See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/301248319

High performance curcumin subcritical water extraction from turmeric

(Curcuma longa L.)

Article  in  Journal of chromatography. B, Analytical technologies in the biomedical and life sciences · April 2016
DOI: 10.1016/j.jchromb.2016.04.021

CITATIONS READS

16 2,290

5 authors, including:

Ghasem Najafpour Mostafa Rahimnejad

Babol Noshirvani University of Technology Babol Noshirvani University of Technology
597 PUBLICATIONS   5,501 CITATIONS    171 PUBLICATIONS   2,215 CITATIONS   

SEE PROFILE SEE PROFILE

Ali Akbar Moghadamnia Meisam Valizadeh Kiamahalleh

Babol University of Medical Sciences University of Adelaide
200 PUBLICATIONS   1,860 CITATIONS    24 PUBLICATIONS   136 CITATIONS   

SEE PROFILE SEE PROFILE

Some of the authors of this publication are also working on these related projects:

Biological treatment of leachate from cattle manure in a hybrid system and instant methane purification View project

Fabrication of ethanol selective membrane for bioethanol production in membrane bioreactor View project

All content following this page was uploaded by Ali Akbar Moghadamnia on 08 October 2017.

The user has requested enhancement of the downloaded file.

 Journal of Chromatography B, 1022 (2016) 191–198

Contents lists available at ScienceDirect

Journal of Chromatography B
journal homepage: www.elsevier.com/locate/chromb

High performance curcumin subcritical water extraction from

turmeric (Curcuma longa L.)
Mohammad Valizadeh Kiamahalleh a , Ghasem Najafpour-Darzi a,∗ , Mostafa Rahimnejad a ,
Ali Akbar Moghadamnia b , Meisam Valizadeh Kiamahalleh c
a
Biotechnology Research Lab, Faculty of Chemical Engineering, Noshirvani University of Technology, Babol, Iran
b
Department of Pharmacology and Physiology, School of Medicine, Babol University of Medical Sciences, Babol, Iran
c
School of Chemical Engineering, The University of Adelaide, Adelaide, SA5005, Australia

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

Article history: Curcumin is a hydrophobic polyphenolic compound derived from turmeric rhizome, which consists about
Received 17 February 2016 2–5% of the total rhizome content and is a more valuable component of turmeric. For reducing the draw-
Received in revised form 1 April 2016 backs of conventional extraction (using organic solvents) of curcumin, the water as a clean solvent was
Accepted 9 April 2016
used for extracting curcumin. Subcritical water extraction (SWE) experimental setup was fabricated in a
Available online 12 April 2016
laboratory scale and the influences of some parameters (e.g. extraction temperature, particle size, reten-
tion time and pressure) on the yield of extraction were investigated. Optimum extraction conditions such
Keywords:
as SWE pressure of 10 bar, extractive temperature of 140 ◦ C, particle size of 0.71 mm and retention time
Curcumin
Extraction
of 14 min were defined. The maximum amount of curcumin extracted at the optimum condition was
Subcritical water 3.8 wt%. The yield of curcumin extraction was more than 76 wt% with regards to the maximum possible
Solvent free curcumin content of turmeric, as known to be 5%. The scanning electron microscope (SEM) images from
Turmeric the outer surface of turmeric, before and after extraction, clearly demonstrated the effect of each param-
eter; changes in porosity and hardness of turmeric that is directly related to the amount of extracted
curcumin in process optimization of the extraction parameters.
© 2016 Elsevier B.V. All rights reserved.

1. Introduction myelodysplastic syndrome) [6] Anti-human immunodeficiency

The turmeric (Curcuma longa L.) plant of ginger family is exten-
sively cultivated in tropical part of south and Southeast Asia. The
root of this plant consisting rhizome which is the most useful part
in particular for culinary and medicinal purposes [1]. Other appli-
cations of turmeric are in dietary spice, foods and textile coloring,
and curing broad ranges of diseases [2]. The orange-yellow crys-
talline powder curcumin, first time isolated from curcuminoids by
Vogel in 1815, is the most active component of turmeric, insoluble
in water and makes up 2–5 wt% of the spice [3]. In 1910, the chem-
ical formula of curcumin (C21 H20 O6 ) was first described by Lampe
and Miłobȩdzka, which is known as diferuloylmethane [4]. The
chemical structures and curcuminoids are summarized in Table 1
[5].
Some of diverse applications of curcumin are in food products
(vegetable and animal products), [3], the treatments of sev-
eral diseases (i.e. multiple myeloma, Alzheimer’s, psoriasis and
∗ Corresponding author.
E-mail address: najafpour8@yahoo.com (G. Najafpour-Darzi).
virus (Anti-HIV) cycle replication, [7,8]. Some more medical appli-
cations of curcumin includes skin and muscle anti-inflammatory,
anti-bacterial, anti-carcinogenic, antifungal, antimicrobial agents
[5,9–11] and recent prominent usage of curcumin is in anticancer
compounds as protective agent to minimize the risk of lung cancer
[12,13], liver, duodenum, and kidneys [14,15].
The curcuminoid and in specific the curcumin is considered
as hydrophobic natural polyphenolic material [12], poorly soluble
in hydrocarbon solvents. Curcumin at acidic condition in prac-
tical is insoluble in water and neutral pH; but soluble in basic
solvent. Different surfactant micellar systems (acetone, methanol,
and ethanol) have been incorporated to the curcumin to make it
water-soluble [1]. It is stable at high temperatures and in acids,
but unstable in alkaline conditions and presence of light. The value
of turmeric products is proportionally related to the curcuminoid
content. Quantitative estimation of curcuminoids can be deter-
mined by photometrically, depends on its absorbance values at
wavelength of 420 nm [3].
The diversity of available techniques for the extraction of
curcumin such as Soxhlet, microwave-assisted extraction (MAE),
maceration, supercritical carbon dioxide extraction, digestion,

http://dx.doi.org/10.1016/j.jchromb.2016.04.021
1570-0232/© 2016 Elsevier B.V. All rights reserved.
192 M. Valizadeh Kiamahalleh et al. / J. Chromatogr. B 1022 (2016) 191–198
Table 1
The structures and properties of curcuminoids.
Curcuminoids compounds Curcumin
Chemical structure
IUPAC name (1E,6E)-1,7-bis (4-hydroxy-3-
methoxyphenyl)-1,6-
heptadiene-3,5-dione
Other names Curcumin, diferuloyl methane,
C.I. 75300, Natural Yellow 3
Chemical formula C21 H20 O6
Molar mass 368.38 g/mol
Appearance Bright yellow to orange
powder
Melting point 183 ◦ C (361 ◦ F; 456 K)
ultra-sonic assisted extraction and enzyme-assisted extraction
were reported in the literature. Amongst all of conventional meth-
ods for extraction of curcumin, Soxhlet extraction (based on organic
solvent) with a heating period of 12 h was desired. The drawbacks
of Soxhlet extraction process are to be a time consuming, too labo-
rious and making use of bulk amount of organic solvents [3,16–20].
In addition, such idea of using organic solvents might be followed
by undesirable impacts towards the environment and on food
components. Hence, several other methods (including solid–liquid
extraction, aqueous alkaline extraction, extraction with vegetable
oils, extraction with aqueous alkanol solutions and supercritical
fluid extraction (SFE)) have been developed to extract efficiently
antioxidants from aromatic plants. Water, due to its unique sol-
vency under the temperature at different conditions can be used
as an effective solvent. The main issue with the above extraction
technique is the isolation of antioxidant constituents which could
be addressed by subcritical water extraction (SWE) [21–23].
Water is known as a cheap, green and an ideal solvent for
the industrial application of extracting pharmaceutical compounds
from the medical plants; but having poor efficiency of extraction for
most of organic compounds, including polychlorinated biphenyls
(PCBs), polycyclic aromatic hydrocarbons (PAHs) and most pes-
ticides at room temperature [22]. However, due to its promising
solvency behavior toward varying the temperature, water has
recently received immense research attentions. It is noteworthy
that one could manipulate the physio-chemical properties of water
in a sealed system by simultaneously adjusting the temperature
and pressure of the system, maintaining water in the liquid phase
while the temperature is significantly rising above the boiling point
of 100 ◦ C. Water can be maintained in liquid phase at critical points,
temperature up to 374 ◦ C and a pressure of 221 bar. Under such
condition, it denoted as “subcritical” or “hot/liquid” water with the
polarity, viscosity, surface tension, and disassociation constant sub-
stantially lower than that of water at ambient conditions, thereby
exhibiting similar chemical properties to those of organic solvents.
Thus, pressurized water with low-polarity and under sub-critical
conditions might be able to solubilize the organic compounds of
either polar (at low temperature) or non-polar (at high temper-
ature), such as phytochemicals which are normally insoluble in
ambient water. Other titles used for SWE are hot water extraction or
pressurized (hot) water extraction, pressurized low polarity water
extraction, high-temperature water extraction, superheated water
extraction or hot liquid water extraction, which is a highly promis-
ing “green” technique based on use of solely solvent of subcritical
water [24–26].
Water, generally known as a highly polar solvent due to having
a large number of hydrogen-bonded structure at ambient tempera-
ture and atmospheric pressure, possesses a high dielectric constant
Demethoxycurcumin Bisdemethoxycurcumin
(1E,6E)-1,6-Heptadiene-3,5-dione, (1E,6E)-1,7-bis
1-(4-hydroxy-3-methoxyphenyl)-7-(4- (4-hydroxyphenyl)hepta-1,6-diene-
hydroxyphenyl) 3,5-dione
4-hydroxycinnamoyl (feroyl)methane, Bis(4-hydroxycinnamoyl)methane,
curcumin II, BHCFM BHCMT
C20 H18 O5 C19 H16 O4
338.35 g/mol 308.33 g/mol
Yellow powder Yellow powder
172 ◦ C (342 ◦ F; 445 K) 222 ◦ C (432 ◦ F; 495 K)
(␧), which makes it unsuitable solvent for extracting non-polar or
organic compounds at such condition [27]. With applying sufficient
pressure (up to ∼50 bar), the water dielectric constant decreases
from ∼80 to ∼27 by elevating the temperature from ambient to
250 ◦ C, whereby retaining water in the liquid phase. Under such
conditions, water similarly behaves to certain organic solvents
which are able to dissolve a wide range of analytic with medium to
low level of polarity [28,29].
Ibáñez et al. extracted numbers of active antioxidant com-
pounds including carnosol, rosmanol, carnosicacid, methyl
carnosate and flavonoids from rosemary using SWE [30]. The SWE
has also served to extract phenolic compounds from pomegranate
[31], dietary fiber from Citrus junos peel [32], ginsenosides from
American ginseng [33]; catechins and epicatechin from tea leaves
and grape seeds [34]; anthraquinones (antibacterial, antiviral and
anticancer compounds) from roots of Morinda citrifolia [35]; and,
flavones, anilines and phenols from orange peels [36]. Recently,
SWE is widely used in determination of organic pollutants in soils,
sludges and sediments, the extraction of spice components from
plants as well as some encouraging applications in the field of
pharmaceutical industry [24].
Use of subcritical water addresses all the environmental issues
and in addition reduces the handling time and solvent con-
sumption, reduces the loss of heat sensitive compounds, and
consequently eliminates the use of any toxic organic solvent, ulti-
mately with the same extraction efficiency to that of organic
solvents. The technique could be adjusted to selectively extract
almost all sorts of polar, moderately polar, and nonpolar organics
by only varying the extraction parameters, such as temperature,
pressure and co-solvent (in case required).
In the present work, the subcritical water extraction apparatus
was fabricated in our research laboratory scale in order to extract
the curcumin from turmeric. As the water is clear solvent, the
extraction by this method can decrease the uses of organic sol-
vent that was the problem for extracting medical component of
turmeric. Also in this work several experimental variables, which
is the novelty of the current work have been tested and optimized.
The influences of some parameters (e.g. extraction temperature,
particle size, retention time and pressure) on the yield of extraction
were also analyzed. This method can increase the yield of extraction
by achieving the best point of parameters. Most part the apparatus
was fabricated in our research laboratory; some controlling units
of the setup were assembled for precise and accurate operation.
2. Materials and method
Turmeric rhizomes were collected from Mashhad (Khorasan
Razavi, Iran) in September 2014. Pure curcumin, for preparing stan-
 M. Valizadeh Kiamahalleh et al. / J. Chromatogr. B 1022 (2016) 191–198
Table 2
Characterization of turmeric by Soxhlet method.
Component wt%
Oil 4.5
Mass of extract 7.0
Moisture 7.0
Mineral 11.5
Organic 70.0
dard solution, was purchased from Merck. The deionized water was
used as extractive solvent. Methanol (HPLC grade) was supplied
by Merck as a carrier of curcumin for quantification and analyti-
cal purposes were used for HPLC analysis. For HPLC mobile phase,
acetonitrile and water (HPLC grade) were also supplied by Merck.
FDU-8624 Freeze dryer was used to prepare powder of extracts.
HPLC KNAUER (Berlin, Germany), were used for analysis the quan-
tities of curcumin.
2.1. Turmeric characterizations
Before the extraction process, the purchased turmeric was eval-
uated and characterized by the Soxhlet method (see the details in
Supplementary materials) that was the ideal process of conven-
tional method to determine the percentage of turmeric component
as shown in Table 2.
2.2. Experimental procedure
2.2.1. Pretreatment of sample
After grinding and classified the turmeric in different particle
size, they were cleaned (oversized unwanted particles were fil-
tered out by sieve) and then put them into n-hexane about 2 h.
The n-hexane pretreatment was conducted in order to eliminate
interaction of oil and fats in curcumin in our analytical methods.
Otherwise for commercial scale this stage could be eliminated.
The n-hexane solvent after the pretreatment was discarded. The
amount of probable dissolved curcumin in n-hexane can be ignored.
2.2.2. Extraction
The SWE apparatus was built in “Biotechnology laboratory of
Babol Noshirvani University of Technology” as shown schemat-
ically in Fig. 1. The sample after weighing embedded in the
extraction cell. Deionized water filled into 5 L feed tank and purged
for 1 h with N2 to remove dissolved oxygen. Dissolved oxygen may
oxidize the analyte. High pressure pump was used to deliver water
Fig. 1. Schematic diagram of lab-scale experimental setup.
193
through the system at a constant flow rate. The output of pump
passes from check valve (on the side way valve). After that, the
water fills the extraction cell via bottom of the pressure vessel. The
extraction cell is made of stainless steel with appropriate thick-
ness that can withstand higher pressure and temperature. There
is a heating jacket around the cell that heats the water and raises
its temperature. Pressure and temperature controllers are adjusted
for the set-point. After the specified retention time the extracts will
exit the cell from the bottom and passed thorough heat exchanger
and cooled with water about 15 ◦ C. Finally the extracts collect into
container and will be analyzed.
2.3. Curcumin HPLC analysis
The collected samples were analyzed by HPLC using UV detec-
tor. The samples were injected and the elution was carried out with
gradient solvent systems with a flow rate of 1.0 mL/min at ambi-
ent temperature. The HPLC column used was C18 (250 × 4.6 mm),
mobile phase were 90% acetonitrile and 10% water; detected at a
wavelength of 420 nm.
To determine the amount of curcumin extracted, pure cur-
cumin with concentrations 1.25, 2.5, 5, 10 and 20 ppm dissolved
in methanol and then injected to HPLC. After the certain retention
time, calibration curve for the HPLC peak from each standard solu-
tion is drawn (see Fig. S1 in Supplementary materials). After that,
from the extracted powder of each run, 5 ppm solution was made
and injected to HPLC. With the aid of calibration curve the cur-
cumin concentration in the sample solutions were determined. The
amount of curcumin (wt%) in turmeric was eventually calculated
as below:
Mass of curcumin (g)
Curcumin (wt%) = × 100
Mass of turmeric (g)
where,
Curcumin concentration (ppm)
Mass of curcumin (g) =
Extract concentration (ppm)
× Mass of extract (g)
In addition, in order to confirm that the extract powder con-
tained curcumin, the HPLC graph of 5 ppm solution of exemplary
extract was qualitatively compared with that of standard curcumin
solution. Comparing Fig. 2a and b, it was evidenced that both exem-
plary extract and standard curcumin solution presented a peak of
similar shape at the same retention time.
194 M. Valizadeh Kiamahalleh et al. / J. Chromatogr. B 1022 (2016) 191–198

Fig. 2. The HPLC curve (a) 2.5 ppm of standard curcumin (b) 5 ppm of extracted curcumin.

2.4. Scanning electron microscopy (SEM)

The SEM imaging was performed on a JEOL 8900 Electron Probe

Microanalyzer to visualize and characterize the surface of turmeric
sample particles. 5 nm coating of Au was applied to surface of the
samples before proceeding with imaging.

3. Results and discussion

For curcumin extraction, the effect of different parameters such

as the temperature, particle size, extraction time and the pressure
in the extraction vessel were investigated.
To determine the effect of temperature on SWE of curcumin
from turmeric, 40.0 g of turmeric was extracted at 120, 130, 140,
150 and 160 ◦ C; with constant particle size of 2 mm for 10 min at
10 bar. The total mass of extraction is determined by weighing the
extracts after freeze drying. The obtained data are shown in Fig. 3. Fig. 3. The effect of temperature on extraction.

The exact amounts (wt%) of curcumin were determined through

HPLC analysis. Results showed that the total mass of extraction
increased while the temperature was raised. The trend of extraction
with increasing temperature was linear. That was due to enhanced
properties of SWE such as solubility and diffusion lead to more
amount of curcumin to be extracted. Increasing the temperature
leads to a steady decrease in water permittivity, viscosity and sur-
face tension, but rise in its diffusivity properties which assisted
the extraction process [28,37]. There was an optimum tempera-
ture for maximum curcumin extraction. That was most probably
due to decrease in dielectric constant of water and tend to dis-
solve more curcumin. In addition, decrease in surface tension and
viscosity of water may assist to dissolve curcumin. The increasing
trend of curcumin extraction continued till the optimum temper-
ature of 140 ◦ C, where the wt% of extracted curcumin reached to
maximum of 1.2%, and after that the extraction was dropping (see
Fig. 3). The decrease in extraction at high temperature could be due
to degradation and hydrolysis of polyphenolic compounds exist in
curcumin.
The effect of particle size in SWE of curcumin was investigated.
A 40 g of turmeric in particle size of 2, 1, 0.71, and 0.6 mm was
extracted at 140 ◦ C, for 10 min at 10 bar. The total mass of extrac-
tion and wt% of curcumin were determined. Fig. 4 shows that the
total mass of extraction increased with the decrease in particle size
due to increase in surface area; while the curcumin wt% reached
to a maximum at a certain particle size of 0.71 mm. The content of
curcumin shows decreasing trends for the particle size larger and
smaller than 0.71 mm as the extraction process for the larger parti-
cles is more controlled by particle mass transfer and decomposition
of curcumin occurs in very small particle size. The best extraction of
curcumin obtained in optimum particle size of 0.71 mm was about
3% curcumin in turmeric.
In next step, it is necessary to define suitable extraction time as
prolonged duration may cause degradation of polyphenolic com-
pounds in turmeric. To determine the effect of time on SWE of
curcumin in turmeric, 40.0 g of turmeric in particle size 0.71 mm
was extracted at 140 ◦ C, for 6, 10, 14, 18 and 22 min at 10 bar.
The extractive mass and curcumint wt% were determined. Fig. 5
depicted that the total mass of extraction increased with respect to
extraction time, but the wt% of curcumin reached to a maximum of
about 3.8 at an optimum extraction time of 14 min, after that the
contents may be degraded. The difference in the polymerization,
 M. Valizadeh Kiamahalleh et al. / J. Chromatogr. B 1022 (2016) 191–198
Fig. 4. The effect of particle size on curcumin extraction.
Fig. 5. The effect of time on curcumin extraction.
Fig. 6. The effect of pressure on curcumin extraction.
solubility and the interaction between the phenolic components
caused to have different equilibrium time.
In order to determine the effect of pressure on SWE of cur-
cumin from turmeric, 40 g of turmeric in particle size of 0.71 mm
was extracted at 140 ◦ C, for 14 min at various pressures (10, 15, 20
and 25 bar). The wt% of curcumin and extractive mass were deter-
mined. Results showed that the total mass of extraction and the wt%
of curcumin had no significant changing and remain constant by
increasing pressure. As shown in Fig. 6, the pressure itself does not
195
Fig. 7. The maximum yield of curcumin extraction after optimizing each parameter.
have significant influence on extraction, but makes the extraction
easier because of the exerted forces on water molecules to pene-
trate inside the matrix of turmeric. Also the pressure is essential to
maintain the water in the phase of liquid.
While investigating all parameters, each parameter was opti-
mized individually; then, the yield, which is extracted curcumin
ratio against the maximum curcumin in turmeric (known to be
5%), was calculated. Fig. 7 shows the increasing trend of maximum
curcumin yield for one parameter to another throughout univari-
ate method. Investigating the temperature effects, the maximum
extraction yield was 24%, while keeping the temperature con-
stant at the optimum point and varying the particle size enhanced
extraction yield up to 60%. In fact, setting the temperature and the
particle size at their optimum level and changing the retention time
yielded 76% of curcumin extraction, whereas varying the pressure,
while other parameters kept constant, showed no enhancement
on the extraction yield. As a result, the parameters of temperature
and particle size and contact time were considerably influential
for improving the extraction yield, whereas the pressure effect
was negligible. Fig. 7 shows how the gradual improvements were
achieved as optimization of SWE of curcumin proceeded.
3.1. SEM analysis
In order to have a better interpretation of the curcumin extrac-
tion from turmeric powder, SEM images from the outer surface of
turmeric before and after extraction at the desired operating con-
dition of operation parameters were captured and compared as in
Figs. 8 and 9, respectively. The SEM images for the surface charac-
teristics such as morphology and porosity relating directly to the
extraction process on the turmeric powder were also analyzed.
Fig. 8 shows the surface of turmeric in two different scale 100×
and 500×. As illustrated from this figure the surface is hard and
without any porosity.
Fig. 9 is the SEM image of turmeric effluent after extraction at
140 ◦ C, particle size of 2.00 mm, 10 min and 10 bar. The surface
image surface shows that the water solution at mentioned con-
dition (at elevated temperature) has affected on the porosity and
hardness of turmeric and made the turmeric looser than before. It
can be proved that curcumin and other composition may easily be
extracted from these porous holes.
Fig. 10 shows the SEM image of turmeric at 140 ◦ C and parti-
cle size of 0.71 which was the optimum particle size at retention
time of 10 min and 10 bar. As compared with Fig. 9, the porosity of
turmeric was improved as curcumin extraction increased. The sur-
face is weakened compared to the previous setup condition. Small
particles size has been affected at high temperature and pressure
196 M. Valizadeh Kiamahalleh et al. / J. Chromatogr. B 1022 (2016) 191–198

Fig. 8. The SEM image of turmeric before extraction in particle size of 2 mm, (a) magnitude of 100× (b) magnitude of 500×.

Fig. 9. The SEM image of turmeric after extraction in the condition of 140 ◦ C, particle size of 2 mm, retention time of 10 min and the pressure of 10 bar: (a) magnitude of
100× (b) magnitude of 500×.

Fig. 10. The SEM image of turmeric after extraction in the condition of 140 ◦ C, particle size of 0.71 mm, retention time of 10 min and the pressure of 10 bar: (a) magnitude of
100× (b) magnitude of 500×.
 M. Valizadeh Kiamahalleh et al. / J. Chromatogr. B 1022 (2016) 191–198 197

Fig. 11. The SEM image of turmeric after extraction in the condition of 140 ◦ C, particle size of 0.71 mm, retention time of 14 min and the pressure of 10 bar: (a) magnitude of
100× (b) magnitude of 500×.

Fig. 12. The SEM image of turmeric after extraction in the condition of 140 ◦ C, particle size of 0.71 mm, retention time of 14 min and the pressure of 25 bar: (a) magnitude of
100× (b) magnitude of 500×.

and the surface changes were in harmonic shape. This structure
allows diffusion of water into the matrix and extracting the cur-
cumin.
After optimizing the temperature and particle size, the retention
time of extraction has been optimized and the curcumin content
increased in 14 min extraction. Fig. 11 shows the SEM image of
turmeric at 140 ◦ C, particle size of 0.71 mm and 14 min extraction
at 10 bar. In comparison with Fig. 10 for 10 min extraction, Fig. 11
shows that any part of turmeric surface change its arrangement
and the surface completely becomes looser than before by the more
sufficient retention time of 14 min. There is not any resistance of
water and after the sufficient time water diffuse into the turmeric
matrix and curcumin easily carrying out of turmeric and dissolved
into the water. This can be one of the reasons of improving curcumin
extraction.
As a result of the investigations of the effects of different param-
eters, pressure has no significant effect on extraction. In order to
observe the pressure effect on the morphology of the turmeric sur-
face, SEM images at a pressure of 25 bar with the other optimized
condition captured (see Fig. 12). Comparing Figs. 11 and 12, the
surface morphology of turmeric did not have any obvious change
while the pressure increased from 10 to 25 bar, confirming the
morphological stability of turmeric even at high pressure. These
similar morphologies could be further related to the similar cur-
cumin extraction yield at the conditions of 10 and 25 bar.
To summarize the work and to rationalize why the technique
used in this work is of the most convenient, the advantages and dis-
advantages of curcumin SWE are evaluated. As mentioned earlier,
SWE is able to extract substantially the water-soluble to liposol-
uble substances. Organic solvents contain some drawbacks, since
they are expensive, potentially harmful to the operator, and need
to be eventually disposed; whereas subcritical water showed to be
highly desirable from environmental and health perspectives.
A further advantage of SWE is that high temperature and pres-
sure produce a high diffusion rate, which promote very efficient
extraction of the raw material. In addition, that rate might vary
according to different chemical structures of organic compounds
being used. Hence, extraction with subcritical water could be selec-
tive with much high extraction rate [23]. On the contrary, SWE
bears some disadvantages which are the high operating pressure
198 M. Valizadeh Kiamahalleh et al. / J. Chromatogr. B 1022 (2016) 191–198
and the requirement to an expensive experimental setup to gener-
ate and handle high pressure system [26]. There are two inevitable
problems of using SWE which need to be considered as well for
the futures studies. Firstly, the nature of reactivity of impure water
that may be harmful for analyte because of the side-reaction in the
process of extraction could be prevented by the use of pure water
and gas separation. Secondly, the relatively high temperatures that
required firstly studies on the thermal stability of the extracted
compounds.
4. Conclusion
SWE unit was fabricated and tested for extraction of curcumin.
The method used in this work is an alternative to classical extrac-
tion as organic solvents are used. Use of water instead of organic
chemical should decrease the side effects of toxic organic sol-
vents. Also water is safe and available recommend for replacement
of organic solvents. In this work, several effective parameters
such as temperature, particle size, extraction time, pressure were
investigated. The results indicated that optimum temperature was
obtained at 140 ◦ C and above from that the curcumin as polyphe-
nolic component was degraded; therefore, the amount of curcumin
extraction decreased. The suitable particle size was defined as
0.71 mm. Also the best retention time and pressure obtained 14 min
and 10 bar, respectively. The HPLC analysis clarified that the purity
of extracted curcumin via SWE was quite high in compare to pure
curcumin sample. As it was assumed form the reference papers,
the maximum weight percent of curcumin is reported as 5%. The
highest percentages of curcumin extracted at optimum condition
was 3.8%. Based on maximum curcumin content, the obtained yield
of extraction was more than 76%. SEM images clearly showed
that the significant changes in morphology of turmeric texture by
influential parameters. Porosity and looseness of turmeric surface
structure had direct relationship with their curcumin extraction.
Acknowledgements
The authors gratefully acknowledge Biotechnology Research
Lab., Noshirvani University of Technology and School of Medicine,
Babol University of Medical Sciences for the facilities provided to
conduct present research work.
Appendix A. Supplementary data
Supplementary data associated with this article can be found,
in the online version, at http://dx.doi.org/10.1016/j.jchromb.2016.
04.021.
References
[1] S. Shishodia, G. Sethi, B.B. Aggarwal, Curcumin: getting back to the roots, Ann.
N. Y. Acad. Sci. 1056 (2005) 206–217.
[2] A.-L. Cheng, C.-H. Hsu, J.-K. Lin, M.-M. Hsu, Y.-F. Ho, T.-S. Shen, J.-Y. Ko, J.-T.
Lin, B.-R. Lin, W. Ming-Shiang, Phase I clinical trial of curcumin a
chemopreventive agent, in patients with high-risk or pre-malignant lesions,
Anticancer Res. 21 (2000) 2895–2900.
[3] H. Vogel, J. Pelletier, Curcumin-biological and medicinal properties, J. Pharma.
2 (1815) 50.
[4] J. Miłobȩdzka, V. Lampe, Zur Kenntnis des curcumins, Ber. Dtsch. Chem. Ges.
43 (1910) 2163–2170.
[5] L. Péret-Almeida, A. Cherubino, R. Alves, L. Dufosse, M. Gloria, Separation and
determination of the physico-chemical characteristics of curcumin,
demethoxycurcumin and bisdemethoxycurcumin, Food Res. Int. 38 (2005)
1039–1044.
[6] H. Hatcher, R. Planalp, J. Cho, F. Torti, S. Torti, Curcumin: from ancient
medicine to current clinical trials, Cell. Mol. Life Sci. 65 (2008) 1631–1652.
[7] A. Vlietinck, T. De Bruyne, S. Apers, L. Pieters, Plant-derived leading
compounds for chemotherapy of human immunodeficiency virus (HIV)
infection, Planta Med. 64 (1998) 97–109.
[8] W. Jordan, C. Drew, Curcumin–a natural herb with anti-HIV activity, J. Natl.
Med. Assoc. 88 (1996) 333.
View publication stats
[9] R. Sinha, D. Anderson, S. McDonald, P. Greenwald, Cancer risk and diet in
India, J. Postgrad. Med. 49 (2003) 222.
[10] C. Martins, D. Da Silva, A. Neres, T. Magalhaes, G. Watanabe, L. Modolo, A.
Sabino, A. De Fátima, M. De Resende, Curcumin as a promising antifungal of
clinical interest, J. Antimicrob. Chemother. 63 (2009) 337–339.
[11] M. Patil, S. Taralkar, V. Sakpal, S. Shewale, R. Sakpal, Extraction, isolation and
evaluation of anti-inflammatory activity of Curcuminoids from Curcuma
longa, Int. J. Chem. Sci. Appl. 2 (2011) 172–174.
[12] M. Abbasi, M. Ilyas, A. Sonia, D. Shahwar, M. Raza, K. Khan, M. Ashraf, I. Afzal,
N. Ambreen, Curcumin and its derivatives: moderate inhibitors of
acetylcholinesterase, butyrylcholinesterase and trypsin, Sci. Iranica 19 (2012)
1580–1583.
[13] Y. Cheng, H. Li, H. Wang, H. Sun, Y. Liu, S. Peng, K. Liu, Z. Guo, Inhibition of
nicotine-DNA adduct formation in mice by six dietary constituents, Food
Chem. Toxicol. 41 (2003) 1045–1050.
[14] M. Vietri, A. Pietrabissa, F. Mosca, R. Spisni, G. Pacifici, Curcumin is a potent
inhibitor of phenol sulfotransferase (SULT1A1) in human liver and
extrahepatic tissues, Xenobiotica 33 (2003) 357–363.
[15] Y. Chen, C. Ho, K. Cheng, Y. Tyan, C. Hung, T. Tan, J. Chung, Curcumin inhibited
the arylamines N-acetyltransferase activity, gene expression and DNA adduct
formation in human lung cancer cells (A549), Toxicol. Invitro 17 (2003)
323–333.
[16] M. Nabati, M. Mahkam, H. Heidari, Isolation and characterization of curcumin
from powdered rhizomes of turmeric plant marketed in Maragheh city of Iran
with soxhlet technique, Iran. Chem. Commun. 2 (2014) 236–243.
[17] V. Mandal, Y. Mohan, S. Hemalatha, Microwave assisted extraction of
curcumin by sample–solvent dual heating mechanism using Taguchi L 9
orthogonal design, J. Pharm. Biomed. Anal. 46 (2008) 322–327.
[18] P. Wakte, B. Sachin, A. Patil, D. Mohato, T. Band, D. Shinde, Optimization of
microwave, ultra-sonic and supercritical carbon dioxide assisted extraction
techniques for curcumin from Curcuma longa, Sep. Purif. Technol. 79 (2011)
50–55.
[19] M.A. Euterpio, C. Cavaliere, A.L. Capriotti, C. Crescenzi, Extending the
applicability of pressurized hot water extraction to compounds exhibiting
limited water solubility by pH control: curcumin from the turmeric rhizome,
Anal. Bioanal. Chem. 401 (2011) 2977–2985.
[20] N. Kurmudle, L.D. Kagliwal, S.B. Bankar, R.S. Singhal, Enzyme-assisted
extraction for enhanced yields of turmeric oleoresin and its constituents,
Food Biosci. 3 (2013) 36–41.
[21] A. Kubátová, A.J. Lagadec, D.J. Miller, S.B. Hawthorne, Selective extraction of
oxygenates from savory and peppermint using subcritical water, Flavour
Fragance J. 16 (2001) 64–73.
[22] S. Rovio, K. Hartonen, Y. Holm, R. Hiltunen, M.L. Riekkola, Extraction of clove
using pressurized hot water, Flavour Fragance J. 14 (1999) 399–404.
[23] M. Hassas Roudsari, Subcritical water extraction of antioxidant compounds
from canola meal, in: M.Sc. Thesis, University of Saskatchwwan, Canada, 2007.
[24] X. Liang, Q. Fan, Application of sub-critical water extraction in pharmaceutical
industry, J. Mater. Sci. Chem. Eng. 1 (2013) 1.
[25] L. Ramos, E. Kristenson, U.T. Brinkman, Current use of pressurised liquid
extraction and subcritical water extraction in environmental analysis, J.
Chromatogr. A 975 (2002) 3–29.
[26] R.M. Smith, Extractions with superheated water, J. Chromatogr. A 975 (2002)
31–46.
[27] K.S. Duba, A.A. Casazza, H.B. Mohamed, P. Perego, L. Fiori, Extraction of
polyphenols from grape skins and defatted grape seeds using subcritical
water: experiment and modeling, Food Bioprod. Process. 94 (2015) 29–38.
[28] C.C. Teo, S.N. Tan, J.W.H. Yong, C.S. Hew, E.S. Ong, Pressurized hot water
extraction (PHWE), J. Chromatogr. A 1217 (2010) 2484–2494.
[29] D. Luong, M.A. Sephton, J.S. Watson, Subcritical water extraction of organic
matter from sedimentary rocks, Anal. Chim. Acta 879 (2015) 48–57.
[30] E. Ibáñez, A. Kubátová, F.J. Señoráns, S. Cavero, G. Reglero, S.B. Hawthorne,
Subcritical water extraction of antioxidant compounds from rosemary plants,
J. Agric. Food Chem. 51 (2003) 375–382.
[31] L. He, X. Zhang, H. Xu, C. Xu, F. Yuan, Ž. Knez, Z. Novak, Y. Gao, Subcritical
water extraction of phenolic compounds from pomegranate (Punica granatum
L.) seed residues and investigation into their antioxidant activities with
HPLC–ABTS+ assay, Food Bioprod. Process. 90 (2012) 215–223.
[32] M. Tanaka, A. Takamizu, M. Hoshino, M. Sasaki, M. Goto, Extraction of dietary
fiber from Citrus junos peel with subcritical water, Food Bioprod. Process. 90
(2012) 180–186.
[33] M.P. Choi, K.K. Chan, H.W. Leung, C.W. Huie, Pressurized liquid extraction of
active ingredients (ginsenosides) from medicinal plants using non-ionic
surfactant solutions, J. Chromatogr. A 983 (2003) 153–162.
[34] Z. Piñeiro, M. Palma, C.G. Barroso, Determination of catechins by means of
extraction with pressurized liquids, J. Chromatogr. A 1026 (2004) 19–23.
[35] A. Shotipruk, J. Kiatsongserm, P. Pavasant, M. Goto, M. Sasaki, Pressurized hot
water extraction of anthraquinones from the roots of Morinda citrifolia,
Biotechnol. Prog. 20 (2004) 1872–1875.
[36] L.J. Lamm, Y. Yang, Off-line coupling of subcritical water extraction with
subcritical water chromatography via a sorbent trap and thermal desorption,
Anal. Chem. 75 (2003) 2237–2242.
[37] S.N. Jha, K. Narsaiah, A.L. Basediya, R. Sharma, P. Jaiswal, R. Kumar, R.
Bhardwaj, Measurement techniques and application of electrical properties
for nondestructive quality evaluation of foods—a review, J. Food Sci. Technol.
48 (2011) 387–411.