
crystalline material by powder X-ray diffraction,
and presence of a glass transition (T
g
) by
differential scanning calorimetry. Amorphous
samples were stored at 208C over phosphorus
pentoxide desiccant until use. The water content
of each sample was found to be below 0.1% (w/w)
by KarlFisher titration.
DSC Measurements
Thermal properties for indomethacin were deter-
mined using a modulated temperature differen-
tial scanning calorimeter (Q1000, TAInstruments
Inc., New Castle, DE). The instrument was
calibrated for temperature and cell constant using
high purity indium. Prior to the measurement of
heat capacity, amorphous indomethacin was
linearly heated at 1K/min to 10 K above its glass
DOI 10.1002/jps JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 99, NO. 3, MARCH 2010
SOLUBILITY ADVANTAGE OF AMORPHOUS PHARMACEUTICALS 1259
transition temperature (T
g
) and rapidly cooled to
50 K below T
g
without recrystallization. For
measurement of heat capacity of either crystalline
or amorphous material, the samples of indo-
methacin were heated at 2K/min with an oscilla-
tion period of 60 s and amplitude of 0.5 K. The
instrument was calibrated for heat capacity using
a sapphire disk. Heat capacities were measured
for crystalline and amorphous indomethacin over
a temperature range from 50K below glass
transition temperature (T
g
) to temperatures just
below the melting point for three individual
samples. While it is difcult to measure absolute
heat capacity accurately using MDSC, it should be
noted that we only require accurate values for the
difference in heat capacity between amorphous
and crystalline solid.
8
Determination of Ionization Constant (pK
a
)
The ionization constant (pK
a
) of indomethacin was
determined by using an aqueous pH solubility
prole method.
9
Briey, the equilibriumsolubility
of indomethacin was determined in aqueous
buffers (0.1 M, phosphate/borate/acetate) over
the range of pH 1.610.5.
The concentration of each sample was analyzed
by reverse-phase gradient HPLC. The HPLC
analysis was performed using Agilent 1100 serial
system (Agilent Technologies, Palo Alto, CA)
equipped with a 3.9 mm150 mm Symmetry-
Shield C18 column (Waters, Milford, MA). The
column temperature was 258C. The wavelength
of detection was set to 210 nm. The mobile
phase consisted of binary gradient of solvent A
(acetonitrile) and solvent B (water with 0.1%
triuroacetic acid). The linear gradient started at
5%Aand increased to 95%Ain 25 min followed by
return to the starting condition within 2 min and
equilibrated at the starting condition for 3min.
The injection volume was 20 mL. The owrate was
1.0 mL/min and total run time was set for 30min.
The pK
a
was then calculated by tting the
solubility versus pH data to a single pK
a
equation,
pK
a
S
o
1 10
pHpK
a

, using Grat
1
software,
where S
o
is the intrinsic solubility measured
at pH 1.6.
Determination of Moisture Sorption Isotherms
The water sorption isotherm of amorphous
indomethacin was determined at 258C using a
symmetrical gravimetric analyzer (Model SGA-
100, VTI, Hialeah, FL) equipped with a controlled
relative humidity vacuum assembly and a
Cahn D-200 microbalance. The amorphous indo-
methacin sample was transferred to the balance
assembly where it was dried at 258C until a
constant mass was attained (i.e., mass change
less than 2mg over 2 h). The moisture sorption
isotherm was measured by increasing percent
relative humidity (RH) in steps by mixing
saturated water vapor and dry nitrogen at a
given ow ratio to achieve the desired RH and
holding at each RH until constant mass was
obtained. The procedure employed RH scans in
steps of 2% below 10% RH and 5% above 10% RH.
Equilibrium was assumed to be established
when the mass change was less than 0.2 mg/
min. Moisture sorption isotherms were generated
on three separately weighed samples, all taken
from the same batch. At the end of the sorption
run, samples were veried to have remained
amorphous by checking for crystallinity by
polarized light microscopy and powder X-ray
diffraction.
Determination of Experimental Solubility
As an experimental approximation to solubility,
we report the maximumof the concentrationtime
curve in a highly controlled dissolution experi-
ment. Since this quantity is controlled in part by
thermodynamics and in part by kinetics of
dissolution and recrystallization, we use the
term apparent solubility when describing this
maximum concentration. Solubility of the crystal-
line form and apparent solubility of the amor-
phous form of indomethacin were measured in
de-ionized water at 258C. Concentrationtime
curves were determined using a USP type II
(paddle) apparatus (Distek dissolution system
2100C, Distek Inc., North Brunswick, NJ). An
excess of powdered drug was added to 250 mL of
water (%2mg/mL). In order to improve the powder
dispersion, a paddle speed of 300 rpm was
used. Both crystalline and amorphous solids were
screened through a 100-mesh screen (150mm size
opening) and retained on 200-mesh screen (75 mm
size opening) in order to avoid nes that dissolve
faster and larger particles that might cause
difculties in dispersing the drug in water,
thereby improving reproducibility of the concen-
tration measurements. At least three solubility
measurements were carried out for both crystal-
line and amorphous indomethacin. The solubility
JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 99, NO. 3, MARCH 2010 DOI 10.1002/jps
1260 MURDANDE ET AL.
samples were analyzed by reverse-phase gradient
HPLC.
An apparent solubility ratio was calculated
from the relationship,
R
s
exp :
C
a
T
C
x
T
where R
s
(exp.) is the apparent solubility enhance-
ment ratio, C
a
T
is the peak concentration of
amorphous indomethacin, or apparent solubility,
and C
x
T
is the solubility of crystalline indomethacin.
RESULTS
Thermal Properties and Ionization Constant (pK
a
)
Thermal properties are summarized in Table 1.
The heat capacities of indomethacin in the
crystalline and amorphous forms are shown in
Figure 1. The heat capacity of the amorphous form
of indomethacin was greater than its crystalline
form at all temperatures, and the heat capacity
increased signicantly as the amorphous form
was heated through the glass transition, as
expected. The thermal properties, including heat
capacity data, were used in Eqs. (19) and (21) to
accurately measure the free energy difference
between amorphous and crystal forms of indo-
methacin. The indomethacin pK
a
was determined
using aqueous pHsolubility prole, and the pK
a
of
indomethacin was calculated to be 4.01 which is
consistent with the literature
10
value of 4.15.
Moisture Sorption Isotherm and Solute Activity
The water sorption isotherms for the amorphous
and crystalline (g form) indomethacin forms at
258C are given in Figure 2. The isotherm for
the amorphous indomethacin shows signicant
sorption of water, whereas the crystalline sample
exhibits negligible sorption relative to the amor-
phous sample. Amorphous indomethacin samples
at the end of sorption run were found to be
amorphous by both polarized light microscopy and
powder X-ray diffraction. Figure 3 shows the
thermodynamic activity of solid amorphous indo-
methacin containing water obtained from numer-
ical integration of the data in Figure 2 using
Table 1. Theoretically Estimated Solubility
Enhancement Ratio for Indomethacin (Amorphous/
Crystal)
Quantity Value
Ionization Acid
c
x
T
1.4810
5
M
T
m
435.2 K
T
g
314.2 K
DC
x;l
p
164.6 J/mol K
a
2
0.716
a
a
0.65
DH
f
36,496 J/mol
g (from C
p
) 0.804
DG
0
x;a
298
11,490 3139 8351J/mol
R
s
(Theory) 7.0
c
x
T
, total concentration (mol/L) of crystalline drug in solution;
T
m
, melting point; DC
x;l
p
, heat capacity difference between
crystal and liquid; a
2
, solute activity (Eq. 14); a
a
, fraction of
ionized amorphous solute (Eq. 23); M, molecular weight; DH
f
,
heats of fusion; DG
0
x;a
298, free energy difference between
amorphous and crystal form at 298K (Eq. 21); R
s
(Theory),
calculated apparent solubility enhancement ratio (Eq. 16).
All data were obtained experimentally.
Figure 1. Constant pressure heat capacities for indo-
methacin in the crystalline (x) and amorphous (D)
solids. DC
x;g
p
DC
x;l
p
DC
g;l
p
.
Figure 2. Water sorption isotherm of indomethacin
in the crystalline (^) and amorphous form (*) at 258C
(n3; standard deviation).
DOI 10.1002/jps JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 99, NO. 3, MARCH 2010
SOLUBILITY ADVANTAGE OF AMORPHOUS PHARMACEUTICALS 1261
Eq. (14). Note that, consistent with the earlier
choice of standard state for the amorphous phase,
we use the pure amorphous component as
the standard state with mole fraction as the
concentration unit.
Experimental Solubilities
Amorphous indomethacin consistently exhibited
signicantly higher concentrations during ap-
parent solubility studies than did the g-crystalline
form of indomethacin at 258C, at least for a period
of about 2 h (Fig. 4). The apparent solubility
enhancement ratio (i.e., ratio of peak concentra-
tion values), was found to be 4.9 for indomethacin
(Table 2). The peak concentration of amorphous
indomethacin always occurred within 1020 min.
During this time, there was no sign of crystal-
lization (polarized light microscopy), indicating
that only the amorphous form was present in
the sample, but the peak concentration was not
sustained beyond 30 min. Therefore, the peak
concentration was perhaps only slightly less than
the true thermodynamic solubility of amorphous
indomethacin. The experimentally measured pH
for the case of crystalline indomethacin was
4.88 at the 2-h time point. In the case of
amorphous indomethacin, the measured pH was
4.40 at 10 min (measured close to the peak
solubility). The fraction of solute ionized
(a) calculated from the pH values was 0.88 in
case of crystalline indomethacin at pH 4.88, and
0.71 for amorphous indomethacin pH 4.40. The
value of 0.71 for amorphous indomethacin is close
to the value 0.65 calculated using Eq. (23).
Investigation of the undissolved solid by powder
X-ray diffraction at the end of 2 h revealed that
the material had partially converted to the
two most common crystalline polymorphs (a and
g form).
DISCUSSION
The challenges in predicting the solubility
enhancement ratio of amorphous solids relative
to their crystalline counterparts arise due to
several factors: (a) a change in thermodynamic
activity of amorphous solute as it absorbs water,
(b) in the case of ionizable compounds in pure
water, the difference in degree of ionization of the
solute at different concentrations, and (c) experi-
mentally, onset of crystallization frustrates
the attempt to determine equilibrium solubility
of the amorphous solid. Also, since an amorphous
solid has thermal history dependent properties,
one might argue that the free energy might
change as the sample anneals below T
g
and undergoes enthalpy relaxation. However,
enthalpy relaxation effects are likely to be
minimal for two reasons. First, the impact of
annealing on heat capacity is very small,
8
and
secondly, after contact with water, the glass
transition temperature will be signicantly
reduced and relaxation will proceed quickly to a
common state regardless of the initial state. We
assume, based on the short times required for
water uptake by an amorphous solid that absorp-
tion of water will be faster than the solubility
equilibration process. As described in this report,
the impact of water sorption on solute activity and
the impact of differing degrees of ionization can be
evaluated from experimental data. The more
serious problem is validation of the theoretical
results since determination of the thermodynamic
Figure 4. Experimental determination of solubi-
lity: concentration:time prole for crystalline (^)
and amorphous (*) indomethacin at 258C (n3;
standard deviation).
Figure 3. Solute activity as a function of function of
water activity for amorphous indomethacin (* solute
activity).
JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 99, NO. 3, MARCH 2010 DOI 10.1002/jps
1262 MURDANDE ET AL.
solubility of the amorphous form is hampered by
the conversion of the solute to a crystalline state
during the course of experimental solubility
measurement. The theoretically estimated solu-
bility enhancement ratio, Eq. (16), for indometha-
cin was found to be 7.0, and was only slightly
greater than the experimentally determined
value of 4.9. As noted earlier, at least with
indomethacin, the apparent solubility of the
amorphous form was expected to be reasonably
close to, but slightly less, than the thermodynamic
value as recrystallization was avoided at least for
a short period of time. This predicted result of
7.0 should be contrasted with that of Hancock and
Parks who reported a prediction of 25104, the
range arising fromthe different assumptions used
to estimate the heat capacity difference between
crystal and amorphous forms.
1
The effects of
ionization and water sorption on the theoretical
solubility enhancement ratio was found to be
highly signicant (Tab. 3). For example, when
only the effects of ionization are ignored, the
theoretical solubility ratio was calculated to be
20.8 (i.e., a factor of 3 larger than with ionization
included). However, if pH is controlled with a
buffer, the fraction ionized is independent of
solute concentration, and ionization does not
impact the solubility enhancement ratio. Alter-
nately, when only the effect of water sorption is
ignored, the theoretically estimated solubility
enhancement ratio is 9.9, about 30% larger
than without the correction. When both effects
of ionization and water sorption are ignored,
the theoretical solubility enhancement ratio is
29.0 (Tab. 3), which is at the low end of the
range of values provided by Hancock and Parks
(25104). Thus, it is not only necessary to employ
accurate heat capacity data, which is not dif-
cult with modulated DSC, the corrections for
water sorption and ionization are critical to the
useful prediction of the solubility enhancement
ratio.
CONCLUSIONS
The approach described in this research provides
a renement in the theoretical prediction of
the enhancement of solubility, relative to the
predictions obtained by Hancock and Parks.
1
It
appears that the calculations are sufciently
accurate to be useful in deciding whether or not
the potential for solubility enhancement via the
amorphous route is sufcient to justify the time
required to pursue this solution to the solubility
problem. The approach presented in this research
requires data on thermal properties, water sorp-
tion, and pK
a
; these data are routinely collected
during preformulation studies. However, this
approach needs to be further validated using a
wide range of compounds representing different
states of ionization and different molecular
properties, and such work is in progress.
Table 3. Contribution of Individual Properties of Amorphous and Crystalline Indomethacin to the Theoretically
Estimated Solubility Enhancement Ratio (Amorphous/Crystal)
Amorphous and Crystalline Properties Considered
Theoretically Estimated
Solubility Ratio (R
s
(Theory))
All: Ionization, accurate heat capacity, and water sorption (solute activity) 7.0
Accurate heat capacity only 29.0
Accurate heat capacity and water sorption (solute activity) only 20.8
Accurate heat capacity and ionization only 9.9
Table 2. Experimentally Determined Apparent Solubility Enhancement Ratio for Indomethacin (Amorphous/
Crystal)
Compound Ionization pK
a
Maximum Conc., mg/mL (M)
R
s
(exp.) Crystal Amorphous
Indomethacin Acid 4.01 5.3 (1.4810
5
M) 25.8 (7.2110
5
M) 4.9
R
s
(exp.), experimentally determined apparent solubility enhancement ratio.
DOI 10.1002/jps JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 99, NO. 3, MARCH 2010
SOLUBILITY ADVANTAGE OF AMORPHOUS PHARMACEUTICALS 1263
NOMENCLATURE
K
a
ionization constant
T
g
glass transition temperature
T
m
melting temperature
X
s
2
mole fraction of solute in the solid phase
a
1
water activity
a
2
solute activity
c
a
T
peak concentration of amorphous solute
c
x
T
solubility of crystalline solute
DC
x;g
p
heat capacity difference between crystal
and glass at constant pressure
DC
x;l
p
heat capacity difference between crystal
and liquid at constant pressure
DG
xa
Gibbs free energy difference between
amorphous and crystalline solute
DH
f
heats of fusion
a fraction of ionized solute
g
s
2
activity coefcient of solute in the solid
phase
m
0
2
Henrys law standard state
m

2
standard state for the solute
m
a
2
chemical potential of amorphous solute
m
a
2
standard state chemical potential for
amorphous solute
m
x
2
chemical potential of crystalline solute
m
x
2
standard state chemical potential for
crystalline solute
m
solid
1
chemical potential of water in the solid
phase
m
vapor
1
chemical potential of water in the vapor
phase
m
solid
2
chemical potential of solute in the solid
phase
m
soln
2
chemical potential of solute in the solution
phase
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