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Water soluble complexes of curcumin with


cyclodextrins: Characterization by FT-Raman
spectroscopy

Article in Vibrational Spectroscopy · September 2012


DOI: 10.1016/j.vibspec.2012.05.002

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Vibrational Spectroscopy
journal homepage: www.elsevier.com/locate/vibspec

Water soluble complexes of curcumin with cyclodextrins: Characterization by


FT-Raman spectroscopy
P.R. Krishna Mohan, G. Sreelakshmi, C.V. Muraleedharan, Roy Joseph ∗
Biomedical Technology Wing, Sree Chitra Tirunal Institute for Medical Sciences and Technology, Satelmond Palace Campus, Thiruvananthapuram 695012, Kerala, India

a r t i c l e i n f o a b s t r a c t

Article history: Many recent reports on curcumin, a polyphenol from Curcuma longa, provide mounting evidence on
Received 31 October 2011 the pharmacological activity of this natural product. However, the pharmaceutical use of this molecule is
Received in revised form 10 April 2012 hampered due to its poor solubility in the aqueous media. Inclusion complex formation with cyclodextrins
Accepted 2 May 2012
has been reported as a means to enhance its aqueous solubility. Most of these studies provide infrared (IR)
Available online xxx
spectroscopic data as an evidence to support inclusion complex formation. However, characterization of
the solid inclusion complexes using IR spectroscopy is hindered due to interfering vibrations of cyclodex-
Keywords:
trin. In this study, fully water soluble complexes of curcumin with three hydroxypropyl derivatives of
Curcumin
Cyclodextrin
cyclodextrins were isolated and characterized. Decrease in the intensity of aromatic ring vibrations and
Freeze dried complex shift in peak position from 1626 cm−1 observed in Raman spectrum provided fresh insights into the type
Water soluble curcumin of interactions occurring in the water soluble complex. A new structure for the inclusion complex has
been proposed. From the results it was demonstrated that Raman spectroscopy would provide clearer
and better evidence of inclusion complex formation.
© 2012 Elsevier B.V. All rights reserved.

1. Introduction complexes with small molecules or fragments of large compounds


[8]. Cyclodextrins have been used extensively in pharmaceutical
Turmeric, derived from the rhizome of Curcuma longa has been research and development, and there are numerous cyclodextrin-
used by the people of Indian subcontinent for centuries with no containing pharmaceutical products marketed worldwide. The
known side effects, not only as a component of food but also to most common pharmaceutical application of CDs is to enhance
treat a wide variety of ailments [1]. Curcumin is the phytochemi- drug solubility in aqueous solutions. CDs are also used for increas-
cal that gives yellow color to turmeric. Extensive research within ing stability and bioavailability of drugs, and other additional
the last half century has proven that most of these activities, once applications [9]. Many studies on enhancement of solubility of
associated with turmeric, are due to curcumin [2]. Curcumin is curcumin with cyclodextrins have been reported [10–13]. Recent
reported to have a number of pharmacological activities including reports on curcumin cyclodextrin complexes show the vast amount
antioxidant, HIV antiproteases activity, anti-inflammatory, anal- of research going on in this field worldwide [14–17]. Solubil-
gesic, anticancer, etc. [3]. Of late, its potent antiamyloidogenic ity enhancement is seen even in physical mixtures of drug and
effects in treating Alzheimer’s disease have ignited widespread cyclodextrin. This is attributed to the self aggregation of cyclodex-
research interest on this drug [4]. Pre-clinical and clinical trials have trins in solution [18]. Formulation with a physical mixture rather
revealed that curcumin is safe even up to a dose level of 8.0 g/kg and than a complex is desirable from a pharmaceutical processing view-
this makes it all the more important [5]. But the pharmaceutical use point, since the steps involved in making the complex could be
of this pharmacologically potential molecule is restricted due to its skipped in the manufacturing process. But studies with physical
poor aqueous solubility, resulting in reduced bioavailability [6]. mixtures have shown that it resulted in an insignificant level or
One way to increase its aqueous solubility is to form inclu- complete lack of bioavailability enhancement [19]. Bioavailability
sion complexes, i.e., to encapsulate curcumin as a guest within the enhancement is reported to be possible for inclusion complexes
internal cavity of a water soluble host [7]. Cyclodextrins (CDs) are rather than physical mixtures [20]. Characterization of the solid
cyclic oligomers of glucose that can form water-soluble inclusion inclusion complexes formed is usually carried out using analyt-
ical techniques such as Fourier transform infrared spectroscopy
(FTIR), differential scanning colorimetry, nuclear magnetic reso-
nance spectroscopy, X-ray diffraction (XRD) spectroscopy, etc. FTIR
∗ Corresponding author at: Sree Chitra Tirunal Institute for Medical Sciences and
spectroscopy is widely used for studying inclusion complex forma-
Technology, BMT Wing, Satelmond Palace Campus, Poojapura, Thiruvananthapuram
695012, Kerala, India. Tel.: +91 471 2520225; fax: +91 471 2341814.
tion [21]. Changes in the peak intensity or shifts in the wave number
E-mail address: rjoseph1965@rediffmail.com (R. Joseph). are indicative of complex formation. In recent papers [15,16] also

0924-2031/$ – see front matter © 2012 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.vibspec.2012.05.002

Please cite this article in press as: P.R.K. Mohan, et al., Water soluble complexes of curcumin with cyclodextrins: Characterization by
FT-Raman spectroscopy, Vib. Spectrosc. (2012), http://dx.doi.org/10.1016/j.vibspec.2012.05.002
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FTIR spectroscopy was used for studying the complex formed above. Influence of method of preparation on the characteristics of
between curcumin and hydroxypropyl beta cyclodextrin. However, the complex formed was studied by preparing the complex by co-
H O H deformation bands present in cyclodextrins mask spectral evaporation technique. For this, HP␤CD and curcumin were taken
bands in the desired regions of 1600–1700 cm−1 . In both reported in the mole ratio 2:1 and individually dissolved in ethanol. Both
studies, only vague details are given as evidence for formation of solutions were mixed together in an amber colored beaker and
inclusion complexes using FTIR spectroscopy. stirred at 45 ◦ C until it turned into a paste. The paste was further
Raman spectroscopy has advanced tremendously in recent years dried at 40 ◦ C under vacuum for a period of 48 h. The dry powder
and has found increased applications in the pharmaceutical indus- obtained was sealed in an aluminum foil to protect it from light and
try. It is suggested to be an excellent tool for studying inclusion stored in a desiccator.
complex formation [22]. Raman spectra of cyclodextrins present
regions free of bands that can be used as windows for moni- 2.2. Characterization techniques
toring relevant guest modes, such as the stretching vibration of
double bonds ((C C), (C O), etc.) and of aromatic C H bonds 2.2.1. FTIR spectroscopy
[23,24]. Spectral responses in the region 1600–1700 cm−1 are quite FTIR spectra of curcumin, cyclodextrins, physical mixtures and
unique indicators for the presence of unsaturation in a molecule complexes were recorded on a Bruker IR spectrophotometer,
and often indicate the presence of drug component within an excip- Model-500. The scanning range used was 450–4000 cm−1 with 32
ient matrix [25]. Although Raman spectral study of curcumin has scans and resolution was set at 4 cm−1 . Diffuse reflectance method
been reported earlier [26,27], it has still not been used for charac- was used here with TGS detectors.
terizing curcumin–cyclodextrin solid inclusion complexes.
Freeze-drying is a widely used complexation technique 2.2.2. FT Raman spectroscopy
especially useful for heat labile guests and water soluble hydrox- The room temperature FT-Raman spectra were recorded on a
ypropylated cyclodextrin complexes [28]. The stoichiometry RFS-100 Bruker FT spectrometer using a Nd:YAG laser with the
between cyclodextrins and curcumin has been studied by Tang et al. excitation wavelength of 1064 nm. Each spectrum is the average
[12] and reported to be 2:1. This has been confirmed by fluorescence of two repeated measurements of 200 scans each and of 2 cm−1
measurement studies [10]. In this study, inclusion complexes of resolution. Laser power of the source was maintained at 100 mW
curcumin with three derivative cyclodextrins were studied using throughout all measurements and a liquid nitrogen cooled Ge
Raman spectroscopy. CDs and curcumin were taken in the molar detector was used.
ratio 2:1 and fully soluble complexes were freeze dried and used
for Raman spectroscopic studies. Complexes were also prepared by 2.2.3. XRD spectroscopy
co-evaporation technique to generate supporting information. The patterns of pure curcumin, HP␥CD and freeze dried complex
of curcumin with HP␥CD were obtained using the X-ray diffrac-
tometer (XPERT PRO) with Cu radiation source. Measurements
2. Experimental
were performed at a voltage of 40 kV and 30 mA. The scanned angle
was set from 10◦ ≤ 2 ≥ 70◦ and the scan rate was 2◦ min−1 .
2.1. Materials
2.2.4. UV–Vis spectroscopy
Curcumin, trade named BiocurcumaxTM , used for this study
Curcumin content in the complex was determined by using
was obtained from M/s. Arjuna Naturals Ltd., Aluva, Kerala State,
a UV–Vis spectrophotometer (model UV-1800, Shimadzu, Japan).
India. Hydroxypropyl derivatives of parent cyclodextrins, namely,
For this 10 mg complex was dissolved in 10 ml of ethanol and the
hydroxypropyl alpha cyclodextrin (HP␣CD), hydroxypropyl beta
absorbance of the solution at 430 nm was recorded. The curcumin
cyclodextrin (HP␤CD) and hydroxypropyl gamma cyclodextrin
content was estimated from a standard X–Y plot of curcumin (dis-
(HP␥CD), were procured from Sigma Aldrich, USA. Ethanol (99.9%
solved in ethanol) vs. concentration. The path length of the quartz
absolute) was procured from M/s. SD fine Chemicals, Mumbai. All
cell was 10 mm. For each measurement the baseline was estab-
the above compounds were used without further purification.
lished by placing blank ethanol in the reference compartment. All
measurements were carried out at 25 ± 1 ◦ C.
2.1.1. Methods used for the preparation of complexes
Water soluble inclusion complexes and physical mixtures of 2.2.5. Differential scanning calorimetry (DSC)
cyclodextrins and curcumin were prepared by taking both com- DSC measurements were carried out in a DSC Q100 V9.6 Build
pounds in the molar ratio 2:1, respectively. Inclusion complexes 290 differential scanning calorimeter (TA Instruments Inc., USA)
were prepared by freeze drying technique according to a method based on ASTM E 537-07 standard. Heating rate employed was
reported elsewhere with slight modification [29]. In brief, 1 g CD 10 ◦ C/min and the samples were scanned in the temperature range
was dissolved in 30 ml water taken in a 250 ml stoppered coni- 25–210 ◦ C. The measurements were carried out under dry nitrogen
cal flask. The solution was stirred for 10 min till a clear solution at a flow rate of 50 ± 5 ml/min.
was obtained. Curcumin was added to this solution, shaken well
and placed in an incubator shaker for 7 days at 37 ◦ C. The reaction 3. Results and discussion
mixture was filtered through 0.45 ␮m filter and the clear solution
was taken. The filtrate thus obtained was freeze dried to obtain Although many studies have been carried out on
the solid complexes. The red colored complex thus obtained were curcumin–cyclodextrin complexes, characterization of fully
ground to a fine powder and kept in a desiccator for characteri- water soluble complexes have not been reported yet. Character-
zation. Physical mixtures of curcumin and CDs were prepared by ization techniques used in earlier studies, especially FTIR, could
thoroughly mixing both compounds in a pestle–mortar for a period not provide adequate evidence for inclusion complex formation
of 1 h. Curcumin–cyclodextrin complexes and physical mixtures [15,16]. The method used for preparing inclusion complexes
were prepared with HP␣CD, HP␤CD and HP␥CD. In order to study is another important factor to be considered. In the present
the effect of mole ratio on the characteristics of the resultant prod- study, the soluble curcumin–cyclodextrin complexes were filtered
uct, HP␤CD and curcumin were taken mole ratio 1:1 and a batch of through 0.45 ␮m filter in order to remove any insoluble curcumin
freeze-dried complex was prepared as per the procedure described present. The filtrate thus obtained, was freeze dried to obtain solid

Please cite this article in press as: P.R.K. Mohan, et al., Water soluble complexes of curcumin with cyclodextrins: Characterization by
FT-Raman spectroscopy, Vib. Spectrosc. (2012), http://dx.doi.org/10.1016/j.vibspec.2012.05.002
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Fig. 1. FTIR spectrum of curcumin. Fig. 2. FTIR spectra of cyclodextrins. (a) HP␣CD, (b) HP␤CD and (c) HP␥CD.

complexes. In all earlier reported studies the investigators had in Table 1. As expected, the IR spectrum of the physical mixture
not filtered out their complexes with curcumin and as a result the (Fig. 3b) contained peaks corresponding to both the components
characterized products were mixtures of uncomplexed curcumin, (Fig. 3a and d) present. IR spectrum of HP␤CD (Fig. 3d) and that
complexed curcumin and uncomplexed cyclodextrin. Due to this of freeze dried complex (Fig. 3c) appear identical. Since the com-
reason FTIR spectral study used in the earlier reported works is not plex is fully soluble in water significant changes in the IR spectra
likely to provide adequate information on the inclusion complex such as shift in peak positions were anticipated. The sharp peak at
formation. 3508 cm−1 due to free hydroxyl (OH) group of curcumin (Fig. 3a),

3.1. FTIR spectroscopy

3.1.1. FTIR spectrum of curcumin


A detailed study on the vibrational spectra of curcumin has been
reported earlier by Kolev et al. [26]. The FTIR spectrum of cur-
cumin is shown in Fig. 1. A broad peak at 3293 cm−1 and the sharp
one at 3508 cm−1 indicate the presence of OH. The strong peak at
1626 cm−1 has a predominantly mixed (C C) and (C O) char-
acter [26]. Another strong band at 1601 cm−1 is attributed to the
symmetric aromatic ring stretching vibrations (C Cring ) [27]. The
1508 cm−1 peak is assigned to the (C O), while enol C O peak
was obtained at 1272 cm−1 , C O C peak at 1023 cm−1 , benzoate
trans-CH vibration at 959 cm−1 and cis CH vibration of aromatic
ring at 713 cm−1 .

3.1.2. FT-IR spectra of cyclodextrins


The overlaid FTIR spectra of HP␣CD, HP␤CD and HP␥CD are
given in Fig. 2. The FTIR peaks of all the hydroxypropyl derivatives
of cyclodextrins are almost identical with slight variations in their
intensities. The OH group stretching vibrations at 3300–3400 cm−1
is a characteristic peak of the cyclodextrins. An intense peak
at 2929 cm−1 due to C H assym/symm stretching is seen. The
H O H deformation bands of water present in cyclodextrins
are seen at 1658 cm−1 . C H overtone stretching vibrations are
seen at 1155 cm−1 and 1082 cm−1 , while at 1082 cm−1 C H, C O
stretching vibrations are present. Absorption at 1300–700 cm−1
corresponds to skeletal C C vibrations and that at 1155 cm−1
corresponds to C O C vibrations. Since there are no significant
variations in the spectra of different derivative cyclodextrins, com-
plexes of curcumin with HP␤CD was selected for detailed analysis.

3.1.3. FT-IR spectra of physical mixture and freeze dried complex


Fig. 3. FTIR spectra of: (a) curcumin, (b) physical mixture (curcumin + HP␤CD),
FT-IR spectra of curcumin, HP␤CD, physical mixture of curcumin
(c) freeze dried complex (curcumin + HP␤CD) and (d) HP␤CD. [*] Indicates that
and HP␤CD and the freeze dried complex of curcumin with HP␤CD peaks due to (C Cring ) and mixed (C C), (C O) are masked due to the OH peaks
are shown in Fig. 3. The peak assignments of all the spectra are given of cyclodextrin.

Please cite this article in press as: P.R.K. Mohan, et al., Water soluble complexes of curcumin with cyclodextrins: Characterization by
FT-Raman spectroscopy, Vib. Spectrosc. (2012), http://dx.doi.org/10.1016/j.vibspec.2012.05.002
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Table 1
IR peak assignments of curcumin [26], HP␤CD, physical mixture (PM) and freeze dried complex (FDC).

Curcumin  [cm−1 ] HP␤CD  [cm−1 ] PM  [cm−1 ] FDC  [cm−1 ] Peak assignment

3508 OH stretching of phenol group, intra-molecular H bond


3360 3360 3360 OH stretching
3293 CH stretching, intermolecular H bond stretching
2972 Asymmetric CH stretch of CH3
2945 CH stretch of OCH3
2929 2928 2929 Asymmetric CH stretching of CH2
1658 1658 HOH of water of crystallization, C C stretching
1626 1626 C O, Cring C C stretching
1601 1601 Aromatic C C stretching
1508 1530 1510 1516 C O stretching, CCC, CC O in plane bending
1429 1429 In plane bending of aromatic (CCC, CCH), enolic (COH), CH in plane
bending due to CH2
1458 1456 In plane bending of CH3
1372 1367 1367 1367 In plane bending of CH, enolic COH, skeletal CCC
1272 1280 CH in plane bending of C CH, aromatic C O stretching
1155 1155 1155 CH overtone stretching, C O C stretching
1150 In plane bending of aromatic CCH, skeletal CCH
1082 1082 1082 C O, C C stretching, in plane bending of OCH
1032 1031 1032 C O, C C, CCO, C O C stretching of glucose units
1023 C O C stretching, out of plane bending of CH3 , in plane bending of
aromatic CCH
959 944 948 950 C O stretching, in plane bending of CCH
852 851 852 851 CH out of plane bending of aromatic CCH and skeletal CCH
753 753 753 Skeletal C C stretching, CH out of plane bending
713 711 711 711 In plane bending of skeletal CCH and aromatic CCH, C C stretching

is found to have merged with broad OH peak of HP␤CD present at reported for characterizing curcumin–cyclodextrin solid inclusion
3360 cm−1 . It may be argued that merging of the peaks is due to complexes. Raman spectrum of curcumin is shown in Fig. 4. Stud-
the interactions that occur with cyclodextrin during complex for- ies suggested that curcumin exists in the enol form rather than
mation. However, it may be counter-argued that only the broad in the diketone form [30]. This is indicated by the absence of
peak of HP␤CD is present and there is no evidence of complex for- (C O) peaks in the region of 1650–1800 cm−1 in our studies. The
mation. Moreover the curcumin content in the complex is very low. bands corresponds to (C C) and (C O) appear to be superposed
Other aspects that are usually taken as evidence of complex forma- at 1626 cm−1 having been reduced from their normal values by
tion are peak shifts of aromatic C C at 1601 cm−1 and that of the conjugation. This band with a mixed (C C) and (C O) charac-
carbonyl C O peaks at 1626 cm−1 . But the molecular vibrations of ter was selected for analysis in the present study. Another strong
cyclodextrins in the range 1700–1550 cm−1 mask these peaks. The band, the 1601 cm−1 due to aromatic ring vibrations (C Cring ) was
(H OH) bending of water molecules attached to cyclodextrins has also selected for studying the inclusion complex formation of cur-
a broad and intense band at 1658 cm−1 and this overlaps with other cumin with cyclodextrin derivatives. Raman spectra of curcumin,
useful vibrations associated with complex formation. The peak at HP␤CD, physical mixture of both curcumin and HP␤CD and freeze
1508 cm−1 due to the (C O), ı(CCC) and ı(CC O) seen in the case dried complex of both were analyzed and compared in Fig. 5. As
of curcumin and physical mixture undergoes a shift to 1516 cm−1 the Raman intensities of HP␤CD (Fig. 5d) are very weak, the spec-
in the case of freeze dried complex and this may be taken as an trum of the physical mixture appeared to be identical to that of
evidence of complex formation but the intensity is too low to be curcumin (Fig. 5a and c). In the case of curcumin–HP␤CD freeze
unambiguously detected. The peaks due to ı(CCH) of the phenol dried complex (Fig. 5b), the peak at 1626 cm−1 has shifted towards
and skeletal ı(CCH) at 1150 cm−1 due to in-plane bending vibra- 1635 cm−1 . This indicates that strong interactions have occurred
tions are masked by the C O vibrations of the cyclodextrin [16,26].
The in-plane bending vibrations of aromatic keto group ı(CCH) and
out of plane vibrations of CH3 , i.e., ı(CH3 ) found at 1023 cm−1 ,
out of plane bending of skeletal CCH and the aromatic CCH at
852 cm−1 of curcumin are masked by the C O C glucose units
and the ring structure C O C peak of cyclodextrin [16,26]. The
peaks at 713 cm−1 of aromatic in plane bending, stretching vibra-
tions of C C and in plane bending of skeletal CCH of curcumin are
masked by corresponding vibrations of cyclodextrin. The peak at
1272 cm−1 due to aromatic keto in plane vibrations ı(CCH), ı(CCC)
and keto-enol ı(COH) are found to have merged with IR vibrations
of HP␤CD molecule. In short all the major peaks of curcumin shown
in Fig. 3 are hidden by the cyclodextrin peaks in the similar regions.
Thus, IR spectroscopy fails to explain inclusion complex formation
of curcumin in detail.

3.2. Raman spectroscopy

Raman spectroscopy has been used in earlier studies for the


characterization of both inclusion complexes [25] and curcumin
[26]. However, use of Raman spectroscopy has not yet been Fig. 4. Raman spectrum of curcumin.

Please cite this article in press as: P.R.K. Mohan, et al., Water soluble complexes of curcumin with cyclodextrins: Characterization by
FT-Raman spectroscopy, Vib. Spectrosc. (2012), http://dx.doi.org/10.1016/j.vibspec.2012.05.002
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Fig. 6. Raman spectrum of curcumin overlaid with curcumin–HP␤CD complexes:


(a) curcumin, (b) freeze dried complex – 1:1 mole ratio, (c) freeze dried complex –
2:1 mole ratio, and (d) co-evaporated complex – 2:1 mole ratio.

Fig. 5. Raman spectra of: (a) curcumin, (b) freeze dried complex of curcumin with complex formation and other is the type of interactions postulated
HP␤CD, (c) physical mixture of curcumin with HP␤CD, and (d) HP␤CD.
by Sanphui et al. for the orthorhombic form of curcumin. In the
presence of cyclodextrin derivatives one end of the curcumin goes
between curcumin and HP␤CD. As a result of inclusion complex into the cyclodextrin cavity [10] and the enolic part of curcumin
formation with HP␤CD, the conjugation of (C C) and (C O) is undergoes OH· · ·O bond with the hydroxyl groups of cyclodextrin.
reduced considerably resulting the peak shift towards 1635 cm−1 . The other end of the curcumin would interact with neighboring
The ring strain of cyclodextrin also contributes to this shift. This curcumin molecule by their respective phenolic OH groups form-
shift in peak position was observed irrespective of the molar ratios ing OH· · ·O linkages. Weak CH· · ·O interactions are also possible
taken for complexation and the method used for complexation. between CH3 O of one molecule with OCH3 of the other. The entry
Fig. 6 compares the spectra obtained from the complexes pre- of one end of curcumin molecule into the cavity of cyclodextrin
pared by taking HP␤CD and curcumin in the mole ratios 2:1 and and the molecular interactions of the type discussed above for
1:1. The Raman spectrum of co-evaporated complex is also over- the orthorhombic polymorph of curcumin may contribute to the
laid for comparison. These results suggest that peak shift is the increased solubility of the complex. The peak shift observed in the
direct result of complex formation. Sanphui et al. [31] reported a case of complex from 1626 cm−1 to 1635 cm−1 is probably due to
similar shift in peak position when commercial curcumin was co- the interaction between the hydroxyl group of cyclodextrin and
crystallized with 4-hydroxypyridine from ethanol solution at room enolic OH of curcumin (Scheme 1). The Raman spectra of commer-
temperature. They found that a different polymorph of curcumin cial curcumin and physical mixture of curcumin with cyclodextrin
that exists in orthorhombic state was formed on co-crystallization shown in Fig. 5 display no peak shift from 1626 cm−1 to 1635 cm−1
with 4-hydroxypyridine. like that is seen in the freeze dried system. It is evident that there
Molecules in the commercial curcumin crystallize in mono- is no polymorph formation or complexation in the physical mix-
clinic lattice and exist in a curved, slightly twisted conformation ture. This confirms that in freeze dried complexes there exists high
whereas in the orthorhombic form the molecules exhibit a linear level of molecular interaction between curcumin and the respec-
and planar conformation [31]. In the monoclinic form there are phe- tive cyclodextrins. Similar effect is seen in the Raman spectrum of
nol O H· · ·O hydrogen bonds on both sides to different molecules HP␣CD complex and HP␥CD complex (Fig. 7).
which are in turn connected to a fourth curcumin molecule in An interesting evidence for inclusion complex formation is
a macrocyclic hydrogen bond ring. However, due to the linear provided by the symmetric aromatic ring stretching vibrations
molecular shape no such ring motif could be dissected in the (C Cring ) of curcumin at 1601 cm−1 (Fig. 7). The intensity of
orthorhombic form [31]. Weak C H· · ·O interactions also play an the 1601 cm−1 peak is considerably reduced suggesting that the
important role in the overall molecular aggregation of both crystal aromatic ring has gone inside the cavity of the cyclodextrin as pos-
forms. Since the curcumin–HP␤CD complex is fully water soluble, tulated by Baglole et al. [10]. Though a decrease in the peak intensity
it is likely that two types of interactions occur here: One is inclusion of (C Cring ) was observed in the case of polymorphs of curcumin

Please cite this article in press as: P.R.K. Mohan, et al., Water soluble complexes of curcumin with cyclodextrins: Characterization by
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Scheme 1. Proposed inclusion complex formation of curcumin with cyclodextrin.

by Sanphui et al. [31], their decrease in peak intensity was asso- in Fig. 7, were compared with the intensities of peak at 1626 cm−1
ciated with a peak shift of about 9 cm−1 . In the present case little (I1626 ) (for curcumin) or that at 1635 cm−1 (I1635 ) (for the com-
shift in peak position was observed and we can expect a different plexes). When the ratio I1626 /I1601 was calculated the value was
type of interaction. It is likely that when the complex is formed the found to be 0.59 for curcumin (Table 2). However, for the com-
benzene ring at one end of the curcumin will move into the cavity plexes the ratio, I1635 /I1601 was ≥1. The decrease in intensity of
of cyclodextrin where the enolic OH of curcumin form OH· · ·O bond (C Cring ) stretching modes of aromatic ring motifs of curcumin in
with the hydroxyl group of the cyclodextrin. This is depicted as a the freeze dried complex suggests the presence of a surrounding
cartoon in Scheme 1. obstacle opposing the Raman active vibrational mode. The reduc-
The cyclodextrin cage may restrict the aromatic vibrations in tion in intensity is apparently due to shielding of the benzene ring
Raman spectra leading to decreased peak intensity. The intensities under the influence of the cyclodextrin cavity. Quenching of the
of the peak at 1601 cm−1 (I1601 ) of curcumin and complexes, shown guest structural electron polarisability occurs due to the interaction
with the frozen localized electron density of the polar carbohydrate
host. This clearly suggests that the aromatic ring of the curcumin
moiety gets included in the cyclodextrin cavity. The aromatic group
at the other end of the molecule is not entrapped in the cyclodex-
trin cavity and this would interact with neighboring molecules in
a linear pattern and exist in a water soluble polymorphic form.
The curcumin content in the complexes was determined by
UV–Vis spectroscopy and the data obtained is shown in Table 2.
Though the Raman spectra of all complexes appear similar, the
curcumin content in the complexes are widely different. Based on
the curcumin content obtained, the complex formation efficiency
of hydroxypropyl derivatives of cyclodextrins may be rated in the
order: HP␤CD < HP␣CD < HP␥CD.

3.3. XRD analysis

The XRD spectra of curcumin, HP␥CD and freeze dried complex


of curcumin with HP␥CD are shown in Fig. 8. Results reveal that the
commercial curcumin exists in a crystalline form whereas HP␥CD
is amorphous in nature. In the case of freeze dried complex, the
spectrum exhibits features of a compound that lacks a well defined
crystalline structure.

3.4. Differential scanning calorimetry

DSC is a very useful tool for collecting qualitative information on


the physico-chemical state of guest in a complex where host–guest
interactions take place [15]. The complex formation with a guest
molecule may result complete disappearance of endothermic peak
or shifting of peak to the other temperatures indicating changes
in crystal lattice, melting, boiling or sublimation points. Since
HP␥CD has shown to have the highest complexation efficiency,
HP␥CD–curcumin complex was used for DSC analysis. Thermo-
grams of curcumin, HP␥CD, physical mixture and freeze dried
complex are overlaid and are shown in Fig. 9. DSC thermogram
of curcumin (Fig. 9a) shows an endothermic peak at 175.26 ◦ C indi-
Fig. 7. Raman spectra of freeze dried complexes of curcumin with ␣, ␤, and ␥
hydroxyl propyl cyclodextrins: (a) HP␣CD complex, (b) HP␤CD complex, (c) HP␥CD cating the melting of curcumin. Thermogram of HP␥CD (Fig. 9b)
complex, and (d) curcumin. shows a broad peak at 84.28 ◦ C due to loss of water. Being

Please cite this article in press as: P.R.K. Mohan, et al., Water soluble complexes of curcumin with cyclodextrins: Characterization by
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Table 2
Raman intensity values and curcumin content in the freeze dried complexes.

Sample details Absorption band Raman Absorption band Raman IA /IB Curcumin content
position ‘A’ intensity IA position ‘B’ intensity IB in the complex/%

Curcumin 1626 3.747 1601 6.304 0.59 –


HP␣CD:Curcumin (2:1) complex 1635 0.022 1603 0.022 1.00 0.47
HP␤CD:Curcumin (2:1) complex 1635 0.021 1604 0.018 1.17 0.30
HP␥CD:Curcumin (2:1) complex 1635 0.081 1601 0.072 1.13 2.39

Complex formation increased the bulk of curcumin molecule and


prevented close packing and crystallization of curcumin. Thus, the
data obtained from DSC supported molecular level interactions and
inclusion complex formation in water soluble cyclodextrin cur-
cumin complexes.

4. Conclusion

Raman spectroscopy provided clear and distinct evidence for


inclusion complex formation of curcumin with derivative cyclodex-
trins. Absence of Raman signals in cyclodextrin in the region
1500–1800 cm−1 provided an opportunity to reveal shift in the
Raman bands of curcumin due to complex formation. Curcumin
absorbed strongly in the Raman region and the molecular inter-
actions occurred in the complex could be studied even when the
curcumin content in the complex was below 1 wt.%. Decrease in
the intensity of aromatic ring vibrations and shift in peak posi-
tion from 1626 cm−1 could be taken as the evidence of inclusion
complex formation. Complexes were found to be formed when
CDs and curcumin were taken in the mole ratio 2:1 and 1:1. Co-
evaporation technique also yielded inclusion complex. XRD and
DSC data showed that water soluble complex formed was amor-
Fig. 8. XRD spectra of: (a) HP␥CD, (b) freeze dried complex of curcumin with HP␥CD phous in nature.
and (c) curcumin.

Acknowledgements
amorphous, it did not show any melting endotherm. Physical mix-
ture shows peaks corresponding to the melting of curcumin and Authors wish to acknowledge Dr. V.K. Krishnan for the provi-
loss of water from HP␥CD (Fig. 9d). However, shift in peak posi- sion of spectroscopy data. They also thank the Director, SCTIMST,
tions are observed in this case and the melting peak is not very Trivandrum for the facilities extended and the kind permission to
sharp. It may be noted that the curcumin content in the physical publish this work.
mixture was only 10 wt.%. The dilution effect by HP␥CD contributed
to the sharp decrease in intensity of the melting peak and its shift References
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FT-Raman spectroscopy, Vib. Spectrosc. (2012), http://dx.doi.org/10.1016/j.vibspec.2012.05.002

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