Available online at www.sciencedirect.
com Current Opinion in
ScienceDirect
Green solvents for sample preparation in analytical
chemistry
Maryline Vian, Cassandra Breil, Léa Vernes,
Emna Chaabani and Farid Chemat
An analytical procedure comprises two steps: sample prepa-
ration mostly extraction (Soxhlet, Dean–Stark, Likens–
Nickerson, Blye & Dyer, Folch, leaching, maceration, percola-
tion) and analysis (spectrophotometry, chromatography, spec-
trometry). Whereas chemical analysis is finished after few
minutes, extraction takes at least several hours and requires
large solvent volumes, energy-consuming for concentration
step by evaporation to recover the final extract. In this current
opinion we provide an instantaneous picture of current
knowledge on alternative and green solvents used in analytical
laboratories for sample preparation and extraction of natural
product in terms of innovation, original procedures, and safe
products. We discussed how bio-based solvents, water in
different forms, deep eutectic solvents, liquefied gas have
already become an important issue in the sample preparation
in analytical chemistry of natural products replacing petroleum
and hazardous solvents.
Addresses
Université d’Avignon et des Pays de Vaucluse, INRA, UMR408,
GREEN Extraction Team, F-84000 Avignon, France
Corresponding author: Chemat, Farid (farid.chemat@univ-avignon.fr)
Current Opinion in Green and Sustainable Chemistry 2017,
5:44–48
This review comes from a themed issue on Green Solvents 2017
Edited by Charlotta Turner and Jianji Wang
http://dx.doi.org/10.1016/j.cogsc.2017.03.010
2452-2236/© 2017 Elsevier B.V. All rights reserved.
Introduction
The ideal solvent for sample preparation in analytical
chemistry is with no doubt “no solvent”. The problem is
that most chemical analysis methods necessitate the
introduction of samples under a liquid form. Sample
preparation has always been regarded as the major
bottleneck and the major time consuming step in the
analytical laboratory when performing analyses, but it is
mostly agreed that it is also the key to accurate analysis.
Sample preparation in not simply dissolution of target
compounds in a solvent, which will be use directly for
analysis. It includes several steps such as sampling, ho-
mogenization, extraction, clean up, and concentration.
The aim is to provide a homogeneous and representative
sample to produce analytically accurate results.
Current Opinion in Green and Sustainable Chemistry 2017, 5:44–48
Green and Sustainable Chemistry
Solvent extraction is a major unit operation for sample
preparation and is obviously a main concern because of
solvent’s toxicity, volatility and inflammability and it
represents a main source of laboratory chemical waste.
Solvents currently employed in extraction are often
classified VOCs (volatile organic compounds) and
possess a negative impact on HSE (Health, Safety and
Environment) [1]. Because of renewed toxicology
standards and the continual extending lists of VOCs and
CMR compounds (Carcinogenic Mutagenic Reprotoxic
in the European Union), substitution of petroleum and
hazardous solvent is being a principal concern for
analytical chemists.
The objective in preparing this current opinion is to
provide an instantaneous picture of current knowledge
on alternative and green solvents used in analytical
laboratories for sample preparation and extraction of
natural product in terms of innovation, original proced-
ures, and safe products (Fig. 1).
Water as solvent with variable geometry
The advantages of using water as alternative solvent for
extraction includes reduced economical cost, environ-
mental impact, use of simple equipment, no hazards,
faster start-up, and simplification of process steps
(Table 1).
Maceration is a very old and simple method for extrac-
tion, which consists in soaking vegetal matrix in a sol-
vent such as water. Used every day, from a simple cup of
tea to the industrial production of several bioactive
compounds such as anthocyanin’s, polyphenols .
Maceration of plant matrix with water can be done with
different procedures regarding temperature and
techniques.
Water is highly polar and a weak solvent for most organic
compounds under ambient conditions but raising the
temperature significantly above ambient, at a pressure
high enough to keep water in a liquid state, permits to
change physical and chemical properties considerably
[7]. This domain is called subcritical water; temperature
and pressure must be respectively between 100 and
374  C, and 1 and 221 bar. Among the varying properties
of water, the dielectric constant ε can heavily impact on
water solvent power. At 250  C and 50 bar, water has
equivalent dielectric constant ε than ethanol (ε = 27),
close to methanol (ε = 33) at 25  C and 1 bar. Under
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 Green solvents for sample preparation Vian et al. 45

Figure 1

Panorama of green solvents.

these conditions, water is able to solubilize more apolar
molecules. Therefore, by tuning water temperature at
high pressure, we can adapt its polarity and target
molecules of interest for extraction.
Enzymes can be used as additives in water extraction
processes to enhance cell wall disruption process [8].
Various enzymes such as cellulases, pectinases and
hemicellulases are often used to disrupt vegetal cell
wall, thereby enhancing extraction of bioactive com-
pounds from plants. To use efficiently enzymes for
extraction applications, it is important to understand
their properties, mode of action and specific operational
conditions. Enzymatic extraction is not the only method
using water additive to enhance natural product ex-
tractions in water. The method of micellar-assisted
extraction uses surfactant additives. This technique
aims to use surfactants properties to form micelles,
creating apolar phases dispersed in water. This allows to
enhance the solubility of hydrophobic molecules in
water [9]. Above cloud point (CP) temperature, mi-
celles formed by surfactant molecules become instable
and break down. Two phases are formed: a surfactant-
rich phase (at a very small volume) and a larger
volume aqueous solution phase (bulk amount) with a
diluted surfactant concentration, which approximates to
its critical micelle concentration (CMC). The hydro-
phobic analytes of the solution are extracted into the
small volume of surfactant-rich phase at higher amount.
While micellar extraction is similar to a two-liquid-phase
extraction method, hydrotropic extraction only involves
one continuous liquid phase. This method uses organic
molecules called hydrotropes. They are short organic
molecules with a structure comparable to surfactants:
they have a lipophilic and a hydrophilic part. However,
unlike surfactants, they cannot form micelles. Hydro-
tropy is described as increase in solubility of

Table 1

Water as green solvent for extraction.

Analytes Materials Techniques Conditions Reference

Polyphenols Dried chokeberry Conventional maceration 170 pm, TA, 50% EtOH, ratio 1:20, 90 min [2]
Lycopene Tomato peels Solanum Enzyme extraction Pancreatine 10 mg ml−1, 1:5 w/l, [3]
peels/water, 37 +C, pH 6.5, 20 min
Phenolic olive mill wastewater Micellar extraction 10% of Triton X-100 at 90  C 30 min [4]
compounds
Polyphenols Vine Shoot Cultivars Hydrotrope extraction 37.7 mg/mL aqueous b-cyclodextrin [5]
solvent, 48 h at 66.6  C
curcumin Curcuma longa Subcritical water 10 bar, 140  C, particle size 0.71 mm, [6]
extraction (SWE) retention time 14 min, pure water

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46 Green Solvents 2017
hydrophobic molecules in water by addition of water-
soluble organic molecules. Hydrotropes may not form
micelles but they can, however, aggregate following
their affinity areas [10]. Hydrotrope-assisted solubili-
zation is done through aggregation of hydrotropes
around hydrophobic molecules. Hydrotropic extraction
relies mainly on the addition of enough hydrotropes in
the solution.
Bio-based solvents: from waste to solvent
Bio-based solvents can be classified in three classes,
according the agro-sector: cereal/sugar, oleo-
proteagineous and wood. Bio-based solvents derived
from cereal/sugar sector are mainly obtained from nat-
ural fermenting of sweet juices contained in plants such
as sugar beet, sugarcane, wheat, corn, etc. Ethanol
derived from vegetable, called bio-ethanol is produced
about 60% from sugarcane and 40% from other crops
[11]. Besides, fermentation of glucose gives many pos-
sibilities to product some molecules, which could be
used as bio-based solvents such as sorbitol, esters of
lactic acid (ethyl lactate), derivatives of succinic acid,
etc. [12]. At least, lignocellulose residues derived from
cereals production, especially from straw, and wood
wastes can be exploited for production of furfural, which
has recently been emphasized as one of the top value-
added chemicals, derived from biomass. Furfural hy-
drogenation can lead to the formation of solvents such as
2-methyl tetrahydrofuran which appears as good po-
tential alternative for the replacement of hexane in the
extraction of vegetable oil [13] and aromas [1].
The oleo-chemistry including all oleo-proteagineous
sector, allows to product solvents from seeds rich in
vegetable oils, such as colza, sunflower and soya. The
main bio-based molecules obtained are esters of fatty
acids and derivatives of glycerol.
The bio-based solvents obtained from wood sector are
principally produced from conifers such as pines or fruit
peels. The main solvents natives from this sector are
terpenes hydrocarbons, C10H16 types, mono-cyclic
terpene (D-limonene) and bi-cyclic terpene (a- and b-
Table 2
Agrosolvents issue from different agro sectors for extraction.
Sector Agrosolvents Materials
Sugar Ethyl lactate Flowers
Ethyl acetate Algae
Wood/fruits Limonene Microorganisms
Vegetable oil Sunflower Carrot
Green chemistry DMCb Yeast
CPMEc Mangostana
a
ASE: Accelerated Solvent Extraction.
b
DMC: DiMethyl Carbonate.
c
CPME: CycloPentyl Methyl Ether.
Current Opinion in Green and Sustainable Chemistry 2017, 5:44–48
pinene). Limonene is most often obtained by steam-
distillation of by-products mostly peels generated by
orange juice industry. a-pinene and b-pinene are pro-
duced by steam-distillation of oleoresins contained in
pines or can be extract from gum turpentine, a kind of
essential oil distilled from pine gum or black liquor from
paper mills (Table 2) [20].
Ionic liquids (ILs) and deep eutectic
solvents (DESs)
Specific mixtures of solid chemicals may become liquid
under certain conditions: that’s the case of Ionic liquids
(ILs) and Deep Eutectic solvents. Ionic liquids (ILs)
are organic salts in the liquid state at ambient conditions
consisting of an organic cation and an organic or inor-
ganic anion, with low melting points, usually below
100  C. ILs are able to dissolve a wide range of polar to
non-polar compounds, have a low vapor pressure, show a
high thermal stability and low combustibility, and some
of them are biodegradable [21]. Many ILs are stable
over wide temperature ranges, and has an almost un-
limited amount of cationeanion combination.
DESs are mixtures of solids that have melting points far
below the melting points of each compound, usually also
below 100  C. DES can be obtained by mixing two
solids, for example a mixture of choline chloride
(melting point 302  C) with urea (melting point
133  C) at a molar ratio of 1:2 results in a DES with a
melting point of only 12  C. The most common DES is
based on choline chloride, carboxylic acids, and other
hydrogen-bond donors, such as urea, citric acid, succinic
acid and glycerol [22]. Natural Deep Eutectic Solvents
(NADES) are DES composed of natural substance,
particularly primary metabolites, such as organic acids,
amino acids, and sugars. They possess better properties
for extraction than DES: liquid state below 0  C,
adjustable viscosity, sustainability and show a very high
solubilization ability of both non-polar and polar com-
pounds. NADES have become more and more popular
for extraction of natural compounds from plants, food
and other natural matrices [23]
Techniques Analytes Reference
a
ASE Polyphenols [14]
Bligh and Dyer Lipids [15]
ASE Lipids [16]
Ultrasound Carotenoids [17]
Maceration Lipids [18]
Heated reflux Polyphenols [19]
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 Green solvents for sample preparation Vian et al. 47

Liquefied gases: from supercritical to Figure 2

subcritical fluid extraction
Liquefied gas such as CO2 is already a promising alter-
native to conventional solvents such as hexane or
dichloromethane. In the last decades, they have gained
increasing attention from chemists because they offer
several operational advantages over traditional tech-
niques. A supercritical fluid is defined as a fluid above its
critical pressure and temperature. Under this condi-
tions, liquid and gas phase of a pure compound are
indistinguishable from each other. The main character-
istics of SCFs are the possibility of modifying the den-
sity of fluid, directly related to solubility, by changing its
pressure and/or its temperature, permitting selective
extraction. In fact, the solvent power of the fluid can be
modified by changing the extraction pressure [5]. The
most commonly used supercritical fluid is carbon diox-
ide (CO2) because of its moderate critical temperature
(31.3  C) and pressure (72.9 atm). Furthermore, it has
significant advantages as to be GRAS (Generally
Recognized as Safe), inert, non-flammable, environ-
mentally sustainable, readily available and inexpensive
solvent [24]. Supercritical-CO2 have been used as sol-
Liquefied gas extraction installation.
vent for the extraction of essential oils, vegetable oils
and animal fats from a variety of sources. It has been
used also for algal lipid extraction.
and the toxicological effect. In this review we have
Others liquefied gases can also be used as solvent for the discussed how bio-based solvents, water in different
extraction of natural products. Compressed or liquefied forms, deep eutectic solvents, liquefied gas have already
gases are able to dissolve natural substances at relatively become an important issue in the sample preparation in
low temperature as normal organic solvents. Volatility of analytical chemistry of natural products.
compressed solvents offer two advantages over the more
conventional organic solvents: low residual solvent and One of the great success stories of green solvents in
less deterioration of the thermally labile components in sample preparation for analytical chemistry has been
the extract. Many studies have been reported the use of the evolution of solvent-free systems that directly
liquefied gases as solvent such as propane [25], dimethyl translate knowledge into technology and commercial
ether [26], and n-butane [27]. Recently, a new process products. The problem is that there is no universal
has been designed by Celsius Sarl (Villette de Vienne, extraction process with universal solvent; the polarity
France) for liquefied gases (Fig. 2). The specificity of and complexity of target molecules need a variety of
this equipment is the absence of pump and compressor solvents. If we could replace only few solvents such as
which reduce consumption of energy. The feasibility of hexane, which is one of the major solvent for Soxhlet
this process has been demonstrated using n-butane as extraction, or toluene used for DeaneStark method, or
alternative solvent to n-hexane for the extraction of methanol and chloroform for Blye & Dyer procedure,
lipophilic compounds [28]. Soares et al. [29] have shown we would reduce the costs of solvent treatment and
that compressed liquefied petroleum gas is more thus avoid adverse effects on humans and the
promising than supercritical-CO2 for the extraction of environment.
rice bran oil.
References
1. Filly A, Fabiano-Tixier AS, Fernandez X, Chemat F: Alternative
Future trends solvents for extraction of food aromas. Experimental and
COSMO-RS study. LWT – Food Sci. Technol. 2015, 61:33–40.
In the 20th century, the choice of which solvent has to http://dx.doi.org/10.1016/j.lwt.2014.11.021.
be used to perform extraction of a desired metabolite   
2. Cuji c N, Savikin K, Jankovi
c T, Pljevljaku
si
c D, Zduni
c G, Ibri
c S:
from a specific plant has to be a result of a compromise Optimization of polyphenols extraction from dried choke-
between the efficiency and reproducibility of extraction, berry using maceration as traditional technique. Food Chem.
2016, 194:135–142. http://dx.doi.org/10.1016/
ease of procedure, together with considerations of cost, j.foodchem.2015.08.008.
time, safety and degree of automation. Nowadays, sol-
3. Lenucci MS, De Caroli M, Marrese PP, Iurlaro A, Rescio L,
vent’s choice is also be related about the origin of sol- Böhm V, Dalessandro G, Piro G: Enzyme-aided extraction of
vent, petroleum or bio-based, the environmental impact lycopene from high-pigment tomato cultivars by supercritical

www.sciencedirect.com Current Opinion in Green and Sustainable Chemistry 2017, 5:44–48

48 Green Solvents 2017
carbon dioxide. Food Chem. 2015, 170:193–202. http://
dx.doi.org/10.1016/j.foodchem.2014.08.081.
4. El-Abbassi A, Kiai H, Raiti J, Hafidi A: Cloud point extraction of
phenolic compounds from pretreated olive mill wastewater.
J. Environ. Chem. Eng. 2014, 2:1480–1486. http://dx.doi.org/
10.1016/j.jece.2014.06.024.
5. Rajha HN, Chacar S, Afif C, Vorobiev E, Louka N, Maroun RG: b-
Cyclodextrin-assisted extraction of polyphenols from vine
shoot cultivars. J. Agric. Food Chem. 2015:3387–3393. http://
dx.doi.org/10.1021/acs.jafc.5b00672.
6. Valizadeh Kiamahalleh M, Najafpour-Darzi G, Rahimnejad M,
Moghadamnia AA, Valizadeh Kiamahalleh M: High performance
curcumin subcritical water extraction from turmeric (Curcu-
ma longa L.). J. Chromatogr. B 2016, 1022:191–198. http://
dx.doi.org/10.1016/j.jchromb.2016.04.021.
7. Plaza M, Turner C: Pressurized hot water extraction of bio-
actives. TrAC – Trends Anal. Chem. 2015, 71:39–54. http://
dx.doi.org/10.1016/j.trac.2015.02.022.
8. Puri M, Sharma D, Barrow CJ: Enzyme-assisted extraction of
bioactives from plants. Trends Biotechnol. 2012, 30:37–44.
http://dx.doi.org/10.1016/j.tibtech.2011.06.014.
9. Kukusamude C, Quirino JP, Srijaranai S: A coacervative
extraction based on single-chain and double-chain cationic
surfactants. J. Chromatogr. A 2016, 1472:10–15. http://
dx.doi.org/10.1016/j.chroma.2016.10.030.
10. Kunz W, Holmberg K, Zemb T: Hydrotropes. Curr. Opin. Colloid
Interface Sci. 2016, 22:99–107. http://dx.doi.org/10.1016/
j.cocis.2016.03.005.
11. Gupta A, Verma JP: Sustainable bio-ethanol production from
agro-residues: a review. Renew. Sustain. Energy Rev. 2015, 41:
550–567. http://dx.doi.org/10.1016/j.rser.2014.08.032.
12. Fine F, Vian MA, Tixier A-SF, Carre P, Pages X, Chemat F: Les
agro-solvants pour l’extraction des huiles végétales issues
de graines oléagineuses. OCL 2013, 20:A502. http://dx.doi.org/
10.1051/ocl/2013020.
13. Sicaire A, Abert Vian M, Fine F, Carre P, Tostain S, Chemat F:
Solvants alternatifs au n-hexane pour l’extraction d’huiles végé-
tales (Poster). 2015.
14. Lores M, Pájaro M, Álvarez-Casas M, Domínguez J, García-
Jares C: Use of ethyl lactate to extract bioactive compounds
from Cytisus scoparius: comparison of pressurized liquid
extraction and medium scale ambient temperature systems.
Talanta 2015, 140:134–142. http://dx.doi.org/10.1016/
j.talanta.2015.03.034.
15. Lu W, Wang Z, Yuan Z: Characteristics of lipid extraction from
Chlorella sp. cultivated in outdoor raceway ponds with
mixture of ethyl acetate and ethanol for biodiesel production.
Bioresour. Technol. 2015, 191:433–437. http://dx.doi.org/
10.1016/j.biortech.2015.02.069.
16. Golmakani M-T, Mendiola JA, Rezaei K, Ibáñez E: Pressurized
limonene as an alternative bio-solvent for the extraction of
lipids from marine microorganisms. J. Supercrit. Fluids 2014,
92:1–7. http://dx.doi.org/10.1016/j.supflu.2014.05.001.
17. Li Y, Fabiano-Tixier AS, Ginies C, Chemat F: Direct green
extraction of volatile aroma compounds using vegetable oils
Current Opinion in Green and Sustainable Chemistry 2017, 5:44–48
as solvents: theoretical and experimental solubility study.
LWT – Food Sci. Technol. 2014, 59:724–731. http://dx.doi.org/
10.1016/j.lwt.2014.05.064.
18. Breil C, Meullemiestre A, Vian M, Chemat F: Bio-based solvents
for green extraction of lipids from oleaginous yeast biomass
for sustainable aviation biofuel. Molecules 2016, 21:196. http://
dx.doi.org/10.3390/molecules21020196.
19. Bundeesomchok K, Filly A, Rakotomanomana N,
Panichayupakaranant P, Chemat F: Extraction of a-
mangostin from Garcinia mangostana L. using alternative
solvents: computational predictive and experimental
studies. LWT – Food Sci. Technol. 2016, 65:297 – 303. http://
dx.doi.org/10.1016/j.lwt.2015.08.036.
20. Dejoye Tanzi C, Albert Vian M, Ginies C, Elmaataoui M,
Chemat F: Terpenes as green solvents for extraction of oil
from microalgae. Molecules 2012, 17:8196–8205.
21. Bera A, Belhaj H: Ionic liquids as alternatives of surfactants in
enhanced oil recovery—a state-of-the-art review. J. Mol. Liq.
2016, 224:177–188. http://dx.doi.org/10.1016/
j.molliq.2016.09.105.
22. Fischer V, Touraud D, Kunz W: Eco-friendly one pot synthesis
of caffeic acid phenethyl ester (CAPE) via an in-situ formed
deep eutectic solvent. Sustain. Chem. Pharm. 2016, 4:40–45.
http://dx.doi.org/10.1016/j.scp.2016.08.002.
23. Owczarek K, Szczepanska N, Plotka-Wasylka J, Rutkowska M,
Shyshchak O, Bratychak M, Namiesnik J: Natural deep eutectic
solvents in extraction process. Chem. Chem. Technol. 2016,
10(4):601–606.
24. Meullemiestre A, Breil C, Vian M, Chemat F: Modern Techniques
and Solvents for the Extraction of Microbial Oils. Springer Inter-
national Publishing AG; 2015.
25. da Silva CM, Zanqui AB, Gohara AK, de Souza AHP, Cardozo-
Filho L, Visentainer JV, Rovigatti Chiavelli LU, Bittencourt PRS,
da Silva EA, Matsushita M: Compressed n-propane extraction
of lipids and bioactive compounds from Perilla (Perilla
frutescens). J. Supercrit. Fluids 2015, 102:1–8. http://dx.doi.org/
10.1016/j.supflu.2015.03.016.
26. Li P, Makino H: Liquefied dimethyl ether: an energy-saving,
green extraction solvent. In Alternative Solvents for Natural
Products Extraction; 2014:91–106. http://dx.doi.org/10.1007/978-
3-662-43628-8_5.
27. Novello Z, Scapinello J, Magro JD, Zin G, Luccio MD, Tres MV,
Oliveira JV: Extraction, chemical characterization and anti-
oxidant activity of andiroba seeds oil obtained from pres-
surized n-butane. Ind. Crops Prod. 2015, 76:697–701. http://
dx.doi.org/10.1016/j.indcrop.2015.07.075.
28. Rapinel V, Rombaut N, Rakotomanomana N, Vallageas A,
Cravotto G, Chemat F: An original approach for lipophilic
natural products extraction: use of liquefied n-butane as
alternative solvent to n-hexane. LWT – Food Sci. Technol.
2016. http://dx.doi.org/10.1016/j.lwt.2016.10.003.
29. Soares JF, Dal Prá V, de Souza M, Lunelli FC, Abaide E, da
Silva JRF, Kuhn RC, Martínez J, Mazutti MA: Extraction of rice
bran oil using supercritical CO2 and compressed liquefied
petroleum gas. J. Food Eng. 2016, 170:58–63. http://dx.doi.org/
10.1016/j.jfoodeng.2015.09.016.
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