chemical engineering research and design 8 8 ( 2 0 1 0 ) 11741181

Contents lists available at ScienceDirect

Chemical Engineering Research and Design

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

A method to crystallize substances that oil out

L. Derdour
AstraZeneca Process R&D, Sdertlje SE-151 85, Sweden

a b s t r a c t
A new approach to crystallize oily substances is described. The tendency for liquidliquid phase separation (LLPS)
is reduced by decreasing the kinetics of self-association via the formation of an intermediate amorphous network.
The path to initial crystal formation followed a sequence of rst freeze-drying an emulsion of solute in the solvent
system followed by suspending the dried solid in water to obtain a hydrated crystalline form. This new procedure
was applied successfully to a pharmaceutical organic substance that was previously isolated only as a viscous oil.
Once isolated, crystals of the drug were utilized as seeds to allow the successful transformation of an emulsion of the
substance into a suspension of crystalline drug solid thus avoiding the freeze-drying step. The isolated crystalline
solid retained its physical and chemical purity at room temperature for at least 3 months.
2010 The Institution of Chemical Engineers. Published by Elsevier B.V. All rights reserved.
Keywords: Oiling out; Liquidliquid phase separation; Freeze-drying; Pharmaceuticals

1.

Introduction

Oiling out, also referred to as demixing or liquidliquid phase

separation (LLPS), is a common phenomenon that occurs
during crystallization development in the pharmaceutical
industry. LLPS is usually unwanted because it impedes crystallization of the drug. Nevertheless, LLPS can be advantageous
for the separation of fatty acids as reported by Maeda et
al. (1997). In the pharmaceutical industry processes involving LLPS are highly undesirable because they lack sufcient
engineering process controls upon scale-up and robustness.
Crystallization processes that exhibit LLPS can create molten
phases that stick to reactor walls, and render the crystallization procedure useless by concentrating impurities in the
solute rich phase which leads to high impurity integration in
the crystals upon nucleation. In recent years, Serajudin and
Pudipeddi (2002) reported that there was an increase in the
number of less hydrophilic and less polar drug candidates
upon discovery of more potent and improved target specic
drug molecules. This likely explains the parallel increase of
case studies involving LLPS reported in recent years in the
pharmaceutical industry (Bonnett et al., 2003; Deneau and

Steele, 2005; Kim et al., 2003; Lafferrre et al., 2004; Lu et al.,

2004; Svrd et al., 2007; Veesler et al., 2003). Less hydrophilic
molecules have an inherent low polarity with a lack of anchoring sites, and therefore they do not easily self-assemble in
an organized manner. Most often, during the early development of drug candidates, seeds of crystalline forms are
not available and one needs to induce nucleation in order
to produce a solid form. Thus, generating supersaturation
is a prerequisite for obtaining a solid form. In early stage
development, limited data are available regarding the solubility of drug candidates in different solvents. Therefore,
supersaturation is usually created by screening solvents and
using different means such as cooling, evaporation or antisolvent addition or a combination of the stated methods. Trial
and error is also utilized at this stage. However, attempts
to crystallize drug substances often lead to a phase separation in which the active molecule is either concentrated in
one phase or is distributed between many phases. This paper
reports a case study of an AstraZeneca developmental drug
that exhibits oiling out and demonstrates a relatively original
approach to produce a solid crystalline phase of that substance.

Abbreviations: LLPS, liquidliquid phase separation; PXRD, powder X-ray diffraction; HNMR, proton nuclear magnetic resonance; DSC,
diffrential scanning calorimetry; TGA, thermal gravimetric analysis; LC/MS, liquid chromatography/mass spectrometry; KF, Karl Fischer
analysis.

Current address: Bristol-Myers Squibb Co., Process Research and Development, 1 Squibb Dr., Bld 50, Rm 232C, New Brunswick,
NJ 08903, USA. Tel.: +1 732 325 7704; fax: +1 732 227 3002.
E-mail address: lot.derdour@bms.com.
Received 10 December 2008; Received in revised form 27 January 2010; Accepted 2 February 2010
0263-8762/$ see front matter 2010 The Institution of Chemical Engineers. Published by Elsevier B.V. All rights reserved.
doi:10.1016/j.cherd.2010.02.001

chemical engineering research and design 8 8 ( 2 0 1 0 ) 11741181

2.

Background

Supersaturation is mandatory to crystallize a substance. In the

pharmaceutical industry, supersaturation is usually achieved
in solution by different means, as mentioned above, depending on the solubility of the substance in the solvent system.
However, crystallization of molecules with low polarity and
low propensity for hydrogen bonding is not easily achieved
because of the following reasons:
- At low supersaturation, nucleation does not occur because
the thermodynamic barrier to overcome is too high, i.e. the
molecules do not self-assemble easily.
- At high supersaturation, the molecules adopt the closest
metastable state, which can be 2 liquids of different compositions. Energetics and kinetics are frequently used to
explain the occurrence of LLPS at high supersaturation as
reported by Deneau and Steele (2005) and Lafferrre et al.
(2004), among others.
Vivares and Bonnet (2004), Liu et al. (1996), Thomson et
al. (1987), and Galkin and Vekilov (2000), among others also
attribute the occurrence of LLPS during the crystallization of
polymers to the large difference in molecular weights between
solutes and solvents.
Theoretically, if the solution concentration is maintained in
the region between the solubility curve and the LLPS curve for
enough time, crystallization should occur. However, in early
stage development, data for solubility and LLPS curves for
drug candidates are usually scarce. Besides, when trying to
crystallize a molecule with a low polarity, the induction time
prior to nucleation can be very high. This can mislead one to
believe that the solution is undersaturated and that the supersaturation needs to be increased. This will ultimately bring
the system beyond the LLPS curve and provoke liquidliquid
separation. LLPS can also occur in the metastable zone of
crystallization because of thermodynamic reasons, i.e. high
activation energy for crystallization as mentioned by Lee et al.
(1992), for the crystallization of polymers. LLPS can also occur
for substances that are relatively straightforward to crystallize as shown by Lee et al. (2008), in which oiling out was
demonstrated for the case of sulfathiazole because of rapid
evaporation of the solvent. The conclusion drawn from such
studies is that crystallization is impossible in such a solvent
system, but in reality it could have been achieved if sufcient time was provided for the onset of nucleation in the
metastable zone. As an alternative technique, ultrasound has
also been used to generate crystals and avoid oiling out as
demonstrated successfully by Kim et al. (2003). Nonetheless,
this approach is still in early development and usually succeeds only when crystallization is not difcult to achieve.
In some cases, oiling out was also found to be a necessary
step before nucleation. Using simultaneous measurements of
solute concentration and turbidity, Groen and Robert (2001)
found that concentrated micro-droplets form rst in a continuous phase of lower concentration followed by nucleation
at the interface of the droplets-continuous phase. Lastly, it is
worth mentioning that modelling was not extensively used to
predict the eventual occurrence of LLPS in the case of crystallization until recently as reported by Kiesow et al. (2008), for
the crystallization of 4,4 -dihydroxydiphenylsulfone.
Therefore, in early development, the unavailability of seeds
of the drug and the lack of reliable modelling tools make

1175

obtaining a crystalline form of a molecule of low polarity quite

arduous and time consuming. From the literature two types of
oiling out are described:
1. Liquidliquid separation occurs where each phase contains
reasonable amounts of solute. According to Veesler et al.
(2003), this occurs usually when a mixture of solvents is
used. In this case, the solute is generally not evenly distributed between the two phases. Lafferrre et al. (2004),
reports that the large difference in afnity of the drug for
each solvent can be the driving force for LLPS which can
be avoided if the right crystallization conditions, particularly the temperature of nucleation, are selected. Deneau
and Steele (2005), gave an explanation of LLPS in terms of
energetics and mentioned that it spontaneously occurs if
the concentration of solute is located between the binodal
points, dened as concentrations with minimal mixing
free energy, and the spinodal points at which spontaneous phase separation occurs. In this type of LLPS, the
crystallization of the drug is possible by adjusting the crystallization condition in order to avoid the LLPS region. One
of the solutions to tackle this problem is seeding which was
used successfully by Veesler et al. (2003), Beckmann (2000),
and Deneau and Steele (2005), to prevent this kind of LLPS
from occurring or to crystallize a solute from an emulsion.
2. Liquidliquid separation occurs where one phase contains
the solvent(s) and the other phase is mainly formed by the
solute in the form of a very heavy viscous oil-like phase,
hence the name oiling out. This phenomenon occurs usually for high solute concentrations at moderately high
temperatures as reported by Svrd et al. (2007), for the
vanillin/water system. These authors attribute the LLPS of
highly concentrated solutions to the closeness of the temperature of separation and the melting point of the solute.
In such cases, lowering the concentration of the solution
should prevent oiling out from occurring (cf. Fig. 1a). However, if phase separation occurs at a low temperature and
for a dilute solution a crystalline form of the drug may
exist but it melts at a temperature lower than the isolation
temperature of the solid, which in most cases lies between
0 C and room temperature. Therefore, the drug separates
from the solvent as a melt at room temperature. In this
case, attempts to crystallize the drug can prove to be a
daunting task requiring a special crystallization procedure
and storage conditions. Thus, such molecules are usually
discarded from further development unless they provide
unique and highly valuable pharmaceutical attributes. As
mentioned above, large differences in molecular weighs
between solutes and solvents as in the case of crystallization of polymers can also lead to LLPS.
In all cases, the determination of the phase diagram can
help greatly in selecting crystallization conditions to avoid
LLPS phenomenon. Phase diagrams can be very different
depending on the system. Fig. 1a, b, c and d are examples of the
main types of phase diagrams encountered in the literature.
These gures show no general trend regarding the occurrence
of LLPS. However, it appears that a critical solute concentration in the metastable region is a prerequisite for causing LLPS.
Fig. 1a is a typical depiction of oiling out from a single solvent
and shows that high temperature and high concentration are
conditions that favour LLPS.
Tendency for LLPS was also studied by Maeda et al. (2001),
who indicate that the width of the metastable zone for LLPS

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chemical engineering research and design 8 8 ( 2 0 1 0 ) 11741181

Fig. 1 (a) Phase diagram, type 1 (Svrd et al., 2007). (b) Phase diagram, type 2 (Bonnett et al., 2003). (c) Phase diagram, type
3 (Veesler et al., 2003). (d) Phase diagram, type 4 (Inspired from Maeda et al., 2001). CLMZ: curve of the limit of the
metastable zone.
is much narrower that the one for crystallization and that
this characteristic can explain the occurrence of LLPS in the
metastable zone for crystallization even for low concentrations. This is depicted in Fig. 1d in which cooling down a
mixture from point B will cross the solute solubility, but then
will cross the liquidliquid solubility and the curve of the limit
of the metastable zone (CLMZ) for LLPS before reaching the
CLMZ for crystallization. If the kinetics of LLPS are fast enough,
this will result in a metastable LLPS. In this case, the emulsion
obtained can in theory be converted into suspension. On the
other hand, cooling a highly concentrated mixture (point A in
Fig. 1d) will result in a stable LLPS because the phase separation occurs outside the metastable zone for crystallization
which excludes the possibility of solid formation. As shown in
Fig. 1d, LLPS can be avoided by lowering the solute concentration, i.e. starting from point C.
If the phase diagram is available, it is straightforward
to select crystallization conditions that should prevent LLPS
from occurring. Unfortunately, constructing the phase diagram is time consuming, labour intensive and is impractical
to complete during drug development in the pharmaceutical industry. Besides, there is no consensus on conditions
that favour LLPS, and the crystallization is system dependent.
Therefore, in industry, where time and labour are major factors, scientists usually rely on experience to select screening
conditions that will produce crystalline materials.

3.

Materials and methods

Crystallization attempts were carried in a 24-well temperature regulated shaker, Eppendorf AG, Hamburg, Germany.

PXRD was determined using a Philips XC

Pert-MPD diffractometer, PANalytical Inc., Westborough, MA. Thermal analysis
was performed using a DSC822e and a TGA/SDTA851e, Mettler Toledo, Columbus OH. Karl Fischer (KF) analyses were
performed using a Metrohm 841 Volumetric Karl Fischer
Titrando, Westbury, NY. Freeze-drying was carried out in a
Heto PowerDry PL9000 Freeze-Dryer, Breda, The Netherlands.
AstraZenecas developmental drug was supplied with a purity
higher than 99.3%. Chemical purity was assessed using a
LCMS-2010EV, Duisburg, Germany. Solution HNMR was determined using a Bruker Ultrashield 400 MHz/54 mm, Bremen,
Germany. All solvents were obtained from SigmaAldrich Sweden AB, Stockholm, Sweden, and were of HPLC grade.

4.

Crystallization procedure development

Numerous attempts were done to crystallize the substance or

at least obtain a solid form. The rst approach was to induce
solidication by solvent evaporation. A multitude of solvents
of varying polarities were used. Using this approach, the drug
substance was isolated as a highly viscous molten phase that
was difcult to process. This rst series of experiments were
benecial in identifying potential solvents and anti-solvents
for the drug substance. From this information, anti-solvent
crystallization schemes were designed in an attempt to crystallize the substance. Several solvent systems were also tested
but liquidliquid separation occurred in all cases as shown
in the photographs in Fig. 2 which shows the formation of
an emulsion upon addition of the anti-solvent. Upon observation under the microscope, the solvent evaporated and a

chemical engineering research and design 8 8 ( 2 0 1 0 ) 11741181

1177

Fig. 2 Emulsion observed under the microscope.

molten phase of the drug was obtained. This observation indicated that a crystalline form of the drug substance could
have a lower melting point than normal conditions, making
attempts to crystallize the substance at room temperature
impossible. Therefore crystallization of the substance as a
solvate/hydrate or a co-crystal was investigated. Attempts to
crystallize a salt form of the drug were ruled out because the
molecule lacked basic or acidic sites. Trials to isolate a cocrystal of the drug substance were also abandoned because the
periphery of the drug molecule lacked reliable H-bond donors
and/or acceptors. The low propensity for H-bond formation
makes self-assembly quite difcult especially at high molecular mobility, i.e. high temperature. Therefore, this explains the
difculty of the drug molecules to create a crystal lattice at
room temperature.
The drug molecule does not have reliable H-bond donors
and acceptors on its periphery but it does contain a reliable Hbond acceptor. However, it is believed to be embedded deep
within the molecule and hence cannot create bonds with
neighbouring drug molecules.

4.1.

Approach to crystallize the drug substance

The approach that we adopted in attempts to obtain a crystalline form for the developmental drug was based on two
fundamental ideas:

1. The presence of an embedded reliable H-bond acceptor

prompted us to investigate the possibilities of isolating a
solvated form of the drug. More particularly, to crystallize
the drug molecule as a hydrate. This choice was based on
the thought that water molecules are small enough to reach
the H-bond acceptor that is embedded in the drug molecule
and could therefore link 2 drug molecules by creating 2 Hbonds with the H-bond acceptors of each drug molecule.
From this rationale, water can link drug molecules through
H-bonding and, consequently, a monohydrate-drug solid
would be possible to isolate. In the subsequent sections,
the drug molecule will be represented by R1 AR2 where

R1 and R2 are 2 bulky parts of the molecule and A is the

reliable H-bond acceptor.
2. Next, we chose to use the strategy of producing an amorphous form of the substance followed by providing a small
amount of energy to slowly relax the substance to a crystalline form (Hancock and Zogra, 1997). One of the main
drivers to LLPS is the self-association of drug molecules via
hydrophobic interactions and self-association of solvent
molecules via dipoles/H-bonding as depicted in Fig. 3 for
the case where the solvent is water. The rationale in forming a crystalline solid through an amorphous structure is
to reduce the molecular mobility of the drug substance, i.e.
to decrease the equilibrium constant for the drug association: ka1 . Freezing the drug molecules in an amorphous
structure should substantially reduce the molecular mobility within the drug substance and decrease the kinetics of
self-association. Fig. 3 shows the proposed mechanism that
occurs in the liquid state which compares high kinetics for
LLPS and crystallization.

The rst attempt to isolate a solid hydrate form of the drug

involved a simple procedure in which the drug is dissolved in
an organic solvent and then water is added as an anti-solvent.
This attempt failed as liquidliquid separation occurred as
mentioned above.
The second approach is depicted in Fig. 4 using the creation
of an amorphous network. Essentially, the drug was dissolved
in a low boiling point organic solvent such as methanol, stage
A, then add water to oil out the drug substance, stage B. The
organic solvent was then removed by vacuum distillation to
obtain an emulsion of the drug and the water, stage C. This
emulsion was then mixed vigorously to create a dispersion of
micro-droplets of the drug in water, stage D. Then, while being
mixed, the emulsion was frozen to 50 C, stage E. Finally the
frozen mixture was lyophilized. The dried material, stage F,
was porous and agglomerated with no distinct faces, and as
shown by microscope images in Fig. 5, it did not exhibit birefringence.

Fig. 3 Molecular association and competition between LLPS and crystallization.

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chemical engineering research and design 8 8 ( 2 0 1 0 ) 11741181

Fig. 4 First attempt to crystallize R1 AR2 .

However, microscope images also revealed that the
lyophilized solid material melted rapidly and a viscous oil was
obtained when warmed to room temperature as depicted in
Fig. 5, stage G. As expected, water content analysis revealed
that all water had sublimed, which meant that the compound
did not form a hydrate. Further solid state analysis could not
be performed on the freeze-dried solid because, as shown in
Fig. 5, it melted quickly. HNMR analysis conrmed that the
drug substance is free of residual solvent. This observation
conrmed that the substance is in a molten phase under normal conditions of temperature and pressure as a neat form.

4.2.
Use of the lyophilized solid to obtain a crystalline
form of a monohydrate-drug
Our next approach was to mix the solid obtained by
freeze-drying with liquid water again to promote favourable
conditions for isolating a hydrated solid form. The amorphous
solid obtained by freeze-drying was highly porous and should
have a lower molecular mobility compared to the molten
phase which would reduce the kinetics of self-association.
It is expected that upon mixing with liquid water, the later
will diffuse into the pores and come in close contact with
the substance for a sufcient time that permits the nucle-

ation of a hydrated form before drug molecules and water

self-association prevent that from happening. Conversely, the
hydrate-drug solid is thermodynamically possible, but it is
blocked by the drug self-association when it is in the liquid state and by the high molecular mobility encountered in
the molten phase. The solid amorphous material has a lower
molecular mobility and a higher relaxation time than the
molten phase and can provide enough time for the molecules
to assemble in an ordered fashion, i.e. crystalline form.
Therefore, the solid obtained by freeze-drying was rapidly
suspended in water at a low temperature in order to slow down
the transformation from the supercooled liquid, i.e. amorphous solid, to the molten phase. The mixture water/drug was
mixed in a multi-well mixer and the solid was monitored periodically for possible form changes. Approximately 1 h after the
start of the mixing some peaks started to appear in the PXRD
pattern. The solid was isolated after 2 h of mixing and after
ltration and drying under vacuum it was analyzed by PXRD
which revealed that it was highly crystalline. Thermal analysis
was also determined and DSC revealed an endothermic event
at the same temperature interval where the TGA displays a
weight loss. Hot-stage microscopy was then used to ascertain the cause of the endotherm and revealed a melting event
at that temperature interval indicating that the endothermic

chemical engineering research and design 8 8 ( 2 0 1 0 ) 11741181

1179

Fig. 5 Melting of solid obtained by freeze-drying observed under microscope.

event corresponds to melting combined with dehydration. The
heat involved during the endotherm displayed by the DSC
event suggests that the lattice energy is low, probably due to
the few H-bonds per unit cell. Crystals initially obtained with
the procedure involving lyophillization were used as seeds in
subsequent crystallization experiments carried out at room
temperature. This seeded crystallization afforded crystalline
material with the same characteristics as the one initially
obtained.
The water content of the solid was also determined by KF
analysis and TGA and it was found to correspond to the level
of the monohydrate which conrmed our initial hypothesis
that a water molecule can act as a link between two drug
molecules. The chemical purity was determined by LC/MS and
HNMR and was found to be identical to the starting material.
The overall procedure to isolate the crystalline form of the
drug is summarized in Fig. 6.

4.3.

Conversion of an emulsion into a suspension

The strategy followed used freeze-drying as a path to produce seeds of crystalline monohydrate. This result indicates
that mixing the drug with water results in a metastable LLPS.
Hence, the possibilities of using the seeds in order drive the
crystallization from an emulsion of the drug substance in
water or a mixture of water and an organic solvent without
performing freeze-drying were investigated. We decided to
introduce the seeds of the hydrate-drug in an emulsion of the
drug in water and to stir the mixture for few hours. Temperature of the mixture was kept in the range 05 C in order to
limit stress to the system. It was observed that after 2 h, the
emulsion turned into a slurry suspension. After ltration and
drying, the solid was analyzed by PXRD and had the same drug
monohydrate form as the seeds. This process proved reliable
as it was repeated several times and produced the crystalline
R1 AR2 H2 O form consistently. Thus, the freeze-drying step

was not needed furthermore and a simple seeded crystallization from an emulsion sufced to produce the highly
crystalline pharmaceutically acceptable crystalline solid drug.
The transformation from the emulsion to a suspension of
crystalline solid indicates that the LLPS obtained is indeed
metastable even though the emulsion remained stable for up
to 3 months without seeding.
Next, the mixture was subjected to temperature uctuations to test the robustness of the procedure. Agitation rate
was also varied. The outcome of the study was always a
crystalline R1 AR2 H2 O with the same PXRD, DSC and water
content. The method thus proved to be robust and reliable.
Also, no issues related to the rheology were observed which
suggest that a priori no potential problems should be expected
upon scale-up.
The stability at room temperature of the solid was also
tested by monitoring the chemical and physical purity for 3
months after isolation using PXRD, KF, LC/MS and HNMR. The
analyses did not show any detectable change proving that the
crystalline R1 AR2 H2 O was stable at room temperature for
up to 3 months.

4.4.
Possible obstacles to nucleation in the liquid
homogeneous phase
It is believed that in the liquid state, nucleation of the crystalline hydrate form is prevented because of the following
reasons:
1. High molecular mobility in molten phase: H-bonds are
continuously formed and disrupted and the duration of Hbonding is short because of the kinetic mobility inherent to
the liquid state. This prevents clusters of hydrate substance
from reaching the critical size for growth.
2. Self-association: water molecules self-associate by Hbonding. Water is known to have a high self-afnity and

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chemical engineering research and design 8 8 ( 2 0 1 0 ) 11741181

Fig. 6 Procedure followed to isolate a solid monohydrate crystalline form of the drug.
tends to form H-bond linked clusters. Hydrophobic drug
molecules also self-associate by hydrophobic interaction
as described by Scheraga (1998).
The strategy used in this study to obtain initial crystals of
R1 AR2 H2 O was to lower the kinetics of self-association of
the drug molecules by freezing the substance in an amorphous
structure and adding water to form the monohydrate.

4.4.1.
phase

Thermodynamic barrier to nucleation in liquid

Self-association also suggests that the energy required to form

a nuclei of R1 AR2 H2 O of a reasonable size is signicantly
higher than the one needed for the formation by LLPS. In
order to create embryos of R1 AR2 H2 O, water molecules have
to self-disrupt their clusters and disrupt the drug molecules
hydrophobic clusters. This task is very energy demanding and
as observed experimentally is not possible without an external aid, i.e. seeding. Crystalline R1 AR2 H2 O has lower energy
than the liquid R1 AR2 because creating H-bonds lowers the
energy much more than hydrophobic interactions do. Consequently, transformation from LLPS to a suspension is possible
once crystals of R1 AR2 H2 O are added to the suspension.

4.4.2.

Kinetic barrier to nucleation in liquid phase

Once clusters of water and drug are disrupted, water has to

form H-bonds with the drug that last long enough to permit
self-assembly with other hydrated drug molecules. Because of
high molecular mobility in the liquid phase, the life time of Hbonds is expected to be short. Thus the size of the embryos
of R1 AR2 H2 O formed in the liquid phase cannot reach the
critical size for nucleation making the latter impossible in the
molten phase. On the other hand, the amorphous matrix lowers the molecular mobility of the drug which can increase the
life time of H-bonds created between water and the drug. As a
result, embryos of R1 AR2 H2 O created can grow larger than
in the liquid phase. According to the concept of sigma cooperativity, as mentioned by Morris (1999), these clusters grow
stronger and larger as more molecules are added to them via

H-bonds. This will ultimately lead to self-sustaining clusters

that will attain the critical size for nucleation and, consequently, nucleate making the crystallization of R1 AR2 H2 O
possible.

5.

Conclusions

A case study of a LLPS encountered during trials to crystallize a developmental drug and a novel method to generate a
crystalline form of that substance were presented. Because
of a lack of reliable hydrogen bond donors and acceptors on its periphery and sterical hindrance, the substance
did not crystallize at room temperature. A crystalline form
was achieved by utilizing water molecules as bridging links
between drug molecules to form a solid hydrate of the substance. Lyophillization was involved in the path to the rst
crystals of a monohydrate crystalline form of the drug. These
crystals were then used as seeds to drive the crystallization
from an emulsion of the drug in water. Drug molecules selfassociation combined with high molecular mobility in the
liquid phase are assumed to block nucleation of the monohydrate in solution. Low molecular mobility in the amorphous
solid created by lyophillization is considered to be the facilitator of nucleation. The solid obtained proved to be stable for
up to 3 months at room temperature.

Acknowledgements
The author is thankful to AstraZenecas senior management
at Sdertlje, Sweden, for assistance and follow-up during the
preparation of this paper. Dr. Sebhathu T. is also sincerely
thanked for determining PXRDs.

References
Beckmann, W., 2000, Seeding the desired polymorph:
background, possibilities, limitations and case studies. Org.
Process Res. Dev., 4: 372383.

chemical engineering research and design 8 8 ( 2 0 1 0 ) 11741181

Bonnett, P.E., Carpenter, K.J., Dawson, S. and Davey, R.J., 2003,

Solution crystallization via submerged liquidliquid phase
boundary: oiling out. Chem. Commun., 698699.
Deneau, E. and Steele, G., 2005, An in-line study of oiling-out and
crystallization. Org. Process Res. Dev., 9: 943950.
Galkin, O. and Vekilov, P.G., 2000, Control of protein crystal
nucleation around the metastable liquidliquid phase
boundary. Proc. Natl. Acad. Sci., 97(12): 62776281.
Groen, H. and Robert, K.J., 2001, Nucleation, growth and
pseudo-polymorphic behaviour of citric acid as monitored in
situ by attenuated total reection Fourier transform infrared
spectroscopy. J. Phys. Chem. B, 105: 1072310730.
Hancock, B.C. and Zogra, G., 1997, Characterization and
signicance of the amorphous state in pharmaceutical
system. J. Pharm. Sci., 86: 112.
Kiesow, K., Tumakaka, F. and Sadowski, G., 2008, Experimental
investigation and prediction of oiling out during
crystallization process. J. Cryst. Growth, 310: 4163
4168.
Kim, S., Chenkou, W. and San Kiang., 2003, Crystallization
process development of an active pharmaceutical ingredient
and particle engineering via the use of ultrasonics and
temperature cycling. Org. Process Res. Dev., 7: 9971001.
Lafferrre, L., Hoff, C. and Veesler, S., 2004, Study of liquidliquid
demixing from drug solution. J. Cryst. Growth, 269: 550557.
Lee, I.S., Lee, A.Y. and Myerson, S.A., 2008, Concomitant
polymorphism in conned environment. Pharm. Res., 25(4):
960968.
Lee, K.H., Myerson, S.A. and Levon, K., 1992, Nonequilibrium
liquidliquid phase separation in crystallisable polymer
solutions. Macromolecules, 25(15): 40024010.
Liu, C., Asherie, N., Lomakin, A., Pande, J., Ogun, O. and Benedek,
G.B., 1996, Phase separation in aqueous solutions of lens
-crystallins: special role of s . Proc. Natl. Acad. Sci., 93:
377382.

1181

Lu, J., Carpenter, K., Li, R.J., Wang, X.J. and Ching, C.B., 2004,
Cloud-point temperature and LLPS of supersaturated
lysozyme solution. Biophys. Chem., 109: 105112.
Maeda, K., Nomura, Y., Fukui, K. and Hirota, S., 1997, Separation
of fatty acids by crystallization using two liquid phases.
Korean J. Chem., 14(3): 175178.
Maeda, K., Aoyama, Y., Fukui, K. and Hirota, S., 2001, Novel
phenomena of crystallization and emulsication of
hydrophobic solute in aqueous solution. J. Colloid Interface
Sci., 234: 217222.
Morris, K.R., 1999, Structural aspects of hydrates and solvates, in
polymorphism in pharmaceutical solids, In Brittain, H.G. (Ed.),
Drugs and the Pharmaceutical Sciences SR (Marcel Dekker, New
York), pp. 125181. (Marcel Dekker, New York).
Scheraga, H.A., 1998, Theory of hydrophobic interactions. J.
Biomol. Struct. Dyn., 16(2): 447460.
Serajudin, A.T.M. and Pudipeddi, M., 2002, Salt selection
strategies, in Handbook of Pharmaceutical Salts: Properties,
Selection And Use, Stahl, P.H. and Wermuth, C.G., Wermuth,
C.G. (eds). (VHCA, Verlag Helvetica Chimica Acta/WileyVCH,
Zrich/Weinheim), pp. 135160
Svrd, M., Gracin, S. and Rasmuson, ., 2007, Oiling out or molten
hydrateLLPS in the system vanillinwater. J. Pharm. Sci.,
96(9): 23902398.
Thomson, J.A., Schurtenberger, P., Thurston, G.M. and Benedek,
G.B., 1987, Binary liquid phase separation and critical
phenomena in proterin/water solution. Proc. Natl. Acad. Sci.,
84: 70797083.
Veesler, L., Lafferrre, L., Garcia, E. and Hoff, C., 2003, Phase
transitions in supersaturated drug solution. Org. Process Res.
Dev., 7: 983989.
Vivares, D. and Bonnet, F., 2004, Liquidliquid phase separations
in urate oxidase/PEG mixtures: characterization and
implications for protein crystallization. J. Phys. Chem. B, 108:
64986507.