J Nanopart Res (2012) 14:757
DOI 10.1007/s11051-012-0757-0
RESEARCH PAPER
Preparation of nanoparticles of poorly water-soluble
antioxidant curcumin by antisolvent precipitation methods
Mitali Kakran • Nanda Gopal Sahoo •
I-Lin Tan • Lin Li
Received: 8 July 2011 / Accepted: 12 January 2012 / Published online: 10 February 2012
Ó Springer Science+Business Media B.V. 2012
Abstract The objective of this study was to enhance
the solubility and dissolution rate of a poorly water-
soluble antioxidant, curcumin, by fabricating its
nanoparticles with two methods: antisolvent precipi-
tation with a syringe pump (APSP) and evaporative
precipitation of nanosuspension (EPN). For APSP,
process parameters like flow rate, stirring speed,
solvent to antisolvent (SAS) ratio, and drug concen-
tration were investigated to obtain the smallest particle
size. For EPN, factors like drug concentration and the
SAS ratio were examined. The effects of these process
parameters on the supersaturation, nucleation, and
growth rate were studied and optimized to obtain the
smallest particle size of curcumin by both the meth-
ods. The average particle size of the original drug was
about 10–12 lm and it was decreased to a mean
diameter of 330 nm for the APSP method and to
150 nm for the EPN method. Overall, decreasing the
drug concentration or increasing the flow rate, stirring
rate, and antisolvent amount resulted in smaller
particle sizes. Differential scanning calorimetry stud-
ies suggested lower crystallinity of curcumin particles
fabricated. The solubility and dissolution rates of the
prepared curcumin particles were significantly higher
than those the original curcumin. The antioxidant
M. Kakran  N. G. Sahoo  I.-L. Tan  L. Li (&)
School of Mechanical and Aerospace Engineering,
Nanyang Technological University, 50 Nanyang Avenue,
Singapore 639798, Singapore
e-mail: mlli@ntu.edu.sg
activity, studied by the DPPH free radical-scavenging
assay, was greater for the curcumin nanoparticles than
the original curcumin. This study demonstrated that
both the methods can successfully prepare curcumin
into submicro to nanoparticles. However, drug parti-
cles prepared by EPN were smaller than those by
APSP and hence, showed the slightly better solubility,
dissolution rate, and antioxidant activity than the
latter.
Keywords Curcumin  Antisolvent precipitation
with a syringe pump  Evaporative precipitation of
nanosuspension  Solubility  Dissolution
Introduction
Curcumin is a hydrophobic polyphenol extracted from
the rhizome of the herb Curcuma longa (Anand et al.
2007) and has been widely investigated for its
pharmacological activities such as anticancer (Aggar-
wal et al. 2003; Sharma et al. 2005), anti-inflammatory
(Jurenka 2009), and antioxidant (Aftab and Vieira
2010) effects. Despite these medicinal benefits, cur-
cumin has a low bioavailability due to its poor aqueous
solubility and dissolution properties (Anand et al.
2007), which greatly restricts its therapeutic use.
Nanoparticles are suitable for delivery of such highly
hydrophobic agents like curcumin and can help
overcome their poor aqueous solubility, slow dissolu-
tion, and/or low bioavailability (Devalapally et al.
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Page 2 of 11
2007). Recently nanocolloids of curcumin for intra-
venous injection have also been formed by Zheng et al.
(2010) for sustained drug release.
Nanoscale can be reached by either ‘‘top down’’ or
‘‘bottom up’’ approach (Pattekari et al. 2011). In the
pharmaceutical industry, the drug nanoparticles are
mainly produced by the ‘‘top down’’ approach using
techniques like milling and high pressure homogeni-
zation (Jacobs and Müller 2002; Keck and Müller
2006). In contrast, the ‘‘bottom up’’ approach, such as
antisolvent precipitation, supercritical fluid technol-
ogy, spray freezing into liquid, etc., is seldom
employed. In comparison to milling and high pressure
homogenization, some ‘‘bottom up’’ techniques, like
antisolvent precipitation are quite simple, cost effec-
tive, and easy for scaling-up (Horn and Rieger 2001;
Rogers et al. 2004). Therefore, in this study, two types
of precipitation processes namely: antisolvent precip-
itation with a syringe pump (APSP) and evaporative
precipitation of nanosuspension (EPN) have been used
to prepare curcumin particles to improve its solubility
and dissolution rate. The final product of the two
methods is the dry curcumin particles for oral drug
delivery. This is the first time EPN, involving the
evaporation of the solvents, has been used to prepare
the curcumin nanoparticles. Besides that the compar-
ison has been made with the APSP method, which
focuses on filtration to obtain the curcumin particles.
Various process parameters have been studied and
optimized to produce the smallest particle sizes by
both the methods and comparison has been made in
terms of the size and quality of the products obtained
by the two methods.
A precipitation process consists of (i) generation of
supersaturation, (ii) nucleation, and (iii) subsequent
growth of nuclei. For antisolvent precipitation of
poorly water-soluble drug, the drug is first dissolved in
a solvent, which is rapidly mixed with a solvent-
miscible antisolvent (e.g., water). The solubility of a
drug in the solvent–antisolvent mixture is lowered
compared to that in the original solvent and in this
way, a driving force for precipitation, called super-
saturation (S), is created, which is defined as:
C0
S¼ ; ð1Þ
C
where C0 is the initial concentration of a drug in the
solution to be precipitated and C* is the solubility of
the drug in the solvent–antisolvent system. Generation
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J Nanopart Res (2012) 14:757
of supersaturation is a prerequisite for the nucleation
to occur. The Gibbs free energy change, DG, is the
thermodynamic parameter that indicates whether
nucleation is possible or not. It is represented as the
energy barrier that a nucleation process must over-
come (Cao and Wang 2011):
16pc3 VS3
DG ¼ ; ð2Þ
3k2 T 2 ðln SÞ2
where c is the surface tension, VS is the molecular
volume (molar volume/Avogadro number), k is the
Boltzmann constant, T is the absolute temperature, and
S is the supersaturation. According to Eq. 2, a higher
supersaturation (S) and a lower surface tension (c)
result in a lower critical energy barrier (DG), which
leads to fast nucleation leading and hence, production
of smaller particles (Rogers et al. 2004; Matteucci
et al. 2006). The kinetic parameter describing how fast
the nucleation occurs is given by the rate of nucleation
per unit volume and per unit time, RN, as (Cao and
Wang 2011):
   
C0 kT DG
RN ¼ exp  ; ð3Þ
3pk3 g kT
where k is the diameter of the growth species and g is
the viscosity of the solution. This equation indicates
that high initial concentration (C0) or a higher
supersaturation (i.e., a large number of nucleation
sites), low viscosity (g), and low critical energy barrier
(DG) favor the formation of a large number of nuclei.
For a given concentration of solute, a larger number of
nuclei mean smaller nuclei.
Figure 1 schematically illustrates the processes of
supersaturation, nucleation, and subsequent growth by
coagulation and condensation. When the concentra-
tion of solute increases as a function of time, no
nucleation would occur even above the equilibrium
solubility. The nucleation occurs only when the
supersaturation reaches a certain value above the
solubility, which corresponds to the energy barrier
defined by Eq. 2 for the formation of nuclei. After the
initial nucleation, the concentration or supersaturation
of the growth species decreases. When the concentra-
tion decreases below this critical concentration, which
corresponds to the critical energy, no more nuclei
would form. The growth will continue until the
concentration of growth species reaches the equilib-
rium concentration or solubility.
J Nanopart Res (2012) 14:757
Fig. 1 Schematic illustration of supersaturation, nucleation,
and subsequent growth (Kakran et al. 2010)
Experimental
Materials
Curcumin was purchased from Sigma-Aldrich, Sin-
gapore. All the reagents used were of technical grade.
N-Hexane (HPLC grade) and ethanol (99.5–99.8%,
Absolute, GR Grade for analysis) were obtained from
Merck, Singapore. The deionized water (Milli-Q,
Millipore, Singapore) was used in experiments.
Methods
For the APSP method, the solutions of original
curcumin were prepared in ethanol at the predeter-
mined concentrations of 5–15 mg/mL. The syringe
was filled with 20 mL of the prepared solution and
secured onto a syringe pump. The drug solution was
quickly injected at a fixed flow rate (2–10 mL/min)
into the deionized water (antisolvent) of a definite
volume under magnetic stirring (200–1,000 rpm).
Different ratios of ethanol to water used were
1:10–1:20 (v/v) corresponding to a volume of
200–400 mL of water for 20 mL of drug solution.
The curcumin nanoparticles formed were filtered and
vacuum dried. For the EPN method, the solution of
original curcumin was prepared in ethanol and then a
nanosuspension was formed by quickly adding hexane
(antisolvent). Drug particles in the nanosuspension
were obtained by quick evaporation of the solvent and
antisolvent, under vacuum using a rotary evaporator
(Kakran et al. 2010). This was followed by vacuum
Page 3 of 11
drying of the nanoparticles to completely evaporate all
the solvents. The drug concentrations used were 5, 10,
15 mg/mL and the solvent to antisolvent (SAS) ratios
were varied from 1:10, 1:15 to 1:20 (v/v). For 20 mL
of the drug solution, 200–400 mL hexane was used.
Characterization
For calculating the S in Eq. 1, the solubility (C*) of
curcumin in the solvent–antisolvent system was
measured by adding an excess amount of the original
curcumin into ethanol antisolvent (water and hexane)
mixture at various ratios (1:10, 1:15, 1:20 by volume).
The mixture was shaken continuously at room
temperature (25 °C) for 24 h and then filtered through
a 0.2-lm polypropylene-reinforced PTFE membrane
and analyzed spectrometrically at 422 nm using UV
spectrometer (UV-3101PC, Shimadzu).
The morphology and size of samples were observed
using a scanning electron microscope (JSM-6390LA-
SEM, Jeol Co., Japan). The particle sizes were
analyzed by the UTHSCSA ImageTool program.
Differential scanning calorimetric (DSC) measure-
ments were carried out using a TA DSC 200 thermal
analyzer in a temperature range of 50–250 °C at a
heating rate of 10 °C/min in nitrogen gas. The melting
point and heat of fusion were calculated using the TA
DSC software.
For the solubility study, an excess amount of the
original curcumin and its nanoparticles prepared by
APSP and EPN were added into 20 mL of deionized
water. The mixture was shaken continuously at room
temperature (25 ± 0. 5 °C) for 24 h at 100 rpm. At
the end of 24 h, samples were filtered through a 0.2-
lm polypropylene-reinforced PTFE membrane (Mini-
start-SRP15, Sartorius, Germany) and diluted imme-
diately with deionized water. The concentration of
drug was determined spectrometrically at 422 nm
using a UV spectrometer.
The in-vitro dissolution of the samples was deter-
mined using the paddle method (USP apparatus II,
Vankel VK 7000 Dissolution Tester) in 900 mL of DI
water. The paddle rotation was set at 100 rpm. The
temperature was maintained at 25 ± 0.5 °C. The
samples containing an equivalent 5 mg of curcumin
were tested for their dissolution. 1 mL of the dissolved
solution samples was collected at 0.25, 0.5, 0.75, 1,
1.5, 2, 3, and 4 h of dissolution time. The dissolution
test for each sample was performed in triplicate and
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the dissolution data was averaged. The concentration
of drug was determined spectrometrically at 422 nm
using a UV spectrometer.
The antioxidant activity was measured by the
DPPH assay. DPPH (200 lM in methanol) was
incubated (in darkness) with 2.0 mL of aqueous
solution of original curcumin and its nanoparticles at
room temperature. After 30 min of incubation at room
temperature, the absorbance was measured at 518 nm
using a UV spectrometer (Mensor et al. 2001). The
percentage inhibition of the samples was calculated
using the formula:
%of inhibition ¼ ½ðcontrol OD  test ODÞ=control OD
 100;
ð4Þ
where OD is the optical density.
Results and discussion
Antisolvent precipitation with a syringe pump
Antisolvent precipitation is a simple and quite effective
approach to produce nanoparticles of poorly water-
soluble drugs by mixing a drug solution and an
antisolvent. As discussed above, the supersaturation is
the prerequisite for nucleation; therefore, the values of
supersaturation (S) are calculated using Eq. 1 and are
listed in Table 1. As discussed earlier, a higher value of
S leads to an increased nucleation rate and a decreased
critical nuclear size. However, it can be seen from
Table 1 that S is not the only parameter that governs the
kinetics of nucleation. Besides the supersaturation that
determines the nucleation, the crystal growth rate is also
a factor that affects the ultimate particle size. Therefore,
in this study, various critical process parameters such as
stirring speed, flow rate, SAS ratio, drug concentration
in solvent, and temperature of the antisolvent have been
examined and discussed below.
Stirring speed
It is observed from Table 1 that with increasing the
stirring speed from 200 to 1,000 rpm, both the diameter
and length of curcumin particles decreased consider-
ably from 550 and 3,800 nm (APSP 1) to 500 and
2,560 nm (APSP 3), respectively. This decrease in
particle size was due to the intensification of micro-
mixing between the multiphases with increasing
stirring speed. The more intense stirring at 1,000 rpm
with high efficiency of micromixing reduced the mass

Table 1 The preparation parameters such as stirring speed properties like the particle size (diameter and length), melting
(rpm), flow rate (mL/min), SAS ratio, drug concentration (DC, temperature (Tm, °C), enthalpy of fusion (DHf, J/g) and
mg/mL); Reynolds number (Re); solubility (C*) of curcumin in solubility (lg/mL) at 25 °C for the curcumin samples prepared
the ethanol–water system (corresponding to the SAS ratio) and by the APSP method
the resulting supersaturation (S) calculated using Eq. 1; and
Samples Process parameters Re C* S Particle size (nm) Tm DHf Solubility
(lg/mL) (°C) (J/g) (lg/mL)
Stirring Flow rate SAS DC Diameter Length
speed (mL/min) (mg/mL)
(rpm)

APSP 1 200 2 1:10 5 145 0.0216 231 550 ± 70 3,800 ± 332 174.0 127.8 2.62
APSP 2 500 2 1:10 5 145 0.0216 231 520 ± 62 3,120 ± 219 173.3 106.4 2.91
APSP 3 1,000 2 1:10 5 145 0.0216 231 500 ± 65 2,560 ± 220 173.2 73.0 3.83
APSP 4 1,000 5 1:10 5 360 0.0216 231 495 ± 50 2,010 ± 166 173.4 96.7 4.34
APSP 5 1,000 10 1:10 5 720 0.0216 231 490 ± 51 1,860 ± 143 173.1 82.1 4.90
APSP 6 1,000 10 1:15 5 720 0.0072 694 420 ± 48 1,340 ± 110 173.5 56.4 5.69
APSP 7 1,000 10 1:20 5 720 0.0021 2380 340 ± 37 930 ± 88 173.5 68.1 6.13
APSP 8 1,000 10 1:20 10 720 0.0021 4761 345 ± 40 950 ± 90 173.2 88.5 6.05
APSP 9 1,000 10 1:20 15 720 0.0021 7142 360 ± 42 965 ± 96 173.1 65.9 6.22
a
APSP 10 1,000 10 1:20 5 720 0.0004 12,500 330 ± 36 860 ± 99 173.7 48.6 7.48
(at 5 °C)
a
Measured at 5 °C, all others at room temperature (25 °C)

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transfer resistance, thereby enhancing the mass trans-
fer and the rate of mixing between the multiphases. As
a result, a homogenous mixing could be achieved in
short time, thus, producing smaller drug particles.
Therefore, all further experiments were performed
with a stirring speed of 1,000 rpm to maintain the
consistent mixing conditions.
Flow rate
At a lower flow rate, the efficiency of solvent/anti-
solvent mixing is lower and hence, there are few
nucleating sites, which give rise to a prolonged crystal
growth process and the formation of larger crystals. It
was observed in our study that as the flow rate of the
curcumin solution into the deionized water was
increased from 2 to 10 mL/min, the length of curcu-
min particles decreased significantly from 2,560 nm
(APSP 3) to 1,860 nm (APSP 5). This is because the
curcumin solution was mixed more rapidly into
deionized water as the flow rate was increased. As
the time was short for allowing the crystal growth,
only smaller crystals were formed. However, it can be
seen that there was no significant decrease in the
particle diameter when increasing the flow rate in our
study. This can be explained based on the fact that the
crystal growth in curcumin occurs along one direction
leading to needle-shaped curcumin crystals as
reported earlier by He et al. (2010). Therefore, in our
study, increasing the flow rate restricted the growth of
curcumin particles in one direction only.
Increasing the flow rate increased the jet velocity,
the Reynolds number (Re, Table 1) and shear forces,
resulting in an increased extent of mixing between the
drug solution and the antisolvent per unit time. The
corresponding Reynolds number Re was calculated
according to the following equation:
VD QD
Re ¼ ¼ ð5Þ
v Av
where V is the flow velocity for the solvent (m/s), D is
the diameter of the pipe (m), v is the kinematic viscosity
(m2/s), Q is the volumetric flow rate (m3/s), and A is the
pipe cross-sectional area (m2). The diameter of the
injection needle, 0.24 mm, and the kinematic viscosity
of ethanol, 1.23 9 10-6 m2/s, were used to calculate the
value of Re. The volumetric flow rate, Q, was varied
from 2 mL/min (3.33 9 10-8 m3/s) to 5 mL/min
(8.33 9 10-8 m3/s), then to 10 mL/min (16.66 9
Page 5 of 11
10-8 m3/s), and the corresponding values of Re
increased from 145 to 360 then to 720 (shown in
Table 1). The decrease in particle size is greater when
Re increased from 145 to 360 than in the case of
360–720. Previously, Dalvi and Dave (2009) also
reported for their antisolvent precipitation system that
controlling the flow rate to increase the Re beyond 900
resulted in a marginal change in particle size.
In light of these experiments, all further experiments
were conducted at the stirring speed of 1,000 rpm and
the flow rate of 10 mL/min. As seen from Table 1, the
values of S were affected by the ratio of the SAS, the
drug concentration in the solution, and the temperature.
From the calculations, a larger SAS ratio, a higher drug
concentration and a lower temperature resulted in
greater S, which principally should lead to more nuclei
with smaller sizes. Following is the discussion about
these factors.
SAS ratio
The average particle sizes in Table 1 and the corre-
sponding SEM images in Fig. 2 suggest an inversely
proportional relationship between the amount of
antisolvent in the SAS ratio and the particle size.
Both the length and the diameter of the curcumin
particles decreased drastically from 1,860 and 490 nm
(APSP 5) to 930 and 340 nm (APSP 7), respectively,
as the SAS ratio was changed from 1:10 to 1:20. There
are a few reasons for this observation. When the
curcumin solution was added to the deionized water,
the curcumin concentration was reduced more quickly
as the amount of antisolvent was increased, which led
to faster precipitation of the drug into nanoparticles.
From Table 1, the S values at the greater antisolvent
amount were higher and according to Eqs. 2, 3 a
greater amount of antisolvent should lead to a greater
nucleation rate and hence smaller nuclei. Once the
nuclei are formed, growth occurs simultaneously. For
the subsequent growth, more antisolvent amount
increases the diffusion distance for growing species
and consequently diffusion becomes the limiting step
for nuclei growth (Cao and Wang 2011).
Drug concentration
The drug concentration affects the size of precipitated
particles in opposing ways: a higher drug concentra-
tion leads to a greater supersaturation (S) as seen from
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Fig. 2 SEM photographs of original curcumin and curcumin particles prepared by APSP and EPN: a original curcumin, b APSP 3,
c APSP 5, d APSP 7, e APSP 10, f EPN 2, g EPN 3, and h EPN 5. The sample codes are described in Table 1

Table 1, which results in a faster nucleation rate and (Matteucci et al. 2006). As shown in Table 1,
thereby smaller particles. However, high supersatura- increasing the drug concentration from 5 to 10 and
tion also speeds up agglomeration through the greater 15 mg/mL resulted in an increase in particle length
chance of particle collision to produce bigger particles from 930 nm (APSP 7) to 950 nm (APSP 8) and

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965 nm (APSP 9), respectively, and a corresponding
minor increase in the particle diameter as well, which
indicated that in our study agglomeration prevailed
over nucleation at high drug concentrations. This
result was governed by two factors: the number of
nuclei formed in the solvent/antisolvent interface, and
the influence of concentration on the viscosity (Gal-
indo-Rodriguez et al. 2004). On one hand, at a higher
concentration, a large number of nuclei were formed at
the interface of two phases which lead to agglomer-
ation and thus, formation of larger particles. Simulta-
neously, those nuclei obstructed the further diffusion
of drug molecules from the SAS. On the other hand,
the viscosity of the drug solution increased with
increasing concentration, which hindered the diffusion
between solution and antisolvent, resulting in non-
uniform supersaturation (Zhang et al. 2009) and then
the larger drug particles.
In this study, a higher antisolvent amount compared
to solvent (SAS ratio) and a lower drug concentration
resulted in smaller particle size for curcumin and
APSP 7. Therefore, the hint to produce smaller
particles is to start with unsaturated initial solutions
(i.e., low drug concentrations), and then create a high
supersaturation by using a greater amount of
antisolvent.
Temperature
The effect of temperature on the size of curcumin
particles was studied. In this experiment, the deionized
cold water was used as antisolvent and kept in an ice
bath to maintain the antisolvent at a temperature of
about 5 °C. The precipitation process is expected to be
influenced by the temperature through several ways.
First, reduction in the antisolvent temperature
decreases the equilibrium solubility of curcumin in
the solute–solvent–antisolvent mixture and hence,
increases the supersaturation. As shown in Table 1, a
change in the antisolvent temperature from 25 to 5 °C
enhanced the value of S from 2,380 to 12,500, so that
the nucleation rate increased and hence the particle
size decreased. Second, the precipitation in the liquid
phase is a diffusion-limited process (Nielsen 1964). At
a lower temperature the diffusion rate is lower and
accordingly the crystal growth rate is lower. The
decrease in the antisolvent temperature from 25 to
5 °C increased the viscosity of water from 0.894 to
1.519 cP, which led to a reduction in particle collision
Page 7 of 11
frequency and resulted in a decrease in particle growth
by preventing coagulation and agglomeration. There-
fore, the smaller particles were formed as a result of
the high nucleation rate and low growth rate at the low
temperature, 5 °C. The particle diameter and length of
sample APSP10 were 330 and 860 nm, respectively.
Evaporative precipitation of nanosuspension
In this process, the antisolvent was changed from
water to hexane to precipitate curcumin particles from
its solution in ethanol. From Tables 1 and 3, with
hexane as antisolvent the S values were lower than that
with water. This was due to the higher solubility (C*)
of curcumin in the ethanol–hexane mixture than the
ethanol–water mixture. According to Eqs. 2, 3, the
lower surface tension and lower viscosity increase
the rate of nucleation, thereby leading to formation of
a large number of smaller sized nuclei. It should be
noted that, although a lower viscosity is desired for a
higher rate of nucleation to facilitate diffusion of drug
from SAS, a higher viscosity would lead to lesser
crystal growth. Table 2 shows the comparison of some
of the physical properties of water and hexane.
Comparing water and hexane, in spite of exhibiting
lower S values, hexane possesses a lower surface
tension and viscosity which facilitate faster nucle-
ation. In the case of using hexane as antisolvent,
because of the lower supersaturation (S) generated, the
chance of particle collision was reduced during the
precipitation process and it resulted in a uniform
nanosuspension independent of the agitation speed
and also the flow rate. Filtering such a nanosuspension
did not yield any particles as they were too small.
Therefore, the EPN process was devised to extract the
curcumin particles from the nanosuspension. The
lower boiling point and heat of vaporization of hexane
(Table 2) make it possible to quickly evaporate the
curcumin–ethanol–hexane nanosuspension using a
Table 2 Physical properties of water and hexane
Properties Water Hexane
Surface tension at 20 °C 72.86 18.4
(dyne/cm or mN/m)
Viscosity (cP) at 25 °C 0.894 0.294
Boiling point (°C) 100 69
Heat of vaporization (kJ/kg) 2,444 366
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rotary evaporator to obtain the curcumin particles.
Water was not used for this method because of its
higher boiling point and greater heat of vaporization
(Table 2). We have shown in our other study that
using water as antisolvent for EPN resulted in big
and non-uniform drug particles (Kakran et al. 2011).
Table 3 lists the curcumin samples prepared by the
EPN method. The similar trends regarding the SAS
ratio and drug concentration were observed for EPN
as in case of the APSP method. The length and
diameter of the curcumin particles decreased from
1,680 and 300 nm (EPN 1) to 850 and 150 nm
(EPN 3) as the SAS ratio was changed from 1:10 to
1:20 due to the increase in the value of S. On the
other hand, increasing the drug concentration from 5
to 15 mg/mL slightly increased the particle length
and diameter from 850 and 150 nm (EPN 3) to 920
and 155 nm (EPN 5), respectively, indicating that
agglomeration dominated over nucleation at high
drug concentrations as discussed earlier for the
APSP method.
As seen from Fig. 2, the original curcumin powder
exhibited particles around 10–12 lm, whereas the
curcumin particles prepared by APSP and EPN were
much smaller in size. For the curcumin particles
prepared by APSP, the APSP 10 exhibited the smallest
particle diameter and length of 330 and 860 nm,
respectively, at the stirring speed of 1,000 rpm, flow
rate of 10 mL/min, SAS volume ratio of 1:20, and
5 mg/mL of drug concentration. For the curcumin
particles prepared by EPN, the EPN 3 exhibited the
smallest diameter and length of 150 and 850 nm,
respectively, at the SAS volume ratio: 1:20 and drug
concentration of 5 mg/mL.
Comparing both the methods, EPN gave particles
with much smaller diameters than those produced by
APSP. The morphologies of the curcumin particles
obtained from both the methods were different from
the original curcumin as observed from Fig. 2. The
external shape, imparted to a crystal by the develop-
ment of its various forms, is referred to as crystal habit,
which is not exclusively controlled by the internal
structure of a crystal, but also by the environmental
conditions during nucleation and growth (Park et al.
2006). In our study, the crystal habit of curcumin
changed from spherical or irregular shape of the
original curcumin to lath-like by the APSP method and
needle-like by the EPN method. This difference in
particle shape should be attributed to the use of
different antisolvents in both the methods. The use of
the organic antisolvent (hexane) favoured the needle-
like morphology, while the polar antisolvent (water)
resulted in the lath-like particles. For the curcumin
particles prepared by APSP, the length of the particles
deceased as the mixing and the nucleation rate
increased as can be seen from Fig. 2b, d. But for
EPN, the shapes of particles were needle like for all the
samples.
DSC analysis
To understand the effect of the two methods, APSP and
EPN, on the melting temperature and the melting
enthalpy of curcumin, DSC was conducted. DSC
thermograms of the original curcumin and its particles
prepared by APSP and EPN are shown in Fig. 3. The
melting temperature (Tm) and heat of fusion (DHf)
obtained are summarized in Tables 1 and 3. The original

Table 3 The preparation parameters like drug concentration using Eq. 1; and properties like the particle size (diameter and
(DC, mg/mL) and SAS ratio; the solubility (C*) of curcumin in length), melting temperature (Tm), enthalpy of fusion (DHf) and
the curcumin–ethanol–hexane system (corresponding to the solubility at 25 °C for the original curcumin and the curcumin
SAS ratio) and the resulting supersaturation (S) calculated samples prepared by the EPN method
Samples Process parameters C* (lg/mL) S Particle size (nm) Tm (°C) DHf (J/g) Solubility
(lg/mL)
SAS DC (mg/mL) Diameter Length

Original – – 12,370 173.8 159.7 0.58

EPN 1 1:10 5 0.7469 6.69 300 ± 34 1,680 ± 90 173.3 112.0 6.11
EPN 2 1:15 5 0.0935 53.47 200 ± 25 1,310 ± 71 172.8 110.3 7.24
EPN 3 1:20 5 0.0134 371.471 150 ± 15 850 ± 36 173.0 68.9 8.23
EPN 4 1:20 10 0.0134 742.94 150 ± 18 905 ± 30 173.1 82.1 8.02
EPN 5 1:20 15 0.0134 1114.41 155 ± 21 920 ± 28 173.9 56.5 8.31

123
J Nanopart Res (2012) 14:757
curcumin used in this study had a sharp melting
endothermic peak at 173.8 °C and a melting enthalpy
of 159.7 J/g. From Table 1, it can be seen that the APSP-
prepared curcumin particles had melting point
*173 °C, very similar to the original curcumin, but
the enthalpy of fusion was lower than that of the original
curcumin. The enthalpy of fusion is proportional to the
degree of crystallinity in the samples. These results
suggested that the crystallinity of curcumin was
decreased after it was prepared by the APSP process.
Table 3 shows that the melting temperatures for the
EPN-prepared samples were also *173 °C and the
enthalpy of fusion was lower than that of the original
curcumin, indicating a certain loss of crystallinity of
curcumin during the EPN process as well. Therefore, it
is seen that decreasing the particle size of the original
curcumin from micro- to nanoscale by APSP and EPN
methods resulted in a reduction in the crystallinity of
Fig. 3 DSC thermograms for original curcumin and particles
prepared by a EPN and b APSP
Page 9 of 11
the particles. Comparing the samples prepared by both
APSP and EPN, it was noted that the crystallinity did not
depend on the particle size but rather on the method of
preparation. There was no significant difference in the
enthalpy of fusion and hence the crystallinity of the
samples by both the methods, in spite of the difference in
particle size. Thus, it is observed that APSP produces
less crystalline particles compared to EPN.
Solubility study
The solubility values at 25 °C measured for the original
curcumin and its particles fabricated by APSP and EPN
are listed in Tables 1 and 3. The solubility of the original
curcumin was extremely low being only 0.58 ± 0.03
lg/mL, and it increased by 14 times to the highest value
of 8.23 ± 0.07 lg/mL for the EPN 3. Among the
APSP-prepared samples, APSP 10 exhibited the max-
imum solubility of 7.48 ± 0.11 lg/mL. The increase in
solubility was because the particles prepared by APSP
and EPN were much smaller as compared to the original
curcumin. It is known that the solubility increases with
decreasing particle size below 1,000 nm (Keck and
Müller 2006). Besides that, an increase in the solubility
and also the dissolution velocity can also be achieved by
changing the crystalline state of a crystalline drug. An
amorphous or metastable form of a drug has a greater
solubility as compared to its crystalline form (Hancock
and Zografi 1997). Therefore, the curcumin nanoparti-
cles exhibited the enhanced solubility because of the
smaller particle size and the lower crystallinity as
compared to the original curcumin. The curcumin
particles prepared by EPN showed the higher solubility
than the ones prepared by APSP because of their
comparatively smaller particle sizes.
Dissolution study
The dissolution tests were performed for the original
curcumin and the selected samples (EPN 3 and 5, APSP
9 and 10) because they all showed the relatively higher
solubility among the samples prepared by EPN and
APSP. As seen in Fig. 4 for the dissolution profiles of
the samples, only about 10% of the original curcumin
dissolved within 4 h as compared to 93–96% of
dissolution for the EPN-prepared curcumin and 75–
81% of dissolution for the APSP-prepared samples.
According to Noyes–Whitney equation (1897), the
dissolution velocity, dc/dt, is expressed as:
123
Page 10 of 11
Fig. 4 Dissolution profiles for original curcumin and particles
prepared by APSP and EPN
dc c  c 
s x
¼AD ; ð6Þ
dt h antioxidant properties of curcumin. Curcumin shows
where A is the surface area, D is the diffusion
coefficient, cs is the saturation solubility, cx is the
bulk concentration, and h is the diffusional distance.
Reducing the particle size to nanoscale can increase
the dissolution velocity or the dissolution rate of a
poorly water-soluble drug by increasing the surface
area (A), the diffusion coefficient (D), and difference
between the saturation solubility and bulk concentra-
tion (cs - cx); or by decreasing the diffusional
distance (h) (Müller et al. 2000). In our study, the
mean particle size of original curcumin was reduced to
a nanometer range by the EPN and APSP methods.
In addition to that an amorphous or metastable form
could dissolve at a faster rate because of its higher
internal energy and greater molecular mobility, as
compared to crystalline materials (Hancock and
Zografi 1997). The DSC study revealed that the
crystallinity of EPN and APSP-prepared curcumin
nanoparticles was lower than that of the original
curcumin. Therefore, reduction in both particle size
and crystallinity led to a faster rate of dissolution for
the curcumin nanoparticles prepared. The higher
dissolution rate could translate into increased bio-
availability upon oral administration.
DPPH free radical-scavenging assay
DPPH is a free radical and stable at room temperature,
which produces a deep violet solution in organic
solvents. It is reduced in the presence of curcumin
molecules, giving rise to uncolored solutions. The use
123
J Nanopart Res (2012) 14:757
Fig. 5 Antioxidant activities for original curcumin and nano-
particles prepared by EPN and APSP
of DPPH provides an easy and rapid way to evaluate
an antioxidative effect which is mainly due to its
phenolic hydroxyl groups. The free radical-scaveng-
ing properties of the original curcumin and curcumin
nanoparticles prepared by EPN and APSP are shown
in Fig. 5. The observed scavenging effect of curcumin
on the DPPH radicals decreased in the following
order: EPN-prepared curcumin nanoparticles C
APSP-prepared curcumin nanoparticles  original
curcumin. From the solubility and dissolution studies,
it was observed that the curcumin nanoparticles
showed the much higher solubility and dissolution
rate in water than the original curcumin, thus showing
a higher free radical-scavenging effect. The previous
studies have also suggested that the nanoparticles of
curcumin (Yen et al. 2010) have an improved antiox-
idant activity in vitro.
Conclusions
This study demonstrated that both APSP and EPN
methods can successfully prepare curcumin nanopar-
ticles. The curcumin nanoparticles prepared by the two
methods exhibited the lower crystallinity as revealed
by the DSC analysis. The EPN process produced
smaller particles, and gave the slightly higher solubil-
ity and dissolution rate than those produced by the
APSP method. The dissolution rate of curcumin was
improved tremendously after it was prepared into
nanoparticles because of the decrease in both the
particle size and crystallinity. The APSP has the
J Nanopart Res (2012) 14:757
advantages of using water as antisolvent and being a
simple pumping, stirring and filtering system, whereas
the EPN uses hexane as antisolvent and is a vacuum
evaporation process to prepare nanoparticles.
Although the EPN method could produce smaller
particles than the APSP, the solubility and dissolution
rates were not significantly different between the drug
particles prepared by the two methods. To conclude,
both the methods represent an efficient means to
obtain curcumin nanoparticles with a higher solubility
and a higher rate of dissolution in vitro. These
nanoparticles are expected to exhibit a higher bio-
availability in vivo, which will lead to considerable
dose reduction for the patients.
Acknowledgments Authors acknowledge Lee Kuan Yew
Postdoctoral Fellowship and SUG grant M58050023, NTU,
Singapore.
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