Hindawi Publishing Corporation

International Journal of Polymer Science

Volume 2014, Article ID 340121, 8 pages
http://dx.doi.org/10.1155/2014/340121

Research Article
Preparation and Characterization of Starch Nanoparticles for
Controlled Release of Curcumin

Suk Fun Chin, Siti Nur Akmar Mohd Yazid, and Suh Cem Pang
Department of Chemistry, Faculty of Resource Science and Technology, Universiti Malaysia Sarawak, 94300 Kota Samarahan,
Sarawak, Malaysia

Correspondence should be addressed to Suk Fun Chin; sukfunchin@gmail.com

Received 21 June 2013; Accepted 28 October 2013; Published 29 January 2014

Academic Editor: Saad Khan

Copyright © 2014 Suk Fun Chin et al. This is an open access article distributed under the Creative Commons Attribution License,
which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

Curcumin was loaded onto starch nanoparticles by using in situ nanoprecipitation method and water-in-oil microemulsion system.
Curcumin loaded starch nanoparticles exhibited enhanced solubility in aqueous solution as compared to free curcumin. Effects of
formulation parameters such as types of reaction medium, types of surfactant, surfactant concentrations, oil/ethanol ratios, loading
time, and initial curcumin concentration were found to affect the particle size and loading efficiency (LF) of the curcumin loaded
starch nanoparticles. Under optimum conditions, curcumin loaded starch nanoparticles with mean particles size of 87 nm and
maximum loading efficiency of 78% were achieved. Curcumin was observed to release out from starch nanoparticles in a sustained
way under physiological pH over a period of 10 days.

1. Introduction have been extensively studied as controlled release nanocar-

riers. For instance, propyl starch nanoparticles loaded with
Curcumin is a natural polyphenol found in the rhizomes of different types of drug (flufenamic acid, testosterone, and
turmeric (Curcuma longa) and exhibited antioxidant, anti- caffeine) showed enhanced effectiveness upon permeation
inflammatory, antiproliferative, anti-invasive, and antiangio- studies on human skin [15]. Dialdehyde starch nanoparticles
genic activities [1, 2]. More recently, curcumin has shown (DASNPs) conjugated with 5-fluorouracil (5-Fu) were found
to have antitumor effects in many cancer cell lines [3, 4] to have enhanced breast cancer cell (MF-7) inhibition in vitro
and the clinical effects of curcumin are being studied in
as compared to free 5-Fu [16].
human clinical trials and animal models on various condi-
tions and numerous myeloma [5–7]. Despite these promising Previously, our research groups have successfully pre-
biomedical properties, free curcumin molecules suffered pared starch nanoparticles by a nanoprecipitation method
from low water solubility, which in turn have resulted in poor [17]. In this present work, we report loading of curcumin in
bioavailability and clinical efficacy [8]. Hence, researchers situ onto native sago starch to form curcumin loaded starch
have attempted to enhance water solubility and bioavailabil- nanoparticles by precipitation in water-in-oil microemulsion.
ity of curcumin by loading of curcumin in biodegradable Water-in-oil microemulsion technique offers many advan-
polymeric nanoparticles. For instance, curcumin loaded tages as it requires no extreme pressure and no special or
poly(lactic-coglycolic acid) PLGA nanospheres were formu- expensive equipment [18]. Besides, preparation of nanopar-
lated for prostate cancer therapy [9]. ticles in water-in-oil microemulsion system affords nanopar-
Starch is a well-known, versatile, and inexpensive ticles with monodispersed particle size distribution [19, 20].
polysaccharide which has received great attention in drug The potential application of these native starch nanoparticles
delivery applications as they are hydrophilic, biodegradable, as controlled release agent for curcumin was studied. For-
and biocompatible with tissue and cells [10–14]. Starch mulation parameters that affected the loading efficiency and
nanoparticles with particle sizes in the range of 10–1000 nm release profiles of curcumin were also investigated.
2 International Journal of Polymer Science

2. Experimental curcumin onto starch nanoparticles was calculated based on

the following equation [21, 22]:
2.1. Materials. All chemicals were of reagent grade and
were used without further purification. Ultrapure water Loading efficiency (%)
(∼18.2 MΩ, 25∘ C) was obtained from a Water Purifying Sys-
[curcumin]tot − [curcumin]free (1)
tem (ELGA, Model Ultra Genetic); cyclohexane and absolute = × 100.
ethanol were obtained from HmbG Chemicals (Germany); [curcumin]tot
oleic acid was obtained from Ajax Chemicals (Australia)
while curcumin powder, polysorbate 80 (Tween 80) surfac- Loading capacity of curcumin onto starch nanoparticles was
tant, and sorbitan monostearate (Span 60) surfactant were calculated based on the following equation:
obtained from Merck (Germany). Native sago starch powder
Loading capacity (mg/mg)
and sunflower oil were obtained from a local grocery store.
[curcumin]tot − [curcumin]free (2)
= × 100,
2.2. Loading of Curcumin onto Starch Nanoparticles. The total weight of nanoparticles
solvent phases essentially consist of absolute ethanol as an where [curcumin]tot is the concentration of curcumin added
organic solvent, oil (oleic acid, sunflower oil, and cyclohex- and [curcumin]free is the concentration of curcumin present
ane) as the continuous phase, and surfactant (Tween 80 or in the supernatant after centrifugation.
Span 60) as an emulsifying agent. About 2.5 × 10−4 M of cur-
cumin was dissolved in 15 mL of ethanol solution containing
of 0.8 × 10−3 M of surfactant, followed by the addition of 5 mL 2.5. Curcumin Release Studies. About 50 mg of curcumin
of oil and the mixture was stirred continuously for 1 h. After loaded starch nanoparticles was placed in 15 mL of buffer
1 h, 1 mL of starch solution (1% w/v) was added drop-wise to solution at pH 7.4 at 37 ± 0.5∘ C [23]. At predetermined time
the mixture and stirred for an addition of 1 h. Curcumin was intervals, the solution was centrifuged, and the supernatant
loaded in situ onto starch nanoparticles as the starch nanopar- was removed and replaced with the same volume of buffer
ticles formed during the precipitation process. The curcumin solution. The amount of curcumin released in the supernatant
loaded starch nanoparticles were collected by centrifugation was determined using a UV/Vis spectrophotometer. The
and the samples were washed several times with ethanol to concentration of curcumin released was calculated with ref-
remove any excess of curcumin that adhered at the surface of erence to the regression equation generated from constructed
the starch nanoparticles. For loading of curcumin onto starch calibration curve of curcumin in PBS (linear plot with slope of
nanoparticles in ethanolic solution without microemulsion 0.006). The percentage of curcumin released at a specific time
system, the same synthesis conditions were employed, except was determined based on the following equation [24, 25]:
that the loading was carried out in the ethanol solution only. [curcumin]rel
Release of curcumin (%) = × 100, (3)
[curcumin]load
2.3. Characterization of Curcumin Loaded Starch Nanoparti-
where [curcumin]rel is the concentration of curcumin
cles. TEM images were obtained using a Transmission Elec-
released and [curcumin]load is the concentration of curcumin
tron Microscope (TEM) (JEOL-2000EX). Starch nanoparti-
loaded into the starch nanoparticles.
cles were deposited onto a formvar-coated copper grid and
dried in a desiccator at room temperature. SEM images
were recorded using a scanning electron microscope (SEM) 2.6. Swelling Studies. The swelling behaviour of the cur-
(JEOL-SM 6390 LA) operating between 10–15 kV with a cumin loaded starch nanoparticle was studied by immersed
filament current of about 60–70 𝜇A. The samples were preweighted dried curcumin loaded starch nanoparticles in
dropped on stainless steel plates, dried at room temperature, 10 mL of buffer solution (pH 7.4) at 37 ± 0.5∘ C. The weights of
and coated with a layer of platinum using JEOL/JFC-1600 the swollen starch nanoparticles were determined at various
Auto Fine Coater. The UV absorbances of the samples were time intervals. The swelling ratio was calculated based on the
measured using a UV/Vis spectrophotometer (Jasco V-630). following equation [26]:
The fluorescence image of curcumin loaded starch nanopar-
ticles was obtained using an Olympus FV300 confocal laser (𝑊𝑤 − 𝑊𝑑 )
Swelling ratio (SR) = , (4)
scanning microscope. 𝑊𝑑
where 𝑊𝑤 and 𝑊𝑑 are the wet and dry weight of the starch
2.4. Loading Efficiency of Curcumin. The curcumin loaded nanoparticles.
starch nanoparticles were separated from the reaction
medium by centrifugation and the UV absorbance of free 3. Results and Discussion
curcumin was measured at a wavelength of 422 nm. The
concentration of curcumin was calculated with reference to a 3.1. Loading of Curcumin into Starch Nanoparticles. Fig-
regression equation (linear plot with slope of 0.023) obtained ure 1(a) shows that the curcumin powder was insoluble in
from constructed calibration curve of curcumin in absolute aqueous media and observed floating on the aqueous media.
ethanol solution. The percentage of loading efficiency of In contrast, Figure 1(b) shows that curcumin loaded starch
International Journal of Polymer Science 3

(a) (b)

Figure 1: Photographic images of (a) free curcumin which were poorly soluble in aqueous media and macroscopic flakes can be seen floating
in the bottle, and (b) curcumin loaded starch nanoparticles were fully dispersible in aqueous media.

the visible green spectrum whereas starch nanoparticles

were transparent [29]. The loading of curcumin onto starch
nanoparticles occurred in situ while the starch nanoparticles
were precipitated at the interface of reaction medium with
curcumin being encapsulated or adsorbed at the same time
[30]. In order to further confirm the loading of curcumin
onto starch nanoparticles, the UV absorbance of curcumin
loaded starch nanoparticles and curcumin solution was
measured.
As shown in Figure 3(b), an aqueous solution of curcumin
loaded starch nanoparticles showed an absorbance peak at
425.5 nm (Figure 3(ii)) which was observed to be very close
80 kV X300000
to the characteristic absorbance peak of curcumin at 422 nm
(Figure 3(i)). The red shift in the UV absorbance peak of cur-
Figure 2: TEM image of curcumin loaded starch nanoparticles. cumin loaded starch nanoparticles was probably attributed to
the formation of intermolecular hydrogen bondings between
curcumin and starch nanoparticles [31, 32].
nanoparticles were completely dispersed in aqueous media
and rendered curcumin in good dispersibility in aqueous 3.2. Optimization of Loading Efficiency of Curcumin
media. The loading of curcumin onto the starch nanoparticles
has increased the solubility of curcumin in the aqueous media 3.2.1. Effect of Reaction Medium. The effects of reaction
as the starch nanoparticles were very hydrophilic in nature medium on the loading efficiency of curcumin onto starch
with small in size and large surface area. It allows for a greater nanoparticles were studied by conducting out the loading
interaction with the aqueous solvent and thus resulted in the process in four different reaction media, namely, ethanolic
increase of curcumin solubility in aqueous media [27, 28]. solution, cyclohexane/ethanol microemulsion solution, sun-
flower oil/ethanol microemulsion solution, and oleic acid/
Figure 2 shows TEM micrograph of curcumin-loaded ethanol microemulsion solution. Figure 4 shows that cur-
starch nanoparticles. Curcumin loaded starch nanoparticles cumin loading efficiency of 78% and 48% was achieved in
were observed to be spherical in shape with mean particle oleic acid/ethanol microemulsion and sunflower oil/ethanol
sizes within the range of 50–80 nm. microemulsion, respectively. However, low loading efficiency
Figure 3(a) shows the confocal microscopy image was obtained in the cyclohexane/ethanol microemulsion
of curcumin-loaded starch nanoparticles. The curcumin- (17%) and ethanolic solution (13%) reaction media.
loaded starch nanoparticles were observed to be fluorescent The microemulsion reaction media have been shown
which confirmed that curcumin had been loaded onto to enhance the loading efficiency of curcumin onto starch
starch nanoparticles. Curcumin was naturally fluorescent in nanoparticles due to the presence of lipophilic domains of the
4 International Journal of Polymer Science

2
(i)

1.5

Absorbance (Abs)
(ii)

0.5

0
350 400 450 500
Wavelength (nm)
(a) (b)

Figure 3: (a) Fluorescent image of curcumin loaded starch nanoparticles and (b) UV spectra of (i) curcumin solution and (ii) curcumin
loaded starch nanoparticles.

100 Table 1: Effects of reaction medium on mean particles diameter of

90 curcumin loaded starch nanoparticles.
Loading efficiency (%)

80
70 Reaction medium Mean particles diameter (nm)
60
50 Ethanolic 171
40 Cyclohexane/ethanol 137
30
Sunflower oil/ethanol 91
20
10 Oleic acid/ethanol 83
0
Reaction medium

Ethanolic solution 3.2.2. Effect of Types of Surfactant. The effect of types of

Cyclohexane/ethanol microemulsion surfactant on the loading efficiency of curcumin onto starch
Sunflower oil/ethanol microemulsion nanoparticles were studied by using two types of nonionic
Oleic acid/ethanol microemulsion surfactant, namely, sorbitan monostearate (Span 60) and
polysorbate 80 (Tween 80). The concentrations of both Span
Figure 4: Effect of reaction medium on loading efficiency of
curcumin onto starch nanoparticles. 60 and Tween 80 were 0.8 × 10−3 M. Nonionic surfactants
were used in this study because they are not ionic strength
dependence and they are nontoxic in nature [36, 37]. Tween
80 was shown to be the more effective surfactant as compared
microemulsion system which could dissolve the hydrophobic to Span 60 for loading of curcumin onto starch nanoparticles
curcumin molecules [33, 34]. Lipophilic domains (hydropho- as evidenced by the higher loading efficiency (78%) obtained
bic) were formed due to the presence of oil phase (cyclohex- when Tween 80 was used (Figure 5).
ane, sunflower oil, and oleic acid) in the system. Preparation
The higher loading efficiency could be due to a more
of curcumin loaded starch nanoparticles in oleic acid/ethanol
microemulsion reaction medium gave rise to the highest stable microemulsion formed in the presence of Tween 80 as
loading efficiency as the highly hydrophobic nature of oleic it was more soluble in water as compared to Span 60 [38, 39].
acid (C18 ) would enable curcumin molecules to be more The hydrophilic polyesters groups of Tween 80 rendered
soluble as compared to in sunflower oil and cyclohexane them water soluble, whereas Span 60 was comparatively
reaction medium. more hydrophobic and only soluble in hot water. Tween
Besides, the smallest mean particle sizes of curcumin- 80 surfactant formed strong hydrogen bonds between its
loaded starch nanoparticles were obtained when oleic oxyethylene groups and hydroxyl groups of water molecules
acid/ethanol microemulsion was used as the reaction resulting in a stable water-in-oil microemulsion system and
medium (Table 1). This was due to the deep penetration of enhanced curcumin loading efficiency. Besides, the longer
oleic acid into the surfactant with its longest alkyl chain hydrophobic chain length of Tween 80 (C64 ) as compared
length and smallest lipophilic domains and thus resulted in to Span 60 surfactant (C24 ) resulted in deeper penetration
the formation of the smallest mean particle sizes of starch of the alkyl chain of the Tween 80 in the oleic acid oil
nanoparticles by controlling the droplet size [35]. (hydrophobic domains), which promoted the formation of
International Journal of Polymer Science 5

100 Table 2: Effects of surfactant concentration on mean particles

90 diameter of curcumin loaded starch nanoparticles.
80
Loading efficiency (%)

70 Surfactant concentration Mean particles diameter (nm)

(×10−3 M)
60
50 0 137
40 0.4 109
30 0.8 83
20 1.5 101
10
2.3 119
0
Types of surfactants

Span 60 0.8 × 10−3 M of Tween 80 concentration, the loading efficiency

Tween 80 started to drop due to the presence of high concentration
of surfactant which could be caused by the highly viscous
Figure 5: Effect of types of surfactant on loading efficiency of
curcumin onto starch nanoparticles.
solution system. As a result, larger particles (Table 2) with
lower surface area and lower loading efficiency of curcumin
were produced [43, 44].
100 The mean particle diameter of curcumin loaded starch
nanoparticles prepared in oleic acid/ethanol microemulsion
Loading efficiency (%)

80 reaction medium with 0.8 × 10−3 M of Tween 80 appeared to

be smaller (83 nm) as compared to sample without surfactant
60
(137 nm) (Table 2). The presence of optimum concentration
40 of surfactant in oleic acid/ethanol microemulsion reduced
the surface tension between the starch nanoparticles, which
20 had prevented the coalescence between starch nanoparticles
in the microemulsion system and led to the formation of
0 smaller particle sizes. In contrast, curcumin loaded starch
0 0.4 0.8 1.5 2.3
nanoparticles of larger particle sizes were formed without the
Concentration of Tween 80 surfactant (×10−3 M) presence of surfactant in the microemulsion system [40, 45].
Figure 6: Effect of surfactant concentration on loading efficiency of
curcumin onto starch nanoparticles. 3.2.4. Effect of Oil/Ethanol Ratio. The effect of oil/ethanol
volume ratios (v/v) on the loading efficiency of curcumin
onto starch nanoparticles was shown in Figure 7. It was found
a stable microemulsion system and enhanced the loading that as the ratio of oleic acid (oil phase) to ethanol increased,
efficiency of curcumin onto starch nanoparticles. the loading efficiency of curcumin onto starch nanoparticles
decreased. This decrease was due to the destabilization of
oleic acid/ethanol microemulsion system by the excess oil
3.2.3. Effect of Surfactant Concentration. The concentration volume which resulted in the formation of an inhomoge-
of surfactant in microemulsion system was observed to affect neous and cloudy solution [46]. The optimum ratio of oil to
the loading efficiency of curcumin onto starch nanoparticles. ethanol (1 : 3) had resulted in the highest loading efficiency of
As could be seen in Figure 6, the highest loading efficiency of curcumin onto starch nanoparticles. This could be attributed
curcumin (78%) was achieved when Tween 80 was used as a to the role of ethanol as a cosurfactant which have reduced the
surfactant in the microemulsion system, whereas the lowest interfacial tension of oleic acid/ethanol microemulsion and
loading efficiency of curcumin (48%) was obtained when no improved the interactions of Tween 80 surfactant monolayer
any surfactant was used in the microemulsion system. This with water and oil at the interface of oleic acid/ethanol
was due to the lower interfacial tension between oleic acid microemulsion system and consequently the formation of a
(oil phase) and water in the presence of surfactant in the stable microemulsion [47].
microemulsion system. This led to the formation of a stable Besides, ethanol also served as a cosolvent to enhance
microemulsion system and facilitated a higher loading of the dissolution of curcumin and Tween 80 surfactant in
curcumin onto starch nanoparticles [40, 41]. the oleic acid/ethanol microemulsion reaction media and
The loading efficiency of curcumin onto starch nanopar- thus improved loading efficiency of curcumin onto starch
ticles was observed to increase when the concentrations nanoparticles. Similar observations have been reported by
of Tween 80 surfactant increased from 0 to 0.8 × 10−3 M. other researchers [48–50], where their studies showed that
This could be due to an increase in Tween 80 surfactant most of the surfactants alone could not produce stable
concentrations that had led to stabilizing the interfacial microemulsion system and the addition of solvent such as
tension of oleic acid/ethanol microemulsion system and alcohol was required to facilitate the formation of a stable
hence increased curcumin loading [42]. However, above microemulsion system.
6 International Journal of Polymer Science

Release of curcumin (%)

100 100 7
90 6
80

Swelling ratio
Loading efficiency (%)

80 70 5
60 4
50
60 40 3
30 2
20 1
40 10
0 0
0 24 48 72 96 120 144 168 192 216 240
20
Time (hours)
0 Curcumin released
Ratio of oil to ethanol Swelling ratio

1:3 Figure 9: Swelling ratio of starch nanoparticles and release profile

1:1 of curcumin from starch nanoparticles as a function of time.
3:1

Figure 7: Effect of oil to ethanol volume (oil : ethanol) on loading

efficiency of curcumin onto starch nanoparticles. are very hydrophilic with plenty of OH group starch nanopar-
ticles swelled due to absorption of the buffer solution and
2.5
curcumin in the swollen part of starch nanoparticles were
diffused out from the starch nanoparticles gradually. Starch
Loading capacity (mg/mg)

2 nanoparticles were observed to afford a sustained release of

curcumin without initial burst release of curcumin being
1.5 observed. In the first 24 h, starch nanoparticles showed a
swelling ratio of 3.66, and a total of about 13% curcumin
1 was released gradually out from starch nanoparticles. As
the swelling ratio of starch nanoparticles increased to 6.15,
0.5
about 93% of curcumin had been released within a period of
0 240 h.
0 1 2 3 4 5 6 7 8
Concentration of curcumin (mg/L)
4. Conclusions
Figure 8: Effect of curcumin concentration on loading capacity of
curcumin onto starch nanoparticles. In conclusion, we have successfully loaded curcumin onto
starch nanoparticles using a simple technique of nanoprecip-
itation in microemulsion system. The loading efficiency can
be optimized by the judicious choice of reaction medium,
3.2.5. Effect of Curcumin Concentration. Figure 8 demon-
types and concentrations of surfactant, oil/ethanol ratios,
strates that the loading capacity of curcumin onto starch
and curcumin concentration. Loading of curcumin by using
nanoparticles increased almost linearly from 0.2 to
2.5 × 10−5 M of curcumin in 1 : 3 (v/v) ratio of oleic acid
2.0 mg/mg as the concentration of curcumin was increased
to ethanol microemulsion system with 0.8 × 10−3 M of
from 0.5 to 2.5 mg/L. A maximum loading capacity of
Tween 80 surfactant and 2 h loading time was found to give
2.0 mg/mg was reached at the curcumin concentration of
rise to highest loading efficiency of curcumin onto starch
2.5 mg/L. This curcumin loading capacity was comparable
nanoparticles. Loading of curcumin onto starch nanoparti-
to other drug carriers such as graphene oxide [51], which
cles affords a controlled release mechanism with about 93%
indicated that starch nanoparticles were indeed promising
of the curcumin being released from starch nanoparticles
drug delivery carriers.
at the physiological pH 7.4 over 10 days. Overall, this study
suggested that starch nanoparticles have the potential to be
3.3. Curcumin Release Studies. The curcumin loaded starch used as controlled release nanocarriers for curcumin.
nanoparticles which were prepared in oleic acid/ethanol
microemulsion reaction medium in the presence of 0.8 ×
10−3 M Tween 80 surfactant were used for release studies. The Conflict of Interests
release studies were performed in buffer solution at pH 7.4 at
37 ± 0.5∘ C as a function of time with continuous stirring over The authors declare that there is no conflict of interests
10 days (240 h). Figure 9 shows the swelling behaviour of the regarding the publication of this paper.
starch nanoparticles and the release profile of curcumin from
starch nanoparticles. Acknowledgments
The release characteristics of curcumin from starch
nanoparticles were observed to be dependent on the swelling Financial support by Ministry of Higher Education (MOHE)
behaviour of the starch nanoparticles. Starch nanoparticles through Fundamental Research Grant Scheme (FRGS), Grant
International Journal of Polymer Science 7

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