Journal of Molecular Liquids 241 (2017) 654–661

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Journal of Molecular Liquids

journal homepage: www.elsevier.com/locate/molliq

Natural deep eutectic solvents from choline chloride and betaine –

Physicochemical properties
Ivo M. Aroso a,b,⁎, Alexandre Paiva c, Rui L. Reis a,b, Ana Rita C. Duarte a,b
a
3B's Research Group - Biomaterials, Biodegradable and Biomimetic, University of Minho, Headquarters of the European Institute of Excellence on Tissue Engineering and Regenerative Medicine,
AvePark, 4805-017 Barco, Guimarães, Portugal
b
ICVS/3B's PT Government Associated Laboratory, Braga, Guimarães, Portugal
c
LAQV-REQUIMTE, Departamento de Química, Faculdade de Ciências e Tecnologia, Universidade Nova de Lisboa, 2829-516 Caparica, Portugal

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

Article history: The preparation of natural deep eutectic solvents (NADESs) from cheap and readily available raw materials is re-
Received 1 February 2017 ported. In this work, we have considered mixtures of choline chloride (CC) or betaine (Bet) with 3 sugar mole-
Received in revised form 5 June 2017 cules (glucose (Glu), xylose (Xyl) and sucrose (Suc)) and 2 carboxylic acids (citric (CA) and tartaric (Tart)
Accepted 11 June 2017
acids). The formation of NADESs was investigated by polarized optical microscopy (POM) and differential scan-
Available online 13 June 2017
ning calorimetry (DSC). The CC mixtures give origin to NADESs for 1:1 M ratio with the sugar molecules and for
Keywords:
2:1, 1:1 and 1:2 with the carboxylic acids, while Bet mixtures only formed NADES with the carboxylic acids. The
Natural deep eutectic solvents effect of water content (up to 5% (wt.%)) and temperature in conductivity and rheology were characterized. The
Thermophysical properties NADESs were found to be non-thixotropic, Newtonian liquids with high viscosity, decreasing with increasing
Rheology temperature and water content. The conductivity is limited by charge carrier mobility, thus increasing with
Conductivity water content and temperature.
Hydrogen bond donor © 2017 Elsevier B.V. All rights reserved.
Hydrogen bond acceptor

1. Introduction proposed as prospective alternatives to industrial solvents that allow

Deep eutectic solvents (DESs) were first proposed by Abbott et al. [1,
2] as potential alternative solvents for ionic liquids (ILs). When the DES
is composed by natural origin components, it is further defined as a nat-
ural deep eutectic solvent (NADES) [3,4]. Therefore, the NADESs can be
defined as mixtures of two or more components of natural origin that,
when at a particular molar ratio, present a strong depression of the
melting point, often becoming liquids at room temperature [5]. The in-
teraction between the compounds occurs through intermolecular hy-
drogen bonds, one acting as the hydrogen bond acceptor (HBA) and
the other as the donner (HBD) [2,6,7]. The preparation process is a
straightforward process, usually achieved after physical mixture of the
components, which can be accelerated by heating and vigorous physical
mixing [6,8,9]. The absence of chemical reactions, combined with the
use of common and readily available natural raw materials such as
sugars, alcohols, and organic and amino acids, results in low cost of pro-
duction and higher safety due to low (or absence of) toxicity for indus-
trial workers and end users [10–13]. Therefore, the NADESs are
⁎ Corresponding author at: 3B's Research Group – Biomaterials, Biodegradables and
Biomimetics, University of Minho, AvePark, Zona Industrial da Gandra, 4805-017 Barco,
Portugal.
E-mail address: ivo.aroso@dep.uminho.pt (I.M. Aroso).
for the development of new processes and for the redesign of current
ones.
The number of potential mixtures is virtually unlimited [5]. The
NADESs properties are a function of composition, making these highly
versatile and easily tunable solvents for a specific application [4]. How-
ever, this also means that it is difficult to predict which mixtures and
which molar ratios will originate a NADES. The composition and physi-
cal, thermal, chemical or biological properties are characterized on a
case by case scenario [8,9,12,14]. Much data correlating composition
and properties is still required before accurate predictive models can
be proposed.
Several recent works have made contributions to overcome this
present situation. In particular, Dai et al. [14] explored the impact of
water content on some of the NADES properties and its solvency
power for natural molecules and Florindo et al. [8], Craveiro et al. [9],
Hayyan, A. et al. [15] and Chemat et al. [16] have reported on the phys-
ical properties such as density, viscosity, polarity and conductivity of
several NADESs systems. The properties are not only dependent of com-
position, but also on temperature and, due to commonly presenting hy-
drophilic character, of water content [8,9,17]. Moreover, the
development of hydrophobic DESs has received recent attention and
its application on processes where the presence of water can be detri-
mental or as substitutes of organic solvents on separation processes
from aqueous streams has been recently proposed [18,19].

http://dx.doi.org/10.1016/j.molliq.2017.06.051
0167-7322/© 2017 Elsevier B.V. All rights reserved.
 I.M. Aroso et al. / Journal of Molecular Liquids 241 (2017) 654–661 655

The potential of NADESs as solvents and or supporting mediums for
different processes has been the focus of an increasing number of stud-
ies. Its solvent properties for DNA [3,4,20,21], plant metabolites (i.e.
phenolics, natural dyes and terpenes) [4,22–25], cellulose and lignin
[26–29] or active pharmaceutical ingredients [30,31] has been demon-
strated, with some molecules exhibiting solubility increases of several
orders of magnitude. They have also been proposed for a multitude of
processes including electrodeposition [32–34], enzymatic based reac-
tions/processes [35–37], polymerizations [38–40] and on improvement
of polymer processing [41,42], curing of resins [43,44] and CO2 capture
[45–47]. The extraction of natural molecules with NADES has gained
special attention [5,23,48]. Many natural compounds show higher solu-
bility in these liquids due to the ability to form hydrogen bonds directly
with the solvent components [23,49]. It is claimed that the plant pheno-
lics present higher stability in these solvents [50].
To be able to successfully choose the right systems for any given ap-
plication it is hence crucial to understand the properties of the systems
alone and how do they behave with operating conditions, such as tem-
perature, the presence of water among others. In this work, the NADES
forming ability of mixtures of choline chloride and betaine with sugar
molecules (glucose, xylose, sucrose) and carboxylic acids (tartaric and
citric acids) were evaluated. The thermodynamic properties were
assessed by differential scanning calorimetry (DSC), and its conductivity
and rheological properties were systematically characterized as func-
tion of temperature, up to 100 °C, and for small water content, up to
5% (wt%).
2.3. Polarized light microscopy (POM)
A droplet of NADES was deposited in a microscope slide for observa-
tion. Optical characterization of the NADES samples was carried out at
22 °C by POM using an Olympus transmission microscope coupled
with a Leica digital camera and Leica Application Suite Software.
2.4. Differential scanning analysis (DSC)
The NADESs mixtures and raw chemicals were characterized by DSC
(model DSC Q100, T.A. instruments, USA) on the temperature range
from −40 °C to degradation temperature, at 5 °C/min, after equilibra-
tion for 5 min at −40 °C. The experiments were performed under nitro-
gen atmosphere (50 mL/min), with 10–15 mg of sample in aluminum
pans with covering lids.
2.5. Conductivity measurements
The conductivity measurements were performed in an Inolab multi-
level 3 (WTW) equipment, equipped with a WTW Tetracon® 325 con-
ductivity probe. The NADESs were enclosed on a sealed chamber, fitted
with the sensors and the temperature was controlled by means of a
thermostatic oil bath. Prior to each measurement point, the tempera-
ture was stabilized for 5 min within ±0.50 K of set point, and monitored
with the probe's built-in temperature sensor (precision 0.2 K). All mea-
surements were performed 3 times with individual NADESs samples.

2.6. Rheological characterization

2. Experimental
The rheological/viscoelastic properties were characterized on a
2.1. Materials Kinexus Prot Rheometer (Kinexus Prot, MAL1097376, Malvern) fitted
with a parallel plate geometry with 20 mm of diameter (PU20 SR1740
All chemical reagents (choline chloride (CC), betaine (Bet), d-(+)- SS) and gap of 1 mm. The samples thixotropic properties were mea-
glucose (Glu), sucrose (Suc), d-(+)-xylose (Xyl), l-(+)-tartaric acid sured at different temperatures, by imposing a continuous shear rate
(Tart) and citric acid (CA) were acquired from Sigma-Aldrich and used ramp cycle from 0.1 to 100 s−1 and decreasing again to 0.1 s−1. The vis-
as received with exception of choline chloride that was kept on a vacu- cosity was measured by equilibrating the sample temperature for 5 min
um oven at 110 °C to prevent water adsorption. and applying a shear rate of 10 s−1 for 1 min. The measurements were
performed from 283 to 373 K, in 10 K intervals. All measurements were
performed 3 times with individual NADESs samples.
2.2. Preparation of NADES
3. Results and discussion
The mixtures were prepared by mixing CC or Bet with Xyl, Glu, Suc,
CA and Tart, in the required amounts, to obtain molar ratios of 4:1, 3:1, 3.1. NADESs preparation, visual aspect and POM results
2:1, 1:1, 1:2, 1:3 and 1:4 (Table 1). For each mixture, the constituents
were transferred to a sealed glass flask and heated to 90 °C, with con- The mixtures were prepared by direct mixing of the constituents.
stant stirring, for the time required until the formation of a clear liquid. The formation of the liquid mixture is dependent of the components na-
The mixture was allowed to naturally cool to room temperature. For the ture and of the molar ratio. A comprehensive list of the mixtures and
mixtures which did not form a liquid, the process was considered con- molar ratios that resulted in formation of liquid mixtures is presented
cluded after 16 h. in Table 1. The formation of the liquid mixture was defined from direct
observation and POM imaging (Fig. 1).
During the preparation process, throughout heating, the mixtures of
Table 1 CC with Glu, Xyl and Suc gradually acquire a liquid aspect. However,
NADES mixtures that formed liquids at room temperature, nomenclature used and the only the mixtures of molar ratio 1:1 present the aspect of transparent
molar ratios of the liquid mixtures. liquid and retain it upon cooling. These observations are supported by
NADES Component 1 Component 2 Molar ratios of mixtures
the POM results (Fig. 1) where the presence of solid crystalline struc-
designation in liquid forma tures are clearly visible for the mixtures of CC and the sugar molecules
for the ratios 2:1 and 1:2. When CC is in excess, the mixtures present
CC:Xyl CC Xyl 1:1
CC:Glu CC Glu 1:1 a milky aspect that is intensified during the cooling to ambient temper-
CC:Suc CC Suc 1:1b ature, whereas when the sugar molecules are in excess, the presence of
CC:CA CC CA 2:1; 1:1; 1:2 large non-soluble crystals are visible during all the preparation process.
CC:Tart CC Tart 2:1; 1:1; 1:2
Conversely, the mixtures of CC with CA and Tart present a distinct be-
Bet:CA:H2Oc Bet CA 1:1:1
Bet:Tart:H2Oc Bet Tart 1:1:1 havior. The liquid aspect is retained for the molar ratios 2:1, 1:1 and
a
1:2 with the presence of solids being directly observable for the mix-
Classified after POM and DSC characterization results.
b
The CC:Suc (1:1) is not completely liquid.
tures 3:1 or 1:3 and further confirmed by the POM results.
c
The betaine based mixtures were prepared as three component mixtures of equal The preparation of the Bet based NADESs was found to be more
molarity, with water (H2O) as the third component. difficult than those based on CC requiring the addition of water to
656 I.M. Aroso et al. / Journal of Molecular Liquids 241 (2017) 654–661

Fig. 1. POM images at 22 °C of prepared mixtures.

promote the formation of the liquid mixture. This behavior for Bet mixtures, Bet with CA or Tart were prepared as three component
mixtures with glucose, urea and small polyol molecules has already systems, of equal molarity, with water as the third component. The
been previously reported [48]. The anhydrous mixtures are very POM characterization confirmed that these mixtures do not contain
viscous, making them very difficult to manipulate. Therefore, these crystals.
 I.M. Aroso et al. / Journal of Molecular Liquids 241 (2017) 654–661 657

3.2. DSC characterization

The mixtures were characterized by DSC to determine the presence

of thermal transitions and compared to the raw materials. The thermo-
grams for CC, Xyl and the CC:Xyl mixtures with molar ratios from 4:1 to
1:4 are presented in Fig. 2.
The DSC of choline chloride is dominated by the presence of a sharp
endothermic peak at 78 °C, that results from a crystallographic arrange-
ment phase transition [51]. For the mixtures where choline chloride is in
molar excess, this peak is shifted to lower temperatures and occurs at 75
°C, irrespective of the molar ratio. The presence of this peak is indicative
that the CC molecules are not involved in molecular interactions with
the corresponding NADES counterpart molecule and, therefore, retain
their original thermal behavior. On the thermograms of CC:Xyl (4:1)
and CC:Xyl (3:1) the heat flow deviates from the baseline towards the
endothermic, starting at, respectively, − 2.8 °C and 9.5 °C due to the
glass transition. The mixture CC:Xyl (2:1) presents a distinct behavior
with the presence of a broad exothermic peak with onset at 13 °C, Fig. 3. DSC thermograms of CC, Glu, Suc and its mixtures at different molar ratios
followed by an endothermic region (heat flow curve above the baseline (thermograms not to scale).
below the 13 °C) which starts at 56 °C. As observed for the mixtures
(4:1) and (3:1), this region is abruptly terminated by the phase transi- to the high viscosity of Suc mixtures, which difficults the complete
tion peak from CC. The exothermic region corresponds to a process of mixing of the components.
cold crystallization, where the molecules upon gaining higher mobility The thermograms of CC mixtures with CA and Tart are shown in
adopt a more ordered structure, releasing energy. It is followed by an Fig. 4. The liquid mixtures (molar ratios (2:1), (1:1) and (1:2)) do not
endothermic transition from the disarrangement of the molecular or-
dering (see insert on Fig. 2 for magnification of this region). This feature
has been previously reported for other eutectic systems [52]. On the
other hand, when xylose is in excess, a broad endothermic peak with
onset at 110 °C and maximum at 141 °C is observed. This peak is
interpreted as the melting of the xylose excess that is not participating
on the NADES complex and compares with onset at 142 °C and maxi-
mum at 152 °C observed for pure xylose. Also a glass transition (Tg) is
observed on the temperature range − 30 °C to −25 °C. The liquid mix-
ture, CC:Xyl (1:1), does not present thermal transitions on the temper-
ature range considered.
The mixtures of CC and Glu (Fig. 3) have an analogous behavior to
that observed for CC and Xyl. Namely, the cold crystallization for the
(2:1) mixture, the absence of thermal processes for the liquid mixture
(1:1) and broad glucose melting peak for the (1:2) mixture occurring
at slight lower temperature (onset at 108 °C and peak maximum at
148 °C) than observed for pure glucose (onset at 144 °C and peak max-
imum at 156 °C). Except for the eutectic mixture, the Tg is observed for
all mixtures at around − 25 °C. Conversely, the CC and Suc mixtures Fig. 4. DSC thermograms of CC, CA, Tart and its mixtures at different molar ratios
present a distinct behavior than observed for Glu and Xyl. For the (thermograms not to scale).
CC:Suc (2:1) mixture, the CC phase transition peak is clearly observed,
but the cold crystallization process, identified for Xyl and Glu mixtures,
is absent. For the (1:1) mixture, albeit of small intensity, the peak corre-
sponding to the CC phase transition is still detected. This is probably due

Fig. 2. DSC thermograms of CC, Xyl and its mixtures at different molar ratios. Insert shows Fig. 5. DSC thermograms of Bet based NADESs and for the CC with Xyl, Glu, Suc, CA and
magnification of CC:Xyl (3:1) and (2:1) for −10 to 90 °C (thermograms not to scale). Tart NADESs, prepared as 3 equi-molar component systems, with water.
658 I.M. Aroso et al. / Journal of Molecular Liquids 241 (2017) 654–661

profile i.e. absence of thermal transitions, with the exception of a Tg on

the region −20 °C to −5 °C (Fig. 5), which was absent for on the anhy-
drous mixtures.

3.3. Conductivity

The conductivity was measured as function of temperature and

Fig. 6. Variation conductivity (σ) with temperature (K) for anhydrous CC based NADES
and Bet based NADES.
present thermal transitions. For representation simplicity, only the
1:1 M ratio curves are presented. When a complete liquid mixture is
not obtained, if CC is in excess, the endothermic peak from its phase
transition is observed at 76 °C. On the other hand, if CA or Tart is in
molar excess, a peak corresponding to the melting process is observed
at 147 °C and 157 °C, respectively. The melting of CA and Tart is prompt-
ly followed by thermal degradation. The non-eutectic mixtures have a
Tg at around − 10 °C, a higher temperature than observed for CC and
sugar molecules mixtures.
The Bet based NADESs with CA and Tart were prepared with water
as an equimolar systems, resulting in the formation of liquid mixtures.
The DSC thermograms are presented in Fig. 5 and, as discussed before,
the eutectic mixtures are characterized by the absence of thermal tran-
sitions other than thermal degradation at high temperatures.
To determine the water content effect, all the eutectic mixtures
discussed before were prepared as equi-molar systems with water.
Despite the new composition, the thermograms maintain the same
water content for selected mixtures: CC:Xyl (1:1), CC:Glu (1:1),
CC:Suc (1:1), CC:CA (1:1), CC:Tart (1:1), Bet:CA:H2O (1:1:1) and
Bet:CA:H2O (1:1:1). Due to the high viscosity of the samples, the mea-
surements were performed for temperatures starting at 313 K up to
373 K. The CC based NADESs were characterized in the presence of
water contents of 0, 1, 3 and 5% (wt%). Due to the high viscosity of an-
hydrous CC:Suc (1:1), it was not possible to measure the conductivity
of this sample.
The results obtained for the anhydrous and Bet based NADESs are
presented in Fig. 6. On the temperature interval considered, among
the CC based NADESs, CC:Xyl (1:1) presents significantly higher con-
ductivity, followed by CC:Glu (1:1), CC:Tart (1:1) and CC:CA (1:1). Sim-
ilarly to the CC based NADESs, despite the different chemical
composition, the conductivity of Bet:Tart:H2O (1:1:1) is higher than of
Bet:CA:H2O (1:1:1). The systems conductivities increase exponentially
with the increasing temperature and the relative trend is maintained
for the entire temperature interval considered.
The change of conductivity with temperature was found to be well
described by the logarithm form of the Arrhenius equation,
Eσ
ln σ ¼ ln σ 0 − ð1Þ
RT

Table 2
Linear regression fitting parameters for conductivity (measured between 313 and 373 K)
(Eq. (1)) and rheology (measured between 283 and 373 K) (Eq. (2)) data.

NADES Water content Conductivity Viscosity

wt%a
Eσ ln σ0 R2 Eη ln η0 R2
(kJ/mol) (kJ/mol)

C:Xyl 0 (0.0) 68.09 21.16 0.996 71.02 −24.71 0.992

(1:1) 1 (0.16) 62.50 19.61 0.996 68.53 −24.04 0.991
3 (0.48) 55.73 17.82 0.996 56.36 −20.40 0.985
5 (0.80) 49.76 16.20 0.997 47.91 −17.55 0.997
CC:Glu 0 (0.0) 78.38 23.36 0.991 91.52 −30.45 0.991
(1:1) 1 (0.18) 71.17 21.35 0.992 77.04 −25.80 0.992
3 (0.53) 61.84 18.94 0.996 60.98 −21.01 0.994
5 (0.89) 55.58 17.40 0.998 52.12 −18.53 0.993
CC:Suc 0 (0.0) – – – 104.50 −30.87 0.982
(1:1) 1 (0.27) 100.93 29.76 0.995 85.21 −24.99 0.982
3 (0.80) 86.36 25.45 0.995 42.06 −10.92 0.910
5 (1.34) 72.13 21.58 0.997 32.18 −8.08 0.921
CC:CA 0 (0.0) 93.45 27.48 0.994 96.15 −30.53 0.994
(1:1) 1 (0.18) 84.95 24.90 0.993 92.17 −29.59 0.992
3 (0.55) 66.96 19.56 0.990 85.45 −28.21 0.992
5 (0.92) 62.82 18.87 0.996 68.74 −23.11 0.984
CC:Tart 0 (0.0) 81.82 24.21 0.995 102.08 −33.4 0.991
(1:1) 1 (0.16) 74.34 22.08 0.993 95.52 −31.6 0.998
3 (0.48) 67.53 20.45 0.997 85.15 −28.7 0.994
5 (0.80) 59.35 18.47 0.997 77.36 −26.9 0.988
Bet:CA:H2O (1:1:1) 76.94 22.68 0.985 97.79 −30.78 0.990
Bet:Tart:H2O (1:1:1) 67.66 20.13 0.991 101.94 −32.60 0.990
Fig. 7. Variation of CC:Xyl (1:1) conductivity with temperature and water content; a)
R2 – coefficient of determination. Eσ and Eη (in kJ/mol) were calculated using R value of measured conductivity between 40 and 100 °C and water content of 0, 1, 3 and 5%
0.008314 kJmol−1 K−1 and σ and η measured in S/m and Pa·s respectively. (wt%) (Insert depicts the linear fitting of ln σ vs. 1/T); b) three dimensional plot of
a
The values for equivalent molar water content is given inside the brackets. conductivity (ln σ) with temperature (1/T) and molar water content (n·H2O).
 I.M. Aroso et al. / Journal of Molecular Liquids 241 (2017) 654–661
where σ0 is a constant, Eσ is the activation energy for electrical conduc-
tion, R the ideal gases constant and T the absolute temperature. The lin-
ear regression of the natural logarithm of conductivity vs. the inverse of
absolute temperature returns a good fitting (R2 N 0.99), including for the
NADESs evaluated with water contents for up to 5% (wt%). The Eσ were
calculated to be in the range of 50 to 100 kJ mol−1, decreasing with the
increase of the water content (Table 2). The conductivity was also found
to increase with the system's water content. For the same water con-
tent, the conductivity trend between the different NADES is preserved
along the entire temperature interval. Therefore, for simplicity and clar-
ity, only the experimental data obtained for the system CC:Xyl is shown
The complete set of experimental results is presented as Supplementary
information. The conductivity of CC:Xyl (1:1), for different tempera-
tures and water content of 0, 1, 3 and 5% and the fitting of Eq. (1) is pre-
sented in Fig. 7a. The combined effect of temperature and water content
is depicted on the three dimensional plot of ln σ as function of 1/T and
molar water content (n·H2O) presented in Fig. 7b. Our results show
that the behavior of conductivity with water content and temperature
is similar for the different systems. Despite the low conductivity of
pure water, 5.5 × 10−5 mS cm−1 (293 K), its presence significantly in-
creases conductivity due to lower viscosity and corresponding higher
ion mobility. The same effect has been observed in ionic liquids [2,7,
53–55]. Nevertheless, in general, the conductivity of NADES is still
lower than that of ionic liquids due to the presence of fewer charge car-
rier species [7,8,14,17].
3.4. Rheology
The rheological properties of the NADESs mixtures were character-
ized by obtaining flow curves. In general, the characterization was per-
formed applying a shear rate ramp from 0.1 to 100 s− 1, on the
temperature range of 283 to 373 K, on steps of 10 K, to the anhydrous
samples and water content of 1, 3 and 5% (wt%). However, due to the
Fig. 8. Flow curves for CC:Xyl (1:1) with water content of 0, 1, 3 and 5% (wt%).
659
high viscosity and equipment limitations, it was not possible to perform
measurements when the viscosity exceeded 10,000 Pa·s. The flow
curves obtained for the NADES CC:Xyl (1:1), with water contents of 0,
1, 3 and 5% (wt%), at different temperatures, are presented in Fig. 8
and are representative of the behavior observed for all NADESs.
All NADESs present Newtonian fluid behavior, i.e., the viscosity is
constant and independent of the value and duration of the shear rate
applied, denoting the absence of molecular structuring or entangle-
ments. The apparent shear thinning observed for low viscosity
(b1 Pa·s) at small shear rates (b0.5 s− 1) is interpreted as a feature
from the system and not a characteristic from the sample. The NADESs
present a rheological behavior similar to that of common liquid sol-
vents. However, these are highly viscous liquids and, with exception
for CC:Xyl (1:1), with typical viscosities at 293 K higher than 1000 Pa·s.
The NADESs viscosities vary for different constituting components,
temperature and water content. Considering the CC based NADES, the
viscosity is higher for CC:Suc (1:1), followed by CC:CA (1:1), CC:Tart
(1:1), CC:Glu (1:1) and CC:Xyl (1:1). This trend is observed on the
whole temperature range and for NADESs with equivalent water con-
tent (Table 2).The viscosity decreases significantly with the increase of
temperature and water content (Fig. 8 and Fig. 9).
As observed for conductivity, the dependence of viscosity with the
temperature was also found to be well described by the logarithmic
form of the Arrhenius equation:
Eη
ln η ¼ ln η0 þ ð2Þ
RT
where η is the shear viscosity, η0 a constant and Eη is the flow activation
energy, R the ideal gas constant and T the absolute temperature. The Eη
for each system was calculated from the slope of ln η vs. 1/T and found
to be approximately 100 kJ mol−1 for the anhydrous systems, decreas-
ing substantially with the presence of water. Dai et al. have shown
660 I.M. Aroso et al. / Journal of Molecular Liquids 241 (2017) 654–661
Fig. 9. Variation of viscosity with temperature and water content; a) Viscosity (η) as
function of temperature (K) for anhydrous CC based NADESs and Bet based NADESs; b)
CC:Xyl (1:1) three dimensional plot of viscosity (ln η) dependence with
temperature (1/T (K−1)) and molar water content (n·H2O).
that the strong H-bond interactions between the NADES components
are gradually decreased when in the presence of water [14]. The results
obtained indicate that the water molecules, due to its small size and H-
bond forming ability, can hence act as plasticizers decreasing the viscos-
ity and the Eη.
The conductivity of a liquid is affected by the number of charged
species and its mobility. If limited by the mobility of the charged
species, the conductivity is proportional to the fluidity (η − 1 ).
Therefore, the plot of σ vs. η− 1 as function of temperature should
return a linear proportionality [17,53,56]. This is indeed the case
for the NADESs, as depicted on Fig. 10a, which shows a strong linear
correlation demonstrating that the conductivity is limited by ionic
Fig. 10. a) Plot of conductivity versus fluidity (η−1) for selected NADES (tendency lines for Bet and CC based NADES are marked with 1 and 2 respectively); b) plot of Eσ versus Eη for the CC
based NADES with different water content.
mobility. Moreover, the values of the slopes for the different CC
anhydrous based NADESs are similar which is indicative that the
charge carrier species is dominated by CC, possibly due to the
presence of chlorine ions within the mixture. The counterpart
NADES molecules have negligible contribution to the overall
conductivity. Likewise, the plots of σ vs. η − 1 for the Bet based
NADESs are also linearly proportional with similar slopes. The rela-
tion between conductivity and viscosity is further stressed by the
proportionality between Eσ and Eη (Fig. 10b). The NADESs behavior
trends with water content and temperature are similar to those
observed for several choline based ILs [53–55].
4. Conclusions
The preparation of NADES that are liquid at room temperature
with the H-bond acceptors CC and Bet and 3 sugar molecules (Xyl,
Glu and Suc) and 2 carboxylic acids (CA and Tart) as bond donors
was thoroughly investigated. NADES formation was characterized
by POM and DSC revealing the potential of these techniques as
tools to rapidly assess if one particular mixture gives origin to a
liquid mixture. The conductivity and rheological properties and its
variation with water content and temperature were assessed. The
conductivity was found to increase exponentially with the tempera-
ture and with the presence of water. The rheological characterization
has shown that these NADESs are very viscous liquids with Newtoni-
an behavior. The viscosity decreases significantly both with temper-
ature and water content. Both the conductivity and viscosity
dependence with temperature are well described by logarithm
form of the Arrhenius equation.
Acknowledgments
Ivo Aroso acknowledges the financial support from FCT through
grant SFRH/BD/42273/2007. Funding was also granted from the Euro-
pean Union Seventh Framework Programme (FP7/2007–2013) under
grant agreement no. REGPOT-CT2012-316331-POLARIS and from Pro-
ject “Novel smart and biomimetic materials for innovative regenerative
Medicine approaches (ref.: RL1 - ABMR - NORTE-01-0124-FEDER-
000016)” co-financed by North Portugal Regional Operational Pro-
gramme (ON.2 – O Novo Norte), under the National Strategic Reference
Framework (NSRF), through the European Regional Development Fund
(ERDF).
Appendix A. Supplementary data
Supplementary data to this article can be found online at http://dx.
doi.org/10.1016/j.molliq.2017.06.051.
 I.M. Aroso et al. / Journal of Molecular Liquids 241 (2017) 654–661
References
[1] A.P. Abbott, G. Capper, D.L. Davies, R.K. Rasheed, V. Tambyrajah, Novel solvent prop-
erties of choline chloride/urea mixtures, Chem. Commun. (2003) 70–71.
[2] A.P. Abbott, D. Boothby, G. Capper, D.L. Davies, R.K. Rasheed, Deep eutectic solvents
formed between choline chloride and carboxylic acids: versatile alternatives to ionic
liquids, J. Am. Chem. Soc. 126 (2004) 9142–9147.
[3] Y.H. Choi, J. van Spronsen, Y.T. Dai, M. Verberne, F. Hollmann, I.W.C.E. Arends, et al.,
Are natural deep eutectic solvents the missing link in understanding cellular metab-
olism and physiology? Plant Physiol. 156 (2011) 1701–1705.
[4] Y.T. Dai, J. van Spronsen, G.J. Witkamp, R. Verpoorte, Y.H. Choi, Natural deep eutectic
solvents as new potential media for green technology, Anal. Chim. Acta 766 (2013)
61–68.
[5] A. Paiva, R. Craveiro, I. Aroso, M. Martins, R.L. Reis, A.R.C. Duarte, Natural deep eutec-
tic solvents - solvents for the 21st century, ACS Sustain. Chem. Eng. 2 (2014)
1063–1071.
[6] Q.H. Zhang, K.D. Vigier, S. Royer, F. Jerome, Deep eutectic solvents: syntheses, prop-
erties and applications, Chem. Soc. Rev. 41 (2012) 7108–7146.
[7] E.L. Smith, A.P. Abbott, K.S. Ryder, Deep eutectic solvents (DESs) and their applica-
tions, Chem. Rev. 114 (2014) 11060–11082.
[8] C. Florindo, F.S. Oliveira, L.P.N. Rebelo, A.M. Fernandes, I.M. Marrucho, Insights into
the synthesis and properties of deep eutectic solvents based on cholinium chloride
and carboxylic acids, ACS Sustain. Chem. Eng. 2 (2014) 2416–2425.
[9] R. Craveiro, I. Aroso, V. Flammia, T. Carvalho, M.T. Viciosa, M. Dionísio, et al., Proper-
ties and thermal behavior of natural deep eutectic solvents, J. Mol. Liq. 215 (2016)
534–540.
[10] B. Kudlak, K. Owczarek, J. Namiesnik, Selected issues related to the toxicity of ionic
liquids and deep eutectic solvents - a review, Environ. Sci. Pollut. Res. 22 (2015)
11975–11992.
[11] I. Juneidi, M. Hayyan, M.A. Hashim, Evaluation of toxicity and biodegradability for
cholinium-based deep eutectic solvents, RSC Adv. 5 (2015) 83636–83647.
[12] F. Cardellini, M. Tiecco, R. Germani, G. Cardinali, L. Corte, L. Roscini, et al., Novel zwit-
terionic deep eutectic solvents from trimethylglycine and carboxylic acids: charac-
terization of their properties and their toxicity, RSC Adv. 4 (2014) 55990–56002.
[13] Q. Wen, J.X. Chen, Y.L. Tang, J. Wang, Z. Yang, Assessing the toxicity and biodegrad-
ability of deep eutectic solvents, Chemosphere 132 (2015) 63–69.
[14] Y.T. Dai, G.J. Witkamp, R. Verpoorte, Y.H. Choi, Tailoring properties of natural deep
eutectic solvents with water to facilitate their applications, Food Chem. 187
(2015) 14–19.
[15] A. Hayyan, F.S. Mjalli, I.M. AlNashef, Y.M. Al-Wahaibi, T. Al-Wahaibi, M.A. Hashim,
Glucose-based deep eutectic solvents: physical properties, J. Mol. Liq. 178 (2013)
137–141.
[16] F. Chemat, H. Anjum, A.M. Shariff, P. Kumar, T. Murugesan, Thermal and physical
properties of (choline chloride + urea + l-arginine) deep eutectic solvents, J. Mol.
Liq. 218 (2016) 301–308.
[17] V.S. Protsenko, A.A. Kityk, D.A. Shaiderov, F.I. Danilov, Effect of water content on
physicochemical properties and electrochemical behavior of ionic liquids containing
choline chloride, ethylene glycol and hydrated nickel chloride, J. Mol. Liq. 212
(2015) 716–722.
[18] D.J.G.P. van Osch, L.F. Zubeir, A. van den Bruinhorst, M.A.A. Rocha, M.C. Kroon, Hy-
drophobic deep eutectic solvents as water-immiscible extractants, Green Chem.
17 (2015) 4518–4521.
[19] B.D. Ribeiro, C. Florindo, L.C. Iff, M.A.Z. Coelho, I.M. Marrucho, Menthol-based eutec-
tic mixtures: hydrophobic low viscosity solvents, ACS Sustain. Chem. Eng. 3 (2015)
2469–2477.
[20] I. Mamajanov, A.E. Engelhart, H.D. Bean, N.V. Hud, DNA and RNA in anhydrous
media: duplex, triplex, and G-quadruplex secondary structures in a deep eutectic
solvent, Angew Chem Int Edit 49 (2010) 6310–6314.
[21] D. Mondal, M. Sharma, C. Mukesh, V. Gupta, K. Prasad, Improved solubility of DNA in
recyclable and reusable bio-based deep eutectic solvents with long-term structural
and chemical stability, Chem. Commun. 49 (2013) 9606–9608.
[22] W.T. Bi, M.L. Tian, K.H. Row, Evaluation of alcohol-based deep eutectic solvent in ex-
traction and determination of flavonoids with response surface methodology opti-
mization, J. Chromatogr. A 1285 (2013) 22–30.
[23] Y.T. Dai, G.J. Witkamp, R. Verpoorte, Y.H. Choi, Natural deep eutectic solvents as a
new extraction media for phenolic metabolites in Carthamus tinctorius L, Anal.
Chem. 85 (2013) 6272–6278.
[24] M.W. Nam, J. Zhao, M.S. Lee, J.H. Jeong, J. Lee, Enhanced extraction of bioactive nat-
ural products using tailor-made deep eutectic solvents: application to flavonoid ex-
traction from Flos sophorae, Green Chem. 17 (2015) 1718–1727.
[25] M. Cvjetko Bubalo, N. Ćurko, M. Tomašević, K. Kovačević Ganić, Redovniković I.
Radojčić, Green extraction of grape skin phenolics by using deep eutectic solvents,
Food Chem. 200 (2016) 159–166.
[26] M. Francisco, A. van den Bruinhorst, M.C. Kroon, New natural and renewable low
transition temperature mixtures (LTTMs): screening as solvents for lignocellulosic
biomass processing, Green Chem. 14 (2012) 2153–2157.
[27] P.D. de Maria, Recent trends in (ligno)cellulose dissolution using neoteric solvents:
switchable, distillable and bio-based ionic liquids, J. Chem. Technol. Biotechnol. 89
(2014) 11–18.
[28] E.P. Zhou, H.R. Liu, A novel deep eutectic solvents synthesized by solid organic com-
pounds and its application on dissolution for cellulose, Asian J. Chem. 26 (2014)
3626–3630.
661
[29] G.C. Xu, J.C. Ding, R.Z. Han, J.J. Dong, Y. Ni, Enhancing cellulose accessibility of corn
stover by deep eutectic solvent pretreatment for butanol fermentation, Bioresour.
Technol. 203 (2016) 364–369.
[30] I.M. Aroso, R. Craveiro, A. Rocha, M. Dionisio, S. Barreiros, R.L. Reis, et al., Design of
controlled release systems for THEDES-therapeutic deep eutectic solvents, using su-
percritical fluid technology, Int. J. Pharm. 492 (2015) 73–79.
[31] J.C. Silva, I.M. Aroso, F. Mano, I. Sa-Nogueira, S. Barreiros, R.L. Reis, et al., Therapeutic
deep eutectic solvents as solubility enhancers for different active pharmaceutical in-
gredients, Tissue Eng. Part A 21 (2015) S106–S107.
[32] A. Abo-Hamad, M. Hayyan, M.A. AlSaadi, M.A. Hashim, Potential applications of deep
eutectic solvents in nanotechnology, Chem. Eng. J. 273 (2015) 551–567.
[33] E.L. Smith, Deep eutectic solvents (DESs) and the metal finishing industry: where
are they now? Trans. Int. Met. Finish 91 (2013) 241–248.
[34] A.P. Abbott, K. El Ttaib, G. Frisch, K.J. McKenzie, K.S. Ryder, Electrodeposition of cop-
per composites from deep eutectic solvents based on choline chloride, Phys. Chem.
Chem. Phys. 11 (2009) 4269–4277.
[35] E. Durand, J. Lecomte, B. Barea, G. Piombo, E. Dubreucq, P. Villeneuve, Evaluation of
deep eutectic solvents as new media for Candida antarctica B lipase catalyzed reac-
tions, Process Biochem. 47 (2012) 2081–2089.
[36] E. Durand, J. Lecomte, B. Barea, E. Dubreucq, R. Lortie, P. Villeneuve, Evaluation of
deep eutectic solvent-water binary mixtures for lipase-catalyzed lipophilization of
phenolic acids, Green Chem. 15 (2013) 2275–2282.
[37] V. Stepankova, S. Bidmanova, T. Koudelakova, Z. Prokop, R. Chaloupkova, J.
Damborsky, Strategies for stabilization of enzymes in organic solvents, ACS Catal.
3 (2013) 2823–2836.
[38] J.D. Mota-Morales, M.C. Gutierrez, I.C. Sanchez, G. Luna-Barcenas, F. del Monte, Fron-
tal polymerizations carried out in deep-eutectic mixtures providing both the mono-
mers and the polymerization medium, Chem. Commun. 47 (2011) 5328–5330.
[39] F. del Monte, D. Carriazo, M.C. Serrano, M.C. Gutierrez, M.L. Ferrer, Deep eutectic sol-
vents in polymerizations: a greener alternative to conventional syntheses,
ChemSusChem 7 (2014) 999–1009.
[40] J.D. Mota-Morales, M.C. Gutierrez, M.L. Ferrer, R. Jimenez, P. Santiago, I.C. Sanchez,
et al., Synthesis of macroporous poly(acrylic acid)-carbon nanotube composites by
frontal polymerization in deep-eutectic solvents, J. Mater. Chem. A 1 (2013)
3970–3976.
[41] A.M.M. Sousa, H.K.S. Souza, J. Uknalis, S.C. Liu, M.P. Goncalves, L.S. Liu, Improving
agar electrospinnability with choline-based deep eutectic solvents, Int. J. Biol.
Macromol. 80 (2015) 139–148.
[42] A.M.M. Sousa, H.K.S. Souza, N. Latona, C.K. Liu, M.P. Goncalves, L.S. Liu, Choline chlo-
ride based ionic liquid analogues as tool for the fabrication of agar films with im-
proved mechanical properties, Carbohydr. Polym. 111 (2014) 206–214.
[43] H. Maka, T. Spychaj, W. Sikorski, Deep eutectic ionic liquids as epoxy resin curing
agents, Int. J. Polym. Anal. Charact. 19 (2014) 682–692.
[44] M.C. Gutierrez, F. Rubio, F. del Monte, Resorcinol-formaldehyde polycondensation in
deep eutectic solvents for the preparation of carbons and carbon-carbon nanotube
composites, Chem. Mater. 22 (2010) 2711–2719.
[45] G. Garcia, M. Atilhan, S. Aparicio, A theoretical study on mitigation of CO2 through
advanced deep eutectic solvents, Int. J. Greenh. Gas Con. 39 (2015) 62–73.
[46] G. Garcia, M. Atilhan, S. Aparicio, Interfacial properties of deep eutectic solvents re-
garding to CO2 capture, J. Phys. Chem. C 119 (2015) 21413–21425.
[47] E. Ali, M.K. Hadj-Kali, S. Mulyono, I. Alnashef, A. Fakeeha, F. Mjalli, et al., Solubility of
CO2 in deep eutectic solvents: experiments and modelling using the Peng-Robinson
equation of state, Chem. Eng. Res. Des. 92 (2014) 1898–1906.
[48] N. Li, Y. Wang, K. Xu, Y. Huang, Q. Wen, X. Ding, Development of green betaine-
based deep eutectic solvent aqueous two-phase system for the extraction of protein,
Talanta 152 (2016) 23–32.
[49] Y.T. Dai, J. van Spronsen, G.J. Witkamp, R. Verpoorte, Y.H. Choi, Ionic liquids and
deep eutectic solvents in natural products research: mixtures of solids as extraction
solvents, J. Nat. Prod. 76 (2013) 2162–2173.
[50] Y.T. Dai, R. Verpoorte, Y.H. Choi, Natural deep eutectic solvents providing enhanced
stability of natural colorants from safflower (Carthamus tinctorius), Food Chem. 159
(2014) 116–121.
[51] R.L. Collin, Polymorphism and radiation decomposition of choline chloride, J. Am.
Chem. Soc. 79 (1957) 6086.
[52] I.M. Aroso, J.C. Silva, F. Mano, A.S.D. Ferreira, M. Dionisio, I. Sa-Nogueira, et al., Disso-
lution enhancement of active pharmaceutical ingredients by therapeutic deep eu-
tectic systems, Eur. J. Pharm. Biopharm. 98 (2016) 57–66.
[53] V.S. Protsenko, L.S. Bobrova, F.I. Danilov, Physicochemical properties of ionic liquid
mixtures containing choline chloride, chromium (III) chloride and water: effects
of temperature and water content, Ionics 23 (2017) 637–643.
[54] C. Du, B. Zhao, X.-B. Chen, N. Birbilis, H. Yang, Effect of water presence on choline
chloride-2urea ionic liquid and coating platings from the hydrated ionic liquid, Sci
Rep 6 (2016) 29225.
[55] L.S. Bobrova, F.I. Danilov, V.S. Protsenko, Effects of temperature and water content
on physicochemical properties of ionic liquids containing CrCl3·xH2O and choline
chloride, J. Mol. Liq. 223 (2016) 48–53.
[56] A.P. Abbott, G. Capper, D.L. Davies, R.K. Rasheed, Ionic liquid analogues formed from
hydrated metal salts, Chem. Eur. J. 10 (2004) 3769–3774.