Vibrational Spectroscopy 62 (2012) 292–298

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

Encapsulation and isomerization of curcumin with cyclodextrins characterized

by electronic and vibrational spectroscopy
E. López-Tobar a , G.P. Blanch b , M.L. Ruiz del Castillo b , S. Sanchez-Cortes a,∗
a
Instituto de Estructura de la Materia (IEM-CSIC), Serrano 121, 28006 Madrid, Spain
b
Instituto de Ciencia y Tecnología de los Alimentos y Nutrición (ICTAN-CSIC), Juan de la Cierva 3, 28006 Madrid, Spain

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

Article history: FT-Raman and UV–visible adsorption spectroscopy are applied for the first time in the structural study of
Received 20 April 2012 the antioxidant, antitumoral polyphenol curcumin and its complexation with ␤- and ␥-cyclodextrin.
Received in revised form 22 June 2012 Additionally, high performance liquid chromatography linked to UV spectroscopy was employed to
Accepted 22 June 2012
monitor the encapsulation yield of curcumin. These techniques indicate that the effectiveness of the
Available online 2 July 2012
encapsulation is higher in the case of ␥-cyclodextrin (␥-CD) likely due to the better fit of the polyphenol
size with the dimensions of the ␥-CD cavity. Raman spectra provided specific structural information from
Keywords:
the ligand which indicates that the encapsulation takes place at the level of the aromatic rings, through
Curcumin
Cyclodextrins
H-bonds, and that a tautomerization from the planar keto enol form to the non-planar diketo form of
Raman spectroscopy curcumin also occurs. These changes may lead to an increase of the chemical stability, the bioavailability
Encapsulation and the biological activity of curcumin.
Isomerization © 2012 Elsevier B.V. All rights reserved.

1. Introduction
Tumeric (Curcuma longa L.) is one of the most popular spices con-
taining natural antioxidants. It is a plant native to tropical South
Asia and it is commonly used in food industry as a curry spice,
food dye (E-100) and preservative. The main active compound
of tumeric is the polyphenol curcumin (1,7-bis(4-hydroxy-3-
methoxyphenyl)-1,6-heptadiene-3,5-dione). Curcumin (Fig. 1A) is
isolated from the dry rhizomes [1] and it has a powerful antioxidant
and HIV antiproteases activity, inflammatory properties and can-
cer preventive properties [2,3]. Despite these advantages, the use
of curcumin as a potent pharmacological agent is hindered by its
poor vascular and oral bioavailability caused by different reasons:
low solubility, degradation in water, photodegradation, high rate
of metabolism and rapid elimination from the body [4,5].
Encapsulation has been recently used to protect unstable com-
pounds in different areas, such as foods, agriculture and pharmacy.
Specifically, the encapsulation of curcumin has been described in
the literature by using different materials. Among others, ␤-casein
micelle [6], poly (lactic-co-glycolic acid) nanoparticles [7], methoxy
poly(ethylene glycol-␤-aromatic anhydride) micelles [8] and 1,3-
␤-glucan isolated from Vietnam Medicinal Mushroom Hericium
erinaceum [9] are some of the most used materials to encapsulate
∗ Corresponding author. Tel.: +34 915616800; fax: +34 915645557.
E-mail addresses: s.sanchez.cortes@csic.es, imts158@iem.cfmac.csic.es
(S. Sanchez-Cortes).
curcumin. In this context, cyclodextrins (CDs, Fig. 1B) for molec-
ular encapsulation offer advantages over the above mentioned
materials. They possess macrocycles that present a torus-shaped
structure with an adaptable hydrophobic cavity in which lipophilic
guest molecules can be hosted [10]. In addition, CDs are non-
toxic, not hygroscopic and stable until 100 ◦ C. The capability of
CDs to form inclusion complexes with a wide variety of hydropho-
bic guest molecules has been tested. This capability is provided
by the distribution of hydrophilic and hydrophobic groups in the
ring. Hydrophilic OH groups are mainly placed in the rims of the
truncated cone which represents the cyclodextrin, and they are
the responsible of the solubility of these molecules in water. On
the contrary, the inside of the cavity is hydrophobic because it is
occupied by C3 H, C5 H, C6 H2 and the ether-like C4 O C bond
(Fig. 1B) [11].
Actually, reports on the encapsulation in CDs of lycopene, aspar-
tame and neotame [12] can be found in the literature. However,
publications about the encapsulation of curcumin with CD are
scarce and mainly focused on the use of ␤-CD [13]. The employ-
ment of ␥-CD as an encapsulating agent has only been described to
study the fluorescence enhancement [14].
FT-Raman spectroscopy is an interesting technique for the study
of polyphenols and carotenes in general [15–17], and curcumin in
particular [18], because of its high sensitivity to this type of com-
pounds without the interference from the fluorescent emission.
In general, Raman spectroscopy has demonstrated to be a pow-
erful technique in the study of encapsulation phenomena since
it can provide valuable and specific molecular information about

0924-2031/$ – see front matter © 2012 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.vibspec.2012.06.008
 E. López-Tobar et al. / Vibrational Spectroscopy 62 (2012) 292–298
Fig. 1. (A) Chemical structure of keto-enol and diketo isomers of curcumin. (B) Cyclic
structure of ␤- and ␥-cyclodextrin.
the interaction mechanism taking place in molecular recognition
problems [19–21]. In previous works, we have employed Raman
spectroscopy in the structural analysis of inclusion complexes
formed by lycopene and ␤-CD [19] providing remarkable informa-
tion about the structural modification of the latter carotenoid. In
particular we deduced a trans-to-cis structural change of lycopene
in this previous work. CDs are normally employed in the encapsu-
lation of water insoluble molecules to increase their bioavailability
[22]. In a recent work it was demonstrated that CDs can stabilize the
structure of curcumin preserving this ligand from the degradation
in aqueous media [23].
The objective of this work was to study the encapsulation into
CD of curcumin by UV–visible absorption spectroscopy and Raman.
The latter is an ideal technique to follow the possible structural
changes induced by the encapsulation with CDs. This is the first
time that Raman spectroscopy is applied to the study of the encap-
sulation of curcumin with CD. To that aim, ␤- and ␥-CD were
employed as encapsulating agents in order to check the effect of
the size of the inner cavity on the complexation effectiveness whilst
Raman spectroscopy was used as a technique to study the stability
of the encapsulated complex.
2. Experimental
2.1. Materials
Curcurmin and emodin (1,3,8-trihydroxi-6-
methylantraquinone-6-methyl-1,3,8-trihydroxiantraquinone)
were purchased from Sigma–Aldrich (Madrid, Spain). ␤-CD and
␥-CD, used without any further purification, were purchased from
Fluka (Madrid, Spain). Methanol (99.9% purity) was acquired from
Lab-Scan Ltd. (Dublin, Ireland) and ethanol (99.8% purity) from
293
Prolabo (Fontenay-sous-Bois, France). Water was obtained from a
Milli-Q purification system (Millipore, Milford, MA, USA).
2.2. Encapsulation of curcumin
The encapsulation procedure was carried out using ␤-CD and
␥-CDs and it was based on that described elsewhere [19,24,25]. Dif-
ferent ␤-CD/curcumin ratios (1/1, 2/1 and 4/1) and ␥-CD/curcumin
ratios (1/1, 2/1, 4/1 and 8/1) were tested. First, each CD was dis-
solved in de-ionized water at 40–45 ◦ C with stirring. The solution
was cold and then curcumin diluted in ethanol was added. The
CD–curcumin reaction was carried out for 24 h at 25 ◦ C, under
nitrogen atmosphere and using stirring. After that, most of the
water was removed by freeze-drying and the resulting precipitate
was washed with ethanol to eliminate all the non-encapsulated
curcumin. Finally, the washed precipitate containing the encapsu-
lated CD with curcumin together with the remaining pure CD was
removed by centrifugation (7600 rpm, 5 min) and dried under high
vacuum. Finally, the encapsulated complex was kept at −8 ◦ C until
its analysis. The encapsulation yield (%) was estimated from the
comparison of the initial weight of curcumin used and the amount
of non-encapsulated curcumin studied determined by HPLC-UV.
These measurements were performed on a Hewlett Packard liquid
chromatograph 1050. The HPLC system was composed of an injec-
tion valve (Rheodyne, model 7125) having a 20 ␮L sample loop and
an ultraviolet (UV) detector operated at 425 nm. The elution of cur-
cumin and emodin, employed as standard, was accomplished on a
C18 column (ACE 5 C18, 4.6 mm × 250 mm, 5 ␮m particle) at 25 ◦ C.
Methanol/water (with 0.1% formic acid) was used as the mobile
phase. Data acquisition was performed by using Agilent ChemSta-
tion (Rev. A 10.02, 1757). The identification of curcumin during
the stability study was made by matching the UV absorption spec-
tra with that of the standard. The quantification of the remaining
curcumin was accomplished at a flow rate of 0.8 mL/min and by
applying a linear gradient within 15 min from an initial eluent com-
position (90/10, methanol/water with 0.1% acetic acid; v/v) to a final
composition (100% methanol), which was kept for 10 min.
2.3. Raman measurements
FT Raman spectra were obtained by using a RFS 100/S Brucker
spectrophotometer equipped with a liquid-nitrogen cooled Ge
detector. The 1064 nm line provided by a Nd:YAG laser was used as
excitation line. The laser power at the sample was 100 mW in the
solid sample and the resolution was set to 4 cm−1 and 180◦ geom-
etry was employed. The Raman spectra displayed in this work are
the average of 200 accumulations.
2.4. UV–vis absorption measurements
UV–vis absorption spectra were recorded with a Shimadzu UV-
2100 by using quartz cells with an optical path of 1 cm. Samples
for the UV–vis spectroscopy in H2 O/EtOH (50%) were prepared by
adding an aliquot of a mix of absolute EtOH and Milli Q water 50%
(1.66 mg/mL). All aliquots were sonicated for several seconds for
the entire dissolution and the final volume was adjusted to 3 mL.
3. Results and discussion
3.1. Absorption UV–visible spectra
The most intense absorption band of curcumin is seen at 432 nm,
which can be assigned to a combination of ␲ → ␲* and n → ␲*
transitions. The band at 265 nm corresponds to a ␲ → ␲* tran-
sition [26–28]. Fig. 2A and B displays the absorption spectra of
294 E. López-Tobar et al. / Vibrational Spectroscopy 62 (2012) 292–298
Fig. 2. (A) Absorption spectra of 1/1, 2/1 and 4/1 ␤-CD/curcumin complexes in
ethanol/water 50% (v/v); (B) absorption spectra of 1/1, 2/1 and 4/1 ␥-CD/curcumin
complexes in ethanol/water 50% (v/v); (C) difference spectrum complex-curcumin
in the case of 4/1 ␤-CD/curcumin and 4/1 ␥-CD/curcumin complexes.
curcumin at different concentrations of ␤-CD and ␥-CD, respec-
tively. As can be seen in Fig. 2B, ␥-CD has a significant effect on the
absorbance spectrum of curcumin in terms of peak position and
width. This confirms the encapsulation of curcumin into the CD. As
the ␥-CD/curcumin ratio increases the curcumin absorption spec-
trum undergoes a continual blue-shift in good accordance, while
a shoulder appearing at 360–370 nm is progressively enhanced.
These changes are attributed to the interaction between curcumin
and the hydrophobic cavity of ␥-CD.
In contrast to the above results, the absorption UV–visible spec-
tra of curcumin show less evident changes in the presence of ␤-CD
(Fig. 2A). The band at 432 nm is slightly weakened in relation to
that at 265 nm, and a weak shoulder about 350 nm appears as the
CD concentration increases.
Difference spectra were obtained by subtracting the curcumin
spectra from the CD/curcumin ones (Fig. 2C) in order to stress the
effect of encapsulation on the electronic spectra. These difference
spectra revealed the existence of a band centred at 369 nm in the
case of ␥-CD/curcumin complex, which is shown at lower wave-
length (353 nm) in the case of ␤-CD/curcumin complex. This band
is a consequence of the interaction of CD with curcumin. Two are
the conclusions from this study: (a) the encapsulation yield of cur-
cumin by ␥-CD is much larger, due to the high intensity of the 369
maximum; and (b) the effect of ␤-CD on the curcumin structure is
higher, due to the smaller size of the cavity (Fig. 1B).
Fig. 3. FT-Raman spectrum of curcumin in the solid state and assignments of the
main bands. Excitation at exc = 1064 nm.
This difference band becomes sharper as the ␤-CD/curcumin
ratio increases. This indicates that a higher thermodynamic bar-
rier exists in the encapsulation of curcumin by ␤-CD in comparison
to ␥-CD because of the smaller cavity of ␤-CD. Thus, a larger
␤-CD/curcumin ratio would be necessary to get a most efficient
encapsulation.
3.2. Raman spectra
The FT-Raman spectrum of solid curcumin is shown in Fig. 3. The
assignment of the main Raman bands was performed on the basis
of already published vibrational studies made for this molecule
[29–31].
Two intense signals at 1627 cm−1 and 1600 cm−1 are attributed
to ␯(C C)/␯(C O) of interring chain and to the aromatic ␯(C C),
respectively [28–30]. A group of three well defined weak bands
appearing in the 1530–1440 cm−1 region are attributed to methyl
groups coupled to ring motions.
The medium band at 1431 cm−1 was assigned by Kolev et al. [29]
to ␦CCC and ␦COH of aromatic rings, while Vu Thi et al. [30] assigned
it to deformation vibration of the methyl groups associated to
vibrations of rings. The band at 1318 cm−1 has been attributed to
␦(C C H) motions of the inter-ring chain, while that at 1249 cm−1
can be attributed to in-plane ␦(C H) vibrations of the aromatic
rings, associated to ␯(C O) of the ether groups linked to these rings
[32]. The strong band at 1184 cm−1 was assigned to ring skele-
tal deformations coupled to the phenolic ␯(C O), while the band
at 1150 cm−1 is rather attributed to ␦(C OH) motions coupled to
␦(C C H) in the enolic group of the inter-ring chain. The band at
962 cm−1 can be attributed to ␯(C OH) of enol group in the inter-
ring chain coupled to ␦(C OH) [29,30]. Finally, the weaker bands
found at 575 cm−1 and 461 cm−1 are due to skeletal vibrations
␦(CCC) of the rings and the inter-ring chain [30]. From the latter
Raman analysis, one can deduced that in solid state curcumin exist
under the keto-enol tautomeric form in solid state as stated as well
by other authors [29,30]. This is also corroborated by the position
of the maximum of UV–visible adsorption spectra at 430 nm [26],
corresponding to absorption of the latter tautomer. In addition,
the keto-enol tautomer is strongly favoured by an intermolecular
H-bonding [33].
Figs. 4 and 5 show the Raman spectra of ␤-CD/curcumin and
␥-CD/curcumin complexes, respectively. No bands of cyclodextrin
are observed by exciting at 1064 nm. This is attributed to the fact
 E. López-Tobar et al. / Vibrational Spectroscopy 62 (2012) 292–298 295

Fig. 4. FT-Raman spectra of solid curcumin (a) and ␤-CD/curcumin complexes at

the following ratios: 1/1 (b); 2/1 (c); 4/1 (d). Excitation at exc = 1064 nm. Fig. 5. FT-Raman spectra of solid curcumin (a) and ␥-CD/curcumin complexes at
the following ratios: 1/1 (b); 2/1 (c); 4/1 (d). Excitation at exc = 1064 nm.
that cyclodextrins have a negligible Raman activity at this excita-
tion wavelength laser line, as reported by other authors, because of
the low polarizability of the carbohydrate with respect the apolar Nevertheless, this interaction also has some influence on the bands
unsaturated conjugated organic guest [21]. As can be seen, cur- of the inter-ring chain, which also suggests the occurrence of a
cumin spectrum was not significantly affected by the presence of deeper structural change, affecting in particular the electron con-
␤-CD at any amount of the host (Fig. 4). This suggests that the dye is jugation throughout the molecule and the possible isomerization
poorly encapsulated by ␤-CD as also deduced from the UV–visible of the inter-ring chain.
absorption spectra [14,34].
In contrast, ␥-CD/curcumin complex (Fig. 5) displays impor-
tant changes in several regions of the Raman spectrum. Fig. 6
shows the difference spectrum obtained by subtracting the cur-
cumin one to that of 4/1 ␥-CD/curcumin complex. These difference
01 spectra reveal that the ␯(C C)/␯(C O) region is highly affected by
the encapsulation since the 1627/1601 cm−1 bands are shifted to
Rupanjali Prasad
1632/1592 cm−1 . Several bands associated to the phenolic rings
underwent slight spectral changes, namely that at 1249 cm−1 ,
which undergo a shift downwards. Nevertheless, the most impor-
tant changes occur in vibrations localized in the inter-ring chain.
This is the case of that at 1150 cm−1 undergoes a dramatic decrease,
and the ␦(C C H) band at 1317 cm−1 of the inter-ring chain, which
shifts to 1307 cm−1 and is strongly intensified. Finally, the band
associated to ␯(C OH) of the enolic group existing in the inter-ring
chain appearing at 962 cm−1 chain decreases.
The above changes indicate a large effect of complexation of cur-
Fig. 6. Comparison between the FT-Raman spectra of solid curcumin (top) and
cumin with ␥-CD on the phenolic groups, thus meaning that the the difference spectrum between ␥-CD/curcumin complex at the 4/1 ratio and the
two aromatic rings are directly interacting with the cyclodextrin. spectrum of curcumin (bottom), showing the most interesting regions.
 296 E. López-Tobar et al. / Vibrational Spectroscopy 62 (2012) 292–298
02
Rupanjali Prasad
Fig. 7. Detail of the FT-Raman spectra in the ␯(C O)/␯(C C) region of curcumin and
its complexes with ␤-CD (A) and ␥-CD (B) at the ratios indicated on the figures.
In order to monitor the structural changes of curcumin by effect
of the encapsulation, Fig. 7 displays in detail the behaviour of bands
falling in the ␯(C C)/␯(C O) region for curcumin complexes with
each cyclodextrin. As can be seen, in the case of ␤-CD (Fig. 7A), these
bands are not significantly modified regarding the spectrum of free
curcumin, while in the case of ␥-CD/curcumin complex (Fig. 7B), a
progressive enhancement and broadening of the band at 1632 cm−1
was seen as the rate of ␥-CD increases. In contrast, the band at
1600 cm−1 , corresponding to ␯(C C) of aromatic rings, also under-
gone a broadening and a shift towards lower wavenumbers, thus
appearing a difference band at 1592 cm−1 . The changes observed
3-4 in ␯(C C) bands corresponding to aromatic rings suggest that the
2 notes:
phenyl groups are directly involved in the interaction with ␥-CD.
However, a number of changes involving the aliphatic chain were
also observed.
On the other hand, the increase of the ␯(C O) band at 1627 cm−1
and the weakening of the bands associated to the enol group in
the inter-ring chain (bands at 1150 and 963 cm−1 ) strongly sug-
gest that an isomerization of curcumin occurs upon formation of
the complex from the keto-enol to the diketo isomers (Fig. 1A).
05 This isomerization leads to deep changes in the electronic conju-
gation degree of curcumin, not only at the level of the interring
Rupanjali Prasad
chain but also in the aromatic rings, which accounts for some of the
wavenumber shifts observed in both the phenolic and the inter-ring
aliphatic moiety bands. In addition, the curcumin isomerization
implies the conformational change from the planar structure in the
keto-enol isomer to the non planar one in the diketo isomer [26].
The diketo isomer is energetically less stable and less polar than the
enol-keto one [26], although the diketo isomer may be stabilized
inside the hydrophobic cavity of ␥-CD.
The formation of the diketo isomer implies the breaking
of the electronic delocalization along the entire molecule that
accounts for the appearance of the absorption maximum at 369 nm
attributed to the diketo structure [26,35].
In order to see in more detail the changes induced by ␥-CD
on the region concerning the double bonds of curcumin we have
performed a deconvolution study of the aromatic ␯(C C) band at
different ␥-CD/curcumin ratios (Fig. 8). The band at 1600 cm−1
can be fitted into two distinct components centred at 1600 and
1592 cm−1 whose areas change as the cyclodextrin amount is
varied (Fig. 8A). In fact, the component appearing at 1592 cm−1
grows as the ␥-CD/curcumin ratio is increased from 1/1 to 4/1 as
a consequence of the bigger interaction degree with the phenyl
rings with the cyclodextrin, and also due to the electron reso-
nance reduction caused by the formation of the diketo isomer. In
Fig. 8B the A1592 /A1600 area ratio is plotted at different ratios for
␤-CD/curcumin and ␥-CD/curcumin complexes. As can be seen a
progressive reduction of the slope in the case of the latter com-
plex, indicating that at a 4/1 ratio the encapsulation yield is almost
maximum. Finally, Fig. 8C displays the I963 /I1630 ratio, which can
be considered as a spectroscopic marker for the keto-enol to diketo
isomerization for the different complexes. While this ratio remains
practically unaltered for ␤-CD/curcumin complexes, in the case
of ␥-CD/curcumin ones a remarkable reduction of this parameter
points out the curcumin isomerization taking place inside the ␥-CD
cavity.
From the UV–vis absorption and the FT-Raman spectra shown
above, we deduced a higher effectiveness of ␥-CD compared to
␤-CD in the encapsulation of curcumin. This effect is obviously
connected to the higher size of the inner cavity of the first cav-
itand. Baglole et al. also reported an isomerization of curcumin
in the presence of ␥-CD and a possible change in the electronic
properties of the polyphenol upon encapsulation with ␥-CD [14].
There are also recent works reporting an isomerization of lig-
ands inside the cavity of CDs due to the stabilization of one of
the molecular isomers observed by UV–visible and Raman spec-
troscopy [36,37]. In addition, the higher solubility reported for
curcumin in the presence of hyroxypropyl ␥-CD [14,33] can be
also associated to the higher effectivity of a cyclodextrin compris-
ing eight glucose rings. The inner cavity of ␤-CD is in the range
6.0–6.5 Å, while that of ␥-CD is 7.5–8.3 Å [11]. Thus, the inner cavity
of the latter cavitand fits very well the dimensions of the ter-
minal aromatic moiety of curcumin as also reported Singh et al.
for curcuminoids bearing a side molecular group [33], since the
approximate width of curcumin in its terminal aromatic part is ca
7.2 Å. The interaction mechanism of curcumin with the cyclodex-
trin can be of two types: van der Waals and/or H-bonding. There
are two clear spectroscopic evidences that suggest the latter as
the main driving force for the encapsulation: (a) the blue shift
of the absorption maximum of curcumin in ␥-CD/curcumin com-
plexes (Fig. 2B), which is a characteristic behaviour of curcumin in
the presence of polar environments [14,26]; and (b) the effect of
the complexation on the Raman bands attributed to the phenolic
OH groups, such as that appearing at 1250 cm−1 , which under-
goes a dramatic weakening likely due to the H-bonding of these
groups with the OH ones of the cyclodextrin. In addition, the latter
H-bonds could explain why ␥-CD is more efficient in the encapsu-
lation of curcumin. At this scenario, the isomerization of curcumin
to the diketo isomer can be considered as a consequence of the
interaction with the cyclodextrin in order to better fit the cavity
of the binder. In fact, the lower delocalization of electrons in the
diketo isomer could favour the H-bonding of phenol groups with
CD.
 E. López-Tobar et al. / Vibrational Spectroscopy 62 (2012) 292–298
Fig. 8. (A) Deconvolution of the 1600 cm−1 band of curcumin at different ␥-
CD/curcumin ratios. (B) Relationship between the areas of the 1592 and 1600 cm−1
bands at different ratios. (C) Relationship between bands at 962 and 1630 cm−1 at
different ratios.
The encapsulation yield (%) was estimated from the compar-
ison of the initial weight of curcumin used and the amount of
non-encapsulated curcumin studied determined by HPLC-UV as is
detailed in Section 2. The encapsulation yields obtained were 20, 30
and 18.8%, for the ␥-CD encapsulation 2/1, 4/1 and 8/1, respectively.
As demonstrated above, the most important spectral changes were
observed in the case of the 4/1 ␥-CD/curcumin complex. For this
complex an encapsulation yield of 30% was attained. Higher ratios
did not lead to an increase of yield, nor Raman changes. Thus, the
4/1 in ␥-CD/curcumin ratio can be considered as the optimal one
297
Fig. 9. Scheme of the 2/1 ␥-CD/curcumin complex.
for the encapsulation of curcumin using this CD. However, due to
the low price of ␤-CD, ratios of 100/1 (␤-CD/curcumin) were also
tested but did not show good results by HPLC neither by Raman.
As concerns the stoichiometry of the complexation, we suggest
the formation of a 2/1 (␥-CD/curcumin) complex model, as already
reported by other authors [14], in 4/1 complexes. Under these con-
ditions two phenyl rings of curcumin are encapsulated by the two
cyclodextrin cavities as depicted in Fig. 9. This 2/1 stoichiometry is
supported by the fact that the ␯(C C) region did not undergo any
more change upon increasing the ratio of the complex above 4/1.
The keto-enol to diketo isomerization of curcumin in the cavity
of CD has a crucial importance, as the chemical structure of cur-
cumin plays a pivotal role in its biological activity [5]. Curcumin is
a relatively unstable molecule which can undergo a degradation in
water which render trans-6-(4 -hydroxy-3 -methoxyphenyl)-2,4-
dioxo-5-hexenal as the major degradation product, while vanillic
and ferulic acid and feruloyl methane are minor degradation
products [38]. This degradation process seems to start from a nucle-
ophilic attack of OH− ions on the carbonyl atom of the keto-enol
isomer [39]. Thus, the isomerization of curcumin in the CD com-
plex could increase its stability and solubility leading to a higher
bioavailability. In addition, the diketo curcumin form was postu-
lated to be the biologically active form of the compound concerning
the antioxidant activity, due to the H-atom transfer reactions from
the CH2 moiety existing in the diketo isomer [40,41]. Con-
sequently, the complexation with ␥-CD could enhance both the
bioavailability and the biological activity of curcumin. Further stud-
ies concerning the influence of CDs on the stability of curcumin are
still in progress in our laboratory.
4. Conclusions
Raman spectroscopy and UV–vis has provided complementary
information regarding the encapsulation of curcumin with ␤-CD
and ␥-CD. These techniques indicate that the effectiveness of the
encapsulation is higher in the case of ␥-CD due to the better fit of
the polyphenol size with the dimensions of the ␥-CD cavity. The
interaction of curcumin seems to occur through the inclusion of
the two phenolic moieties inside the cavity leading to a 2/1 (␥-
CD/curcumin) stoichiometry and implies an interaction through
the formation of H-bond between phenolic OH and the OH groups
of the cyclodextrin. The encapsulation leads to the isomerization
of curcumin from the keto enol form to the diketo one, which
strongly affects the electron conjugation throughout the molecule.
298 E. López-Tobar et al. / Vibrational Spectroscopy 62 (2012) 292–298
Consequently, the encapsulation leads to an increase of the
bioavailability of curcumin due to the lower reactivity of the diketo
isomer. In addition, the formation of this isomer increases the
biological activity of the polyphenol, since the diketo isomer is con-
sidered to be the actual biologically active form of curcumin. The
spectroscopic markers and the analysis of the encapsulation yield
revealed that the optimal ratio for the encapsulation is 4/1 for the ␥-
CD/curcumin complex, since at this ratio the maximum structural
change is induced at the highest encapsulation yield.
Acknowledgements
The authors thank A.R Santiago Clérigo for her help in part
of the experimental work. This research was financial supported
by ANALISYC-II and MICROSERES II networks (CAM grants S-
2009/AGR1464 and S2009/TIC-1476), and also by the grants of the
Spanish Ministerio de Ciencia e Innovación NATURAGE (AGL2010-
1779) and POEMS (FIS2010-15405).
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Annotations

Encapsulation and isomerization of curcumin with cyclodextrins

characterized by electronic and vibrational spectroscopy
López-Tobar, E; Blanch, G P; Ruiz Del Castillo, M L; Sanchez-Cortes, S

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