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Characterization, physicochemical and photo-stability of a co-crystal
involving an antibiotic drug, nitrofurantoin, and 4-hydroxybenzoic acid†
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Venu R. Vangala,*a Pui Shan Chowa and Reginald B. H. Tan*ab

Received 26th October 2010, Accepted 22nd November 2010
DOI: 10.1039/c0ce00772b

Pharmaceutical co-crystallizing ability of an antibiotic agent,

nitrofurantoin, is screened with three isomeric monohydroxybenzoic
acids. 1 : 1 co-crystal of nitrofurantoin-4-hydroxybenzoic acid has
been prepared and structurally characterized. Further, the co-
crystal displays superior physicochemical and photo-stability
compared to nitrofurantoin.
Co-crystals are an emerging class of solid forms of pharmaceutical
materials.1 Co-crystallization of active pharmaceutical ingredient
(API) with pharmaceutically acceptable co-formers via a molecular
recognition process offers a promising approach to alter the melting
behavior,2 solubility,3 dissolution,4 bioavailability5 and mechanical
properties6,7 of an API. Recent advancements have indicated a great
potential of cocrystals for improving the physicochemical properties
of API. For example, Jones and co-workers addressed the hygro-
scopicity problem of caffeine and theophylline through co-crystal
formation with oxalic acid.8 McNamara et al. showed an improve-
ment in the physical stability of 2-[4-(4-chloro-2-fluorphenoxy)-
phenyl]pyrimidine-4-carboxamide by forming co-crystal with glutaric
acid.5a Rodrıguez-Hornedo and co-workers improved the chemical
stability of carbamazepine by co-crystallization with saccharin or
nicotinamide.9 In this contribution, 1 : 1 co-crystal involving nitro-
furantoin and 4-hydroxybenzoic acid has been prepared and struc-
turally characterized. Our investigations further demonstrated that
the physicochemical and photo-stability of the co-crystal was superior
to that of API.
Nitrofurantoin (NF) is a well known antibacterial drug extensively
used as an oral treatment for urinary tract infections (Scheme 1).10 It
has been reported that the dissolution rate and bioavailability of
nitrofurantoin in commercial tablets decreased upon storage at
Scheme 1 Molecular structures of nitrofurantoin (NF) and 4-hydroxy-
benzoic acid (4HBA).
different relative humidities and elevated temperature.11 Otsuka and
co-workers suggested that the physicochemical stability of NF could
be one of the important factors for controlling the bioavailability of
commercial preparations.12 NF exists in both anhydrous (a- and b-)
and hydrate polymorphic forms (Forms I and II).10 Besides, it is
a photosensitive drug.13 NF (b-form) is commercially available and
was utilized for the study.
A screening of NF (b-form) with simple regioisomeric mono-
hydroxybenzoic acids namely, 2-, 3-, and 4-hydroxybenzoic acid
(HBA) (Scheme 1) was conducted by solid-state grinding (neat and
solvent-drop) and in solution. The HBA co-formers14 were chosen
because of their ability to form co-crystals and these isomers represent
a systematic positioning of functional groups on the aromatic ring.15
Products from solid-state (neat and solvent drop) grinding experi-
ments were analyzed by PXRD and there was a matching of PXRD
patterns with physical forms of starting materials except for NF-
4HBA under the tapered conditions (ESI†).
Evaporative crystallization of NF (b-form, 238 mg, 1 mmol) and
4HBA (138 mg, 1 mmol) from acetonitrile (35 mL) yielded good
quality yellow blocks whereas 3HBA/2HBA provided orange red

a
Crystallisation and Particle Science, Institute of Chemical and
Engineering Sciences, A*STAR (Agency for Science Technology and
Research), 1, Pesek Road, Jurong Island, Singapore 627833. E-mail:
venugopal_vangala@ices.a-star.edu.sg; Fax: +65 6316 6183; Tel: +65
6796 3862
b
Department of Chemical & Biomolecular Engineering, National University
of Singapore, 10 Kent Ridge Crescent, Singapore 117576. E-mail:
reginald_tan@ices.a-star.edu.sg
† Electronic supplementary information (ESI) available: Experimental,
grinding, thermal, spectroscopy, H-bond geometries, physicochemical
and photostability details. CCDC reference number 787039. For ESI
and crystallographic data in CIF or other electronic format see DOI: Fig. 1 (a) DSC traces for NF, 4HBA and NF-4HBA, and (b) the PXRD
10.1039/c0ce00772b pattern of NF-4HBA is unique from either NF or 4HBA.

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needles of NF–H2O (Form II)‡ in a day at ambient conditions.

Thermal analysis by DSC and TGA of yellow blocks were carried
out. DSC trace showed one major transition at 243.58  C, which is
attributed to a melting event (Fig. 1a). The melting point is in between
that of the API and 4HBA. TGA profile showed the absence of
solvent molecules.† The powder X-ray diffraction pattern of NF-
4HBA was distinctly different from NF and 4HBA (Fig. 1b), con-
firming that it is a non-solvated new crystalline phase. In fact, IR and
Raman analyses substantiate the same conclusions.†
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Single crystal X-ray diffraction analysis confirmed that NF-

4HBAx is indeed a co-crystal with 1 : 1 molar ratio crystallizing in the
monoclinic system with P21/c space group. The asymmetric unit
consists of one molecule each of API and co-former. NF and 4HBA
molecules were held together by a primary synthon of N–H/O–H/
 q/ : 1.89, 171; 1.70, 171) between imide N–H and phenolic
O (d/A,
O–H along the b-axis (Fig. 2a). This building block was supported by
significant C–H/O interactions.† In the next dimension, 4HBA
molecules interact among themselves via O–H/O homosynthon to
form centrosymmetric acid-acid dimers (d/A,  q/ : 1.62, 172).16 The
overall crystal packing was layered type. The layers were stabilized by
p–p stacking from 4HBA-furanyl ring of NF and also within 4HBA
 3.25, 3.45). These layers were further
in an antiparallel fashion (d/A:
assisted by methylene C–H/O bonds to complete the third
dimension.†
Intriguing supramolecular synthon concept formed the basis for
co-crystal engineering and retrosynthetic analysis.17 While NF (b-
form) structure was predominantly stabilized by imide catemer of N–
H/O synthons,10 in 1 : 1 co-crystal of NF-4HBA, however, that was Fig. 3 PXRD patterns of (a) NF (b-form), and (b) NF-4HBA incubated
replaced by what appears to be the advantageous N–H/O–H/O at 24  C and 97% RH. While NF changed to NF–H2O (Form II) by
catemer type of synthon, conserving a two fold screw axis. 7 weeks, the co-crystal remained intact.
The physical stability of NF-4HBA (1 : 1) co-crystal was tested
against that of NF (b-form) at various relative humidity (RH) and
temperature conditions recommended by the ICH guidelines for it occurred within one week.† Fig. 3 shows the PXRD patterns
pharmaceutical stability testing.18 Incubated samples were analyzed analyzed for stored samples of NF (b-form) and NF-4HBA at 24  C
by PXRD at designated timepoints and for selected samples DTA/ and 97% RH up to 13 weeks and compared against native samples
DSC was used. The representative results are shown in Fig. 3 and 4. and pure NF–H2O (Form II). The start of phase transformation was
Table 1 shows a summary of the experimental findings and more observed at 7 weeks for NF (b-form) in PXRD and the results
details are included in ESI.† There were no detectable changes with complemented well with DTA/DSC data (Fig. 4). Interestingly, NF-
NF (b-form) stored at 24  C and RH (<10, 33, 57, 75%) and at 40  C 4HBA was robust and was found to remain unchanged in all the
and 75% RH as per the PXRD. However, NF anhydrous (b-form) tested stress conditions (Table 1).
converted to NF hydrate (Form II) at 24  C and 97% RH in 7 weeks Preliminary chemical stability investigations were carried out for
whereas at 40  C and 96% RH, phase transformation was rapid and NF (b-form) and NF-4HBA stored at 40  C and 75% RH and 24  C

Fig. 2 (a) Crystal structure of NF-4HBA (1 : 1) view down the c-axis. Note the key N–H/O–H/O synthons, and the insertion of dimer units of 4HBA
within NF molecules. (b) For comparison, crystal packing of NF (b-form) with the N–H/O catemers is provided.10

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Fig. 4 DTA/DSC curves showed that NF (b-form) transformed to NF–

H2O (Form II) by 7 weeks at 24  C, 97% RH, however, under similar
conditions the co-crystal was stable for 13 weeks. For comparison, DTA Fig. 5 UV irradiation of NF and NF-4HBA samples up to a week, and
profiles of NF (b-form) and NF–H2O (Form II) before storage are also the photodegradation curves with error bars show the enhanced photo-
provided. stability of 1 : 1 co-crystal of NF-4HBA.

(Fig. 5), demonstrating some degree of stabilization achieved through

Table 1 RH stability studies at ambient and 40  C temperatures
co-crystallization.†
Conditions (T, RH) Results In conclusion, a systematic crystal engineering study was per-
formed on the model API, NF. A 1 : 1 co-crystal of NF-4HBA has
1a 24  C, <10, 33, 57, No phase change in 13 weeks been prepared and characterized. The physicochemical stability with
75%RH; 40  C, 75%RH
respect to temperature and RH was evaluated. The cocrystal
24  C, 97% RH In 7 weeks, NF (b-form) / NF$H2O
(Form II) exhibited significant stability to varying temperature and humidity
40  C, 96% RH In 1 week, NF / NF$H2O over a period of 13 weeks. It was also demonstrated that the photo-
stability of co-crystal is superior over the API.
2a 24  C, <10, 33, 57, 75, 97%RH No dissociation or phase changes
40  C, 75, 96% RH during 13 weeks of storage in all We gratefully acknowledge the support for this research by SERC
the specified storages. of A*STAR, Singapore. We thank Yeo Hui Hui Melissa and Tan Li
a Teng for their technical support and Drs. Srinivasulu Aitipamula and
1 and 2 represents NF and NF-4HBA (1 : 1) co-crystal respectively.
Parijat Kanaujia for helpful discussions.

Table 2 Chemical degradation studies at selected conditions Notes and references

‡ Orange red needles melted at 272  C and the unit cell determinations
% Degradation
shown to be matching with NF–H2O (Form II, pbca space group).
Conditions (T, RH) 1a 2a x Crystallographic data for NF–4HBA (C8H6N4O5$C7H6O3, M ¼
376.29, Rigaku Saturn CCD, 110 K): monoclinic, a ¼ 10.298 (2), b ¼
 b ¼ 108.08(3) , V ¼ 1548.6(6) A
6.4096(13), c ¼ 24.680(7) A, 3, D ¼ 1.614
24  C, 57%RH 7.4 0.3
40  C, 75%RH 11.2 0.7 g cm3, space group ¼ P21/c, Z ¼ 4, m(Mo-Ka) ¼ 0.134 mm1, size 0.40 
0.18  0.13 mm. 12212 total reflections of which 4303 were independent,
a
1 and 2 represents NF and NF-4HBA (1 : 1) co-crystal respectively. 4023 observed [I > 2s(I)]. Refinement against F2 with 244 parameters, R1
[I > 2s(I)] ¼ 0.0552, wR2 ¼ 0.1546.

1 (a) W. Jones, W. D. S. Motherwell and A. V. Trask, MRS Bull., 2006,

and 57% RH in open vials for 13 weeks. The assay content of NF was
analyzed by HPLC. Results tabulated in Table 2 indicated that NF-
4HBA showed a greater stability towards chemical degradation in
these two storage conditions.
Finally, co-crystal photo-stability was tested against the API as NF
is known to be a photosensitive drug.13 Irradiation with artificial light
was performed by exposure to 315–400 nm UV lamp. Time depen-
dent change in the NF content under UV irradiation was moni-
tored.19 NF and NF-4HBA (1 : 1) in powder form were distributed in
watch glass aliquots independently and analyzed at various intervals
during exposure (1, 3, 6, 12, 18, 24, 48, 72 and 168 h). While 27.7%
(0.1, n ¼ 3) NF were degraded after 168 h of UV irradiation, 16.7%
(0.4, n ¼ 3) of NF was degraded in 1 : 1 co-crystal of NF-4HBA
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