Trends in Analytical Chemistry 105 (2018) 225e239

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Trends in Analytical Chemistry

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

Extraction techniques with deep eutectic solvents

Sara C. Cunha**, Jose

O. Fernandes*

cia, Universidade do Porto, Rua de Jorge Viterbo Ferreira 228, 4050-313,

rio de Bromatologia e Hidrologia, Faculdade de Farma
LAQV-REQUIMTE, Laborato
Porto, Portugal

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

Article history: In last years, a plethora of extraction techniques has emerged as environmental-friendly alternatives to
Available online 9 May 2018 conventional extraction procedures. In this particular field, a novel class of solvents known as deep
eutectic solvents (DES) has arisen as a new and very promising tool. Compared with conventional organic
Keywords: solvents, DES as well as the so-called natural deep eutectic solvents (NADES) have attracted considerable
Sample preparation attention due to the fact that they not only are eco-friendly, non-toxic, and biodegradable organic
Deep eutectic solvents
compounds but also have a low cost, being easy to produce in the own laboratory. The present review
Natural deep eutectic solvents
provides a critical and organized overview of novel extraction techniques using DES as extracting sol-
Microextraction
Chemical analysis
vents that were applied in food, biological and environmental sample analysis. An evaluation of how
these DES/NADES could improve extraction yields of a variety of analytes and advantages and limitations
of each proposal will be discussed and compared with earlier studies.
© 2018 Elsevier B.V. All rights reserved.

1. Introduction different techniques characterized by using low amounts of sample

Sample preparation is a common step in trace analytical
methods, involving the extraction of an analyte or a class of ana-
lytes from its original matrix into a solvent in order to allow its
further analysis by a sensitive and selective way. Conventional
sample preparation techniques such as multi-step liquid-liquid
extraction, Soxhlet extraction and solid-phase extraction, among
others, are usually time-consuming, labor intensive and involve the
use of a large volume of organic solvents, which is expensive and
generate a considerable amount of waste harmful for human health
and the environment. To overcome these drawbacks, numerous
efforts have been made over the last decades, to streamline sample
extraction procedures and also to reduce or eliminate the use or
generation of hazardous substances in agreement with the princi-
ples of the green chemistry [1]. New techniques miniaturizing
solid-phase extraction or liquid-liquid extraction have appeared
such as SPME and LPME, respectively. SPME is a solventless
extraction technique very popular in recent years that use different
fiber materials in various configurations for the extraction of a wide
range of volatile analytes [2]. LPME comprises a range of slightly
* Corresponding author. Fax: þ351 226093390.
** Corresponding author. Fax: þ351 226093390.
E-mail addresses: sara.cunha@ff.up.pt (S.C. Cunha), josefer@ff.up.pt
(J.O. Fernandes).
matrices and small volumes of organic solvents. Often it can
employ different types of kinetic energy such as ultrasound (UAME)
or microwave (MAE) in order to improve extraction efficiency,
enrichment factor and simultaneously reduce extraction time.
Progresses made in the last decade (2006e2017) in the field of
sample extraction are confirmed by the increasing number of
published scientific articles on SPME, LPME, UAME and MAE. In the
ISI Web of knowledge there are references for that period of about
11,133 published scientific papers on SPME, 1351 on LPME, 1790 on
UAME and 2803 on MAE for determination of different organic and
inorganic compounds, of which more than 60% were published in
the last five years. The keywords used for this research were: “solid-
phase microextraction”, “liquid phase microextraction”, “ultra-
sound assisted extraction” and “microwave-assisted extraction”.
Until some years ago, solvents used in LPME techniques were
always associated with a certain degree of toxicity, but the recent
development of a remarkable generation of new extracting sol-
vents, i.e. DES is changing this scenario. These solvents are
commonly composed of two non-toxic components, one of them
with capacity to be a HBA (quaternary ammonium, tetralky-
lammonium or phosphonium salts) while the other (acids, alcohols,
amines or carbohydrates) possess HBD properties [3]. Generally,
DES have much lower melting point than that of any of its indi-
vidual components, which is due to the formation of intramolecular
hydrogen bonds [4], and possess some noteworthy chemical
properties, such as low vapor pressure, relatively wide liquid-range,

https://doi.org/10.1016/j.trac.2018.05.001
0165-9936/© 2018 Elsevier B.V. All rights reserved.
226 S.C. Cunha, J.O. Fernandes / Trends in Analytical Chemistry 105 (2018) 225e239

Abbreviations MDES magnetic deep eutectic solvents

MIPs molecularly imprinted polymers
AA-LLME air-assisted liquid-liquid microextraction MMWCNT magnetic multi-walled carbon nanotube
AA-DLLME air-assisted dispersive liquid-liquid microextraction nanocomposite
AAS atomic absorption spectrometry NIPs non-imprinted polymers
CPEs carbon paste electrodes NMR nuclear magnetic resonance
DEHP di(2-ethylhexyl)phthalate OCPs organochlorine pesticides
DLLME dispersive liquid-liquid microextraction Ox oxalic acid
DNA deoxyribonucleic acid PAHs polycyclic aromatic hydrocarbons
ELISA enzyme-linked immunosorbent assay PDA photodiode array
ELLME emulsification liquid-liquid microextraction PCH 1,2-propanediolecholine chloride
ETAAS electrothermal atomic absorption spectrometry PT-SPE pipette-tip solid-phase extraction
FAAS flame atomic absorption spectrometry ROS rosmarinic acid
FT-IR fourier-transform infrared spectroscopy RSD relative standard deviation
GA-DLLME gas air-dispersive liquid-liquid microextraction SDME single drop microextraction
GC-ECD gas chromatography-electron capture detector SPME solid-phase microextraction
GC-FID gas chromatography-flame ionization detector SPE solid-phase extraction
HBA hydrogen-bond acceptor S-SIL-MSPD silica-supported ionic liquid-based matrix solid
HBD hydrogen-bond donor phase dispersion
HPLC-DAD high performance liquid chromatography-diode UAE ultrasound-assisted extraction
array detection UHPLC-UV ultra-high performance liquid chromatography with
HPLC-UV high performance liquid chromatography-ultraviolet ultraviolet detection
HS-SME headpsace solid-single microextraction UAME ultrasound assisted microextraction
HF-LPME hollow fiber-liquid phase microextraction UALPME ultrasound assisted liquid phase microextraction
IAA Indole-3-acetic acid USAEME ultrasound-assisted emulsification microextraction
IBA 3-indolebutyric acid UPLC-TOF-MS ultra performance liquid chromatography-time-
4-IPOAA 4-iodophenoxyacetic acid of-flight mass spectrometry
LGH lactic acideglucose UV-VIS ultraviolet-visible spectrophotometry
LIT lithospermic acid TIIA tanshinone IIA
LPME liquid phase microextraction THF tetrahydrofuran
MAE microwave-assisted extraction VWD variable wavelength detector

non-flammability and unreactivity towards water [5]. Moreover,
DES are easy to be prepared not requiring purification steps, are
made from low cost compounds, present low or negligible toxicity
and are biodegradable and easily recyclable. These features make
DES preferable over the conventional solvents used in extraction
procedures.
Similar properties to DES have the so-called NADES, solvents
which are prepared using natural components produced by cell
metabolism. Based on the hypothesis that the existence of NADES
in living organisms can explain many biological processes, several
NADES composed of simple molecules always present in living cells
(urea, amino acids, sugars and choline) have been synthesized and
studied in what respect their solvent properties [6].
Indeed, DES and NADES showed to have excellent potential as
extracting solvents in several sample preparation procedures such
as LPME, UAME and MAE. Literature research in ISI Web of knowl-
edge indicates a total of 1303 references about DES and 56
regarding NADES, of which 321 and 31, respectively, are related
with extraction procedures. Since 2011 the number of publications
has increased considerably, arising 462 in 2017, 443 of which
related to DES and 19 to NADES (Fig. 1). According to the Web
Science Categories, most of these references are within the scope of
multidisciplinary chemistry, followed by chemistry physical or
chemistry analytical, respectively. The reason for the increase
number of publications in the analytical field can be attributed to
the unique features of these new liquids such as high thermal
stability, low thermal conductivity, low volatility and adjustable
miscibility as well as the capacity to be combined with advanced
separation techniques like HPLC and GC. The present review, cover
applications reported until the end of 2017 and is mainly focused on
microextraction techniques using DES/NADES as extracting sol-
vents for food, biological and environmental sample analysis. The
potential of each modern extraction technique will be discussed
and compared with earlier studies and an evaluation of how these
techniques could improve extraction efficiency for a variety of
analytes will be made.
2. Deep eutectic solvents (DES): a brief overview
In the year of 2001, Abbot and co-workers reported that a
mixture of a choline chloride and a metal salt (zinc chloride) could
form a liquid, at temperatures below 100 C [7]. Two years later, the
same group developed a mixture of ChCl with an hydrogen-bond
donor (urea) [8] designating it as DES. In the subsequent years
other DES have been reported namely that formed by mixing ChCl
with different carboxylic acids (oxalic, malonic, and succinic acids)
[9]. Another major family of DES that have been widely explored are
those that combine a carbohydrate (or a reduced derivative as is the
case with sorbitol and mannitol), an urea derivative (N,N0 -dime-
thylurea), and a chloride salt (ammonium chloride) [10]. Generally,
the melting point of the DES is lower than the melting points of
each of its starting components. One of the attractive features of
these novel solvents is the possibility of having, simply by changing
one or both components, a huge number of eutectic mixtures with
different chemical properties. Recently, a new class of DES have
been synthetized, based on combinations of decanoic acid with
 S.C. Cunha, J.O. Fernandes / Trends in Analytical Chemistry 105 (2018) 225e239 227

DES NADES
500 20
450 18

Number of publicaons
Number of publicaons

400 16
350 14
300 12
250 10
200 8
150 6
100 4
50 2
0 0
2006 2007 2008 2009 2010 2011 2012 2013 2014 2015 2016 2017 2006 2007 2008 2009 2010 2011 2012 2013 2014 2015 2016 2017

year year

Web of Science Categories Web of Science Categories

ENERGY FUELS Chemistry AppliedCHMEISTRY APPLIED
ELECTROCHEMISTRY BIOCHEMISTRY MOLECULAR BIOLOGY
CHEMISTRY ANALYTICAL PLANT SCIENCES
THERMODYNAMICS FOOD SCIENCE TECHNOLOGY
PHYSICS ATOMIC MOLECULAR CHEMICAL CHEMISTRY MEDICINAL
MATERIALS SCIENCE MULTIDISCIPLINARY GREEN SUSTAINABLE SCIENCE…
GREEN SUSTAINABLE SCIENCE TECHNOLOGY PHARMACOLOGY PHARMACY
ENGINEERING CHEMICAL ENGINEERING CHEMICAL
CHEMISTRY PHYSICAL CHEMISTRY ANALYTICAL
CHEMISTRY MULTIDISCIPLINARY CHEMISTRY MULTIDISCIPLINARY
0 5 10 15 20 25 30 35 0 5 10 15 20 25 30
% of publicaon from a total of 1303 % of publicaon from a total of 56

Fig. 1. Time-trend (2006e2017) representation of the DES and NADES application and their distribution in the different Web Science Categories.

various quaternary ammonium salts [11] or DL-menthol with properties of DES including freezing point, density, viscosity, con-
several natural acids [12]. This new class of DES showed a high ductivity, and polarity are briefly discussed below in this section
hydrophobic behavior in contrast to the previous hydrophilic DES and some DES are highlighted in Table 1. More detailed information
[11]. Many extraction procedures can become more effective by on the properties of DES can be found in recent reviews by Zang
using these hydrophobic DES due their capability of extracting et al. [3], Tang and Row [16], Smith et al. [5], and references cited
apolar analytes from aqueous solutions. Additionally, these new therein.
solvents have the ability to extract both dissociated and undisso- The DES freezing point is dependent on DES components (type
ciated forms of acidic compounds, broadening the application of of HBD and HBA) and its molar ratio. For instance, the freezing point
hydrophobic DES for different pH environments [11]. of a choline salt-derived DES combined with urea decreases in the
DES are easily produced by just mixing two or more compounds order F > NO3- > Cl > BF4- indicating a correlation with the
and heating them to around 80 C [8,13] or freeze-drying [14], hydrogen bond strength [3,17]. The organic salt/HBD molar ratio
without the subsequent need of any complex purification step, thus has also significant effect on the freezing point of a DES. For
reaction conditions are accessible for any laboratory. However,
owing to the extreme hygroscopicity of the HBA (e.g. ChCl, tetra-
butylammonium, DL-menthol), attracting moisture to prepara-
Table 1
tions, a careful manipulation through using vacuum conditions is Density, viscosity, conductivity and spectroscopic polarity index (EN
T )of some DES at
advised. Although ChCl is the most employed quaternary ammo- specific temperatures [3e5,16].
nium salt because of its low cost and biodegradability (it is indeed
DES (molar ratio) Density Viscosity Condutivity EN
T
produced in large scale and added to the chicken feed as nutrient),
3 2 1
many other halides (methyltriphenylphosphonium bromide, ben- g/cm mm /s (mS cm )
zyltriphenylphosphonium chloride, acetylcholine chloride, tetra- Urea:ChCl (2:1) 1.25 (25 C) 750 (25 C) 0.20 (40 C) 0.84
methylammonium chloride among others) are suitable for making Ethylene Glycol: ChCl (2:1) 1.12 (25 C) 37 (25 C) 7.61 (20 C) 0.8
DES [5]. Glycerol: ChCl (2:1) 1.18 (25 C) 359 (25 C) 1.05 (20 C) 0.86
Malonic: ChCl (1:1) 1.25 (25 C) 721 (25 C) 0.55 (25 C) e
One of the main features of DES is their possibility to be used as 1,4- butanediol: ChCl (3:1) 1.06 140 (20 C) 1.64 (20 C) e
extracting solvent for a wide range of chemical different solutes [3]. Urea: ethylammonium 1.041 128 (40 C) e e
Their uses as solvents in extraction procedures depends on their chloride (1.5:1)
physical properties, such as viscosity, density, miscibility and po- Acetamide: ethylammonium 1.14 64 (40 C) 0.69 (40 C) e
chloride (1.5:1)
larity. It is convenient to select solvents with low viscosity to
2,2,4-trifluroacetamide: ChCl (2:1) 1.342 77 (40 C) 0.286 (40 C) e
facilitate mixing but with a large density difference from the matrix Water 0.992 1 250 1
in order to easy the separation of phases [15]. The main physical
()not found.
228 S.C. Cunha, J.O. Fernandes / Trends in Analytical Chemistry 105 (2018) 225e239

instance, when ChCl was mixed with urea in a molar ratio of 1:1 Table 2
and 1:2, the resulting DES showed a freezing point >50 C and 12 C, Density, viscosity, decomposition temperature (Td) and glass transition temperature
(Tg) of some NADES at specific temperatures [4,20].
respectively [3].
DES exhibit in general higher density values than water with NADES (molar ratio) Density (40 C) Viscosity (40 C) Td/ C Tg/ C
levels ranging from 1.041 g cm3 to 1.63 g cm3 [3]. This feature g/cm3 mm2/s

helps the rapid settling of DES phase in phase separation devices Fructose: ChCl: water (2:5:5) 1.2078 280.8 160 84.58
used in DLLME techniques. Glucose: ChCl: water (2:5:5) 1.2069 397.4 170 83.86
Lactic acid: glucose: water 1.2497 37 135 77.06
Most of the DES exhibit a relatively high viscosity at room
(5:1:3)
temperature (>100 cP) [3]. This can be of substantial benefit when Levulinic acid: ChCl (2:1) 1.14 226.8 176.6 11.9
carrying out SDME, as the high viscosity facilitates the suspension Malic acid: ChCl: water (1:1:2) 1.2303 445.9 201 71.32
of larger drops at the tip of a needle. However, in some extraction Sorbitol: ChCl: water (2:5:6) 1 1854 138.4 >200 89.62
Sucrose: ChCl: water (1:4:4) 1.2269 581 >200 82.96
procedures such as DLLME this characteristic can affect negatively
Xylitol: ChCl: water (1:2:3) 1 17,841 86.1 >200 93.33
the diffusion of analytes. To surpass this drawback some authors, Water 0992 1
increase the temperature during extraction or, alternatively, in-
Td -decomposition temperature; Tg -glass transition temperature; ENR ¼ hcNA/
crease the ChCl concentration, which are reported in literature to
lmax ¼ 28,591/lmax.
reduce the viscosity of some eutectic mixtures.
In general, DES show poor conductivity (lower than 2 mS cm1
at room temperature) [3] due to their high viscosity. However, on 1,2-ethanediol and glycerol [25]. Despite this data, detailed in-
conductivities of DES increase significantly as the temperature in- formation about chemical and physical properties is still scarce, and
creases due to a decrease of the respective viscosity [3,17,18]. only few applications involving NADES in the analytical field, e.g.
Furthermore, the successive addition of ChCl to glycerol lowers the extraction of organic compounds, are reported. Therefore, a deep
viscosity and increases the conductivity (from 0.74 mS cm1 for a research on the preparation of NADES for specific applications is
molar ratio of 1:4 ChCl/glycerol to 1.30 mS cm1 for a molar ratio still required.
of 1:2 ChCl/glycerol) [18] due to more available charge carriers in
an increasingly less viscous solvent. 4. Liquid-phase microextraction
Polarity is one of the most important distinguishing character-
istics of DES, in view of their extraction ability and their miscibility LPME extraction procedures correspond to efficient alternatives
with other solvents. Nevertheless, few studies related to DES po- to traditional liquid-liquid extraction, offering numerous advan-
larity are published. Abbott and co-workers reported that the po- tages such as a high degree of concentration (or enrichment factor),
larity scale of ChCl-glycerol was similar to RNHþ  þ 
3 X , R2NH2 X , and and low consumption of organic solvents while requesting minute
imidazolium ionic liquids [17,18]. In DES composed of an ammo- amounts of samples.
nium salt and carboxylic acids the acidity is mainly provided by the In LPME techniques, usually few microliters of the extracting
organic acid present in the mixture, and an increase of the alkyl side solvent (usually designated as acceptor phase) are placed directly
chain of both compounds leads to a lower ability of the solvent to into the aqueous sample containing analytes (donor phase) or in its
donate protons [19]. Carbohydrate derivative DES present higher headspace and later the extracting solvent is collected.
polarity than those observed for short chain alcohols (e.g. ethanol, In the last few years numerous LPME procedures have been
2-propanol) and some polar aprotic solvents (e.g. dimethylsulf- developed such as SDME, HF-LPME and DLLME. In SDME a drop of
oxide and dimethylformamide) [10]. extractive solvent is suspended in the tip of a syringe which can be
immersed in the aqueous phase of the sample or exposed to the
3. Natural deep eutectic solvents: a brief overview respective headspace (HS-SME). In HF-LPME the extracting phase is
placed inside the lumen of a porous polypropylene hollow fiber, to
Recently, Choi and collaborators have explored natural products improve the stability and reliability of extraction [26]. In DLLME a
as a source of DES solvents, including primary metabolites common cloudy solution of small droplets of extracting solvent (microliters
in living cells (sugars, sugar alcohols, organic acids, amino acids, of a water-immiscible high density organic solvent) is formed and
amines) as well as water [6]. NADES can be obtained by heating a dispersed throughout the aqueous phase using a dispersive solvent,
mixture of two or three components in certain molar ratios in the miscible in both extracting and aqueous phases. Currently some
presence of water, which decrease their viscosity and allows the DLLME procedures do not require the use of dispersive solvent
occurrence of extensive inter molecular interactions e.g. H-bonds being in those cases the cloud solution obtained either by the in-
or ionic bonds [20](Table 2). The usually components of NADES as jection of air bubbles (GA-DLLME), formation of air bubbles using a
HBA are amines (ChCl, ammonium chloride) or amino acids vortex, or by the mechanic action of sucking and injecting the
(alanine, proline, glycine, betain) while as HBD the more common aqueous solution with a syringe (AALLME and AA-DLME). Overall,
are organic acids (oxalic acid, lactic acid, malic acid, etc.) or car- in all these procedures, extraction efficiency is dependent of many
bohydrates (glucose, fructose, maltose, etc.). factors such as partition coefficient, type and volume of extracting
As a whole NADES possess excellent properties as solvents [20], and dispersive solvents, sample volume, analyte properties, agita-
e.g., negligible volatility, a very low melting point (they are liquid tion, ionic strength, extraction time, temperature, etc. A detailed
even below 20 C), a broad polarity range and high solubilization discussion of the importance of each parameter can be found in the
power of a wide range of compounds, especially poorly water- literature [27].
soluble compounds [6,20]. The high solubility of scarce water- LPME techniques using DES or NADES as extracting solvent had
soluble metabolites and macromolecules (e.g. DNA, proteins, cel- been used for extraction of several polar volatile and no-volatile
lulose, and amino acids) has been demonstrated [20e22] as well as compounds from food and water matrices (Table 3).
their suitability as media for enzymatic reactions [6,23] and bio- Tang et al. [28] optimized a two phase HS-SME technique where
transformations [24]. It was verified that NADES composed by a DES (ChCl and ethylene glycol, 1:4 M ratio) drop suspended in a
1 mol of ChCl and 2 mol of 1,2-ethanediol, glycerol, malonic acid, or tip of a microsyringe needle was exposed for 30 min to the head-
urea, the two first present a more dipolar nature than the last ones, space of leaf samples to extract bioactive terpenoids. The use of a
which can be attributed to the presence of alcohol functionalities 2 mL DES drop was enough to allow the adsorption of the target
Table 3
Summary of the application of DES and NADES in extraction techniques from 2006 to 2017.

Techniques DES - NADES Sample Extraction Other features Analyts extracted Instrumental LOQ LOD Ref.
Time analysis
Composition Molar Volume Type amount
ratio

LPME ChCl-1,4-butanediol 1:5 2.0 mL Leaves (Chamaecyparis 200 mg 40 min Heating 70 C-30% vol Flavonoids (myricetin, HPLC-UV Myricetin-0.07 mg/mL; [68]
obtusa) water amentoflavone) amentoflavone-0.09
mg/mL
LPME ChCl-xylitol 2:1 14 g Olive oil 14 g 1h Heating 40 C Phenolic compounds HPLC-DAD [66]
(hydroxytyrosol,
tyrosol, oleacein,
oleocanthal,
oleuropein agylcon,
ligstroside aglycon)
DLLME ChCl-urea 1:2 50 mL Water samples (farm 50 mL 1 min 5 mg MMWCNTs, 150 mL Pesticides (alpha-HCH, GC-ECD alpha-HCB 0.83 ng/L; alpha-HCH 0.25 ng/L; [31]
water, rural water, lake acetonitrile, ultrasonic beta-HCH, gamma- beta-HCH 0.20 ng/L; beta-HCH 0.06 ng/L;
water and river water) bath HCH, delta-HCH, gamma-HCH 0.50 ng/ gamma-HCH 0.15 ng/L;

S.C. Cunha, J.O. Fernandes / Trends in Analytical Chemistry 105 (2018) 225e239
heptachlor, aldrin, L; delta-HCB-0.17 ng/ delta-HCH- 0.05 ng/L;
heptachlor-endo- L; heptachlor- 0.89 heptachlor- 0.27 ng/L;
epoxide, ng/L; aldrin 0.73 ng/L; aldrin- 0.22 ng/L;
alphaendosulfan, heptachlor heptachlor
dieldrin, 4,40 eDDE, endoepoxide 0.46 ng/ endoepoxide 0.14 ng/
endrin, L; alpha-endosulfan L; alpha-endosulfan-
betaendosulfan, 4,40 0.13 ng/L; dieldrin- 0.04 ng/L; dieldrin-
eDDD, 0.10 ng/L; 4,40 -DDE- 0.03 ng/L; 4,40 -DDE
endrinaldehyde, 0.17 ng/L; endrin- 0.05 ng/L; endrin- 0.05
endosulfan-sulfate, 4,40 0.17 ng/L; ng/L; betaendosulfan
eDDT, endrin-ketone betaendosulfan 0.17 0.05 ng/L; 4,40 -DDD
and methoxychlor) ng/L; 4,40 -DDD- 0.17 0.05 ng/L; endrin-
ng/L; endrin- aldehyde 0.11 ng/L;
aldehyde 0.36 ng/L; endosulfan-sulfate
endosulfan-sulfate 0.13 ng/L; 4,40 -DDT-
0.43 ng/L; 4,40 -DDT- 0.22 ng/L;
0.73 ng/L; endrinketone- 0.19 ng/
endrinketone- 0.63 L; methoxychlor- 0.11
ng/L; methoxychlor- ng/L
0.36 ng/L
AG-DLLME ChCl-Chlorophenol 1:2 190 mL Fruit and vegetable 5 ml 6 min pH 2; water bath at 40 C Pesticides GC-FID Diazinon 4.2 mg/L; Diazinon 1.4 mg/L; [33]
juice for 3 min, ice and NaCl (Penconazole, penconazole 0.75 mg/ penconazole 0.25 mg/L;
bath hexaconazole, L; haloxyfop-R- haloxyfop-R-methyl
diniconazole, methyl 1.4 mg/L; 0.45 mg/L;
tebuconazole, hexaconazole 0.84 mg/ hexaconazole 0.28 mg/
diazinon, fenazaquin, L; diniconazole 1.3 L; diniconazole 0.43
clodinafoppropargyl, mg/L; mg/L;
and haloxyfop- R- clodinafoppropargyl clodinafoppropargyl
methyl) 3.9 mg/L; 1.3 mg/L; tebuconazole
tebuconazole 1.9 mg/ 0.64 mg/L;
L; bromopropylate bromopropylate 0.24
0.71 mg/L; fenazaquin mg/L; fenazaquin 0.90
2.7 mg/L mg/L
AAELLME ChCl-5,6,7,8- 1:2 100 mL Water, urine and Urine and plasma 3 min pH 10; 100 mL THF; 10 Methadone GC-FID 0.70 mg/L 2.30 mg/L [34]
Tetrahydro-5,5,8,8- plasma (1 mL) times pulling and
tetramethylnapthalen- pushing
2-ol
HPLME ChCl-ethylene glycol 1:3 10 mL Gasoline, diesel fuel, 1 ml 3 min Ultrasonic bath Phenolic (phenol, r HPLC-UV e phenol-0.025 mg/L; [29]
kerosene -cresol, b-naphthol) r -cresol-0.05 mg/L;
b-naphthol
(continued on next page)

229
Table 3 (continued )

230
Techniques DES - NADES Sample Extraction Other features Analyts extracted Instrumental LOQ LOD Ref.
Time analysis
Composition Molar Volume Type amount
ratio

HPLME ChCl-ethylene glycol 1:4 2 mL Leaves of 300 mg 30 min Heating 100 C ultrasonic Bioactive terpenoids GC-FID Linalool- 6.687 ng/ Linalool- 2.006 ng/mL; [28]
Chamaecyparis obtusa irradian 70 W (Linalool, a- terpineol, mL; a-terpineol- a-terpineol- 3.150 ng/
terpinyl-acetate) 10.50 ng/mL; mL; terpinylacetate-
terpinylacetate- 7.099 2.129 ng/mL
ng/mL
LPME Lactic acid-glucose- 5:1:3 1.5 mL Orange petals of 50 mg 30 min Stirring at 40 C Anthocyanins UPLC-TOF- MS [39]
water Catharanthus roseus

LPME Sucrose-ChCl:water 1:4:4 1.5 mL Safflower (Carthamus 100 mg 1h Stirring at 40 C Phenolic compounds HPLC-DAD [37]
tinctorius L.) (hydroxysafflor Yellow
A, cartormin, and
carthamin)
LPME Sucrose-ChCl:water 1:4:4 3 mL Safflower (Carthamus 50 mg 30 min Stirring at 40 C Colorants (carthamin) HPLC-DAD [38]
tinctorius L.)

S.C. Cunha, J.O. Fernandes / Trends in Analytical Chemistry 105 (2018) 225e239
UAME ChCl-ethylene glycol 1:2 50 mL Olive, almond, sesame 2 mL1(mLoil 1 mL 5 min Ultrasonic bath Phenolic acids (ferulic, HPLC-UV Ferulic acid 1.30- Ferulic acid 0.39-0.56 [44]
and Cinnamon oils hexane) caffeic, and cinnamic 1.86 mg/L; caffeic acid mg/L; caffeic acid - 0.43
acids) - 1.43e2.10 mg/L; e0.63 mg/L; cinnamic
cinnamic acid 1.60 acid 0.48e0.63 mg/L
e2.10 mg/L
UALME ChCl-phenol 1:4 500 mL Tea Residue resulted of 2 min Ultrasonic bath; pH > 6; 1 Cobalt (II) FAAS 3.60 mg/L 1.1 mg/L [51]
wet digestion (0.5 g) ml nitroso-2-naphthol;
added with 10 mL 0.5 ml THF
water
UALME ChCl-phenol 1:3 450 mL Waters (tap, lake, 10 ml 2 min Ultrasonic bath; 0.375 Chromium (III/VI) FAAS 18.2 mg/L 5.5 mg/L [52]
waste) mL 0.5 M H2SO4; 0.4 mL
0.125% sodium
diethyldithiocarbamate;
0.45 mLTHF
UALME ChCl:phenol:FeCl3 1:2:1 25 mL n-Heptane 1.5 ml 5 min Ultrasonic bath (35 kHz) Thiophene GC-FID [47]
25 C
UALME ChCl-phenol 1:2 100 mL Waters (tap and 1.5 ml 20 min Ultrasonic bath; 100 ml Benzene, toluene, HPLC-UV Benzene- 21.0 mg/L; Benzene- 6.2 mg/L; [48]
industrial wastewater) THF ethylbenezene, PAHs toluene- 22.0 mg/L; toluene- 6.8 mg/L;
(biphenyl, fluorene, ethylbenzene-2.90 ethylbenzene-0.8 mg/L;
phenanthrene, mg/L; biphenyl- 2.30 biphenyl- 0.7 mg/L;
anthracene, pyrene, mg/L; fluorene- 2.20 fluorene- 0.7 mg/L;
chrysene and benzo [a] mg/L; phenanthrene- phenanthrene- 0.09
pyrene) 0.30 mg/L; mg/L; anthracene- 0.02
anthracene- 0.06 mg/ mg/L; pyrene- 0.07 mg/
L; pyrene- 0.26 mg/L; L; chrysene- 0.21 mg/L;
chrysene- 0.70 mg/l; benzo [a]pyrene- 0.08
benzo [a]pyrene- 0.28 mg/L
mg/L
UALME ChCl-phenol 1:4 500 mL Waters (farmed and 2 ml 3 min Ultrasonic bath; pH ¼ 3; Malachite green UV-VIS 11.8 mg/L 3.6 mg/L [49]
ornamental aquarium 500 ml THF
fish)
UALME ChCl-Ox 1:1 1 mL Grape skin 100 mg freeze dried 50 min Ultrasonic bath: 65 C, 35 Phenolics HPLC-UV (þ)-Catechin- 1.24 (þ)-Catechin- 0.37 mg/ [42]
skin kHz; 25% water ((þ)-Catechin, mg/L, delphinindin-3- L, delphinindin-3-O-
delphinindin-3- O- O- glucoside- 0.60 glucoside- 0.18 mg/L,
glucoside, Cyanidin-3- mg/L, Cyanidin-3-O- cyanidin-3-O-
O-glucoside, glucoside- 0.71 mg/L, glucoside-0.21 mg/L,
petunidin-3-O- petunidin-3- petunidin-3-O-
 glucoside, peonidin-3- Oglucoside- 0.80 mg/ glucoside- 0.24 mg/L,
Oglucoside, malvidin- L, peonidin-3- O- peonidin-3-O-
3-O-glucoside, glucoside-0.65 mg/L, glucoside- 0.19 mg/L,
quercetin-3-O- malvidin- 3-O- malvidin-3-O-
glucoside) glucoside - 0.90 mg/L, glucoside - 0.30 mg/L,
quercetin-3-O- quercetin-3-O
glucoside 0.46 mg/L glucoside 0.05 mg/L
UALME tetramethylammonium 1:3 30 mL Saflower, 1 ml (10% oil - 90% 7 min Ultrasonic bath 50 C Plant growth HPLC-UV IAA -0.05 mg/mL; IBA [45]
chlorideeethylene olive,camellia, colza hexane) regulators (IAA, IBA, 4- -0.06 mg/mL; 4-IPOAA
glycol and soybean oils IPOAA) 0.75 mg/mL
UALME ChCl-Laevulinic Acid 1:2 1 mL Platycladi Cacumen 25 mg 30 min Ultrasonic bath: 50 C, Flavonoid glycosides HPLC-UV [43]
200 W; 75% water and aglycones
(myricitrin, quercitrin,
amentoflavone,
hinokiflavone)
UAME ChCl-malic acid 1:1 1 mL Wine lees 33.3 mg 30.6 min Ultrasaound bath: 341.5 Anthocyanins HPLC-DAD Delphinidin-3-O Delphinidin-3-O [57]
W, 37 kHz, 35 C; 35.4% glucoside- 0.87 mg/L; glucoside- 0.28 mg/L;
water cyanidin-3-O cyanidin-3-O

S.C. Cunha, J.O. Fernandes / Trends in Analytical Chemistry 105 (2018) 225e239
glucoside- 0.52 mg/L; glucoside- 0.17 mg/L;
petunidin-3-O petunidin-3-O
glucoside- 0.44 mg/L; glucoside- 0.15 mg/L;
peonidin-3- O peonidin-3- O
glucoside- 0.62 mg/L; glucoside- 0.2 mg/L;
malvidin- 3-O malvidin-3- O
glucoside- 0.81 mg/L glucoside- 0.25 mg/L
UAME ChCl-glycerol 1:1 1 mL Tartary buckwheat 40 mg 1h Ultrasaound bath: 20 Flavonoid (rutin) HPLC-UV [58]
hulls kHz, 200 W, 40 C; 20%
water
UAME Fructose-citric acid 1:1 1 mL Raw and processed 25 mg 15 min Ultrasound bath (100 w Gluten Enzyme Linked [55]
food (spaghetti, biscuts, 42 kHz) 40% amplitude; ImmunoSorbent
and ham) 20% water Assay
UAME ChCl:citric acid 1.1 600 mL Soy products 200 mg 60 min Ultrasonic bath: 440 W, Isoflavones (GT, DA, UHPC-UV DAGL- 0.40 mg/g; DAGL- 0.12 mg/g; [56]
(soybeans, flour, pasta, 60 C; 30% water GTGL, daidzin DAGL) GTGL- 0.45 mg/g; GTGL- 0.14 mg/g; DA-
breakfast cereals, DA- 0.25 mg/g; 0.08 mg/g; GT 0.06 mg/g
cutlets, tripe, soy drink, GT 0.20 mg/g
soy nuts, soy cubes and
three different dietary
supplements)
MAE ChCl-1,2-propanediol 1:1 10 mL Radix Salviae 50 mg 11.11 min Microwave: 800 W, 70 C Bioactive compounds HPLC-DAD ROS- 0.80 mg/mL; LIT- ROS- 0.24 mg/mL; LIT- [60]
miltiorrhizae (ROS, LIT, SAB, SAA, 1.37 mg/mL; SAB- 1.96 0.49 mg/mL; SAB- 0.62
TIIA) mg/mL; SAA- 0.87 mg/ mg/mL; SAA- 0.31 mg/
mL; TIIA- 1.45 mg/mL mL; TIIA- 0.48 mg/mL
UAME ChCl-ethylene glycol 1:2 50 mL Olive, almond, sesame 2 mL (1 mL oil 1 5 min Ultrasonic bath Phenolic acids (ferulic, HPLC-UV Ferulic acid 1.30- Ferulic acid 0.39-0.56 [44]
and Cinnamon oils mL hexane) caffeic, and cinnamic 1.86 mg/L; caffeic acid mg/L; caffeic acid - 0.43
acids) - 1.43e2.10 mg/L; e0.63 mg/L; cinnamic
cinnamic acid 1.60 acid 0.48e0.63 mg/L
e2.10 mg/L
UALME ChCl-phenol 1:4 500 mL Tea Residue resulted of 2 min Ultrasonic bath; pH > 6; 1 Cobalt (II) FAAS 3.60 mg/L 1.1 mg/L [51]
wet digestion (0.5 g) mL nitroso-2-naphthol;
added with 10 mL 0.5 mL THF
water
UALME ChCl-phenol 1:3 450 mL Waters (tap, lake, 10 mL 2 min Ultrasonic bath; 0.375 Chromium (III/VI) FAAS 18.2 mg/L 5.5 mg/L [52]
waste) mL 0.5 M H2SO4; 0.4 ml
0.125% sodium
diethyldithiocarbamate;
0.45 mL THF
UALME ChCl:phenol:FeCl3 1:2:1 25 mL n-heptane 1.5 mL 5 min Ultrasonic bath (35 kHz) Thiophene GC-FID [47]
25 C
(continued on next page)

231
Table 3 (continued )

232
Techniques DES - NADES Sample Extraction Other features Analyts extracted Instrumental LOQ LOD Ref.
Time analysis
Composition Molar Volume Type amount
ratio

UALME ChCl-phenol 1:2 100 mL Waters (tap and 1.5 mL 20 min Ultrasonic bath; 100 mL Benzene, toluene, HPLC-UV Benzene- 21.0 mg/L; Benzene- 6.2 mg/L; [48]
industrial wastewater) THF ethylbenezene, PAHs toluene- 22.0 mg/L; toluene- 6.8 mg/L;
(biphenyl, fluorene, ethylbenzene-2.90 ethylbenzene-0.8 mg/L;
phenanthrene, mg/L; biphenyl- 2.30 biphenyl- 0.7 mg/L;
anthracene, pyrene, mg/L; fluorene- 2.20 fluorene- 0.7 mg/L;
chrysene and benzo [a] mg/L; phenanthrene- phenanthrene- 0.09
pyrene) 0.30 mg/L; mg/L; anthracene- 0.02
anthracene- 0.06 mg/ mg/L; pyrene- 0.07 mg/
L; pyrene- 0.26 mg/L; L; chrysene- 0.21 mg/L;
chrysene- 0.70 mg/L; benzo [a]pyrene- 0.08
benzo [a]pyrene- mg/L
0.28 mg/L
UALME ChCl-phenol 1:4 500 mL Waters (farmed and 2 mL 3 min Ultrasonic bath; pH ¼ 3; Malachite green UV-VIS 11.8 mg/L 3.6 mg/L [49]

S.C. Cunha, J.O. Fernandes / Trends in Analytical Chemistry 105 (2018) 225e239
ornamental aquarium 500 mL THF
fish)
UALME ChCl-Ox 1:1 1 mL Grape skin 100 mg freeze dried 50 min Ultrasonic bath: 65 C, 35 Phenolics HPLC-UV (þ)-Catechin- 1.24 (þ)-Catechin- 0.37 mg/ [42]
skin kHz; 25% water ((þ)-Catechin, mg/L, delphinindin-3- L, delphinindin-3-O-
delphinindin-3- O- O- glucoside- 0.60 glucoside- 0.18 mg/L,
glucoside, Cyanidin-3- mg/L, Cyanidin-3-O- cyanidin-3-O-
O-glucoside, glucoside- 0.71 mg/L, glucoside-0.21 mg/L,
petunidin-3-O- petunidin-3- petunidin-3-O-
glucoside, peonidin-3- Oglucoside- 0.80 mg/ glucoside- 0.24 mg/L,
Oglucoside, malvidin- L, peonidin-3- O- peonidin-3-O-
3-O-glucoside, glucoside-0.65 mg/L, glucoside- 0.19 mg/L,
quercetin-3-O- malvidin- 3-O- malvidin-3-O-
glucoside) glucoside - 0.90 mg/L, glucoside - 0.30 mg/L,
quercetin-3-O- quercetin-3-O
glucoside 0.46 mg/L glucoside 0.05 mg/L
UALME tetramethylammonium 1:3 30 mL Saflower, 1 ml (10% oil - 90% 7 min Ultrasonic bath 50 C Plant growth HPLC-UV IAA -0.05 mg/mL; IBA [45]
chlorideeethylene olive,camellia, colza hexane) regulators (IAA, IBA, 4- -0.06 mg/mL; 4-IPOAA
glycol and soybean oils IPOAA) 0.75 mg/mL
UALME ChCl-Laevulinic Acid 1:2 1 mL Platycladi Cacumen 25 mg 30 min Ultrasonic bath: 50 C, Flavonoid glycosides HPLC-UV [43]
200 W; 75% water and aglycones
(myricitrin, quercitrin,
amentoflavone,
hinokiflavone)
UAME ChCl-malic acid 1:1 1 mL Wine lees 33.3 mg 30.6 min Ultrasaound bath: 341.5 Anthocyanins HPLC-DAD Delphinidin-3-O Delphinidin-3-O [57]
W, 37 kHz, 35 C; 35.4% glucoside- 0.87 mg/L; glucoside- 0.28 mg/L;
water cyanidin-3-O cyanidin-3-O
glucoside- 0.52 mg/L; glucoside- 0.17 mg/L;
petunidin-3-O petunidin-3-O
glucoside- 0.44 mg/L; glucoside- 0.15 mg/L;
peonidin-3- O peonidin-3- O
glucoside- 0.62 mg/L; glucoside- 0.2 mg/L;
malvidin- 3-O malvidin-3- O
glucoside- 0.81 mg/L glucoside- 0.25 mg/L
UAME ChCl-glycerol 1:1 1 mL Tartary buckwheat 40 mg 1h Ultrasaound bath: 20 Flavonoid (rutin) HPLC-UV [58]
hulls kHz, 200 W, 40 C; 20%
water
UAME Fructose-citric acid 1:1 1 mL Raw and processed 25 mg 15 min Ultrasound bath (100 w Gluten Enzyme Linked [55]
food (spaghetti, biscuts, 42 kHz) 40% amplitude; ImmunoSorbent
and ham) 20% water Assay
 S.C. Cunha, J.O. Fernandes / Trends in Analytical Chemistry 105 (2018) 225e239 233

compounds volatilized from the samples heated at 100 C and

[56]

[60]
ultrasonicated at 70 W. Once stopped the procedure, the suspended
drop was retracted back into the microsyringe and injected into a
0.08 mg/g; GT 0.06 mg/g

0.49 mg/mL; SAB- 0.62

ROS- 0.24 mg/mL; LIT-
GTGL- 0.45 mg/g; DA- GTGL- 0.14 mg/g; DA-

mg/mL; SAA- 0.31 mg/

mL; TIIA- 0.48 mg/mL
GC-FID.
DAGL- 0.12 mg/g;

In the same year 2014, Gu et al. [29] employed a similar HS-SME

procedure for the extraction of phenolic compounds from crude
oils; they also used ChCl and ethylene glycol (1:3, molar ratio) as
DES and ultrasonication to reduce the extraction time and to
improve the yields [29].
ROS- 0.80 mg/mL; LIT-
1.37 mg/mL; SAB- 1.96
mg/mL; SAA- 0.87 mg/
mL; TIIA- 1.45 mg/mL

A three phase HS-SME procedure using DES (methyl-

0.25 mg/g; GT 0.20

triphenylphosphonium iodide and ethylene glycol, 1:4 M ratio) plus

DAGL- 0.40 mg/g;

20% v/w methanol as an acceptor phase and n-dodecane as

extraction solvent was applied to extract steroidal hormones
(dydrogesterone and cyproterone acetate) from biological samples
mg/g

(urine and plasma) [30]. The analytes were further analyzed by LC-
UV, and the obtained performance in what respect range of line-
arity and LOD have compared positively to those reported in liter-
HPLC-DAD
UHPC-UV

ature using more sensitive detectors.

Recently, Yousefi et al. [31] developed a two phase HS-SME
procedure using an hydrophobic magnetic bucky gel formed by
combining DES (ChCl:chlorophenol, 1:2 M ratio) and magnetic
11.11 min Microwave: 800 W, 70 C Bioactive compounds
GTGL, daidzin DAGL)
Isoflavones (GT, DA,

(ROS, LIT, SAB, SAA,

multiwalled carbon nanotubes for the extraction of volatile hydro-

carbons from water and urine samples. The extraction was suc-
cessfully completed in 10 min using a drop of 20 mL that was placed
in the headspace of the vial containing the aqueous sample. The
extraction temperature was kept at 30 C and the vial was stirred at
TIIA)

1200 rpm. The use of a large drop compared with other HS-SME
Ultrasonic bath: 440 W,

based DES procedure showed to provide a higher sensitivity.

When compared with conventional solvents DES showed more
60 C; 30% water

adequate to form stable drops for HS-SME due to its higher thermal
stability, higher viscosity, lower volatility and adjustable miscibility.
Concerning to DLLME conventional process using DES as
extracting solvent it was used just by Ferrone et al. [32] for deter-
mination of oxyprenylated phenylpropanoids in vegetable oils. The
extraction was performed with 45 mL of DES (phenylacetic acid:
60 min

betaine, 2:1 M ratio) and isopropanol (200 mL) as dispersive sol-

vent; following agitation and centrifugation, 5 mL of the bottom
phase were injected in a UHPLC-PDA system. This method allowed
the achievement of good limits of detection, linearity and repro-
ducibility similar or better to those reported in literature for the
same analytes.
200 mg

50 mg

A novel rapid and efficient GA-DLLME procedure characterized

by the absence of dispersive solvent was developed by Farajzadeh
cutlets, tripe, soy drink,
soy nuts, soy cubes and

et al. [33] for the extraction of 9 pesticide residues from fruit and
(soybeans, flour, pasta,

three different dietary

vegetable juices. In the optimized extraction procedure, air was

breakfast cereals,

bubbled into a test tube to disperse the extracting DES (ChCl: 4-

supplements)
Radix Salviae
Soy products

miltiorrhizae

chlorophenol, 1:2 M ratio) into the aqueous solution, in order to

obtain a cloudy solution. After centrifugation, an aliquot of the
sedimented phase is injected into GC-FID for the separation and
determination of the enriched pesticide residues [33].
600 mL

Another new type of DLLME is the AA-LLME. Similar to GA-

10 mL

DLLME, in this technique the cloud solution is obtained by suck-

ing and injecting several times the mixture of extracting solvent
1:1
1.1

and aqueous sample. Lamei et al. [34] used this technique

employing a mixture of ChCl and 5,6,7,8-tetrahydro-5,5,8,8-
ChCl-1,2-propanediol

tetramethylnaphthalen-2-ol (1:2 M ratio) as DES extractor and

THF as a demulsifier agent, for the extraction of methadone from
ChCl:citric acid

water and biological samples. Therefore, the dispersion of the

aggregated DES droplets into aqueous phase was achieved by the
effect of sucking and injecting (10 times) the mixture of sample,
extracting solvent and demulsifier agent. After a brief centrifuga-
tion the uplayer was directly analyzed by GC-FID. A pronounced
advantage in comparison to the other LPME described in the liter-
UAME

ature for methadone extraction is the low toxicity and low cost of
MA

the extracting solvent used.

234 S.C. Cunha, J.O. Fernandes / Trends in Analytical Chemistry 105 (2018) 225e239
AA-DLLME was developed and optimized by Ge et al. [35] for
pre-concentration and separation of 6 benzophenones from
different types of waters. Using a hydrophobic DES (mixture of DL-
menthol and decanoic acid, 2:1 M ratio) as extractor solvent, which
was several times injected into the aqueous solutions, the authors
were able to extract efficiently the benzophenones without the use
of dispersive solvents.
In a similar way, Zheng et al. [36] used a hydrophobic DES with
1-decyl-3-methylimidazolium chloride and 1-dodecanol (1:2, M
ratio) for the extraction of benzoylureas from waters, followed by
HPLC-UV analysis. Various conditions were optimized, namely the
type and volume of extractor, salt addition, vortex time, tempera-
ture of extraction and pH of the aqueous sample. The authors
concluded that 8.0 ml of sample solution with 450 mg sodium
chloride and pH at 4e6 provided the best recoveries when
extracted with 40 ml of DES during 3 min in vortex at 40 C.
Since the discovery of NADES by the Choi group [6] several
works have been published by the same research group exploring
the potential of NADES as solvents to extract bioactive compounds
such as antocyanins, and phenols from several solid samples
(Table 3) [20,37,38]. In general, a low amount of sample (<100 mg)
is extracted with a small volume of the chosen NADES (<3 mL) at
controlled temperature (40 C) using vortex stirring. A pronounced
advantage of the use of NADES in LPME in comparison to the
conventional extracting solvents is the higher stability of the
extract obtained and, of course, the use of a solvent sustainable and
environmental-friendly.
In 2013, Dai et al. [20] tested different NADES and a multivariate
data analysis to demonstrate that the extractability of a wide range
of phenolic compounds (hydrophilic and hydrophobic metabolites
of safflower) were greater with NADES than with conventional
solvents. This high extractability was attributed to H-bonding in-
teractions between molecules of NADES and phenolic compounds.
In 2014 the same group continued the previous work, exploring
the ability of different NADES to stabilize unstable phenolic com-
pounds extracted from safflower such as carthamin [38]. They
found that carthamin is more stable in glucose-ChCl and sucrose-
ChCl than in acidic NADES such as proline-malic acid or lactic-
acid-glucose and the stabilization aptitude of NADES increases
with increasing viscosity (low water content). Lately, Dai et al. [39]
tested diverse NADES for the extraction of anthocyanins from
purple and orange petals of Catharanthus roseus. Among the
NADES evaluated, LGH and PCH showed to present a similar
extraction power for anthocyanins as conventional organic sol-
vents, with the additional advantage exhibited by LGH to provide
at least three times higher stability capacity for cyanidins than
acidified ethanol, facilitating their extraction and further analysis
by UPLC-TOF-MS.
These works clearly showed that both DES and NADES have a
high potential as extracting solvents for a wide range of analytes of
low to medium polarity and demonstrated a bright future for the
application of these novel classes of extracting solvents in LPME
field.
5. Ultrasound assisted microextraction
In UAME techniques the extraction procedure takes place under
ultrasonic energy (typical systems use a frequency energy of
40 kHz), which favors the contact between sample and extracting
solvent. Ultrasounds radiation can be applied by means of water-
baths or ultrasonic probes, being the last ones able to deliver
higher energy input per volume due to a more focused and uniform
power input. Conversely, ultrasonic probes provide an entire con-
trol over the most important sonication parameters, leading to
more reproducible results [40].
In general, extraction efficiency of UAME techniques depends on
different factors such as extracting solvent (type, pH, volume), ul-
trasonic conditions (temperature, amplitude of sonication, soni-
cation time), and sample features (matrix, amount, particle size)
[40,41].
The main advantage of using UAME compared with conven-
tional DLLME is that no dispersive solvent is needed to achieve a
higher surface area of contact between the extracting solvent and
sample. Additionally, UAME can promote a greater penetration of
the extracting solvent in solid samples matrices.
DES based UAME have been applied mostly for extraction of
organic compounds from liquid samples, although their application
to solid samples or to the extraction of inorganic analytes has also
been proposed (Table 3). Some works have explored the use of
magnetic DES or NADES as extracting solvents with the purpose to
simplify the process and improve yields. In general, UAME involves
lower volumes of DES (less than 500 mL) and shorter times of
extraction (typically less than 15 min), than the previous
mentioned LPME techniques. Like LPME, UAME also did not allow
automation of the extraction process.
Cvjetko Bubalo et al. [42] used a ChCl-based DES containing
oxalic acid as a hydrogen bond donor with 25% of water, for the
extraction of grape skin phenolic compounds. To increase extrac-
tion yield, samples were sonicated in a water bath during 50 min at
65 C. According to the authors, the extraction efficiency obtained
by UAME was higher than those obtained with microwave-assisted
extraction or conventional extraction methods.
Recently, Zhuang et al. [43] combined DES-UAME with HPLC-
UV for the simultaneous determination of flavonoid glycosides
and respective aglycones in Platycladi Cacumen, a traditional
Chinese herbal medicine. In the optimized conditions, 1 ml of
DES (ChCl-laevulinic acid with 75% of water) was mixed with
25 mg of sample during 30 min at 50 C under an ultrasound bath
(200 W, 40 kHz); the extract was further diluted 8 times with
acetonitrile before HPLC-UV analysis. Additionally, the authors
evaluated the possibility to recover the target analytes from the
DES extracts, in order to enable their further application in
pharmaceutical or food industry. For that purpose, several mac-
roporous resins were evaluated, being the best results for both
flavonoid glycosides and aglycones obtained with LX-38.
Khezeli et al. [44] have reported a simple and highly repro-
ducible UAME method for the determination of ferulic, caffeic and
cinnamic acids in olive, almond, sesame, and cinnamon oil samples,
using ChCl and ethylene glycol (1:2, M ratio) as DES extracting
solvent. The solvent was added to the sample dissolved in n-hex-
ane, then the mixture was placed in an ultrasound bath for 5 min, to
form an emulsion of microdroplets, and consequently increase the
contact surface between DES and sample. Once completed the
extraction, the DES phase containing the analytes was separated by
centrifugation, and submitted to HPLC-UV analysis. The results
indicated that the novel approach have shown the advantages of
good sensitivity (limits of detection between 0.39 and 0.63 mg/L),
reproducibility (RSD <5.1%), convenience (extraction time less than
15 min), and high accuracy (recoveries from 95 to 105%). Tan et al.
[45] employed also UAME-DES followed by HPLC/UV for determi-
nation of plant growth regulators from several vegetable oil sam-
ples, using as DES a tetramethylammonium chlorideeethylene
glycol mixture (1:3 M ratio). When compared with other tech-
niques such as SDME, SPME and S-SIL-based MSPD the UAME-DES
proposed in the works above referred are more sensible, fast and of
simpler execution [39,40].
The applicability of hydrophobic DES for extraction of analytes
dissolved in raw waters such as UV-filters have been evaluated by
Wang et al. [46]. After optimization of diverse parameters (e.g.
sample volume, salt addition, sample pH, ultrasonic time) the
 S.C. Cunha, J.O. Fernandes / Trends in Analytical Chemistry 105 (2018) 225e239
authors proposed the use of a mixture of tri-
octylmethylammonium chloride with decanoic acid (1:3 M ratio)
as DES, using sonication for 5 min in an ultrasonic bath (150 W;
40 kHz) for pre-concentration and separation of three benzo-
phenones. This method has the great advantage of enabling the
extraction and purification in one step without the use of toxic
solvents.
The use of a magnetic deep eutectic solvent (MDES) formed by
the mixture of ChCl with phenol and anhydrous FeCl3 (1:2:1 M
ratio) was recently proposed by Khezeli et al. [47] for the extraction
of thiophene from a heptane solution. The MDES was dispersed into
the solution by ultrasounds during 5 min, in order to enhance the
mass transfer of thiophene from n-heptane to MDES phase. After
that, microdroplets of MDES were collected by a magnet and the
remained concentration of thiophene in n-heptane was analyzed
by GC-FID. The method provided very good results in the elimi-
nation of thiophene from the initial solution (close to 100%), with
the great advantage of eliminating the centrifugation step usually
required in any LPME extraction. According to the authors, the
developed MDES are able to be reused after four runs without
losses of extraction efficiency.
A novel UAME application based on DES, named by the authors
as emulsification liquid-liquid microextraction (ELLME-DES) was
introduced by the group of Khezeli [48] for the simultaneous
extraction of BTE (benzene, toluene, and ethylbenzene) and seven
polycyclic aromatic hydrocarbons from water samples. Here, the
ultrasound helps both the mass transfer between phases and the
formation of tiny emulsified droplets, increasing the contact area.
Other novelty was the use of a water miscible aprotic solvent, THF
for instance, able to decrease the tendency of water molecules to
interact with DES and thus promoting the self-aggregation of DES
microdroplets. Briefly, 100 mL of DES (mixture of ChCl:phenol in
1:2, 1:3, and 1:4 M ratios) were added to 1.5 mL of sample and a
homogeneous solution was formed immediately. The further in-
jection of 100 mL of THF followed by sonication for 20 min in an
ultrasonic bath provided a turbid state in which DES micro-
droplets are entirely disperse along all the water phase, so
enhancing the transference rate of the compounds from the water
to the DES phase. After a centrifugation step (10 min at 3000 rpm)
the DES upper phase was withdrawn through a micro-syringe and
analyzed by HPLC-UV. Very good results were obtained in what
concerns linearity, precision, and accuracy. These approach could
be very useful for the multi-analysis of hydrocarbons in envi-
ronmental water samples, with the advantages of simplicity, low
cost, and high sensitivity providing by the high enrichment factor
of the extraction procedure, while avoiding the use of chlorinated
solvents.
Similar approach was used by Aydin et al. [49] for the
extraction of malachite green from farmed and ornamental
aquarium fish water samples followed by a simple UVeVIS
spectrometry analysis. The developed method presented good
extraction recoveries (95%) and high precision (RSD < 4% for real
samples) for evaluation of this anti-parasitic which its illegally
used in the aquaculture industry to prevent fish diseases caused
by external parasites and fungus. Moreover, the method showed a
good selectivity for malachite green along with other advantages
such as simplicity, low cost, rapid separation and ease of opera-
tion. The same authors proposed a ELLME-DES combined vortex
for separation and pre-concentration of curcumin in herbal tea
samples followed by UV-Vis determination using 400 mL of DES
(ChCl: phenol, M ratio 1:4) as extracting solvent, 400 mL of THF as
emulsifier agent and 2 min of ultrasonication to achieve the best
extraction yields [50].
The same principles were at the base of the application of DES as
extracting solvent on UAME procedures for extraction of inorganic
235
compounds, namely cobalt (II) and chromium (III/VI) ions from
aqueous matrices, performed by the group of Soylak [51,52]. In both
works, the metals were previously complexed by adding as ligands
1-nitroso-2-naphthol and sodium diethyldithiocarbamate, respec-
tively, allowing the formation of very stable and hydrophobic
complex and consequently promoting the extraction efficiency. The
DES solvents were mixtures of ChCl with phenol (4:1, and 3:1 M
ratios, respectively) and THF was added to allow the self-
aggregation and full separation of the DES molecules from the
water phase. Following sonication for 2 min and centrifugation, the
authors were able to recollect a DES aggregate containing the
complexed metals, suitable for quantification by AAS. Several pa-
rameters affecting the extraction performance of the analytes were
optimized, namely sample amount, pH, type and volumes of DES,
complexing agent, and emulsifier solvent, and ultrasonication time.
The results obtained under the optimized conditions are compa-
rable or even better to those reported in literature for the same
analytes, apart from presenting a high enrichment factor and
consequently low detection limits.
A comparable approach to those previously described was used
by Panhwar et al. in two works aiming the pre-concentration of
inorganic compounds from water and food samples, namely hy-
drophobic chelates of Se(IV) with 3,30 -diaminobenzidine [53] and
Al (III) with 8-hydroxyquinoline [54] before ETAAS analysis. In both
works mixtures of ChCl:phenol were used as extractors with a
molar ratio of 1:3 and 1:4, respectively, and THF was the emulsifier
agent. The authors compared the proposed methods with other
ETAAS procedures previously used, in which the pre-concentration
was done with LLE, SPE or conventional DLLME, and found no
significant differences between the performance data achieved,
with the advantages already indicated that characterize the use of
DES.
The use of NADES in UAME is been increasing in last years as
mentioned for other microextraction techniques. Lores et al. [55]
used a fructose-citric acid NADES mixture in combination with
UAE for fast and green gluten determination by ELISA. The NADES-
UAME developed allows the replacement of the ethanol-water
solution commonly used for gluten extraction with success, once
good recoveries (62e135%) and high precision RSD < 15%) were
obtained. In another work, Bajkacz and Adamek [56] used NADES
combined with UAME for the isolation of isoflavones (daidzin,
genistin, genistein, daidzein) from soy products followed by UPLC-
UV analysis. The best results were obtained using a mixture of
ChCl:citric acid (1:1 M ratio) with a water content of 30%, and a
ratio of NADES volume to sample amount of 3 The extraction time
was 60 min at 60 C and the ultrasonic power was 616 W. Bosiljkov
et al. [57] used chemometric tools, namely a response surface
methodology to optimize NADES-UAME/HPLC-DAD method for the
quantification of anthocyanins in wine lees. The maximum amount
of extracted compounds were achieved using ChCl:malic acid
(1:1 M ratio) with 35.4% (w/w) of water in a ultrasound bath at
341.5 W during 30.6 min. Recently, Huang et al. [58] used a NADES-
UAME/HPLC-UV method for the extraction of rutin from tartary
buckwheat hull. The authors proved that the solubility of rutin
increased by 660e1577 times in ChCL-glycerol-based NADES when
compared to water.
The increasing number of works using DES and NADES in UAME,
in recent years make us believe that this novel class of extracting
solvents can be used in removal and extraction of a wide range of
analyte in environmental and food matrices. Despite all the ad-
vantages, the majority of the DES extracting solvents used in UAME
described procedures requires the use of HPLC for organic com-
pounds, or ETAAS and AAS for inorganic compounds, as quantita-
tive final technique, due to the incompatibility with GC systems
caused by DES low volatility.
236 S.C. Cunha, J.O. Fernandes / Trends in Analytical Chemistry 105 (2018) 225e239
6. Microwave-assisted extraction
MAE employs non-ioning electromagnetic irradiation (in a fre-
quency range of 0.3e300 GHz) to heat both solvent and samples by
movements of ions and rotation of molecular and atomic dipoles, in
order to increase extraction kinetics [59].
MAE may be performed in closed vessels under pressure
(pressurized MAE) or in open vessels at atmospheric pressure
(focused MAE). The first ones are most used in food analysis since
usually provide enhanced extraction yields. The extraction effi-
ciency is dependent on the solvent (nature and solvent/sample
ratio), temperature and pressure, extraction time, radiation power,
sample composition (moisture mainly), and particle size (prefer-
ably 0.1e2 mm) [41,59].
MAE using DES as extracting solvent was employed by Chen
et al. [60] to extract 5 active ingredients namely rosmarinic acid,
lithospermic acid, salvionalic acid B, salvionalic acid A, and tan-
shinone II A from Radix Salviae miltiorrhizae followed by HPLC-VWD
quantification. Among twenty-five DES tested, the authors
concluded that ChCl-1,2-propanediol (1:1, M ratio) showed the best
yields. Other extraction factors, including temperature, time, power
of microwave, and solid/liquid ratio, were systematically investi-
gated by response surface methodology. The hydrophilic and hy-
drophobic compounds were extracted simultaneously under the
optimized conditions: 20 vol% of water in ChCl-1,2-propanediol as
solvent, microwave power of 800 W, temperature at 70 C, time at
11.11 min, and solid/liquid ratio of 0.007 g/mL. The proposed DES
combined with the MAE provided a prominent advantage for fast
and efficient extraction of active compounds compared with con-
ventional procedures. Moreover, in comparison to the techniques
previously used, MAE allows the automation of the extraction (40
samples per batch). The main limitations of MAE based techniques
are the higher costs of the equipment and the usual presence on the
extracts of many interfering compounds due to the exhaustive
extraction process, which requires a cleanup step prior to the
analysis.
7. Other applications of DES/NADES in extraction techniques
DES have been also demonstrated to be an interesting alterna-
tive as reaction solvents in the production of new sorbents [61] or
polymeric phases [62] used in solid-phase extractions (SPE), which
enhance the range of possible DES applications in environmentally
friendly extraction procedures. Another application is using DES as
dissolution solvent [63] as a new alternative to the conventional
Soxhlet extraction.
7.1. DES as solvents on liquid-liquid extraction
Guo et al. [64] used a DES based on tetrahylammonium chloride
and phenol (0.8:1, M ratio) to extract phenol (99.9%) from model oil
(toluene). The extraction was achieved by mixing directly the DES
with the matrix by 30 min at room temperature, avoiding the use of
alkali and acids and the production of phenol containing waste
water, common in the traditional methods. The same group pre-
viously has verified that the addition of ChCl allowed a formation of
DES due to the presence of phenols (phenol, cresols) in model oils
(hexane, toluene, and p-xylene) [65]. The extraction of phenolic
compounds from virgin olive oil using DES as extracting solvent
was evaluated by García et al. [66]. The best results for the two most
abundant secoiridoid derivatives (oleacein and oleocanthal) were
obtained with ChCl: xylitol (2:1, M ratio) and chlorine chloride:1,2
propanediol (1:1, M ratio) as DES solvents. The better conditions
were achieved by mixing the extractive solvent directly with the
samples inside a bath at 40 C with agitation for 1 h. Excellent
extraction yields were obtained with the tested DES compared with
the commonly used mixture of methanol/water. Despite all the
advantages due to the use of DES such as low cost and low toxicity,
the extraction time was not reduced.
The group of Ghanemi [67] used DES (ChCleOx, 1:2 M ratio) at
55 C for 30 min to dissolve fish samples as a preliminary step for
determination of PAHs by HPLV-UV. After dissolution, PAHs were
quantitatively extracted from ChCl-Ox solution with 5 mL of
cyclohexane and stirring for 20 min [67]. These procedures provide
some operational advantages when compared with the conven-
tional Soxhlet extraction such as simplicity, low-cost and eco-
sustainability of the eutectic solvent, relatively high-speed sample
preparation and low consumption of both sample and organic
solvents. Similar advantages were also described by Bi et al. [68]
who applied a mixture of ChCl and 1,4-butanediol (1:5 M ratio)
with 35% of water at 70 C for 40 min for the extraction of antiox-
idant flavonoids (myricetin and mentoflavone) from leave plants,
using a sample/volume ratio of 1 g/10 mL.
7.2. DES as sorbent on solid-phase extraction
Solid-phase extraction (SPE) is one of the most popular extrac-
tion techniques, making use of a wide range of supporting packings
commercially available with different sorbents. Depending on the
nature of the sorbent, a number of mechanisms of solute retention
can be assumed to rule the process. Partitioning of analytes be-
tween a liquid solution (sample matrix or extract) and a viscous
liquid that is immobilized on a solid support (sorbent phase) is the
most usual retention mechanism for SPE process. Liquid/solid
adsorption as well as ion exchange or size exclusion are also
possible mechanisms in various separations [69]. SPE extraction is a
safe, efficient and reproducible separation technique that can be
automatized.
Wang et al. [61] employed DES (ChCl and urea, 1:2 M ratio) as
reaction solvent to prepare a new nitro-substituted tris(indolyl)
methane modified silica phase. This new sorbent was used in the
extraction of organic acids (benzoic, p-anisic, salicylic and cinnamic
acids) from grape juice and mineralized drinking water followed by
HPLC-DAD analysis. Owing the multiple intermolecular forces that
might occur on the new phase, such as pep, hydrogen bonding and
hydrophobic interactions, good extraction efficiency (recoveries
>68%) was achieved for the selected organic acids. Another
important advantage of this SPE sorbent is the possibility to be
reused, which decrease the cost of the analysis.
Liu et al. [70] used a DES (ChCl: ethylene glycol) to modify
graphene. Compared with conventional graphene, DES-graphene
had a large winkle and formed a structure with a high specific
surface area because linked groups were introduced into the parent
graphene structure that gives it a higher adsorption ability. Two
milligrams of DES-graphene were employed to pipette-tip SPE of
sulfamerazine from river waters followed by HPLC analysis. The
optimization of PT-SPE included the evaluation of volume of
washing and elution solvent. Under the optimum conditions this
method showed good recoveries (91e97%) and higher precision
intraday (RSD range from 1.6 to 3.5%) and interday (RSD range from
0.7 to 3.8%).
7.3. DES as dissolution solvent
DES have been used by the group of Ghanemi [63] for the
dissolution of marine biological samples, which facilitated the
quantitative extraction of some metals under study with small
volumes of diluted nitric acid for determination by FAAS. Briefly, in
this method, 100 mg of the sample were dissolved in ChCleOx (1:2,
M ratio) at 100 C for 45 min. Then, after addition of 5.0 mL HNO3
 S.C. Cunha, J.O. Fernandes / Trends in Analytical Chemistry 105 (2018) 225e239
(1.0 M) the mixture was centrifuged and the supernatant analyzed.
This procedure provided some clear advantages when compared
with other techniques (microwave or ultrasonic acid digestion)
such as low-cost and simplicity of the sample preparation while
avoiding the formation of highly carcinogenic nitrous vapors.
7.4. DES as carbon paste electrode
Carbon paste prepared through a mixture of carbon (graphite)
powder and a binder (pasting liquid), has been used in the prepa-
ration of various electrodes, sensors, and detectors [65]. Currently,
CPEs represent a popular type of electrode, because carbon pastes
are easily obtainable at minimal costs and can be modified simply
to obtain quantitatively new sensors with the desired properties.
The basic requirements for a pasting liquid are its practical insol-
ubility in the solution under measurement, a low vapor pressure to
ensure both mechanical stability and long lifetime, and further, in
the case of voltammetric and amperometric applications, its elec-
trochemical inactivity in the potential window of interest [71].
Zhu et al. [62] used a NADES composed of ChCl and urea (2:1 M
ratio) mixed with graphite power to prepare a CPE, which was used
to directly extract DEHP from polymer films. The microextraction
process follows an exponential association function with the
apparent first order rate constant of 6.35  104 s1.
7.5. DES as modified molecular imprinted polymers (MIP)
MIPs are crosslinked polymers containing specific recognition
sites (involved in various types of interactions-covalent, non-co-
valent and semi-covalent) with a predetermined selectivity for a
target analyte. This technique provides several advantages, such as
high selectivity and great stability to heating and pH shifts when
compared with SPE and LLE. MIPs have been commonly employed
in the preconcentration of analytes, acting as the selective adsor-
bent of SPE [72]. However, MIP's preparation can pose some
problems, like inconsistent molecular recognition, polymer
swelling in unfavorable solvents, slow binding kinetics, and po-
tential sample contamination by template bleeding [72].
Li et al. [73] compared the efficiency of a DES (synthesized with
ChCl and glycerol, 1:2 M ratio) modified molecular imprinted
polymer a DES modified non-imprinted polymers (without tem-
plate), a MIP (without DES) and a NIP (without DES and without
template) for the purification of chlorogenic acid from honey-
suckles. The extract of honeysuckles was previous obtained using
ultrasound (20 min), ethanol at 60% and a ratio of liquid to material
of 15 ml/g. Among the polymers proposed the DES-MIPs used as
SPE showed the highest adsorption capacity owing to its specific
imprinted recognition and the DES improved affinity, selectivity
and adsorption in purification.
A similar approach was applied more recently by the same
group using DES formed by ChCl and glycerol (1:2 M ratio) to
synthetize a novel MIP/SPE. The application was successfully
applied in the purification of chloromycetin and thiamphenicol
from milk [74].
8. Combination of analytical technologies with
microextraction techniques
A broad range of analytical extractive and microextractive
techniques have been developed using DES/NADES as extracting
solvents for many quantitative analytical purposes such as ETAAS
determination of inorganic compounds, and HPLC and GC deter-
mination of organic compounds. In the majority of the cases, when
DES/NADES are employed as extracting solvent in various DLLME,
UAME, MAE and other microextractions procedures, HPLC is
237
preferred to GC as final quantitative technique since the low vola-
tility of the DES/NADES hinder the GC analysis. Nevertheless, Tang
et al. [28], Farajzadeh et al. [33], Lamei et al. [34] and Khezeli et al.
[47] have successfully employed GC-FID for quantification of
several organic compounds using the direct injection of DES
extract. The possible contamination of the GC system namely col-
umn and injector due to insufficient volatilization of DES extracts
was not mentioned in these works. Therefore, the advantages of the
use of DES with microextraction techniques, avoiding the toxicity
associated with the use of organic solvents, are added to the
analytical possibilities of GC. Furthermore, DES as extractor possess
an extensive range of polarity/volatility which can be useful in
many pre-concentration and separation processes.
9. Outlook of DES/NADES solvents within green analytical
chemistry
According to the twelve principles that govern the concept of
Green Chemistry proposed by Anastas and Warner [1], analytical
methods should reduce or eliminate hazardous substances used in
or generated by a method. The use of DES/NADES as extracting
solvents in sample preparation techniques fits perfectly with this
approach due to their low toxicity and high biodegradability.
Additionally, strategies that combine the use of low volumes of
these solvents such as LPME, MAE and UAME with green analytical
techniques like thermogravimetric, electrochemical or immuno-
assays analysis [75,76] are regarded as very interesting green
analytical approaches. Despite most of the described sample
preparation methodologies presented in the previous sections can
be fully considered as environmental-friendly, especially due to low
green solvent consumption, the further use of a chromatographic
analytical determination commonly associated increases energy
costs of the analysis which is somehow a drawback for the
analytical method greenness (Table 3). According the existing green
analytical evaluation tools (National Environmental Methods Index
and Eco-scale) [75,76] the most energy-consuming techniques are
NMR, GC-MS, LC-MS, and X-ray diffractometry as opposed to
immunoassay, spectroscopy, and electrochemical techniques that
are the more energy saved. Additional penalty points on the scales
of assessment of analytical methods greenness comes from the
requirements of calibration and validation of the methods, since
the use of calibration solutions, internal and external standards,
standard reference materials or isotope dilution enhance reagents
consumption and waste generation [76]. In summary, ideal green
analytical methods will be those that eliminate or minimize the use
of hazardous solvents, reduce energy use and generate minute
quantities of wastes. Therefore, clear efforts are still needed to be
made to encourage the development of sustainable analytical
methods based on the use of DES and NADES solvents. Information
regarding the performance of (micro)extraction techniques based
on application of DES/NADES are highlighted in Table 4.
10. Conclusions and future perspectives
In sample preparation, the choice of the right extracting
solvent is essential to achieve a near-complete extraction of
the analytes of interest, simultaneously with minimizing the
amount of interferents. For this purpose, a new class of alter-
native and environmental-friendly DES and NADES solvents have
been employed in several novel microextraction techniques such
as LPME, UAME and MAE as well as in more conventional
extraction procedures like SPE. Indeed, by replacing conventional
solvents with DES the main merits of microextraction techniques
such as simplicity of operation, low cost, and environmental
safety were enhanced. Additionally, the selectivity and
238 S.C. Cunha, J.O. Fernandes / Trends in Analytical Chemistry 105 (2018) 225e239
Table 4
Overall performance of extraction techniques based on the use DES/NADES solvents.
Technique Time Simplicity Low Solvent Volume
LPME e þ þ þ
DLLME þþ þþ þ þ
AG-DLLME and AA-LLME þ þþ þþ
HPLME þ þ þþ
UAME þ þþ þþ
MAE þ þþ þþ
(), less favorable; (þ), favorable; (þþ), more favorable.
LPME, liquid-phase microextraction; DLLME, dispersive liquid-liquid microextraction; AG-DLLME, gas air-dispersive liqui-liquid microextraction; AA-LLME, air assisted liquid-
liquid microextraction; HPLME, hollow fiber-liquid-phase microextraction; UAME, ultrasound-assisted microextraction; MAE, microwave-assisted Extraction; HPLC, high
performance liquid chromatography; GC, gas chromatography.
sensitivity achieved by the new analytical methods designed for
food and environmental analysis were often better than those
obtained by conventional extraction techniques.
There is no doubt that the application of DES and NADES sol-
vents in food and environmental analysis will grow in a near future.
Forthcoming studies and developments utilizing DES may focus on
the following areas; (I) synthesis of new DES and NADES solvents
with different polarities and their exploitation in the development
of novel extraction techniques for multi-analyte food and envi-
ronmental analyses; (II) enhancement of the performance and
selectivity of the several microextraction techniques to help the
execution of the extraction and separation procedure in the
analytical laboratories; and (III) employment of new DES and
NADES as extracting solvents, sorbents or selective binding agents
for the trace analysis of contaminants in both food and environ-
mental samples.
Acknowledgments
Sara C. Cunha and Jose
O. Fernandes thanks REQUIMTE, FCT
(Fundaça~o para a Cie
^ncia e a Tecnologia) and FEDER through the
project UID/QUI/50006/2013 e POCI/01/0145/FEDER/007265 with
financial support from FCT/MEC through national funds and co-
financed by FEDER, under the Partnership Agreement PT2020.
Sara C. Cunha acknowledges FCT for the IF/01616/2015 contract.
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