Food Chemistry 336 (2021) 127660

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Food Chemistry
journal homepage: www.elsevier.com/locate/foodchem

Solubilization and extraction of curcumin from Curcuma Longa using green, T

sustainable, and food-approved surfactant-free microemulsions
⁎
Pierre Degot ,1, Verena Huber1, Evamaria Hofmann, Maximilian Hahn, Didier Touraud,
⁎
Werner Kunz
Institute of Physical and Theoretical Chemistry, University of Regensburg, D-93040 Regensburg, Germany

A R T I C LE I N FO A B S T R A C T

Chemical Compounds in this article: Curcumin is a powerful coloring agent widely used in the food industry. Its extraction from the plant Curcuma
Ethanol (PubChem CID: 702) longa is commonly done with aqueous solvent solutions. In contrast to the conventional extraction methods, the
Triacetin (PubChem CID: 5541) present study aimed to compare two different green and bio-based surfactant-free microemulsion (SFME) ex-
Diacetin (PubChem CID: 66021) traction systems, which are approved for food and yield a higher extracting power of curcuminoids. Two SFMEs,
Curcumin (PubChem CID: 969516)
water/ethanol/triacetin and water/diacetin/triacetin, were investigated via dynamic light scattering. Curcumin
Demethoxycurcumin (PubChem CID: 5469424)
Bisdemethoxycurcumin (PubChem CID:
solubility in binary mixtures consisting of ethanol/triacetin or diacetin/triacetin was studied both experimen-
5315472) tally and theoretically using UV–Vis measurements and COSMO-RS. The SFMEs were further examined and
compared to a common ethanol/water (80/20) extraction mixture with respect to their extracting ability using
Keywords: high performance liquid chromatography. The SFMEs containing ethanol were found to extract ~18% more
Curcumin curcuminoids than the SFMEs containing diacetin and ~53% more than the ordinary ethanol/water mixture.
Surfactant-free microemulsions
Extraction
COSMO-RS
Green chemistry
Food

1. Introduction Nowadays, various methods to extract curcuminoids from different

1.1. Curcumin
Curcumin has widely been used for centuries in Asia, particularly in
India, as a spice, preservative, food colorant and therapeutic agent
(Nelson et al., 2017; Wilken, Veena, Wang, & Srivatsan, 2011; Wilson
et al., 2005). Curcumin is the principal curcuminoid of the turmeric
Curcuma Longa (C. Longa), a member of the ginger family (Zingiber-
aceae). Curcuminoids are responsible for the bright yellow color of the
plant’s rhizomes. Three different curcuminoids can be found in C.
Longa: curcumin I, known as curcumin, curcumin II, known as de-
methoxycurcumin and curcumin III, known as bisdemethoxycurcumin
(Ali, Haque, & Saleem, 2014; Geethanjali, Lalitha, & Firdhouse, 2016;
Taylor & McDowell, 1992). They can be distinguished from each other
by the number of methoxy groups, as shown in Fig. 1. The curcuminoids
are highly soluble in oil and insoluble in water.
varieties of the plant C. Longa such as Soxhlet, solvent, ultrasonic, su-
percritical carbon dioxide, and microwave extraction are found in lit-
erature. What they all have in common is the use of conventionally non-
sustainable solvents. Amongst others, methanol and aqueous solutions
thereof, as well as acetone and petroleum ether can be found in lit-
erature as extracting solvents. Since a lot of them are not edible and
toxic for the human body, it is necessary to remove them after extrac-
tion, e.g. via evaporation. A contemporarily used, healthier alternative
in the industry are ethanol and aqueous ethanol solutions (here espe-
cially the mixture ethanol/water 8:2). These mixtures with ethanol on
the other hand are not harmful to humans. However, the extraction
yield (as can be seen in Section 3.3) needs improvement. Therefore, the
addition of triacetin to an ethanol water mixture can be of benefit for
the extraction yield, while still being food approved (Baumann,
Rodrigues, & Viana, 2000; Chassagnez-Méndez, Corrêa, França,
Machado, & Araújo, 2000; Chemat et al., 2017; Jansirani, Saradha,

⁎
Corresponding authors.
E-mail addresses: pierre-emile-lucien.degot@ur.de (P. Degot), verena1.huber@ur.de (V. Huber), evamaria.hofmann@ur.de (E. Hofmann),
maximilian.hahn@ur.de (M. Hahn), didier.touraud@ur.de (D. Touraud), werner.kunz@ur.de (W. Kunz).
1
Authors contributed equally to the experimental work and to writing the paper.

https://doi.org/10.1016/j.foodchem.2020.127660
Received 27 May 2020; Received in revised form 21 July 2020; Accepted 21 July 2020
Available online 03 August 2020
0308-8146/ © 2020 Elsevier Ltd. All rights reserved.
P. Degot, et al.
Fig. 1. Structure of curcumin I (bottom), II (middle), and III (top).
Salomideborani, & Selvapriyadharshini, 2014; Mandal, Mohan, &
Hemalatha, 2008).
HPLC, sometimes combined with electrospray ionization and mass
spectrometry, spectrophotometric methods, capillary electrophoresis or
simple nuclear magnetic resonance methods have been established to
detect and/or determine the content of curcumin, demethoxycurcumin
and bisdemethoxycurcumin (Geethanjali et al., 2016; Jayaprakasha,
Rao, & Sakariah, 2002; Marczylo, Steward, & Gescher, 2009; Nugroho,
Rohman, Lukitaningsih, Rakhmawati, & Sudjadi, 2015; Sun, Gao, Cao,
Yang, & Wang, 2002; Taylor & McDowell, 1992; Tayyem, Heath, Al-
Delaimy, & Rock, 2006; Wichitnithad, Jongaroonngamsang,
Pummangura, & Rojsitthisak, 2009). In the present study, it was at-
tempted to develop an energy efficient, simple, one-step solvent ex-
traction with a novel extraction system. The idea was to replace the
potentially harmful organic solvents by food agreed SFMEs, which can
yield even better results when extracting curcuminoids from the ground
turmeric rhizomes while being more energy efficient and effortless than
the conventional methods. To validate this hypothesis the extracts were
analyzed via HPLC.
The development of natural dyes is more and more important and
crucial for food industry. One reason is that the use of synthetic dyes
does not match with the current consumers’ perception of safety and
health of food formulations. Especially tartrazine, one of the most
prominent representatives of synthetic azo-dyes, often faces criticism
due to its effects on human health. It can be responsible for pseudo
allergic reactions either via oral or cutaneous intake. The synthetic azo-
dyes, however, were declared passably safe for food as long as the daily
intake does not exceed 0–7.5 mg/kg body weight. Thus, they are still
permitted in food formulations, which have to carry a warning label
(Amchova, Kotolova, & Ruda-Kucerova, 2015; Choulis, 2010; Food
Standards Agency, 2009) Curcumin on the other hand is basically non-
toxic, biodegradable and its degradation products are inactive
(Schneider, Gordon, Edwards, & Luis, 2015). The major problem with
curcumin is its insolubility in water and its instability against en-
vironmental impacts like UV radiation and elevated temperatures
amongst others, which are limiting factors for its extraction and for-
mulation (Chignell et al., 1994; Tønnesen, Karlsen, & Henegouwen,
1986).
1.2. Surfactant-free microemulsions (SFME)
Generally, microemulsions consist of oil, water, surfactant, and
2
Food Chemistry 336 (2021) 127660
sometimes a cosurfactant. They are macroscopically homogenous li-
quids and thermodynamically stable solutions due to the presence of a
surfactant and/or a co-surfactant. In a non-conventional way, micro-
emulsions can also be formed without surfactant or co-surfactant. Those
microemulsions are called SFME and consist of oil, water, and a co-
solvent or hydrotrope, which is miscible with water and oil. The
structuring of SFME close to the liquid–liquid phase separation border
is an intriguing phenomenon and can be used to perform extractions.
Indeed, as will be shown in this paper, the structuring in the extraction
media could improve the yield of the extraction of the target compound
curcumin. Green SFMEs with only food-admitted components have a
considerable industrial potential, particularly for the food industry.
Zhang, Song, Chai, Cui, and Wang (2020) tried to replace the com-
monly used petrol-based extraction solvents by SFMEs consisting of 1-
octen-3-ol, ethanol (EtOH), and water (Zhang et al., 2020). However,
the oil phase they employed can only be used to a small extent since it is
only approved as a synthetic flavoring or adjuvant by the FDA (US Food
and Drug Administration) (Burdock & Fenaroli, 2016). Due to the new
consumers’ perception of safety and health of food formulations, the use
of naturally derived SFMEs as an extraction system and/or formulation
is a big advantage. Thus, the constituents of the SFMEs for this study
were chosen regarding the regulations in the food industry. The natural
triglyceride triacetin (TriA) was used as the oil phase of the SFME. This
component has thoroughly been investigated by the FDA and EFSA
(European Food Safety Authority) and eventually was generally re-
cognized as safe human food ingredient by the FDA (European Food
Safety Authority, n.d.). Further assessment of the potential toxicity of
TriA led to the conclusion that it most certainly degrades to acetic acid
and glycerol by hydrolysis and is further broken down to carbon di-
oxide. Even upon ingestion, the human body is able to easily metabolize
it within the β-oxidation of fatty acids, highlighting the biocompat-
ibility of TriA (Quinn & Ziolkowski, 2015). Same is true for its deri-
vative DiA, which is used as a co-solvent in this study. Ethanol, which is
freely available and consumable in alcoholic beverages, was the second
co-solvent investigated in this study. It can easily be metabolized in the
human body and is nowadays mainly made from biomass, guaranteeing
a sustainable production (Patel, Crank, Dornburg, Hermann, & Roes,
2004). Comparing this assessment of the used components with the
principles of green chemistry postulated by Anastas, P. (Anastas &
Eghbali, 2010) and the six principles of green extraction as specified by
Chemat, Vian, & Cravotto (2012), we may classify our SFMEs as green,
biocompatible and renewable extraction media.
2. Materials and methods
2.1. Chemicals
The synthetic phytochemical curcumin (purity > 97%) was bought
from TCI (Eschborn, Germany), demethoxycurcumin and bisde-
methoxycurcumin from Sigma Aldrich (Darmstadt, Germany). The
dried rhizome powder of C. Longa. was purchased from Kwizda (Linz,
Austria). The solvents, acetone (purity > 99%), ethanol (EtOH,
purity > 99%), acetonitrile (purity > 99%, HPLC grade), acetic acid
(purity > 99%), triacetin (TriA, purity > 99%, food-grade), and
diacetin (DiA, purity > 99%) were purchased from Merck (Darmstadt,
Germany). All chemicals were used without further purification.
2.2. Methods
2.2.1. Ternary phase diagrams
To obtain the phase diagram of water/EtOH/TriA and water/DiA/
TriA, binary mixtures composed of either water/EtOH or EtOH/TriA or
water/Dia or DiA/TriA with different mass ratios were prepared in a
total amount of 3 g in borosilicate tubes. The third component (re-
spectively TriA and water) was added dropwise to the binary mixture
until the solution became turbid. The needed amount of the third
P. Degot, et al.
component was recorded in order to determine the miscibility gap.
2.2.2. UV–Vis spectroscopy
Optical density measurements were carried out via UV/Vis, using a
Lambda 18 UV/Vis spectrometer by Perkin Elmer (Waltham, USA).
Solutions saturated with synthetic curcumin were examined to find the
optimum binary solvent and SFME compositions. Before the measure-
ment, all samples were diluted 1000-fold with acetone.
2.2.3. Conductor-like screening model for real solvents
2.2.3.1. COSMO-RS theory. The COSMO-RS theory (COnductor like
Screening MOdel for Real Solvents) links quantum chemistry with fast
statistical thermodynamics and thus enables a great variety of
physicochemical properties to be predicted. In a nutshell, COSMO-RS
utilizes the state of a molecule embedded in a virtual conductor with an
infinite dielectric as a new reference state. After an initial calculation of
the screening charge density σ on the surface of a molecular shaped
cavity around each molecule (COSMO surface), COSMO-RS divides this
COSMO surface into smaller segments and applies fast statistical
thermodynamics. Following the assumption that all relevant
molecular interactions can be displayed by local pairwise interacting
COSMO surface segments, a broad range of physicochemical properties
can be computed (Klamt, Krooshof, & Taylor, 2002; Klamt, 2005). As
shown in Eq. (1), the chemical potential of a solute in a solvent μsi can
also be derived from the σ–potentials μs (σ ) and σ–profiles pi (σ ) of the
molecules. The σ–profile is a probability distribution that is
characteristic for each molecule and evaluates the amount of COSMO
surface segments with a specific polarity. The σ–potential describes the
affinity of the system to surface segments with a specific polarity. The
combinatorial term μci, s takes size and shape differences of the present
molecules into account (COSMOlogic GmbH Co. KG., 2019; Klamt,
2005).
μsi = ∫ pi (σ ) μs (σ ) + μci,s (1)
2.2.3.2. COSMO-RS calculations. The COSMOthermX (COSMOlogic
GmbH & Co. KG - a Dassault Systèmes company, 2019) software
(version 19.0.4 by COSMOlogic), which is based on COSMO-RS
theory (Klamt, Volker, Bürger, & Lohrenz, 1998; Klamt, 1995), was
applied to compute the chemical potential of curcumin in binary
mixtures consisting of EtOH and TriA in different ratios. For this
purpose, calculations of the chemical potential at infinite dilution
have been performed on TZVPD-FINE level. Molecules were taken
from the COSMObase TZVPD-FINE 19.0 database.
2.2.4. Dynamic light scattering
Dynamic light scattering (DLS) measurements were performed with
an ALV/CGS-3 goniometer with an ALV/LSE-5004 correlator at room
temperature (25 °C). To remove dust and any impurities, the solutions
were filtered with 0.2 μm PTFE membrane filters before the measure-
ment. All measured solutions were recorded three times for 300 s.
2.2.5. HPLC method
The chemical composition of the rhizome extracts of C. Longa was
analyzed by HPLC using a Waters HPLC system with two Waters 515
HPLC pumps, Waters 717 autosampler and Waters 2487 UV/Vis de-
tector and an ACE Equivalent 3 C18-Column (300 Å, 150 × 2.1 mm).
10 μL of the samples were injected and eluted at a flow of 0.4 mL/min
with a gradient method at 40 °C. Each sample was made in triplicate
and eluted three times. The mobile phase consisted of water with 0.3%
of acetic acid (A) and acetonitrile (B). Table S1 of the supplementary
shows the used gradient program (Curcuma longa (rhizome) - AHPA
Botanical Identity References Compendium, n.d.).
A calibration curve of the three different curcuminoids was done in
the concentration range of 0.04–0.2 mg/mL, as shown in Figure S 1 in
3
Food Chemistry 336 (2021) 127660
the supplementary material (S.M.). The elution was carried out as de-
scribed in Table S1 in the S.M. and the calibration curve was obtained
by plotting the concentration of curcumin, demethoxycurcumin, and
bisdemethoxycurcumin versus the peak area (average of three runs of
three calibration curves for each type of curcumin).
2.2.6. Determination of the curcumin content
2.0 g of turmeric powder was extracted with 50–60 mL of acetone as
extracting solvent using a Soxhlet apparatus. The extraction was carried
out in triplicates. The extract solutions were then put in a volumetric
flask, topped off with acetonitrile, diluted 50-fold, and eluted by HPLC
(three times). The curcumin content was determined using a calibration
curve.
2.2.7. Determination of the best solvent composition
Firstly, the maximum solubility of curcumin in the binary system
(EtOH/water, EtOH/TriA and DiA/TriA) was examined to find the best
binary composition. Then water was added to this optimum binary
mixture and the maximum solubility of curcumin was examined again.
Curcumin was added to the solvent mixtures (5 g each sample) under
constant stirring at 750 rpm and room temperature, until saturation
was reached. Then the saturated solutions were filtered and analyzed
via UV/Vis.
2.2.8. Determination of the best ratio of turmeric powder to extraction
mixture
4 g of turmeric powder was extracted with varying amounts of ex-
tracting solvent at room temperature under constant stirring at 750 rpm
for one hour. Different weight ratios from 1:2 to 1:7 (turmeric
powder / extraction mixture, w/w) were investigated with the binary
mixture EtOH/TriA 40/60 (w/w). All the extract solutions were cen-
trifuged (4200 g 10 min), filtered, put in a volumetric flask, topped off
with acetonitrile, diluted 25-fold, and then eluted by HPLC. All samples
were made in triplicates and each sample was eluted three times. The
curcumin content of each extract solution was determined.
2.2.9. Extraction procedure
4 g of turmeric powder was extracted with 16 g of different ex-
traction mixture compositions (ratio 1:4) under constant stirring at
750 rpm at room temperature for 1 h. The extract solutions were cen-
trifuged, filtered and then put in a volumetric flask, topped off with
acetonitrile, diluted 25–fold and then eluted by HPLC. All samples were
made in triplicates and each sample was eluted three times. The cur-
cumin content of each extract solution was determined.
3. Results and discussion
3.1. Ternary diagrams and determination of the best solvent composition
The systems examined in this study were chosen concerning their
edibility, sustainability, and low toxicity. They were investigated fo-
cusing on their mixing behavior and also to check, if they possess
structuring close to the phase border. TriA, which is used as the oil
phase, is slightly miscible with water. The realms of complete mis-
cibility of TriA with water solubilized by the co-solvents EtOH and DiA
are depicted in Figure
Fig. 2a) and b). In weight ratios (black dotted line), the EtOH system
shows a much smaller two-phase region. However, regarding the mis-
cibility gap in the molar ratio (orange dotted line) the biphasic region in
both systems covers approximately the same area. Therefore, EtOH and
DiA can be assumed to be equally potent in solubilizing water and TriA.
In literature, mainly non-green solvents like methanol or petroleum
ether are used for the extraction of curcumin (Jansirani et al., 2014;
Priyadarsini, 2014). To start, the solubility of curcumin in binary sol-
vent mixtures of hydrotrope/TriA in comparison to the pure solvents
was examined, followed by the solubility in ternary mixtures of water/
P. Degot, et al.
Fig. 2. Ternary Phase Diagrams of the Systems a) H2O/EtOH/TriA and b) H2O/DiA/TriA in wt% (black) and mol% (orange) containing the examined data points
(blue for the EtOH and red for the DiA systems) and qualitative determination of the maximum solubility in the c) binary and d) ternary EtOH (blue) and DiA (red)
systems starting from a weight ratio of 4:6 EtOH/TriA and 6:4 DiA/TriA. (For interpretation of the references to color in this figure legend, the reader is referred to
the web version of this article.)
hydrotrope/TriA.
Fig. 2c) shows the qualitative solubility of curcumin I in the binary
mixtures. Remarkably, the mixing of EtOH with TriA showed a strong
solubilizing synergy. Thus, the solubility of curcumin could be in-
creased 3-fold in a 4:6 mixture of EtOH/TriA, respectively to pure TriA
and even more with respect to pure ethanol. Therefore, and for reasons
of viscosity, 4:6 was determined as the optimum binary composition in
the EtOH/TriA system. The different binary mixtures of DiA and TriA,
on the contrary, did not show a pronounced synergy effect.
The qualitative UV findings are verified by COSMO-RS calculations
(see Fig. 3a). The predicted chemical potential of curcumin at infinite
dilution in the binary mixtures depicts a minimum with a composition
of 2:8 EtOH/TriA, which indicates a high affinity of the solute curcumin
for this mixture. This is in good agreement with experimental results
showing maximum absorbance in the mixtures of 4:6 to 2:8 EtOH/TriA.
Considering the individual σ-profiles (see Fig. 3b), it is noticeable that
TriA (blue curve) has a broad peak in the non-polar area around 0 e/Å2.
A further peak at the right side of the diagram represents the highly
polar and potentially hydrogen bonding surface segments and hence,
the hydrogen bond acceptor abilities of the molecule. The negatively
charged segments of the oxygen atoms appear at positive σ due to the
sign inversion from the molecular polarity to the screening charge
density σ. Since there is a lack of surface segments within the σ-range
from −0.010 to −0.020 e/Å2 that could interact properly with the
corresponding peak on the right side, an electrostatic misfit is given in
pure TriA. The rather symmetrical σ-profile of ethanol provides highly
4
Food Chemistry 336 (2021) 127660
polar and potentially hydrogen bonding surface segments of opposite
polarities in a balanced ratio. Accordingly, the addition of ethanol to
TriA reduces the electrostatic misfit. Strong hydrogen bonds between
ethanol and TriA molecules can be formed since peaks beyond ± 0.01
e/Å2 (dashed lines) are sufficiently polar. The molecule curcumin
mainly consists of a non-polar surface leading to significant Van-der-
Waals interactions with TriA. In addition, the curcumin molecule de-
picts not only hydrogen bond acceptor abilities but also hydrogen bond
donor abilities. Hence, curcumin and TriA can interact via hydrogen
bonding as well. The existence of hydrogen bonding and significant
Van-der-Waals interactions results in good solubility of curcumin in
pure TriA. In the presence of curcumin, the addition of ethanol not only
reduces the electrostatic misfit of TriA, further hydrogen bonds between
curcumin and ethanol molecules are possible as well. A significant
decrease in the chemical potential of curcumin can be noticed with
increasing hydrogen bond donor/acceptor ratio due to the addition of
ethanol. It thus seems reasonable that an optimum interplay of Van-der-
Waals, electrostatic, and hydrogen bonding interactions in a 2:8 mix-
ture of EtOH/TriA results in such a strong synergetic effect in solubi-
lizing curcumin. Besides, the small size of the ethanol molecules may
allow faster penetration to separate, and therefore, dissolve individual
curcumin molecules.
Although there is a free hydroxy group, as it is the case with EtOH,
DiA does not deliver the same effect. The molecule has significantly
more hydrogen bond acceptor than hydrogen bond donor abilities and
besides, is too bulky to allow the same rapid penetration as ethanol.
P. Degot, et al. Food Chemistry 336 (2021) 127660

Fig. 3. (a) Predicted chemical potential of the solute curcumin at infinite dilution in mixtures of EtOH/TriA with different weight ratios (black line) compared to
experimentally determined maximum absorbance of curcumin. (b) Predicted σ-profiles of curcumin (orange curve), TriA (blue curve), and EtOH (green curve). The
contributions of the individual conformers are weighted by their weight factor given in the mixture at the minimum of the chemical potential. (c) σ-surfaces of the
corresponding molecules in their C0 conformation. Green segments indicate segments with non-polar σ, while the red segments exhibit positive and the blue ones
exhibit negative σ. (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.)

Further, COSMO-RS calculations (see S.M., Fig. S 3) confirm the ex-
perimental observations. Only negligible synergy is predicted, which
may not be present in experiments due to the difficult handling of the
highly viscous samples and due to the lower purity of DiA. A compo-
sition of 6:4 DiA/TriA was thus chosen as examining composition.
Starting from the chosen compositions in the binary mixtures, the
solubility of curcumin was investigated upon addition of water. As
expected, the solubility in both systems followed a trend as can be seen
in Fig. 2d). The solubility of curcumin decreased with a rising amount
of water. However, in the DiA system, it was found that the solubility of
curcumin firstly increased and then decreased with more water, which
is particularly interesting, since curcumin is not soluble in water
(Hartmann-Schreier, n.d.). This find gave the idea that extraction with
an addition of water in the solvent system could be beneficial for the
yield. Therefore, extraction experiments with different amounts of
water were conducted in the progress to find the optimum composition
of the extraction solvent (cf. Section 3.3, Extraction Process).
correlating curve could be detected. This hinted at some loose asso-
ciations between EtOH and TriA, but no real, fully-grown aggregates.
Starting from the optimum binary compositions, the solvent beha-
vior of the ternary solutions was investigated. And as can be seen in
Figure S 2 c + d of the S.M., clear correlation functions could be de-
tected upon addition of water. The structuring in the DiA system was
way more pronounced than in the EtOH system. The detection of such
pronounced correlation functions indicated the generation of SFMEs in
both ternary systems. With an increasing amount of water, the x-in-
tercept of the correlation curves was shifted to longer lag times, in-
dicating bigger structures close to the miscibility gap. Unsurprisingly,
the correlation curves of the ternary DiA system were shifted to even
longer lag times as compared to the EtOH system. This can easily be
explained since DiA is bulkier than EtOH. Therefore, the aggregates of
DiA/TriA have to be larger than for EtOH/TriA. These findings suggest
that these ternary systems are indeed SFMEs.

3.3. Extraction process

3.2. Dynamic light scattering for the determination of SFMEs
The behavior of the optimum extraction compositions of 4:6 EtOH/
TriA and 6:4 DiA/TriA were examined via DLS measurements. The
corresponding correlation curves can be seen in the S.M. First, the
binary hydrotrope/oil mixtures were investigated upon structuring.
However, no real indication of any structuring could be detected (cf. SI,
Fig. S 2 a + b). For the EtOH/TriA system, depicted in Fig. S 2 a) of the
S.M., only a slight deviation of the symmetric shape of the non-
A total of 17.13 mg of total curcumin (curcumin, demethox-
ycurcumin, and bisdemethoxycurcumin) per gram of the purchased C.
Longa powder was confirmed by Soxhlet extraction. This value was set
to be 100% yield. However, curcumin degrades by the impact of heat,
therefore, the determined total curcumin amount is undervalued. This
is especially true for bisdemethoxycurcumin, which is the most heat
sensitive curcuminoid. It is also the most polar constituent of the cur-
cuminoids, which makes it even harder to determine its total content

5
P. Degot, et al. Food Chemistry 336 (2021) 127660

the most polar and water-soluble curcuminoid. The different solubilities

Fig. 4. Yield of the different forms of curcumin for varying powder to solvent
ratios 100% yield of the Soxhlet extract is depicted on the right bar.
via Soxhlet extraction with acetone. This is confirmed in Fig. 4, as the
amount of bisdemethoxycurcumin in our newly developed, more gentle
extraction mixture, was up to a quarter higher than in the Soxhlet
samples.
In a first attempt of extraction, the optimum powder to solvent ratio
was examined and evaluated qualitatively and quantitatively by HPLC.
These experiments were conducted exemplarily with a binary extrac-
tion mixture of EtOH/TriA 4:6. The highest yield could be achieved by a
powder to solvent ratio of 1:6. However, this ratio only achieved a 7%
increase in the extraction yield in comparison to a ratio of 1:4.
Therefore, a ratio of 1:4 was chosen to be the most profitable and green,
as the benefit of the highest yielding ratio was not sufficient regarding
the waste of solvent. Thus, all further extraction experiments were
conducted using a 1:4 powder to solvent ratio. The results for the binary
mixture were used as a basis for all following extraction experiments
with varying amounts of water.
In Table 1, the yield of every curcuminoid, as obtained by HPLC, is
shown. The amount obtained with the Soxhlet method was used as a
reference. It provided the highest results for curcumin and demethox-
ycurcumin. However, it cannot be guaranteed that the amount of cur-
cuminoids obtained by Soxhlet meets the total content of curcuminoids
in the rhizomes, as they are sensitive to elevated temperatures and start
to degrade during the process.
The addition of water to the binary mixtures of EtOH/TriA resulted
in an increase in the extraction yield of the three different curcumi-
noids. This is especially the case for bisdemethoxycurcumin, which is
of the three forms of curcumin are also predicted by COSMO-RS cal-
culations (see SI, Fig. S 4 and 5). Surprisingly, DiA as a pure solvent was
very potent in extracting curcumin. However, the extraction media
containing EtOH showed superior extraction yields compared to the
mixtures with DiA or pure DiA. This is due to different reasons: first of
all, the maximum solubility of curcumin is higher in the SFMEs and
binary mixtures with EtOH, secondly the viscosity is much higher when
using DiA compared to EtOH. The best extraction medium surprisingly
is the SFME of TriA/EtOH/water with 40 percent of water, although
curcumin is completely water-insoluble. This mixture is also the most
structured SFME according to dynamic light scattering measurements.
The addition of water helps to swell the rhizomes to open the plant
matrix. Here again the SFMEs with EtOH showed a better extraction
yield of the curcuminoids than the SFMEs with DiA did, probably be-
cause of viscosity reasons. All the SFMEs and the binary mixtures
showed a better extraction yield than the pure solvents or the common
extraction medium EtOH (80)/water (20) (Sasikumar, 2001). Figure S 6
of the Supplementary shows that the maximum solubility of curcumin
in aqueous solutions of ethanol is of course lower than its solubility in
the pure solvent, as curcumin is not water soluble. However, by adding
20 wt% of water to ethanol the extraction yield of the total curcumi-
noids can be increased. This proves that the extraction is not solely
dependent on the solubility of the targeted compound in the extraction
solvent, but is driven by various other factors. As has already been
investigated by Jacotet-Navarro, M. et al., the extraction can be influ-
enced by the penetration of the plant matrix by the solvent, the deso-
rption of the target compounds, and the mass transfer of the solute from
the plant matrix to the solvent (Jacotet-Navarro et al., 2018). These
finds can also be applied to the SFME extractions. Here we found that
the mixture of EtOH and TriA already results in an extraction yield that
is twice as high as the extraction yield for the pure compounds and the
EtOH/water mixture. Combining these results with the results of Ja-
cotet-Navarro et al., we found that the extraction ability of the oil
phase, consisting of TriA and EtOH, can even be improved by ~8% by
simply adding water. Therefore, the SFME makes for an optimum, su-
perior extraction solvent, which is food approved and not harmful to
the human body. Additionally, double the amount of water is used in
the SFME than compared to the EtOH/water 8:2 mixture. This again
agrees with the postulations of green extraction by Chemat et al., who
called for water-based extraction systems (Chemat et al., 2012).
Regarding the six principles of green extraction, as specified by
Chemat et al. (2012), our SFME extraction system may be classified as
green and biocompatible. Indeed, Curcuma L. has been used as raw
material, which is a renewable plant resource. The developed SFMEs
contain only food approved, non-toxic and bio-degradable solvents and
especially water. The organic part of the SFME, consisting of EtOH and
TriA, is responsible for the formation of the SFME and especially for the

Table 1
Curcumin extracted from C. longa with varying solvent compositions in mg/g rhizome.
Extraction System Curcumin (mg/g) Demethoxycurcumin (mg/g) Bisdemethoxycurcumin (mg/g)

Soxhlet 11.64 ± 0.70 3.82 ± 0.21 1.67 ± 0.26

TriA/EtOH 60/40 8.82 ± 0.11 2.78 ± 0.04 2.19 ± 0.04
TriA/EtOH/H2O 57/38/5 8.98 ± 0.16 2.85 ± 0.04 2.29 ± 0.04
TriA/EtOH/ H2O 51/34/15 8.97 ± 0.21 2.97 ± 0.07 2.58 ± 0.10
TriA/EtOH/ H2O 42/28/30 8.96 ± 0.25 3.07 ± 0.09 2.83 ± 0.06
TriA/EtOH/ H2O 36/24/40 9.21 ± 0.32 3.18 ± 0.23 2.89 ± 0.38
TriA/DiA 40/60 7.19 ± 0.13 2.25 ± 0.04 1.67 ± 0.06
TriA/DiA/ H2O 38/57/5 7.54 ± 0.06 2.36 ± 0.02 1.81 ± 0.01
TriA/DiA/ H2O 34/51/15 6.97 ± 0.28 2.42 ± 0.11 2.15 ± 0.10
TriA/DiA/ H2O 30/45/25 7.1 ± 0.34 2.42 ± 0.11 2.32 ± 0.14
EtOH/H2O 80/20 4.36 ± 0.14 1.43 ± 0.05 1.03 ± 0.10
EtOH 4.34 ± 0.08 1.34 ± 0.05 0.92 ± 0.04
DiA 7.41 ± 0.04 2.28 ± 0.02 1.61 ± 0.01
TriA 3.13 ± 0.21 0.94 ± 0.06 0.51 ± 0.06

6
P. Degot, et al.
solubilization of the curcuminoids in the mixture. The combination
thereof is by far more efficient in terms of solubilization and extraction
than the mixture of EtOH and water on their own. The used extraction
procedure is performed at room temperature for 1 h, with no need to
heat or to stir for hours (high energy consumption) in order to have a
better extraction yield than common solvents or common solvent
mixtures. The principle and big advantage of this extraction procedure
is the use of a food approved SFME. This means, there is no need to
remove the solvent as the SFME is completely food grade and edible.
Moreover, the only produced waste is the remaining plant material
(Curcuma L.) which again is bio-degradable.
4. Conclusion
Two different food-approved and edible SFMEs consisting of water/
hydrotrope/TriA were examined for their ability to extract curcumin
from C. Longa. The hydrotropes in question were EtOH and DiA. It was
found that both hydrotropes have a similar solubilization power of
water with oil and by using DLS measurements the existence of SFMEs
close to the miscibility gaps for both systems was confirmed.
UV/Vis measurements were employed to examine the hydrotrope/
TriA binary mixtures. In doing so, a solubilizing synergy of curcumin
was found in the binary EtOH/TriA mixture resulting in a threefold
increase of curcumin solubility in a range of 2:8–4:6 EtOH/TriA as
compared to pure TriA. This find was validated by COSMO-RS simu-
lations and can be explained due to the polarity of the solvent mixtures
and the curcuminoids. COSMO-RS also predicted a less pronounced
synergy for the DiA/TriA systems, which could not be found in the UV/
Vis experiments due to viscosity reasons.
Upon addition of water, a decrease in curcumin solubility was found
for both ternary systems. However, an increasing amount of water in
the extraction systems led to an improved extraction power of curcu-
minoids from C. Longa. With a rising amount of water, the extraction
yields of the curcuminoids increased in the following way:
Curcumin < Demethoxycurcumin < Bisdemethoxycurcumin. This
can be explained by taking the COSMO-RS data into account, as the
polarity of the curcuminoids increases in the same order. Hence, the
more polar curcuminoids are better extracted in extracting mixtures
made more polar by addition of water. This explanation is also true for
the comparison of EtOH and DiA as hydrotropes. The solubility and
extraction power of the EtOH systems is superior to the DiA systems, as
EtOH is more polar than DiA.
Comparing the extraction yields of the employed SFMEs with the
mixture of 2:8 water/EtOH this study found that the addition of the
third compound TriA made a large advance in extraction power.
Taking all the findings into account, the investigated SFME extrac-
tion method employing water/hydrotrope/TriA as a solvent is a sig-
nificant improvement of the common, industrially used extraction with
an aqueous solvent system. The extraction using a Soxhlet apparatus
still gives superior yields for curcumin and demethoxycurcumin.
However, this method consumes more energy, due to the permanent
heating, and hazardous solvents which have to be removed afterwards.
With the SFME extraction we managed to come close to the same ex-
traction power in way shorter amounts of time and with less practical
effort. Additionally, it is not necessary to remove the SFME solvent
mixture as it is completely food-agreed and edible.
In upcoming studies, we will determine the purity of the obtained
extracts. Further, we will investigate the utility of the high solubiliza-
tion power of the EtOH/TriA mixtures to obtain even more con-
centrated extracts.
CRediT authorship contribution statement
Pierre Degot: Methodology, Validation, Formal analysis,
Investigation, Writing - original draft. Verena Huber: Methodology,
Validation, Formal analysis, Investigation, Writing - original draft.
7
Food Chemistry 336 (2021) 127660
Evamaria Hofmann: Software, Data curation, Writing - original draft.
Maximilian Hahn: Software, Supervision. Didier Touraud:
Conceptualization, Supervision. Werner Kunz: Conceptualization,
Supervision.
Declaration of Competing Interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influ-
ence the work reported in this paper.
Acknowledgments
We thankfully acknowledge Prof. Dr. Heilmann and his group at
Regensburg University for providing us access to their HPLC-systems.
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://
doi.org/10.1016/j.foodchem.2020.127660.
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