Enantiotropically Related Albendazole Polymorphs

MARCO B. PRANZO,1 DYANNE CRUICKSHANK,2 MASSIMO CORUZZI,1 MINO R. CAIRA,2 RUGGERO BETTINI1
1
Department of Pharmacy, University of Parma, Viale G.P. Usberti 27/A, 43124 Parma, Italy
2
Department of Chemistry, University of Cape Town, Rondebosch 7701, South Africa

Received 9 October 2009; revised 23 November 2009; accepted 27 November 2009

Published online 28 January 2010 in Wiley InterScience (www.interscience.wiley.com). DOI 10.1002/jps.22072

ABSTRACT: In the present study we report the solid-state properties of albendazole (ABZ)
re-crystallized from different solvents for comparison with the commercially available form.
Crystalline phases were characterized as to thermal behavior, X-ray diffractometry, both on
powder and single crystal, and solubility in methanol or 0.1 N HCl. The relevant thermodynamic
parameters were calculated from solubility measurements at different temperatures. The re-
crystallization of ABZ both from methanol and N,N-dimethylformamide afforded a new stable
polymorph form (Form II) enantiotropically related to the commercially available ABZ (Form I),
the latter being the metastable form at ambient temperature. Both forms proved to be physically
quite stable, likely due to a high-energy barrier for the activation of the interconversion. ABZ in
the solid state represents a rather complex system in which the molecular structural differences
that could be associated with the polymorphism are of at least four possible types, or combina-
tions of these: (a) tautomeric; (b) different conformations of either or both of the side-chains
attached to the bicyclic ring system; (c) the occurrence of molecular disorder or its absence; (d) no
essential difference in molecular structure but different hydrogen bonding arrangements in
the two polymorphs. ß 2010 Wiley-Liss, Inc. and the American Pharmacists Association J Pharm Sci
99:3731–3742, 2010
Keywords: albendazole; polymorphism; enantiotropy; crystal structure; solid state

INTRODUCTION of ABZ to class II or IV of the Biopharmaceutics

Albendazole (ABZ), methyl [5-(propylthio)-1-H-ben-
zimidazol-2-yl] carbamic acid methyl ester, is among
the most effective broad-spectrum anthelmintic
agents.1,2 The molecule was patented in 19753 and
the drug is described as occurring as colorless crystals
with melting point between 208 and 2108C, and being
practically insoluble in water.4
ABZ therapy is very important in systemic cestode
infections especially in inoperable or disseminated
cases of hydatidosis1 and neurocystercosis1,5,6 both in
human and veterinary medicine.7
ABZ is undetected7–10 or present at extremely low
concentrations2,7,9,11–13 in blood plasma after oral
administration in various animal species and in man,
due to an extensive first-pass metabolism occurring in
the enterocytes and in liver cells as well as the very
low intestinal absorption stemming from the unfa-
vorable aqueous solubility.5 Both these effects result
in a low and erratic bioavailability. The assignment
Correspondence to: Ruggero Bettini (Telephone: 390521905089;
Fax: 390521905006; E-mail: bettini@unipr.it)
Journal of Pharmaceutical Sciences, Vol. 99, 3731–3742 (2010)
ß 2010 Wiley-Liss, Inc. and the American Pharmacists Association
Classification System is still a matter of debate.14
Irrespective of this, the insufficient aqueous solubility
is an issue that has to be addressed by pharmaceutical
formulators.
In this respect, solid dispersions with polyvinyl-
pyrrolidone,5 liquid formulations with Transcutol115
and ABZ b-cyclodextrin complexes16 were prepared to
improve the aqueous solubility and dissolution
characteristics of ABZ.
On the other hand, it is known that solid-state
properties may play a crucial role in dissolution rate
and solubility, especially when polymorphs with
different thermodynamic stability are involved.
These properties deeply influence all steps of drug
product development from the drug candidate dis-
covery, through its processability and production of
the dosage form, up to the in vivo performance.17–19 In
fact, differences in crystal packing as well as in lattice
energy and entropy often result in significant changes
in physical properties, such as density, hardness,
tablettability, refractive index, melting point,
enthalpy of fusion, vapor pressure, solubility, dis-
solution rate, as well as other thermodynamic and
kinetic properties and even color between different
polymorphs of the same drug substance.18

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3732 PRANZO ET AL.

Despite the fact that ABZ was discovered more than Single Crystal X-Ray Analysis of Form II of ABZ
35 years ago and its use in therapy is well established,
Microscopic examination of many batches of ABZ
no reports are presently available on its solid-state
obtained by re-crystallization from methanol and
characterization nor on its possible polymorphism.
DMF revealed a constant crystal morphology, namely
The aim of the present work was to study the solid-
poorly formed laminae with typical thickness only
state properties of ABZ re-crystallized from different
10 mm. A specimen viewed normal to the thin
solvents for comparison with the commercially
section is shown in Figure 1. Eventually, a specimen
available form.
with thickness 0.06 mm was identified and cut to a
Crystalline phases were characterized as to ther-
more equant shape (0.18 mm  0.18 mm  0.06 mm)
mal behavior, X-ray diffractometry, both on powder
to minimize X-ray absorption error and ensure suffi-
and single crystal, and solubility in methanol or 0.1 N
cient volume for adequate X-ray diffraction intensity.
HCl. The relevant thermodynamic parameters were
This was mounted on a cryoloop with Paratone oil
calculated from solubility measurements at different
(Exxon Chemical Co., Houston, TX), placed on a
temperatures.
Nonius Kappa CCD four-circle diffractometer and
cooled in a stream of nitrogen vapor to 213(2) K using
MATERIALS AND METHODS an Oxford Cryostream cooler (Oxford Cryosystems,
Oxford, UK) to ensure a rigid mount. No phase change
ABZ Crystallization accompanied the cooling process.
Data collection (COLLECT software20) with Mo
An amount corresponding to about 100 mg of ABZ
Ka-radiation (l ¼ 0.71073 Å) involved a combination
(Sigma, Steinheim, Germany) was dissolved in
of f- and v-scans of 1.008 and 0.508, respectively. All
100 mL of methanol, (HPLC grade, 99.9% minimum
data were corrected for Lorentz-polarization effects.
assay, Carlo Erba Reagents, Milan, Italy) or N,N-
Absorption effects were negligible for the specimen
dimethylformamide, DMF, (99.8% assay, Fluka
used (transmission range 0.9555–0.9858). Unit cell
Chemie GmBH, Buchs, Switzerland) and stirred at
refinement and data-reduction were performed using
408C until a clear solution was obtained. The filtered
DENZO-SMN and SCALEPACK.21 The monoclinic
solution was allowed to spontaneously evaporate
crystal system was established from the Laue
under ambient conditions until ABZ crystallization
symmetry (2/m) and the space group C2/c from
was complete. Colorless crystals formed after a few
systematic absences, centrosymmetry being indicated
days from methanol, whereas light brown crystals
by the hE2
1i value of 0.922.
were collected after about 1 month from DMF.
The structure was solved using direct methods
(program SHELXS-9722) and refined by full-matrix
Thermal Analysis
least-squares against F2 (program SHELXL-9723).
Differential scanning calorimetry (DSC) was per- After location of the nonhydrogen atoms of the
formed on an indium calibrated Mettler DSC 821e asymmetric unit (a single molecule of ABZ), it was
(Mettler Toledo, Columbus, OH) driven by STARe noted that the propylthio-chain was disordered over
software (Mettler Toledo). DSC traces were recorded two chemically equivalent positions. Each component
by placing precisely weighed quantities (1–5 mg) in a was modeled with a fixed site occupancy of 0.5, based
sealed and pierced 40 mL aluminium pan. Scans were on very similar electron densities for the atoms
performed between 30 and 2258C at 5 K min
1 or comprising the two side-chains as well as intermo-
between 30 and 2708C at 40 K min
1 under a flux of lecular distance constraints. Following treatment of
dry nitrogen (100 mL min
1). Each powder sample the non-H atoms, special care was exercised in the
was analyzed at least in triplicate. location of hydrogen atoms, given the possibility of
Thermogravimetric analysis (TG 50, Mettler
Toledo) was carried out on ABZ samples placed in
70 mL alumina pans with a pierced cover. Samples
were heated under a flux of dry nitrogen
(100 mL min
1) at 5 K min
1 in the 30–2258C tem-
perature range.

X-Ray Diffraction Studies

X-ray diffraction patterns on powder were recorded
on a bench-scale diffractometer (Rigaku, Tokyo,
Japan) using a Cu Ka radiation source at 30 kV
voltage over the scanning range (2u) 5–358 (scanning Figure 1. Typical morphology of a crystal of albendazole
speed of 0.058/min). obtained by re-crystallization from methanol or DMF.

JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 99, NO. 9, SEPTEMBER 2010 DOI 10.1002/jps
 ENANTIOTROPICALLY RELATED ALBENDAZOLE POLYMORPHS 3733

tautomerism for this molecule. All H atoms were Table 1. Crystal Data and Refinement Parameters for
located unequivocally in difference Fourier electron Form II of Albendazole
density maps. In particular, both N atoms of the
Parameter Form II
imidazole ring were found to bear H atoms, while no
electron density attributable to a hydrogen atom was Chemical formula C12H15N3O2S
found on the exocyclic N atom. Figure 2a shows the Formula weight 265.33
Dcalc (g cm
3) 1.401
conventional structural diagram for ABZ and
Crystal system Monoclinic
Figure 2b the structure of the tautomer found in Space group C2/c
the crystal of Form II. The disordered model was a (Å) 23.934(5)
completed with half-atoms of hydrogen added at b (Å) 5.440(1)
positions C7, C8 of the phenyl ring. Except for one of c (Å) 19.586(6)
a (8) 90.0
the atoms of a disordered chain component (C18A), all
b (8) 99.29(1)
non-H atoms refined anisotropically. H atoms were g (8) 90.0
generally added in a riding model at idealized V (Å3) 2516.6(10)
positions based on their stereochemistries, clearly Formula units (Z) 8
established from electron density maps, and were m (Mo Ka) (mm
1) 0.255
T (K) 213(2)
assigned isotropic thermal parameters 1.2–1.3 times
Reflections 2157
those of their parent atoms. Crystal data and Parameters 204
refinement parameters are reported in Table 1. Completeness (%) 97.8
Molecular parameters were calculated with program R1 (on F > 4s ( F)) 0.0723
PLATON.24 wR2 (on F2, F > 4s ( F)) 0.1547
Goodness-of fit (S) 1.033
Final D/s <0.001
Solubility Determination Dr min.; max. (e Å
3)
0.257; 0.259

The solubility of both commercially available and

re-crystallized ABZ was determined at 258C in
CH3OH or HCl 0.1 N. The latter solubility determina-
tion was also performed in the 25–1008C interval. An
excess of drug was suspended in the selected solvent
and stirred with a Vortex three times within 1 h.
Thereafter, the suspension was allowed to settle for
48 h, then it was filtered (0.45 mm) and the filtrate was
diluted and analyzed by HPLC to determine the ABZ
concentration. Each determination was performed in
triplicate.
High-Performance Liquid Chromatography (HPLC)
ABZ in solution was quantified according to the
method described in the European Pharmacopoeia25
using an isocratic HPLC system (LC-10 ATvp,
Shimadzu, Tokyo, Japan) equipped with a Spher-
isorb1 column (5 mm, 4.6 mm  250 mm, ODS2,
Waters, Milford, MA) and a DAD detector (SPD-
M10Avp, Shimadzu) set between 250 and 260 nm.
The volume of the injected samples was 20 mL
(Rheodyne injector 7010).
As mobile phase a mixture consisting of 30% (v/v)
of ammonium dihydrogen phosphate solution
(1.67 g L
1) and 70% (v/v) of CH3OH at a flow rate
of 0.7 mL min
1 was used. ABZ retention time was
6 min.
The method was validated for linearity (R2 ¼
0.999), limit of detection (1.1 mg mL
1), limit of
quantification (3.6 mg mL
1) and relative standard
deviation (<1.5%).

RESULTS AND DISCUSSION

Figure 2. The conventional structural diagram for alben-
dazole (a) and the structure of the tautomer occurring in the
stable crystal Form II (b). The latter also shows the disorder
of the propylthio-chain established in this study, each com-
ponent being present at 50% site occupancy.
For ABZ re-crystallization, two polar solvents were
selected, one protic and one aprotic, differing mainly
in their volatility (boiling points 64.8 and 1538C for
methanol and DMF, respectively).

DOI 10.1002/jps JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 99, NO. 9, SEPTEMBER 2010
3734 PRANZO ET AL.

As stated above the microscopic examination of

ABZ crystals obtained from both methanol and DMF
revealed a laminar morphology. The different solvent
volatility resulted in a larger average dimension for
crystals obtained from DMF than from methanol
likely due to the slower crystal growth: as the
supersaturation was attained slowly, as in the case
of DMF, crossing of the metastable region was slow;
then the process of particle growth overtook the
process of nucleation, eventually leading to large
crystals.
The commercial product consisted of very small
crystals showing a strong tendency to aggregation
(Fig. 3).
TGA analysis carried out at the scanning rate of
5 K min
1 on the different ABZ specimens indicated a
weight loss of about 1.5% and about 13% in the 30–
1508C and 175–2258C intervals, respectively. No
significant differences were observed among the
samples tested. This indicates that the re-crystal-
lization process did not result in the formation of
solvate forms. In fact the theoretical content of
solvent for a ABZ monosolvate form would be
21.6% and 10.8% (w/w) for DMF and methanol,
respectively, whereas the water content by weight in
a hypothetical monohydrate form would be 6.3%.
DSC traces recorded from the commercial ABZ in
the 30–2258C range at 5 K min
1 (Fig. 4A) showed a
single broad endothermic peak at 202.3  2.28C (onset
189.1  2.98C) likely ascribable to final melting with
decomposition. This interpretation is supported by
the literature data4 and by the above mentioned
weight loss recorded in the melting interval by
TGA.
The traces relevant to the ABZ re-crystallized both
from methanol and from DMF were not different from
that of the commercial form, except for a small
endothermic phenomenon at 1508C (onset 1428C)
immediately followed by an exotherm.
The DSC traces recorded upon scanning at higher
rate (40 K min
1) (Fig. 4B) put in better evidence Figure 3. Pictures taken at the polarized light micro-
some interesting features: the curve for the commer- scope of (a) commercial albendazole crystals and re-crystal-
cial material, again, presented just one endothermic lized albendazole (b) from methanol, (c) from DMF. Original
peak around 2208C (onset 212.8  2.28C), whereas the magnification 20.
re-crystallized materials, in addition to the peak
around 2208C, presented also a clear endothermic
peak at about 1608C, closely followed by an exother- alone or in various carriers26–29 this is the first time
mic one. Although the two phenomena could not be that a thermal event other than final melting is
separated, the enthalpy change associated with the presented for ABZ.
first one was measured. The relevant data are We hypothesized that the endo–exo thermal events
reported in Table 2. observed between 140 and 1808C in the re-crystal-
It should be emphasized that the differences lized ABZ could be ascribed to the melting of a crystal
between the enthalpies associated with both the first form not yet described in literature, followed by a re-
and the second peak of the studied materials were not crystallization leading to a crystal phase correspond-
statistically significant (Student’s t-test: p > 0.05). ing to the commercial one that eventually melted with
Despite the large number of papers available in decomposition. This new crystal phase stems from the
literature presenting the thermal behavior of ABZ re-crystallization process.

JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 99, NO. 9, SEPTEMBER 2010 DOI 10.1002/jps
 ENANTIOTROPICALLY RELATED ALBENDAZOLE POLYMORPHS
Figure 4. DSC traces of commercial albendazole (curves
a), and albendazole re-crystallized from DMF (curves b) and
from methanol (curves c). Scan carried out at 5 K min
1
(panel A) and at 40 K min
1 (panel B).
The hypothesis was verified by X-ray diffraction on
powder. Figure 5 reports the diffraction patterns
obtained from the commercial and re-crystallized
ABZ crystals. The comparison between the pattern of
the commercially available material and that of the
ABZ re-crystallized from DMF (Fig. 5a) evidenced
some differences related to the shape of the peaks at
78 and around 258 2u and the presence of three peaks
(10.58, 158, and 308 2u) in the pattern of the re-
crystallized product that are not observed in that of
the commercial one. Moreover, the latter shows a
peak around 11.58 2u not present in the re-crystallized
ABZ.
On the other hand, the diffraction patterns of the
re-crystallized materials were practically superimpo-
Table 2. Peak Temperatures and Relevant Enthalpies of ABZ Samples
1st Peak, 8C
Commercial product — —
Re-crystallized from DMF 159.0 (0.9)
Re-crystallized from methanol 160.3 (0.5)
Standard deviation in parenthesis (n ¼ 3).
DOI 10.1002/jps
3735
sable (Fig. 5b). The only noticeable difference was
represented by the higher intensity of the diffraction
peaks of ABZ re-crystallized from DMF, most likely
ascribable to different extents of crystallinity and
preferred orientation.
On the basis of these findings it can be affirmed that
the re-crystallization of ABZ at ambient temperature
both from methanol and DMF leads to a new
polymorph (Form II) having a melting temperature
at 1608C which re-crystallizes during the DSC scan to
give rise to the polymorph melting at 2208C (Form I).
The fusion and re-crystallization of the low melting
polymorph can be better evidenced when a high
scanning rate is used in the thermal analysis.
The remarkable similarity between the DSC curves
of the two re-crystallized products indicates that the
polymorphs obtained upon solvent evaporation may
depend on the crystallization conditions, in particular
the temperature, rather than on the type of solvent
used in the re-crystallization process.
These observations allow us to assume that the two
polymorphs likely constitute an enantiotropic pair
with a transition temperature lower than 1408C; in
this respect, the lower-melting crystal would repre-
sent the stable form at ambient temperature.
The solubility in methanol and 0.1 N HCl aqueous
solution was determined at 258C, both for investigat-
ing possible differences between commercial and re-
crystallized ABZ and to serve as a coarse indicator of
the relative thermodynamic stability of the two
crystal forms.
The values obtained are reported in Table 3. ABZ
Form I (commercial) proved to be more soluble than
Form II (re-crystallized from DMF). The difference
was low but statistically significant in both solvents
( p < 0.01 by Student’s t-test).
It is worth stressing that the accurate determina-
tion of equilibrium solubility of two polymorphs is
often a difficult task as solution-mediated conversion
may occur in the timeframe of the measurements.
Furthermore, we are aware that the approach is
conceptually incorrect since, by definition, only the
stable form is ‘‘thermodynamically allowed’’ to
achieve equilibrium solubility.
With respect to this concern DSC analysis was
carried out on the solid phases recovered at the end
of the equilibrium solubility experiments. For both
materials no change of phase was noticed, thus
it was concluded that ABZ Form I, although
DH 1st Peak, J g
1 2nd Peak, 8C DH 2nd Peak, J g
1
220.7 (0.1)
133.5 (2.1)

10.6 (2.6) 220.4 (0.6)
119.7 (18.9)

8.4 (0.2) 218.7 (1.78)
137.2 (5.6)
JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 99, NO. 9, SEPTEMBER 2010
3736 PRANZO ET AL.

Figure 5. X-ray diffraction patterns on powder of albendazole samples: (panel a)

commercial and re-crystallized from DMF; (panel b) re-crystallized from DMF and
re-crystallized from methanol.

JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 99, NO. 9, SEPTEMBER 2010 DOI 10.1002/jps
 ENANTIOTROPICALLY RELATED ALBENDAZOLE POLYMORPHS 3737

Table 3. Solubility of Commercial or DMF

Re-Crystallized ABZ in Methanol and in HCl 0.1 N
Aqueous Solution at 258C

Solubility in Solubility in
Methanol HCl 0.1 N

Commercial product (Form I) 1.45 (0.47) 0.54 (0.17)

Re-crystallized from DMF (Form II) 0.72 (0.05) 0.30 (0.07)

Standard deviation in parenthesis (n ¼ 3).

thermodynamically unfavorable at 258C, was kineti-

cally stable enough for allow the determination of a
reliable solubility value.
The differences in solubility observed are useful to
gain some insights into the thermodynamic relation-
ship between the two polymorphs. Form II displays
the lower solubility in both solvents; therefore, it
should be considered the more stable form at 258C.
Furthermore, the possibility to determine the
temperature range of thermodynamic stability by
measurements of equilibrium solubility of different
polymorphs is well established.30–32 Therefore, in
order to verify the correctness of our assumption
about the enantiotropic relationship, the ABZ II ! I
transition temperature was estimated by measuring
the ABZ solubility in aqueous HCl solution at
Figure 6. Solubility of albendazole polymorphs in the
different temperatures in the 25–1008C interval. 25–1008C range in 0.1 N aqueous HCl solution represented
This solvent was selected because of the ease of in mass per unit volume (panel A) and as van’t Hoff type plot
analysis. (panel B). Form I square, Form II circle. The bars represent
The obtained data are reported in Figure 6 both as the standard deviation.
concentration (mass per unit volume) versus tem-
perature (panel A) and as classical van’t Hoff plot
(panel B). The values obtained were 21.2 and 10.4 kJ mol
1 for
For both plots in Figure 6 the regression lines Form II and Form I, respectively. From these figures
(second order polynomial and linear regression for the enthalpy of transition from Form I to Form II,
panel A and B, respectively) relevant to the solubility DHI ! II, was calculated as
10.8 kJ mol
1.
data of the two forms intersect at around 808C. This At constant temperature and pressure the free
point represents the solid–solid transition tempera- energy difference between the two Forms, DGT, can be
ture and this observation confirms that ABZ Form I calculated from the logarithm of the ratio between the
represents the metastable polymorphs at ambient relevant solubility values
temperature. Csmetastable
It can be noted from Figure 6 panel B that in the DGT ¼
RT ln (2)
Csstable
temperature range studied a linear relationship
exists between the logarithm of the solubility and
the inverse of the absolute temperature (R2 ¼ 0.9 and At 258C the free energy change computed from the
0.94 for Form I and Form II, respectively). Therefore, solubility data obtained in HCl aqueous solution was
the apparent enthalpies of solution of the two
1.5 kJ mol
1. This value is not significantly different
polymorphs can be calculated from the relevant slope from that computed from solubility data obtained in
according to the equation methanol at the same temperature (
1.7 kJ mol
1),
thus justifying the calculation of the thermodynamic

DHs 1000 relationships on the basis of the assumption that
lnCs ¼ þb (1) Henry’s law applies and the obtained values are
R T
independent of the solvent used.
where Cs is the molar solubility, DHs is the heat of The values of DGT are reported in Figure 7 as a
solution at saturation, R is the gas constant, T is the function of the absolute temperature in the 25–1008C
absolute temperature, and b is a constant. temperature interval. In this plot the transition

DOI 10.1002/jps JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 99, NO. 9, SEPTEMBER 2010
3738 PRANZO ET AL.
Figure 7. Gibbs free energy variation as a function of the
absolute temperature for the albendazole Form I and Form
II system.
temperature can be graphically evaluated by looking
at the point where DGT becomes zero.
Finally, the entropy change for the transition of
Form I to Form II at a particular temperature T, was
calculated from the following equation
DHI!II
DGT
DST ¼
T which a scanning rate of 5 K min
1 was high enough
At 258C DST is
31.5 J K
1 mol
1. At the transition
point (350 K) DGT is 0 and Eq. (3) affords the entropy
change as
30.9 J K
1 mol
1.
Thermodynamic relationships are summarized in
Figure 8 that provides a semi-quantitative energy
versus temperature representation and highlights
the relevant transition points as well as the relative
stability of the two ABZ Forms.
Figure 8. Semi-quantitative energy versus temperature
diagram for the albendazole Form I and Form II pair. G,
Gibbs free energy; H, enthalpy; Ttrans, transition tempera-
ture; TmI and TmII, melting temperature for Form I and
Form II, respectively.
JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 99, NO. 9, SEPTEMBER 2010
These data put into evidence that the newly
isolated ABZ crystal phase and the commercially
available one constitute an enantiotropic pair, the
commercially available Form being metastable at
ambient temperature and thermodynamically stable
above 350 K.
It is worth emphasizing that for pharmaceutical
applications attention to these new findings is
advised. In fact, the use of a metastable form may
be advantageous (e.g., for exploiting higher solubility
in the GI tract) only when the kinetics of conversion
would be slow, namely when the energetic barrier
Form I $ Form II cannot be overcome during specific
storage conditions. In this respect, the metastable
phase should be kept cool and dry and should not be
too finely subdivided.31 On the other hand, one should
be aware that many pharmaceutical operations may
cause an undesired change from the metastable to the
stable Form, especially those in which the conversion
could be mediated by solubilization.33
As a general consideration, ABZ Form I is evidently
physically (kinetically) stable, being a commercia-
lized product (although with the warning: keep under
refrigeration). Form II proved to be physically quite
stable as well, as indicated by DSC experiments in
(3)
to allow it to ‘‘pass through’’ the transition tempera-
ture and to put into evidence the melting and re-
crystallization of Form II (see, Fig. 4A).
We also heated Form II at 1108C in a dry nitrogen
atmosphere under static conditions for 48 h without
detecting any phase change. A progressive Form
II ! I transition was instead observed upon heating
the ABZ Form II at 1308C. This condition afforded
complete conversion in <20 h.
X-Ray Structure of Form II
The molecular and crystal structure reported here is
the first for any ABZ crystal modification, despite the
longevity of this compound. This is likely due to the
practical difficulty of obtaining suitable single crys-
tals. Figure 9 shows the molecular structure occur-
ring in the crystal of Form II. Despite the fairly severe
limitations of crystal quality and size mentioned
above, as well as significant molecular disorder, the
Figure 9. Structure and atomic numbering of the alben-
dazole molecule in Form II. Thermal ellipsoids are drawn at
the 50% probability level.
DOI 10.1002/jps
 ENANTIOTROPICALLY RELATED ALBENDAZOLE POLYMORPHS 3739

Table 4. Selected Molecular Parameters for Form II of Table 5. Hydrogen Bond Data for Form II of Albendazole
Albendazole
D–H  A D–H (Å) H  A (Å) D  A (Å) D–H  A (8)
Bond Distances (Å) a
N1–H1  N10 0.87 1.96 2.816(5) 168
N1–C2 1.348(5) N3–C2 1.344(5) N3–H3  O12b 0.87 2.20 2.928(4) 140
N10–C2 1.330(5) N3–C4 1.396(5) N3–H3  O12 0.87 2.25 2.735(5) 115
N1–C5 1.394(5) N10–C11 1.345(5) a
1/2
x, 5/2
y, 2
z.
C11–O12 1.221(5) C11–O13 1.364(5) b
1/2
x,
1/2 þ y, 3/2
z.
Bond angles (8)
C5–N1–C2 109.0(3) C4–N3–C2 109.2(3)
N1–C2–N3 108.2(4) N1–C2–N10 120.9(4)
N3–C2–N10 130.8(4) C2–N10–C11 118.7(3) due to formation of a six-membered ring as a result of
intramolecular hydrogen bonding (N3–H3  O12,
Tab. 5). Atom H3 is in fact involved in an inter-
structural refinement proceeded reasonably well, all molecular H-bond as well, as described below. The
parameters converging satisfactorily and no abnor- formal bonding pattern C2 – N10–C11 occurring in
mal residual electron density being evident. this tautomer is also easily distinguished from the
The most significant molecular features are the alternative tautomeric possibility C2–N10(H)–C11 by
locations of the H atoms on N1 and N3, and the the value of the bond angle subtended at N10
twofold disorder of the propylthio-chain. As expected, (118.7(3)8 in the present case), which is the same as
the atoms of the side-chains display significantly the reported value of 118.5(4)8 in N-(2-benzimidazo-
higher thermal vibration than the rest of the lyl)-O-methyl carbamate. Instead, in benzimidazole
molecule. The five-membered ring is symmetrical, analogues in which N10 bears a hydrogen atom, and
the bond distances N1–C2 and N3–C2 being equal either one or both endocyclic N atoms bear a hydrogen
and longer than N10–C2 (Tab. 4), in accord with atom, the angle subtended at N10 is significantly
the formal bond orders indicated in Figure 2b. The larger (range 121.8–123.78 in the closely related
corresponding N–C and N – – C distances in N-(2- molecules BEWLOF,35 BMCBIB,36 QESBOG37).
benzimidazolyl)-O-methyl carbamate (refcode SED- Thus, there is ample evidence from the present X-
ZUW34), which has the same formal bonding arrange- ray structure that the ABZ tautomer in Form II
ment, are respectively 1.352(4), 1.364(6), and corresponds to that shown in Figure 2b.
1.328(5) Å. These values do not differ significantly Molecular association via intermolecular hydrogen
from their counterparts listed in Table 4. Further ring bonding is extensive, with two well-defined motifs
symmetry is indicated in the present study by the occurring, one finite and the other infinite. The
equality of the endocyclic angles at N1 and N3. The former is a centrosymmetric dimer with graph set R22
bond angle N3–C2–N10 exceeds N1–C2–N10 by 108 (8),38 shown at the top of Figure 10, in which the

Figure 10. Stereoview showing details of the primary hydrogen bonding motifs and
the generation of nonparallel molecular stacks in the crystal structure of albendazole,
Form II.

DOI 10.1002/jps JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 99, NO. 9, SEPTEMBER 2010
3740 PRANZO ET AL.
unique H-bond is N1–H1  N10a (Tab. 5). The second
is a continuous spiral array of molecules parallel to
the crystallographic twofold screw axis (unique H-
bond N3–H3  O12b). The net effect of these interac-
tions is to produce infinite stacks of dimers, alternate
stacks along the c-axis being rotated by 808 relative
to each other. This is evident in the packing diagram
projected down the c-axis while the full hydrogen
bonding scheme is evident in the b-axis projection
(Fig. 11). Nearly isostructural hydrogen bonding
networks occur in the crystal structure of N-(2-
benzimidazolyl)-O-methyl carbamate6. Other inter-
actions such as p-stacking and C–H  O interactions
do not play a significant role in the present crystal
structure.
The experimental and computed PXRD patterns for
Form II of ABZ are shown in Figure 12, together with
the experimental PXRD pattern of the commercial
material, Form I. The three most intense reflections
in the computed pattern39 are those occurring at 2u
values 7.498, 24.968, and 25.958. The primary
contributors to these three characteristic peaks (with
their relative intensity contributions in parentheses)
are reflections from the crystal planes 200 (1.00), 510
(0.55), and 511 (0.56), respectively. The high inten-
sities of the first two peaks may be rationalized with
Figure 11. Crystal packing diagrams projected down the
c-axis (top), the b-axis (bottom).
JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 99, NO. 9, SEPTEMBER 2010
Figure 12. PXRD patterns for albendazole: experimental
(a) and computed (b) of Form II, and the experimental
pattern of the metastable Form I (c).
reference to the packing diagrams (Fig. 11), as
follows. In addition to having a high inherent
multiplicity, the 200 planes (d ¼ 11.810 Å) are
straddled by the ‘‘heavy’’ sulfur atoms (Fig. 11,
bottom). The 510 planes are populated by the closely
spaced sheets (d ¼ 3.567 Å) running diagonally (/) in
Figure 11 (top) while the geometrically equivalent
planes, with indices
510 coincide with sheets
inclined along the other diagonal (\). These striking
packing features can thus easily be reconciled with
the X-ray data. (In the case of the third strong peak,
rotation of Fig. 11 (top) around the b-axis is necessary
to view the relevant planes 511; these coincide with a
set of closely stacked molecular sheets [d ¼ 3.434 Å]
crystallographically distinct from those described
above.)
The 2u values 7.498, 24.968, and 25.958 for the three
most intense reflections in the computed pattern
correspond with those at 2u ¼ 7.308, 24.658, and
25.558, respectively, in the experimental PXRD
pattern. Peak locations at slightly higher angles in
the former case are due to unit cell shrinkage at the
lower temperature of the single crystal X-ray analysis
compared with ambient temperature for the experi-
mental PXRD record. Relatively poor agreement
between corresponding peak intensities in the com-
puted and experimental PXRD patterns indicates
significant preferred orientation in the sample. This
is not surprising given the typical morphology shown
in Figure 1.
As regards Form I, the commercial material whose
structure is unknown, the presence of strong peaks
in the experimental PXRD pattern around 2u 7.08
and 24–258 (Fig. 12) suggests that similar crystal
DOI 10.1002/jps
 ENANTIOTROPICALLY RELATED ALBENDAZOLE POLYMORPHS
packing features to those described above for Form II
might obtain.
CONCLUSIONS
From the presented data it can be concluded that the
re-crystallization of ABZ both from methanol and
DMF affords a new stable polymorphic form enantio-
tropically related to the commercially available ABZ,
the latter being the metastable form at ambient
temperature.
Both forms proved to be physically quite stable,
likely due to a high-energy barrier for the activation
of the interconversion.
ABZ represents a rather complex system in which
the molecular structural differences that could be
associated with the polymorphism are of at least four
possible types, or combinations of these: (a) tauto-
meric (with e.g., both N atoms of the 5-ring
protonated [as in the crystal structure determined
in the present work], or with only one of these two N
atoms protonated [as in the conventional structural
depiction of the ABZ molecule]); (b) different con-
formations of either or both of the side-chains
attached to the bicyclic ring system; (c) the occurrence
of molecular disorder (as we have for the propylthio-
side chain) or its absence; (d) no essential difference in
molecular structure but different hydrogen bonding
arrangements in the two polymorphs.
The findings reported in the present paper clearly
put into evidence the need both for ABZ manufac-
turers and pharmaceutical formulators to carefully
consider the crystallization procedure and to perform
specific test to characterize the obtained crystal phase
as well as to pay particular attention to possible
undesired phase change (from the metastable to the
stable) stemming from pharmaceutical operations
implying heating, mechanical stress, or partial
solubilization.
SUPPORTING INFORMATION
The CIF file for the single crystal X-ray structure
described has been deposited at the Cambridge
Crystallographic Data Centre (CCDC 736654).
ACKNOWLEDGMENTS
This work was supported by a grant of the Italian
Ministry of Education University and Research
(MIUR) through the PRIN 2006 program. MRC and
RB also wish to acknowledge financial support of the
Executive Programme of Scientific and Technological
Co-operation between the Italian Republic and
Republic of South Africa 2008–2010.
DOI 10.1002/jps
3741
REFERENCES
1. Cook GC. 1990. Use of benzimidazole chemotherapy in human
helminthiases: Indications and efficacy. Parasitol Today 6:133–
136.
2. Marriner S. 1986. Anthelmintic drugs. Vet Rec 118:181–184.
3. Gyurik RJ, Theodorides VJ. 1975. Methyl 5-propylthio-
2-benzimidazolcarbamate. US patent, ed.: SmithKline
Corporation.
4. O’Neil MJ, editor. 2006. The Merck Index. 40th edition. White-
house Station, NJ: Merch Research Laboratoires. p 39.
5. Torrado S, Torrado S, Torrado JJ, Caerdoniga R. 1996. Pre-
paration, dissolution and characterisation of albendazole solid
dispersions. Int J Pharm 140:247–250.
6. Del Brutto OH, Sotelo J, Roman GC. 1993. Therapy for neu-
rocysticercosis: A reappraisal. Clin Infect Dis 17:730–735.
7. Batzias GC, Theodosiadou E, Delis GA. 2004. Quantitative
determination of albendazole metabolites in sheep spermatozoa
and seminal plasma by liquid chromatographic analysis with
fluorescence detection. J Pharm Biomed Anal 35:1191–1202.
8. Swarnkar CP, Sanyal PK, Singh D, Khan FA, Bhagwan PS.
1998. Comparative disposition kinetics of albendazole in sheep
following oral and intraruminal administration. Vet Res Com-
mun 22:545–551.
9. Mirfazaelian A, Dadashzadeh S, Rouini MR. 2002. Effect of
gender in the disposition of albendazole metabolites in humans.
Eur J Clin Pharmacol 58:403–408.
10. Mirfazaelian A, Rouini MR, Dadashzadeh S. 2002. Dose depen-
dent pharmacokinetics of albendazole in human. Biopharm
Drug Dispos 23:379–383.
11. Alvarez LI, Sanchez SF, Lanusse CE. 1997. Modified plasma
and abomasal disposition of albendazole in nematode-infected
sheep. Vet Parasitol 69:241–253.
12. Prichard RK, Hennessy DR, Steel JW, Lacey E. 1985. Meta-
bolite concentrations in plasma following treatment of cattle
with five anthelmintics. Res Vet Sci 39:173–178.
13. Garcia JJ, Bolas-Fernandez F, Torrado JJ. 1999. Quantitative
determination of albendazole and its main metabolites in
plasma. J Chromatogr B 723:265–271.
14. Lindemberg M, Kopp S, Dressman JB. 2004. Classification of
orally administered drugs on the World Health Organization
Model list of Essential Medicines according to the biopharma-
ceutics classification system. Eur J Pharm Biopharm 58:265–
278.
15. Torrado S, Torrado S, Caerdoniga R, Torrado JJ. 1996. For-
mulation parameters of albendazole solution. Int J Pharm
140:45–50.
16. Castillo JA, Palomo-Canales J, Garcia JJ, Lastres JL, Bolas F,
Torrado JJ. 1999. Preparation and characterization of alben-
dazole beta-cyclodextrin complexes. Drug Dev Ind Pharm 25:
1241–1248.
17. Haleblian J, McCrone W. 1969. Pharmaceutical applications of
polymorphism. J Pharm Sci 58:911–929.
18. Grant DJW. 1999. Theory and origin of polymorphism. In:
Brittain GH, editor. Polymorphism in pharmaceutical solids,
ed. New York: Marcel Dekker. pp. 1–33.
19. Vippagunta SR, Brittain HG, Grant DJW. 2001. Crystalline
solids. Adv Drug Del Rev 48:3–26.
20. Nonius BV. 2002. COLLECT Nonius. Delft, the Netherlands:
Nonius BV.
21. Otwinowski Z, Minor W. 1997. Processing of X-ray diffraction
data collected in oscillation mode. In: Carter CW, Sweet RM,
editors. Methods in enzymology, ed. New York: Academic
Press. pp. 307–326.
22. Sheldrick G. 1990. Phase annealing in SHELX-90: Direct
methods for larger structure. Acta Crystallogr A46:467–473.
23. Sheldrick GM. 1997. SHELX-97: Program for the refinement of
crystal structures. University of Göttingen, Germany.
JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 99, NO. 9, SEPTEMBER 2010
3742 PRANZO ET AL.
24. Spek AL. 1990. PLATON: A multipurpose crystallographic tool.
Acta Crystallogr A46:C34.
25. 2008. Monograph 01/2008:1386. European Pharmacopoeia 6.0,
ed. Strasbourg, France: EDQM. pp. 1122–1123.
26. Alanazi FK, El-Badry M, Ahmed MO, Alsarra IA. 2007.
Improvement of albendazole dissolution by preparing micro-
particles using spray-drying technique. Sci Pharm 75:63–79.
27. Moriwaki C, Costa GL, Ferracini CN, de Moraes FF, Zanin GM,
Pineda EAG, Matioli G. 2008. Enhancement of solubility of
albendazole by complexation with b-cyclodextrin. Brazilan J
Che Eng 25:255–267.
28. Kalaiselvan R, Mohanta GP, Manna PK, Manavalan R. 2006.
Studies on mechanism of enhanced dissolution of albendazole
solid dispersions with crystalline carriers. Indian J Pharm Sci
68:599–607.
29. Kalaiselvan R, Mohantha GP, Manna PK, Manavalan R. 2006.
Multicoponent system of albendazole with cyclodextrins and
hydroxyacids. Acta Pharm Sci 48:19–33.
30. Aguiar AJ, Zelmer JE. 1969. Dissolution behavior of poly-
morphs of chloramphenicol palmitate andmefenamic acid.
J Pharm Sci 58:983–987.
31. Carstensen JT. 2001. Polymorphism. In: Carstensen JT,
editor. Advanced Pharmaceutical Solids, ed. New York: Marcel
Dekker. pp. 117–131.
32. Brittain HG, Grant DJW. 1999. Effect of polymorphism and
solid-state solvation on solubility and dissolution rate. In:
Brittain HG, editor. Polymorphism in pharmaceutical solids,
ed. New York: Marcel Dekker. pp. 291–329.
JOURNAL OF PHARMACEUTICAL SCIENCES, VOL. 99, NO. 9, SEPTEMBER 2010
33. Bettini R, Bonassi L, Castoro V, Rossi A, Zema L, Gazzaniga A,
Giordano F. 2001. Solubility and conversion of carbamazepine
polymorphs in supercritical carbon dioxide. Eur J Pharm Sci
13:281–286.
34. Sokol VI, Davydov VV, Porai-Koshits MA, Zaitsev BE, Palish-
kin MV, Sheban GV, Pakhomov VI, Kukalenko SS. 1988.
Crystal and molecular structure of N-(2-benzimidazolyl)-O-
alkyl carbamates. Izv Akad Nauk SSSR Ser Khim (Russ) (Russ
Chem Bull) 37:1306–1311.
35. Iriepa I, Villasante FJ, Galvez E, Bellanato J, Martin A,
Gomez-Sal P. 2004. Synthesis, spectroscopic and crystallo-
graphic study of some carbamates from an azabicyclic chlor-
oformate and primary heterocyclic amines. New J Chem
28:618–624.
36. Blaton NM, Peeters OM, De Ranter CJ. 1980. (5-Benzoyl-1H-
benzimidazol-2-yl)carbamic acid methyl ester hydrobromide
(Mebendazole.HBr), C16H14BrN3O3. Cryst Struct Commun
9:181–186.
37. Sepassi K, Nichol GS, Yalkowsky SH. 2006. The napsylate salt
of carbendazim. 2-(Methoxycarbonylamino)benzimidazolium
naphthalene-1-sulfonate. Acta Crystallogr Sect E: Struct Rep
Online E62:o5172–o5173.
38. Etter MC. 1990. Encoding and decoding hydrogen bond
patterns of organic compounds. Acc Chem Res 12:120–
126.
39. Yvon K, Jeitschko W, Parthe E. 1977. LAZY PULVERIX: A
computer program for calculating X-ray and neutron diffrac-
tion powder patterns. J Appl Crystallogr 10:73–74.
DOI 10.1002/jps