Journal of Crystal Growth 76 (1986) 645-~-655 645

North-Holland, Amsterdam

GROWTH AND CHARACTERIZATION OF SMALL MOLECULE ORGANIC CRYSTALS

Margaret C. ETTER *, Donald A. JAHN and Brian S. DONAHUE

Department of Chemistry, University of Minnesota, 207 Pleasant St. SE, Minneapolis, Minnesota 55455, USA

and

Ruth B. JOHNSON and Charles OJALA

Roche/Ic Crystal Corporation, St. Paul, Minnesota 55116, USA

Received 30 September 1985; manuscript received in final form 28 April 1986

High qualilty organic crystals are needed for X-ray diffraction analyses and for use as solid-state materials. Current routine and
nonroutine methods of growing organic crystals are discussed here and comparisons are made between these methods and those that
are more suitable for inorganic or protein crystal growth. Examples of growth of crystals with controlled morphology, controlled
growth of metastable crystals, and growth of polymorphic crystal forms are given.

1. Introduction 2. Background

The need for readily available high quality
organic crystals for X-ray diffraction analyses and
for use as solid-state materials has grown dramati-
cally in the last decade. Organic chemists now
routinely use X-ray structure analyses to identify
their new compounds and many organic solids
have been shown to have technologically im-
portant properties such as electrical conductivity
[1] and nonlinear optical behavior [2]. Neverthe-
less, studies on organic crystal growth processes
have been ignored by comparison to the efforts
devoted over the years to the study of inorganic
crystals and, more recently, to the study of protein
crystal growth phenomena [3]. In this article, we
compare crystal growth conditions for organic
compounds to those of inorganic and protein
crystals, discuss the commonly used techniques for
growing small-molecule organic crystals, and pre-
sent representative examples of crystal growth ex-
periments carried out in our laboratories.
* Address correspondence to this author.
It is common lore among chemists and crys-
tallographers that organic * crystals are easier to
grow than protein crystals, but harder to grow
than inorganic crystals. While there are many
exceptions to this rule, notably the relative ease
with which lysozyme can be grown compared to
the difficulties encountered when growing crystals
of oxalates and citrates [4], in general it is a useful
rule. Organic compounds are usually insoluble in,
and may be reactive with, water while proteins
and inorganic compounds are usually recrystal-
lized from aqueous solutions. It is an advantage
for organic crystals that they can be grown from a
wide variety of solvents, but since these solvents
are non-aqueous, pH and ionic strength cannot be
used as recrystallization variables. A common
organic solution recrystallization technique is to
saturate a boiling solution of the compound and
then allow it to slowly cool to room temperature.
* Protein molecules are organic compounds, but the term
organic is used here to mean small-molecule organic cam-.
pounds ( <100 atoms).

0022-0248/86/$03.50 © Elsevier Science Publishers B.V.

(North-Holland Physics Publishing Division)
646 M.C. Etter et al. / Growth and characterization of small molecule organic crystals

Table I
Comparison of general crystal growth characteristics ‘° of small-molecule organic compounds. proteins, and inorganic compounds

Property Organic compounds Proteins Inorganic compounds

Chemical nature Covalently bonded. Macromolecules composed of Salts, composed of
neutral small molecules, covalent bonds, charged inorganic ions
<100 atoms charged groups. metal ions, and water

Soluhility Soluble in a wide variety Soluble in water Soluble in water

of organic solvents
Sensitivity to pH and NA Vet-v sensitive Sensitive
ionic strengths
Thermal properties Thermally stable up to Readily denatured by heating above Highly thermally
melting point (common room temperature stable
melting points are 50—150°C)
3, from common < 0.5 mm3, by any method No size limit for flianv
Crystal size 0.1—10 mm
crystal growth methods inorganics

X-ray diffraction Good—moderate Poor—good Excellent

quality

Mechanical strength Fair—good Extremely poor Extremely good

The characteristics given in this table are qualitative observations that may be useful when designing growth procedures for
compounds in one of the given classes. There are exceptions to each of the above descriptions. This list should be used as a guide.
not as a definitive analysis. Organometallics have not been treated as a separate class, hut they have some of the characteristics of
both inorganic and organic crystals. A notable characteristic of organometallic crystals is that they may he extremely air and
moisture sensitive.

This procedure could not be used for proteins
which are rapidly denatured at elevated tempera-
lures. On the other hand, the extremely high tern-
peratures used for melt recrystallizations of many
inorganic compounds cannot be used for organics
which usually melt below 200°C, and char at
higher temperatures. Organic crystals, if obtained
at all, are usually in the size range 0.1—10 mm3,
while many protein crystals seem to have a limit-
ing dimension of about 0.5 mm. Some inorganic
crystals can be grown several feet long [5], with
the availability of material and the patience of the
operator being the limiting factors! Inorganic
crystals frequently have superb X-ray diffraction
qualities (high scattering power, high degree of
order, low thermal motion, and good stability in
the X-ray beam), while organics and proteins may
not scatter well and may exhibit a mosaic spread
in the diffraction spots due to crystal disorder and
impurities. Protein crystals are exceedingly fragile,
sometimes containing up to 70% or 80% water.
Inorganic crystals are characteristically strong, and
organic crystals, although somewhat softer, are
mechanically strong enough to be stored safely in
glass containers or handled with a needle pointer.
In table 1, the differences and similarities between
protein, organic and inorganic crystal growth con-
ditions are summarized. Features which all these
types of crystals have in common include char-
acteristic crystal properties such as optical bire-
fringence, X-ray diffraction, polymorphism (the
presence of more than one crystal form), the pos-
sibility of habit modification, and the possibility
of undergoing single crystal phase transforma-
tions.
3. Preparation of organic crystals
The three most common procedures used for
growing organic crystals are: (1) evaporation of a
dilute solution at room temperature; (2) boiling
then cooling a supersatured solution; (3) sublima-
tion. For each of these techniques, preparation of
high quality single organic crystals for X-ray dif-
fraction analyses requires that one start with a
 M. C. Etter et at / Growth and characterization of smallmolecule organic crystals
sample which is pure. It is not usually necessary to
remove impurities that are present at less than
about 0.1% concentration. An indication that a
compound is pure enough to try to grow crystals
is that it looks homogeneous, that particles appear
microcrystalline under the microscope (sharp
edges, reflective faces, or birefringent) and that it
has a sharp melting point and a clean infrared or
solution NMR spectrum. For methods 1 and 2
above, the next step is to determine what solvent
would be appropriate. If the compound is known,
some of its solubility characteristics will be given
in the Handbook of Chemistry and Physics [6].
For the first crystal growth method, room tern-
peralure evaporation, any solvent which will dis-
solve the compound can be used. Since the rate of
evaporation is directly related to the vapor pres-
sure of the solvent, solvents such as DMSO (di-
methylsulfoxide), HMPA (hemamethyiphosphor-
amide), DMF (dimethylformamide), and glycerol
are not good choices because of their low volatil-
ity. A sample of 10—100 mg is dissolved in the
solvent of choice and a filtration is done if the
solution contains visible particles such as dust or
insoluble residue. The solution is placed in a crys-
tallizing dish (Pyrex petri dish with sides about 2
inches high) and covered with the glass top, which
does not seal the sample but does significantly
lower the rate of evaporation and inhibits forma-
tion of a surface layer of crystals. This dish is then
set aside and periodically examined visually or
under the microscope for crystal formation. When
a good crystal is observed, it is immediately re-
moved from the solution so that it cannot trans-
form or aggregate. A stainless steel needle pointer
or a fine spatula is used to remove the crystal,
which is then laid on a piece of absorbant paper to
wick off solvent, and placed on a microscope slide
for further examination and mounting. The origi-
nal solution is left to allow growth of additional
crystals.
The second recrystallization technique, saturat-
ing a boiling solution and then allowing it to cool,
is commonly used as a means for purifying bulk
quantities of samples. This technique can also be
adapted for growing good single crystals. With
this method, choice of solvent is more critical,
since the sample should have a large solubility
647
differential between the boiling point and room
temperature (or freezing temperature). We usually
test five to six different solvents in which the
compound should be partially soluble. Using small
test tubes, we put about 50 mg of sample in six
different test tubes and add ten drops of solvent
to each one. Solvents which completely dissolve
the sample under these conditions are not satisfac-
tory. For solvents which show partial solubility or
no solubility, we heat the samples to boiling by
passing them over a gentle bunsen burner flame.
If they are not completely dissolved at the boiling
point, then more solvent is added. The samples are
cooled in air, cold water, or an ice bath to induce
crystal formation. If no crystals are obtained, a
seed is introduced or the side of the test tube is
scratched with a glass rod to introduce nucleation
sites. The crystals can be removed by decanting
the supernatant solvent, and gently transferring
the crystals to several pieces of filter paper.
Crystals which stick to the sides of the test tube
can be removed with a needle pointer or can be
shaken out of the tube. The procedure can readily
be scaled up to quantities of 500 mg to 1 g.
Samples larger than that will result in too large a
mass of crystals and aggregation and manupula-
tion of the crystals becomes a problem.
The third common recrystallization technique,
sublimation, is an alternative for thermally stable
compounds. A simple test to see if a compound
will sublime readily is to put 10—20 mg of sample
on a petri dish and cover the dish with an inverted
watch glass. The petri dish can be heated gently
on a hot plate and the presence of crystals grow-
ing on the watch glass, indicating that sublimation
is occurring, can be readily observed. One can
sometimes observe the same phenomenon simply
by heating a small sample between cover slips on
a Fisher—Johns melting point apparatus. In many
cases high temperatures are needed to obtain sub-
limation, and in order to prevent sample degrada-
tion, the sublimation can be carried out under
vacuum. A commercial sublimator can be used or
a simple vacuum sublimator can be made in the
lab. The sample (50—100 mg) is placed in the
bottom of a 5—6 inch long thick walled tube,
attached to a vacuum line, and the sample is
gently heated under vacuum until crystals grow on
648 M. C. EOer et al. / Growth and characterization
the sides of the tube. Many sophisticated modifi-
cations of this set-up are possible, including in-
troduction of a cold finger into the tube, or setting
up a controlled temperature gradient along the
sides of the tube. Crystals can be removed by
cutting the tube and carefully removing the crystals
with a pointer. Unless it is known ahead of time
that your crystals can be obtained by sublimation,
this technique is not the best choice for obtaining
good single crystals. The sublimed crystals are
frequently thin and fragile, or they may be stuck
to the sides of the wall and be difficult to remove
without incurring fractures. Also, the sample in
the bottom of the sublimation tube is frequently
thermally degraded and must be discarded. Nev-
ertheless, this method is an alternative to solution
methods and may result in new polymorphs,
crystals with modified habits, or highly purified
crystals. The reader is referred to several books on
crystal growth (mostly dealing with inorganic
crystals) for other common laboratory methods
[7].
4. Alternatives to routine methods
In actual practice, if the techniques described
above do not give suitable crystals, there are two
practical choices available to the organic chemist,
The simplest one often is to synthesize another
derivative of the compound, hoping that it will
crystallize more easily. The second alternative is to
modify the crystal growth methods described
above using knowledge about the crystal chem-
istry and properties of the particular compound of
interest. Several examples from our recent re-
search work serve to illustrate this point.
Acetanilide crystals are readily obtainable by
recrystallization from boiling water and this pro-
cedure has been used for years in undergraduate
chemistry laboratories across the country as a
safe, reliable example of crystal growth methods.
The crystals obtained are pure but of poor quality
by X-ray diffraction standards. Crystal structure
analysis of acetanilide, I, shows that that a hydro-
gen-bonded chain of molecules is aligned along
the needle axis of the crystals [8]. This kind of
pattern is characteristic of secondary amides which
of small molecule organiv crystals
o
‘.jl
~ ~ (I)
I
,.—. 0
\
H
I
always crystallize in a trans-conformation so the
carbonyl acceptor group and the —NH hydrogen
bond donor are anti— to one another [9j, as shown.
We can control the morphology of acetanilide
crystals by choosing solvents which promote or
inhibit the formation of this hydrogen-bond chain.
Hydrophobic solvents like benzene and CC!4 will
not participate in hydrogen-bond formation, so
they will induce the formation of rapidly growing
chains of hydrogen-bonded amides. This mecha-
nism is reflected in the observed crystal mor-
phology, where crystals grown from benzene or
Cd4 by evaporation methods are seen to be long
needles extending from one end of the petri dish
to the other (fig. Ia). Solvents that are proton
donors or proton acceptors should inhibit chain
formation by competing with amide molecules for
hydrogen-bonding sites. Thus acetone should in-
hibit chain growth at the —NH end, and methanol
should inhibit chain growth at the carbonyl end of
the chain. Both solvents result in the growth of
short rod-like crystals of acetanilide (fig. ib), as
expected. These morphologies are reproducibly
observed from multiple recrystallizations. A sam-
pie grown from benzene and acetone gave a hy-
brid crystal form (fig. Ic), which is a rod-shaped
crystal with fine needles growing on the ends. The
entire crystal is one single crystal which cx-
tinguishes uniformly under polarized light, show-
ing no phase boundary between rod and needles.
Three crystal structures have been reported for
dibenzoylmethane (DBM), II, but none refer to
/ \ c —CH2--—- c / \ (II)
— —
 M. C. Etter et al. / Growth and charaiter,zation of small molecule organic crystals 649

Fig. 1. Modification of acetanilide crystal morphology by crystallization from solvents with hydrogen-bond properties that favor or
retard growth of the crystals along the needle axis. (a) Aeetanilide crystals grown from benzene as long needles that extend from one
edge of a crystallizing dish to the other. (b) Acetanilide crystals grown from either proton-donating solvents (alcohols) or
proton-accepting solvents (DMSO, acetone), showing retarded growth along the needle axes of the crystals. (c) Hybrid acetanilide
crystals grown from mixed solvents such as benzene and acetone.

the existence of a second polymorphic form. We unusual induction mechanism. A diamond-shaped

found a reference in Chem. Berichte [10] that gave
two different melting points for DBM, suggesting
that a second metastable form of DBM exists. We
found that DBM crystallizes readily from a rapidly
cooled ethanol solution to give long morphologi-
cally well-developed single crystals (fig. 2a), which
transform within minutes to the stable form by an
crystal suddenly appears at the tip of the needle
causing neighboring regions of the needle to
change into diamonds in a chain-type mechanism
proceeding down the length of the crystal to give a
rock-candy type array, shown in fig. 2b. If the
original needles are removed from solution, they
immediately transform by a solvent-mediated
650 MC. Etter etal. / Growth and characterization of small molecule organic crystals

mechanism [11] into a polycrystalline pseudo- cold dry nitrogen to evaporate the solvent, the
morph (fig. 3). If the kinetically favored needle crystals remain clear indefinitely and are suitable
crystals are first cooled in the mother liquor to for X-ray diffraction analysis [12]. By heating the
about —20°C. then removed under a stream of crystals to 50°C on a hot stage, a solid-state

~A.
I
lig. 2. Dihenzoylmethane ( DB!tl cr~stalpols.niorphs grown frotri ethanol. .i) Kiiietieall~ fasored, thermally metastable, needle
polymorph of dibenzoylmethane. h Stable, thermodsnamically favored polymorph of dihenzoylmethane obtained by solvent-media-
ted phase transformation of a metast,ible crystal.
 MC. Etter et a!. / Growth and characterization of small molecule organic crystals 651

‘ 0~

\N~
V I, a~uE
~L.3c5
E~.
01~
Sina
652 M. C. Euer vi al. / Growth and characterization of small molecule organic crvstal.s
transformation, much like the solvent-mediated
process, takes place.
~ ~
CH3
One of the most challenging crystal growth
problems we have encountered involves an organic
dye~—dye complex, called a cyanine-oxonol dye,
III. This compound crystallizes in at least 14
polymorphic and pseudopolymorphic forms. dis-
tinguishable by brilliant color differences, chem-
ical stability differences, and morphology dif-
ferences. The more effort we put into learning
how to preferentially grow one or the other crystal
form, the more polymorphs we find. For one
six-month period we were able to grow only gold
needles. For the next six-month period we ob-
tamed exclusively red plates or purple diamonds,
The only consistent result was that spaghetti-
shaped crystals would form from a transient gel-
Fig. 4. Polymorphic forms of esanine—oxonol d~esobtained from chloroform solutions oh a cs.ininc tosylate and an oxonol tetraethyl
ammonium salt.
like phase when the two components were mixed
together too fast or in concentrations that were
(III)
0 CH3
too high. Fig. 4 shows three of the polymorphic
forms that we commonly obtained. The crystal
structures of the gold and red plates have been
solved, showing that they are indeed polymorphs,
and that they each contain one molecule of ebb-
roform which is hydrogen-bonded to a carbonyl
group of the oxonol anion [13].
We are presently studying the possibility of
using gel-growth techniques to control and limit
diffusion rates of solvents and solutions. Inorganic
chemists have used silica gels successfully for many
years as crystal-growing media hut relatively little
work has been done with organic based gels for
growing organic crystals. We are currently using a
Sephadex LH-20 or LH-60 preparation described
 M C. Etter ci a!., Grim ih and i hurat ti’r/:aj,o,
7 of .rnw/l ini’lecul~’organic cry ital~ 653

a
Fig. 5 Benzil crystals grown from (a) ethanol Solution and (hi cihanol Sephadex gels. Although crystal morphologic.s are distinLils
different. X-ras diffraction pattern.5 of the gel-grown crystals match patterns of solution-growi~crystahs so these are not pohsmorphic
forms.

previously by Curtin and coworkers [14] and are nique involves swelling the gel with a solution of
testing the crYstal-growth characteristics of a benzil. IV. and then adding a nonsolvezIt such as
variety of solvents and organic compounds in
these gels. Sephadex is available as heads which 0
can he swollen in organic solvents to form a thick / \ ~t ~ / \ iv
slurry. A second solution phase is placed o~er(or
under) the gel and two componetits can slowly
diffuse together with crystals forming in the gel water to the top layer. After about 24 h. cr~~tals
phase. A straightforward application of this tech- of henzil can he seen grossing in the gel phase.
6s4 .51.C Liter ci a!. , (,roo i/i and i liaroi ier,zai,on o/ anal! p110/ti u/i organic i rrsia/s

7 ~

— --

Fig. 6. A polar ers stal of I I organic complex (a) grow 1 from soh ution and ib I grow n in a Sephade \ gel. The p~‘I ar morphiohogs is
apparent in gel—grown crsstals, hut a markedh~different morphologs is reproducihhs obtained under these conditions oh diffusion
controlled crsstal growth.

Fig. 5 shows a picture of a henzil cr~stalobtained [crences for crystals of a 3 : I organic complex of
from a solution experiment, compared to the un— 4.4’—dinitrohiphen~I and phenviphenol. V. gross n
usual morphology obtained for crystals gross n ill irom solution or from Sephadex gels, fig. 6. [he
Sephadex. Similaris, we found morphology dif— picket—fence morpholog~is frequentis observed in

~ • ~—~‘0H (\)
 M. C. Etter et a!. / Growth and characterization of small molecule organic crystals
gel preparations of V, but from solution only
single arrow-head shaped crystals grow. Each indi-
vidual crystal from the gel is one single crystal,
giving single crystal X-ray diffraction patterns
identical to those obtained from solution-grown
crystals.
5. Conclusion
Much work needs to be done to improve cx-
isting methods for organic crystal preparation.
The effort needs to be interdisciplinary, involving
organic chemists, crystallographers and materials
scientists. As scientists, we believe that crystal
growth mechanisms are rational and ultimately
understandable. As crystal growth practitioners
we know that the art of crystal growth is much
more highly developed than the science. Innova-
tive, interdisciplinary approaches are needed to
make advances in this field, and the future utility
of organic solids as technologically important
materials depends on such advances.
655
References
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(1984) 690.
[3] A. McPherson, Preparation and Analysis of Protein
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[4] R. Job, P.J. Kelleher, W.C. Stalhings, Jr., CT. Monti and
J.P. Glusker, lnorg. Chem. 21(1982) 3760.
[5] K.A. Jackson, Crystal Growth Kinetics, presented at 1st
Intern. Conf. on Protein Crystal Growth, Stanford. CA,
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[6] CRC Handbook of Chemistry and Physics, 66th ed. (CRC
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[11] PT. Cardew and R.J. Davey, Proc. Roy. Soc. (London)
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[12] MC. Etter, Z. Urbauiczyk-Lipkowksa and D.A. Jahn, Am.
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