Ecotoxicology and Environmental Safety 99 (2014) 1–12

Contents lists available at ScienceDirect

Ecotoxicology and Environmental Safety

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

Review

A brief overview of the potential environmental hazards

of ionic liquids
Marina Cvjetko Bubalo a, Kristina Radošević a, Ivana Radojčić Redovniković a,
Jasna Halambek b, Višnja Gaurina Srček a,n
a
Laboratory for Cell Culture Technology, Application and Biotransformations, Faculty of Food Technology and Biotechnology, University of Zagreb, Pierottijeva
6, HR-10000 Zagreb, Croatia
b
Laboratory for Physical Chemistry and Corrosion, Faculty of Food Technology and Biotechnology, University of Zagreb, Pierottijeva 6,
HR-10000 Zagreb, Croatia

art ic l e i nf o a b s t r a c t

Article history: Over past decades ionic liquids, a promising alternative to traditional organic solvents, have been
Received 18 July 2013 dramatically expanding in popularity as a new generation of chemicals with potential uses in various
Received in revised form areas in industry. In the literature these compounds have often been referred to as environmentally
15 October 2013
friendly; however, in recent years the perception of their greenness dramatically changed as the scientific
Accepted 18 October 2013
Available online 6 November 2013
community began to proactively assess the risk of their application based on the entire life-cycle. This
review gives a brief overview of the current knowledge regarding the potential risks linked to the
Keywords: application of ionic liquids – from preparation to their disposal, with special emphasis on their potential
(Bio)degradation environmental impacts and future directions in designing inherently safer ionic liquids.
Ionic liquids
& 2013 Elsevier Inc. All rights reserved.
Environmental fate
Synthesis
Toxicity

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
2. Issues concerning the preparation of ILs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 3
3. Methods of ILs degradation. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 4
4. Environmental fate and toxicity of ILs . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 5
5. Future directions – is it possible to design environmentally acceptable ILs? . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 7
6. Conclusion . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
Acknowledgments . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 9

1. Introduction friendly synthetic procedure, while also meeting technological

and economical demands (Matzke et al., 2010). Over past decades
A growing area of research in the development of green ionic liquids (ILs), organic salts consisting entirely of ions and with
technologies is devoted to designing new, more environmentally melting points lower than 100 1C, have been in the spotlight of the
friendly solvents (Alfonsi et al., 2008; Brennecke and Maginn, 2001). scientific and industrial community as a promising alternative to
From the point of green chemistry, solvents should be non-toxic, traditional organic solvents, from both the environmental and
readily biodegradable and synthesized by an environmentally technological perspectives. Regarding the structure of ILs, cations
are usually variously substituted bulky organic molecules of low
symmetry containing a positively charged nitrogen, sulfur or
n
Corresponding author. Fax: þ 385 1 46 05 065. phosphorus atom (e.g. N, N′-dialkylimidazolium, N-alkylpyridinium,
E-mail address: vgaurina@pbf.hr (V. Gaurina Srček). alkylammonium, alkylphosphonium, alkylsulphonium and tiazolium

0147-6513/$ - see front matter & 2013 Elsevier Inc. All rights reserved.
http://dx.doi.org/10.1016/j.ecoenv.2013.10.019
2 M. Cvjetko Bubalo et al. / Ecotoxicology and Environmental Safety 99 (2014) 1–12
Fig. 1. Possible applications of ILs (Adapted from Pham et al., 2010).
cation), while typical anions are usually inorganic or organic
species such as halides (e.g. Br  , Cl  ), tetrafluorborate (BF4  ),
hexafluorphosphate (PF6  ), bis(trifluoromethylsulfonyl)imide
((CF3SO3)2N  ), acetate (CH3CO2  ) and dicyanamide (N(CN)2  )
(Fig. 1) (Earle and Seddon, 2000; Petkovic et al., 2011). These
solvents are referred as non-flammable compounds with negligi-
bly low vapor–pressure (often 10  11–10  10 mbar) and excellent
thermal stability (the thermal decomposition temperature of ILs is
generally in range from 250 1C to 450 1C). However, since ILs are
heterogeneous group of compounds with diverse physicochemical
properties, mentioned features of ILs should not be generalized
since no general ILs properties exist, a part of that included into
the definition of this compounds (salt with melting point below
100 1C). Additionally, some attributes used to describe ILs are even
partially wrong. For example, Smiglak et al. (2006) observed that
imidazolium, pyridinium and phosphonium ILs should not be
necessarily considered safe when working with or near a heat or
ignition source, while Meine et al. (2010) showed that 10 percent
of 1-butyl-3-methylimidazolium bromide degraded upon heating
at 200 1C for 24 h.
ILs are intensively studied for application in a variety of
different areas, such as organic synthesis and (bio)catalysis,
electrochemistry, analytical chemistry, separation technology,
nanotechnology, renewable resource utilization, and in use as
functional fluids (e.g. lubricants, heat transfer fluids, corrosion
inhibitors) (Plechkova and Seddon, 2008: Pham et al., 2010). It is
important to emphasize that the possible combinations of differ-
ent cations and anions are enormous and it is estimated that
roughly 1018 ILs are accessible. This fact ensures the possibility of
tuning the chemical and physical properties (e.g. melting point,
solubility, acidity, hydrophobicity, density and viscosity) in order to
design an optimal IL for a specific purpose (Earle and Seddon, 2000).
Additionally, the possibility of using IL mixtures to increases
synthetic flexibility and tunability for specific purpose has recently
mobilized researches to explore their properties (Niedermeyer
et al., 2012). Namely, Taige et al. (2010) have demonstrated that
imidazolium and pyridinium ILs could be combined to give binary
mixtures with reduced viscosity and enhanced conductivity, while
Annat et al. (2012) showed that in a number of the pyridinium and
phosphonium IL mixtures the crystallization of both components
was completely suppressed. Recently, a new class of ILs with an
entirely new cationic substructure and a completely different
charge distribution, so-called Tunable Aryl Alkyl Ionic Liquids
(TAAILs), have been developed (Ahrens et al., 2009; Stolte et al.,
2013). These ILs distinguished from classical imidazolium ILs by
the substitution pattern of the imidazolium core, whereby electro-
nic interaction between the aromatic substituent and the core
allows fine and more precisely tuning of properties than is
possible for currently available ILs (Stolte et al., 2013).
Even though ILs are not currently widely used for commer-
cial purposes, some companies have started their industrial use.
In 1998, the French Petroleum Institute approved the commer-
cial use of ILs in the preparation of polybutene (Difasol process),
important in the production of plastics, rubber or similar
materials (Olivier, 1999). However, the first large-scale indus-
trial application of ILs was in 2003 through BASF's BASIL process
(Biphasic Acid Scavenging Utilizing Ionic Liquids), in which
N-alkylimidazole was used to remove acid from a particular
process in situ, forming ILs that could be easily removed from
the reaction mixture (Masse and Massonne, 2005). In 2004,
this process was awarded the prestigious Innovation Award
and is now conducted in multi-tone scale (Plechkova and
Seddon, 2008).
To date, introducing ILs into various processes has resulted in
process improvements in terms of yield and productivity, and also
in improvements from the economical perspective if ILs reuse and
recycle is included. To our knowledge, there are no data confirm-
ing the presence of ILs in natural environment, although rising
interest for these compounds and great number of possible
applications would result in their uncontrolled transfer in the
environment someday in the near future. Thus, in the case of
larger production (over one metric tonne per year) safety informa-
tion on the ILs should be provided, as current legislations such as
European Union Regulation REACH demand (REACH, 2006). So far
the properties of ILs which gained them attribute of being
“environmentally friendly” are non-volatility (reduced air pollu-
tion), non-flammability (process safety) and excellent stability
(recycling and reusing potential). Regardless ILs' declarative green
properties, in recent years the perception of their greenness
dramatically changed as the scientific community began to care-
fully assess the risk of their application based on the entire life-
cycle, including preparation methods, methods of their degrada-
tion after use and their impacts on the ecosystem (Fig. 2) (Coleman
and Gathergood, 2010; Siedlecka et al., 2011; Pham et al., 2010).
 M. Cvjetko Bubalo et al. / Ecotoxicology and Environmental Safety 99 (2014) 1–12
Fig. 2. Ionic liquids (ILs): from preparation to disposal.
Jastorff et al. (2003) were the first to stress the importance of using
multidimensional risk analysis based on ecotoxicological indica-
tors (release, spatiotemporal range, bioaccumulation, biological
activity and uncertainty) in order to designate the intensity of risk
posed by ILs. It was observed that insufficient data on degradation,
aqueous solubility, biotic/abiotic transformation, sorption onto
solid phases and toxicity identified them as harmful chemicals
with respect to the uncertainty indicator. The first comprehensive
cradle-to-gate life cycle of 1-butyl-3-methylimidazolium tetra-
fluoroborate used for cyclohexane production (including the
synthesis of IL used) was reported by Zhang et al. (2008).
Comparison with traditional solvents on a mass basis indicated
that ILs may simply shift associated pollution to earlier stages of
the life cycle than some common organic solvents and may cause
even more severe pollution. In the past 10 years a vast number
of scientific publications dealing with the environmental aspects
of ILs have been published, indicating that research in the field
of preparation, application and environmental fate of ILs is a
true example of proactive hazard assessment. A comprehensive
discussion is far beyond the scope of this paper, however same
major facts about the potential hazards relating to the use of ILs,
with emphasis on greenness of some common ILs' preparative
methods, their toxicological effect and criteria for the design of
inherently safer ILs, are presented and discussed herein.
2. Issues concerning the preparation of ILs
Generally, the preparation of ILs is relatively simple and it is
estimated that synthetic procedures have been reported for about
103 different ILs. Standard procedures start with the formation of
the halide salt with the desired cation via a quaternization
reaction of heterocyclic compounds (e.g. N-methylmidazole, pyr-
idine) with specific alkyl halides, followed by the reaction for
3
anion exchange with metal salts (e.g. KPF6, NaBF4) or acid (HPF6,
HBF4) to form the IL with the corresponding anion (e.g. HBF4, HPF6,
respectively) (Bonhôte et al., 1996; Deetlefs and Seddon, 2010;
Huddleston et al., 1998).
In order to evaluate the efficiency and potential environmental
impact of ILs synthesis, or any other synthetic procedure, a series
of green chemistry metrics of different categories are often used
(Curzons et al., 2001) among which the Environmental factor
(E-factor), representing ratio of the weight of waste per unit of
product, and Atom economy, representing the ratio between the
mass of the atoms making up the final product(s) and the mass
of the atoms that are incorporated in all reactants, have
been widely used (Sheldon, 2007). Deetlefs and Seddon (2010)
assessed the greenness of some common laboratory-scale IL’
preparative methods and concluded that the preparation of
1-alkyl-3-methylimidazolium halide salts via quaternization of
1-methylimidazole with alkyl halide is 100 percent atom efficient
since no by-products are formed. Meanwhile, metathesis reactions
with metal salts or acid are o100 percent atom efficient since a
stoichiometric amount of inorganic halide salt as waste is gener-
ated (e.g. LiBr or NH4Br). However, atom economy calculation does
not take into account that some reactions with favorable stoichio-
metry require excesses of reagents, as it is case for quaternization.
As for the quaternization reaction E-factor, during the preparation
of 1-alkyl-3-methylimidazolium halide salts within organic sol-
vent and using a excess of alkyl halide leading to highly undesir-
able value, while the purification of 1-alkyl-3-methylimidazolium
halide salts also yields poor E-factors (Deetlefs and Seddon, 2010).
For example during the conventional laboratory synthesis of 3 g
1-alkyl-3-methylimidazolium halide about 50 ml of toluene or
acetonitrile (reaction solvent) and about 50 ml of ethyl acetate
(product washing) is required (Cvjetko et al., 2012), resulting in
the formation of about 100 ml of waste per 3 g of product, and an
E-factor of about 30 g waste/g product, which puts the greenness
4 M. Cvjetko Bubalo et al. / Ecotoxicology and Environmental Safety 99 (2014) 1–12
of ILs synthesis in the range of environmentally unfavorable pre-
paration of fine chemicals. To reduce the E-factor, it is crucial to
minimize solvent consumption or to recycle used solvent. So far, IL
synthesis has been successfully performed in solvent-free systems
(Aupoix et al., 2010; Namboodiri and Varma, 2002), although the use
of solvents for the purification step is still mandatory since chemical
equilibrium of quaternization usually does not exceed 95 percent
(usually 80–90 percent) (Bonhôte et al., 1996; Cvjetko et al., 2012;
Deetlefs and Seddon, 2010; Huddleston et al., 1998). It is important to
emphasize that the E-factor is an oversimplification and does not
include certain environmentally important properties of the chemi-
cals used (e.g. origin, toxicity, biodegradability, bioaccumulation and
environmental fate) and the possibility of their recycling. For
example, raw materials for ILs synthesis, such as imidazole and
haloalkanes, come from petroleum feedstock that are neither green
nor sustainable, while for the preparation of, for example 1-butyl-3-
methyilimidazolium dycianamide or 1-butyl-3-methyilimidazolium
bis(trifluoromethylsulfonyl)imide, the highly toxic AgN(CN)2 and LiN
(CF3SO3)2 salts were used (Li, 2005). Righi et al. (2011) analyzed
environmental impacts of 1-butyl-3-methylimidazolium chloride
synthesis (discontinuous three-step batch process followed by down-
stream processing) through Life Cycle Assessment (LCA), a method
that addresses the environmental aspects and potential environ-
mental impacts throughout a product life cycle (origin of raw
material, production, use, end-of-life treatment, recycling and final
disposal) or just selected life cycle stages (ISO, 2006). They found
that global warming, freshwater aquatic ecotoxicity and human
toxicity potentials were the most significant environmental
threats associated with the synthesis of this IL, whereby the major
impacts in all categories are resulted from 1-methylimidazole
synthesis. This conclusion agreed with Kralisch et al. (2005)
findings which emphasized high cumulative energy demand and
medium human toxicity of 1-methylimidazole as well as medium
ecotoxicity of 1-chlorobutane for aquatic organisms.
Several authors studied the preparation of ILs from renewable
sources such as nicotinic acid (Harjani et al., 2010), cholin chloride
(Hou et al., 2012; Liu et al., 2012; Petkovic et al., 2010; Weaver
et al., 2010), sugars (Handy et al., 2003; Chiappe et al., 2010) and
amino acids (He et al., 2009; Liu et al., 2012; Tao et al., 2005). ILs
containing biocompatible cholinium as the cation and amino acids
as the anions are currently intensively studied for selective
removal of lignin from lignocellulosic biomass (Hou et al., 2012;
Hou et al., 2013a; Liu et al., 2012). These ILs can be synthesized
from renewable starting materials via a simple neutralization
reaction, with water as the only byproduct. Furthermore, new
generations of liquid salts based on mixtures of nontoxic quatern-
ary ammonium salt and an uncharged hydrogen-bond donor with
a much lower melting point than that of any of its individual
components, called deep eutectic solvents, have recently gained
attention. In literature, deep eutectic solvents are sometimes
referred as the fourth generation of ILs, even though they could
not be considered as true ILs as they are not entirely composed of
ionic species. The syntheses of these compounds proceed simply
by mixing together two safe components (cheap, renewable and
biodegradable), which are capable of forming an eutectic mixture,
without the use of additional solvents and formation of by-
products (desirable Atom economy and E-factor), and results in
easily biodegradable products (Zhang et al., 2012).
Another important factor that should be considered when
discussing the greenness of ILs preparation is energy consumption.
Namely, conventional procedures for the quaternization step take
24–48 h at elevated temperatures (50–80 1C) requiring consider-
able energy input (Bonhôte et al., 1996; Huddleston et al.,1998;
Cvjetko et al., 2012) which is unsuitable from economical and
environmental point of view (Deetlefs and Seddon, 2010). First
reports on non-conventional procedures for ILs' syntheses using
microwave and ultrasound-assisted processes were given by
Varma and Namboodiri (2001) (Namboodiri and Varma, 2002),
claiming a drastic decrease in the reaction time in both cases.
Since then, a number of reports on successful microwave- and
ultrasound-assisted ILs' syntheses confirmed the benefit of their
application at the laboratory scale which have resulted in a
significant reaction time reduction compared to conventional
synthesis and consequently to reduced energy consumption
(Lévêque et al., 2002). However, disadvantages of implementation
of ultrasound- or microwave- assisted technologies into large-
scale syntheses in general include the lack of quantitative energy
efficiency data, the cost of the apparatus, as well as inability, via
design constraints, to incorporate in situ reaction monitoring
(Deetlefs and Seddon, 2003). Furthermore, strong colourization
of imidazolium IL prepared by quaternization under ultrasound or
microwave treatment, as a consequence of decomposition or
polymerization of imidazolium species due to ineffective heat
release, were reported (Cvjetko Bubalo et al., 2013; Oxley et al.,
2003). This is serious problem from the economical aspect and
thus from the aspect of green chemistry, since unfavorable cost-
and time-intense procedures have to be employed (Earle et al.,
2007). To eliminate this problem, microreactor technology was
introduced to ILs' synthetic procedures by Renken et al. (2007) and
Waterkamp et al. (2007). The use of microfluidic devices for IL
synthesis revealed that microreactor systems were advantageous
as compared to conventional processes enabling high yields and
high purity of product (non-colored) due to isothermal conditions
within channels (Cvjetko and Žnidaršič-Plazl, 2011). In order to
explore efficiency of microreactor technology for ILs synthesis, a
comparative study of ultrasound, microwave and microreactor-
assisted imidazolium-based IL synthesis via quaternization of 1,2-
dimethylimidazole with 1-bromoheptane on the lab-scale was
recently done by Cvjetko Bubalo et al. (2013). Based on the
compromise between productivity, specific power consumption
and product purity, a continuous synthesis within microreactor
was shown to be the most interesting approach in terms of
industrial application, both from environmental and economic
point of view. Possibility of large scale production within such
microreactor system is confirmed by the fact that microreactor
technology has already been introduced in industrial production
of ILs (mostly halides) in several kg/day quantities, with ambitions
to increase the capacity up to 1 t/year (Reisinger, 2007).
Overall, to reduce the amount of harmful row materials
and generated waste, as well as energy input, development of
alternative procedures and novel synthetic routes mentioned
above, is mandatory. As presented, a great improvement in ILs'
synthesis methods have already been achieved with the applica-
tion of new raw materials and improved reaction conditions (e.g.
solvent-free synthesis and reduced energy input) so the green-
ness of synthetic procedures should not be a matter of significant
concern. Additionally, reduced energy requirements and renew-
able raw materials for ILs preparation would greatly reduce the
price of ILs, which is currently 5–20-fold higher than traditional
organic solvents, an issue that still prevails their use in large
quantities.
3. Methods of ILs degradation
The most significant environmental and economic benefit of using ILs as
solvents, resulting from their low vapor pressure and excellent stability, is their
recycling potential. Numerous authors have reported successful ILs' recovery and
reuse after specific processes for several consecutive cycles (Abu-Eishah, 2011).
These properties of ILs meet the demands of the waste hierarchy developed by
European Union Directive 2008/98/EC (European Union, 2008) and make them
good candidates for environmentally benign and sustainable processes. However,
ILs reuse is limited to a certain number of cycles when the contaminant
concentration makes them unusable for a particular purpose and turns them into
 M. Cvjetko Bubalo et al. / Ecotoxicology and Environmental Safety 99 (2014) 1–12
waste (Siedlecka et al., 2011). Therefore, it is very important to find an adequate
solution for the issue of disposal of such stabile waste and to examine adequate
degradation methods for ILs that exhibit low biodegradability and/or high toxicity
with particular emphasis on the possibility of technological wastewater pre-
treatment by chemical methods and further degradation by sewage treatment
micro-organisms.
Numerous authors have studied advanced oxidation processes (Berthon et al.,
2006; Itakura et al., 2009; Li et al., 2007; Morawski et al., 2005; Siedlecka et al.,
2012, 2008a, 2008b; Siedlecka and Stepnowski, 2009; Stepnowski and Zaleska,
2005) and biodegradation (Docherty et al., 2007; Garcia et al., 2005; Gathergood
et al., 2006, 2004; Liwarska-Bizukojc et al., 2013; Markiewicz et al., 2009; Neumann
et al., 2012; Stolte et al., 2010, 2008) of ILs in aqueous media. Generally, advanced
oxidation processes are a particularly attractive treatment option for non-
biodegradable or toxic wastewaters. Stepnowski and Zaleska (2005) and
Morawski et al. (2005) studied ILs decomposition via common advanced oxidation
processes and showed that the degradation was achievable by a combination of UV
light and catalytic oxidants (e.g. hydrogen peroxide or titanium dioxide) treatment.
It was also reported that phosphonium ILs are more degradable than imidazolium
and pyridinium ILs, while elongating the 3-methyl substituent on imidazolium
strongly influence IL stability. Li et al. (2007) studied the oxidative degradation of
1,3-dialkilimidazolium ILs with the corresponding Cl  , Br  , BF4  and PF6 
counter-anions in a mixture of hydrogen peroxide/acetic acid under ultrasonic
activation and observed that under appropriate conditions it is possible to degrade
99 percent of the initial feedstock, with acetoxyacetic acid and biurea as the final
kinetically stable degraded products. They also studied mechanism for the
degradation of these ILs and concluded that three hydrogen atoms in the
imidazolium ring are the first preferably oxidized sites, followed by cleavage of
the attached alkyl groups, while the alkyl chain length on the imidazolium ring and
the type of counter anion does not affect the degradation process. Several authors
reported that oxidative degradation of several imidazolium and pyridinium halides
in the Fenton system (1 mM Fe (III) and 100 mM H2O2) shows levels of degradation
from 68 percent to 97 percent (Siedlecka et al., 2008a, 2008b; Siedlecka and
Stepnowski, 2009). Furthermore, Itakura et al. (2009) demonstrated detoxification
of several imidazolium based ILs with PF6  , BF4  and Br  in aqueous media by
combining the hydrothermal mineralization method with Ca (OH)2 mineralizer
for effective removal of the inorganic anion part, and a photocatalytic decomposi-
tion method for degradation of the organic cation part. The electrochemical
oxidation of imidazolium ILs on a boron-doped diamond (BDD) anode was carried
out by Stolte et al. (2008) and Siedlecka et al. (2013). Their studies showed that
almost complete degradation of 1-butyl-3-methylimidazolium chloride is achiev-
able, whereby products formed electrochemically were easily accessible to biolo-
gical degradation. A high cost and a short lifetime of BDD anode intrigued Siedlecka
et al. (2012) to explore possible application of low cost and quite stable PbO2 anode
for the elimination of imidazolium-based ILs from water. During electrolysis all the
parent compounds were completely decomposed and the efficiency of chemical
oxygen demand removal on completion of the process was from 69 percent to 97
percent.
Though chemical and electrochemical degradation methods, as pre-treatment
methods for biological treatment of wastewater streams, have been successfully
demonstrated, microbial breakdown, as a much friendlier process, has not proven
to be as effective. Minimal or no degradation of 1-alkyl-3-methylimidazolium-
based ILs was confirmed by several authors (Docherty et al., 2007; Garcia et al.,
2005; Gathergood et al., 2006, 2004) and according to the Organization for
Economic Cooperation and Development (OECD) standard tests (OECD, 1992),
almost none of the commonly used ILs could be classified as readily biodegradable
(defined as at least 60 percent of compound mineralization within a 10-day period
falling within 28 days of incubation with the microbial inoculate). The influence of
ILs' structure on biodegradation potential was intensively studied and it was
concluded that the degree of degradation is strongly influenced by the length of
the alkyl side-chain, core ring structure and by the presence of functionalized
groups (e.g. ester, amide) (Docherty et al., 2007; Gathergood et al., 2004; Stolte
et al., 2008), while the impact of the anion structure is less pronounced (Stolte
et al., 2010). For instance, Stolte et al. (2008) studied biodegradation of 27 ILs with
different head groups and attached side-chains and they found that imidazolium
ILs proved less degradable than pyridinium, while longer side-chains were
more susceptible to biodegradation. However, it was also observed that ILs
with long alkyl chain were also more toxic for activated-sludge microorganisms.
This observation was also reported by Markiewicz et al. (2009), who showed that
primary biodegradation of 1-methyl-3-octylimidazolium chloride can occur at
concentrations below 0.2 mM, while higher concentrations led to sorption onto
sewage sludge floccules and inhibition of cell function. Though some major
guidelines for designing biodegradable ILs have been established, Docherty et al.
(2007) and Gathergood et al. (2004) pointed out that despite of the fact that
majority of imidazolium ILs containing longer chains can be partially mineralized,
they do not achieve the OECD pass levels for biodegradability, as well as
imidazolium ILs substitution with side-chain ester groups. Recently, Liwarska-
Bizukojc et al. (2013) studied biodegradability of new peralkylated imidazolium ILs
by standard OECD methods and none of IL studied could be classified as readily or
inherently biodegradable. Biodegradability of five technologically relevant fluor-
oorganic and cyano-based ILs anions was determined in activated sludge over a
5
period of around 60 days under aerobic and anaerobic conditions by Neumann
et al. (2012) and none of these anions could be biodegraded under either aerobic or
denitrifying conditions.
To our knowledge, of all the imidazolium and pyridinium ILs tested, only a
few could be designated as readily biodegradable: 3-methyl-1-octylpyridinium
bromide (activated sludge, dissolved organic carbon Die-Away test) (Docherty
et al., 2007); 3-methyl-1-(alkyloxycarbonyl)imidazolium octyl sulfates (n¼ 1,3,5)
(Gathergood et al., 2006) and N-(alkyloxycarbonyl)pyridinium octyl sulfates
(Harjani et al., 2008) (wastewater microorganisms, CO2 Headspace test, ISO); and
1-alkoxymethyl-3-hydroxypyridinium saccharinate (n ¼11) (wastewater microor-
ganisms, 3010D test, OECD) (Stasiewicz et al., 2008). However, drawbacks of the
OECD method is that no direct chemical analyses or further toxicity measures are
required to assess readily biodegradability (Kulacki et al., 2008). In case of ILs
biodegradation potential some results could be misleading as the percentage of
degradation closely corresponds to the oxidizable C content of the biodegradable
parts of the side chain, and of the anion, meaning that the core structure could not
degraded and be resistant to further biodegradation (Matzke et al., 2010). Such
incomplete degradation may result in intermediary molecules of differing toxicities
compared to the parent compound (Kulacki et al., 2008). For example, Docherty
et al. (2007) showed by 1H NMR analysis that the partial catabolism measured is
due to degradation of the alkyl chain substitution and not of the imidazolium ring
itself. All these facts strongly indicate that the biodegradation of ILs in wastewater
streams is of great concern when discussing the large scale application of ILs and
further methods for their degradation should be investigated. Additionally, under-
standing the further transformation of degradation products, and their environ-
mental impact, is essential for designing (bio)degradable ILs.
4. Environmental fate and toxicity of ILs
Though ILs are unlikely to act as air contaminants or inhalation
toxins, they can be potential water and soil contaminants resulting
from accidental spills or lack of wastewater treatment effective-
ness, leaching of landfill sites or via effluents. Therefore, their
persistence in air, water, and soil; (bio)degradation; migration in
groundwater; bioaccumulation in aquatic or terrestrial organisms
and the (eco)toxicity of ILs play a very important role in their
environmental impact and fate (Coleman and Gathergood, 2010;
Romero et al., 2008; Pham et al., 2010; Zhu et al., 2009).
As for xenobiotics in general, bioavailability of ILs is highly
influenced by their sorption onto soil. Several studies (Beaulieu
et al., 2008; Gorman-Lewis and Fein, 2004; Matzke et al., 2009;
Mrozik et al., 2012; Stepnowski et al., 2007; Studzińska et al.,
2009) showed that the imidazolium and pyridinium cations can
adsorb onto different types of soils and marine sediments,
whereby hydrophobic long-chain ILs adsorb much more strongly
than those with short alkyl side-chains and hydroxylated deriva-
tives that would be minimally retained by geological adsorption in
non-interlayer clay systems, resulting in unrestricted transport
through soils/sediments and thus, might cause a danger of con-
tamination of surface or ground waters. These results indicate that
the transport of ILs in the environment would be determined by
their hydrophobicity: hydrophobic ILs could be attenuated by
sediments and become persistent contaminants in environment,
while more mobile hydrophilic ILs are likely to enter aquatic
ecosystems (Matzke et al., 2009; Stepnowski et al., 2007). More-
over, studies on the influence of key environmental variables (e.g.
organic matter, clay salinity and pH) on the bioavailability of ILs
showed that strength of sorption depends on both ILs structure
(Stepnowski, 2005) and physicochemical properties of various
sorbates, such as cation exchange capacity, organic matter content
and clay minerals, and pure water properties like pH and ionic
strength (Gorman-Lewis and Fein, 2004; Matzke et al., 2009;
Stepnowski, 2005). For example Studzińska et al. (2009) showed
that the higher total organic carbon value in soil, the stronger the
bonding of the analyzed compounds to the soil matrices and the
lower the quantity of the solutes migrating to the soil solution.
Nevertheless ILs become attenuated by sediments or trans-
ported further into ecosystem they are likely to get in contact with
soil- and water-microorganisms or be subjected to abiotic trans-
formation processes. According to OECD, the degradation of
6 M. Cvjetko Bubalo et al. / Ecotoxicology and Environmental Safety 99 (2014) 1–12
organic chemicals in the environment influences exposure and,
hence, it is a key parameter for estimating the risk of long-term
adverse effects on biota (OECD, 2005). Therefore, various tests are
used to simulate the biodegradation of organic chemicals under
environmentally realistic conditions in soil, sediment or surface
water. Data on ILs' biodegradability by wastewater organisms, as a
first line of protection prior to disposal into the environment, were
discussed above. In general, mode of degradation by water micro-
organisms normally occurring in specific environment correlates
to that of wastewater microorganism, although it should take into
account that certain microorganisms are normally present in
much lower densities as compared with wastewater treatment
plants (Kulacki et al., 2008). As for microbial breakdown of ILs by
microorganisms naturally occurring in soil the data are scarce.
However, Kumar et al. (2006) investigated the fate of 1-butyl-3-
methylimidazolium tetrafluoroborate when in contact with Pseu-
domonas putida and Escherichia coli, and it was observed that
P. putida was able to break down 1-butyl-3-methylimidazolium
tetrafluoroborate. Modelli et al. (2008) studied the aerobic
biodegradation processes of 1-butyl-3-methylimidazolium and
1-methoxyethyl-3-methylimidazolium cations combined with
the BF4  and N(CN)2  counter anions in soil and observed that
the biodegradability rate ranged from 17 percent to 52 percent for
1-butyl-3-methylimidazolium and 0.1–3.6 percent for 1-methox-
yethyl-3-methylimidazolium, with N(CN)2  and BF4  respectively,
while in both cases the biodegradation rate did not exceed 10
percent in the 28 day test duration. Abiotic transformation
processes, such as hydrolysis, oxidation and photolysis, was
investigated by several authors (Baker and Baker, 2005; Steudte
et al., 2012; Villagran et al., 2004). Baker and Baker (2005) and
Villagran et al. (2004) showed that the hydrolysis of BF4  and
PF6  anions results in the release of corrosive and toxic HF.
However, Matzke et al. (2010) pointed out that ecotoxicity and
bioaccumulation of evolving compounds (fluoride, boric acid and
phosphoric acid) is of minor concern because they are not harmful
to the environment in moderate concentrations. Steudte et al.
(2012) investigated the hydrolytic stability of the most common
anions, N(CN)2  , C(CN)3  , B(CN)4  , (CF3SO2)2N  , (C2F5)3PF3  , H
(C2F4)SO3  in 1-ethyl-3-methylimidazolium based ILs and the
results showed that all anions were stable under neutral and
slightly basic conditions. However, in strongly acidic and basic
solutions B(CN)4  , (CF3SO2)2N  , (C2F5)3PF3  and H(C2F4)SO3 
were hydrolytically stable, whereas N(CN)2  and C(CN)3  were
not. These findings strongly indicate that once ILs enter the
environment their mode of degradation is determined by numer-
ous biotic and abiotic factors. Therefore only an interdisciplinary
approach which is yet to be applied, could resolve the degradation
pathways that occur in complex natural environment.
As for the impact of ILs on living organisms, toxicity studies
were carried out by performing a series of tests on bacteria (e.g.
Vibrio fischeri, E. coli, Staphylococcus aureus, Photobacterium phos-
phoreum and Bacillus subtilis) (Cornmell et al., 2008; Docherty and
Kulpa, 2005; Matzke et al., 2007; Ranke et al., 2004; Ventura et al.,
2012); yeast Saccharomyces cerevisiae (Docherty and Kulpa, 2005;
Zhu et al., 2013); algae (e.g. Oocystis submarine, Pseudokirchneriella
subcapitata and Cyclotella meneghiniana) (Cho et al., 2008; Latała
et al., 2005, 2010); multicellular organisms such as Caenorhabditis
elegans (nematode) (Swatloski et al., 2004), Physa acuta (water
snail) (Bernot et al., 2005a), Daphnia magna (water flea) (Bernot
et al., 2005b), Danio rerio (zebra fish) (Pretti et al., 2006), green
seaweed (Ulva lactuca) (Kumar et al., 2011); soil invertebrates
(spring tail Folsomia candida, Lemna minor (duckweed), Lepidium
sativum (watercress), Hordeum vulgare (spring barley), Triticum
aestivum L. (wheat) and Raphanus sativus (radish) (Liu et al., 2010;
Matzke et al., 2007; Wang et al., 2009; Zhang et al., 2013);
mammals (Yu et al., 2008); as well as in different mammalian
(García-Lorenzo et al., 2008; Ranke et al., 2004; Stepnowski et al.,
2004, Stolte et al., 2007; Wang et al., 2007) and fish cells
(Radošević et al., 2013). In general, depending on the model
system for acute toxicity testing and chemical structure (e.g. type
of cation and the type and length of alkyl substituents bonded to a
heterocyclic ring), ILs showed moderate to high toxicity, in some
cases, equal to or even two to four orders of magnitude higher
than conventional organic solvents such as benzene, methanol,
dimethylformamide or propan-2-ol (Bernot et al., 2005b; Cho
et al., 2008). According to the current knowledge, it appears that
the toxicity of ILs is much more affected by the structure of the
cation than the anion (Matzke et al., 2007; Radošević et al., 2013;
Ranke et al., 2004, 2007), whereby the alkyl chain length, which
determines IL hydrophobicity, seems to be the main factor
influencing its toxicity. For example, Ranke et al. (2004, 2007),
found a good correlation of the alkyl chain lengths in 1,3-
dialkylimidazolium salts with their cytotoxicity in two different
cell cultures and the marine bacteria V. fischeri (longer alkyl chains
exhibit stronger inhibitory effect). Matzke et al. (2007) investi-
gated acute toxicity of 1-alkyl-3-methylimidazolium ILs associated
with different inorganic and organic cations (e.g. Cl  , BF4, (CF3SO2)
N  , (CF3)2N  ) on aquatic and terrestrial organisms at different
trophic levels including mammalian cells (IPC-81), marine bacteria
V. fischeri, limnic unicellular green algae (Scenedesmus vacuolatus),
wheat (T. aestivum), watercress (L. sativum), duckweed (L. minor)
and a soil invertebrate (spring tail F. candida) and confirmed that
lengthening the alkyl chain adversely affects viability, while the
anion effects were not as distinctive as the side-chain length effect.
Moreover, in several studies, it was demonstrated that the (eco)
toxicity of an IL can be modulated over several orders of magni-
tude by altering the hydrophobicity of the side-chain (Matzke
et al., 2010). Additionally, Samori et al. (2010 and 2011) showed
that the introduction of ethoxy moieties within the side-chain of
imidazolium cations reduced the biological effects in D. magna,
V. fischeri, marine diatoms (Skeletonema marinoi and Phaeodactylum
tricornutum) and rat pheochromocytoma PC12 cells. To explain
these differences, Domanska et al. (2003) and Ropel et al. (2005)
measured the octanol–water partition coefficients of some hydro-
phobic ILs and reported relatively high values, which implies
that these ILs can accumulate in organisms. Furthermore, Stolte
et al. (2007), when studied the influence of the seven different
head groups (dimethylamino)pyridinium, pyridinium, imidazo-
lium, morpholinium, piperidinium, pyrrolidinium and quaternary
ammonium) containing the same fixed side-chain and halides as
counter ions on promyelotic leukemia rat cell line IPC-81 viability,
showed that the IL’ head group had a minor effect on toxicity. As
mentioned, the anion influence on ILs’ toxicity is subordinated to
the cation effect and so far the observed effects of different anionic
moieties on toxicity were heterogeneous and varied between the
different studies (Matzke et al., 2007; Radošević et al., 2013).
However, several authors point out that fluorine containing ILs
present bigger threats to the environment than ones that are
fluorine-free (Fatemi and Izadyian, 2011; Radošević et al., 2013;
Stolte et al., 2006). Recently, Stolte et al. (2013) performed an
initial evaluation of the hazard potential of TAAILs. In comparison
with dialkylimidazolium ILs, the acute toxicity toward IPC-81cell
line, bacteria and algae was similar to 1-alkyl-3-methylimidazolim
chloride. However, it was observed that a minor change in
structure can change log10IC50 values by one to two orders of
magnitude, whereby hydrophobicity of a cation does not necessa-
rily leads to higher toxicity, as is observed for classical ILs.
The toxicity mechanism of ILs is not yet fully understood, but it
has been proposed that the mode of toxic action takes place
through membrane disruption due to the IL structural similarities
to detergents, pesticides and antibiotics (Docherty and Kulpa,
2005; Li et al., 2012). Specifically, the long alkyl chains often
 M. Cvjetko Bubalo et al. / Ecotoxicology and Environmental Safety 99 (2014) 1–12
present in the cation increase the lipophilic nature of these
compounds and thus increases the possibility of their interaction
with cell membrane phospholipid bilayers and hydrophobic
domains of membrane proteins, leading to disruption of mem-
brane physiological functions and, consequently, to cell death
(Latała et al., 2005; Ranke et al., 2007; Stepnowski et al., 2004).
To confirm the interactions of ILs with cell membrane, several
authors performed specific investigations on that topic (Cornmell
et al., 2008; Petkovic et al., 2012; Radošević et al., 2013; Samori
et al., 2010). Cornmell et al. (2008) analyzed subcellular fractions
of E. coli after exposure to trihexyltetradecylphosphonium bis
(trifluoromethylsulfonyl)imide and detected that the tested ILs
accumulated specifically in the membrane fraction of the cells.
Petkovic et al. (2012) and Radošević et al. (2013) studied the
influence of various alikyltributylphosphonium chlorides and
1-alkyl-3-methylimidazolium ILs on conidia of the filamentous
fungus Aspergillus nidulans and the membrane permeability of fish
CCO cells by fluorescence dying after exposure. The uptake of
fluorescent dye, which readily crosses the damaged plasma mem-
brane, was observed in the case of all tested ILs, indicating
membrane integrity loss. As expected, enhanced membrane per-
meability and evident membrane disruption was more pro-
nounced for ILs with the longer alkyl chain on the imidazolium
ring. Furthermore, Samorì et al. (2011) tracked lactate dehydro-
genase release into the culture medium, as a clear end-point of
membrane disruption, after exposure of rat pheochromocytoma
PC12 cell lines to oxygenated imidazolium ILs, and dehydrogenase
release was observed for all tested ILs at concentrations
41.66 mM. Furthermore, Liu et al. (2010) showed that the toxic
effect of 1-methyl-3-octylimidazolium bromide on wheat seed-
lings is also associated with membrane deterioration induced by
lipid peroxidation as a consequence of free radical formation.
Oxidative stress and membrane distortion in plants induced by ILs
was also reported by Kumar et al. (2011), who showed that in
green seaweed Ulva lactuta long-chained imidazolium ILs expo-
sure triggered the generation of reactive oxygen species, damage
of the membrane and DNA, and inhibition of antioxidant systems,
while Liu et al. (2010) and Wang et al. (2009) reported a decrease
in photosynthetic activity due to the disruption of chloroplast
membranes in wheat seedlings. Oxidative stress in invertebrates
(D. magna) after exposure to 1-alkyl-3-methylimidazolium bro-
mides was also reported by Yu et al. (2009) while Yu et al., 2008
showed that the solution of 1-octyl-3-methylimidazolium bro-
mide caused acute toxicity on the antioxidant enzymes present in
the mouse liver. Morphology and metabolic activity of yeast S.
cerevisiae after treatment by 1-butyl-3-methylimidazolim bromide
was studied by Zhu et al. (2013) and the single cellular morphol-
ogy of the yeast remained unchanged at all IL concentrations,
however, its reproduction rate by budding and metabolic activity
decreased with the increase of IL concentration. So far the toxicity
mechanism of the anionic part is less understood, though several
authors explained the higher toxicity of fluorinated anions, such as
PF6  , BF4  and (CF3SO2)2N  , due to their hydrolysis and forma-
tion of fluorides that act as potential inhibitors of Na þ K þ ATPase,
the enzyme involved in the maintenance of static electric potential
stations and transport and the regulation of cell volume (Fatemi
and Izadyian, 2011).
Though some major facts regarding environmental fate of ILs
have been established, in order to understand it more accurately,
some novel multidisciplinary approaches that involve ILs sorption
mechanism both with different natural environmental surfaces, as
well as biodegradation and transformation in complex natural
environment must be included. Besides, more complex multi-
species bioassays and predictive models to estimate complex
effects of ILs on multiple organisms from entire food chains as
well as the interactions among should be used since standard tests
7
may be inadequate to fully predict the (eco)toxicity and biode-
gradability of given ILs in the environment. Furthermore, chronic
toxicity caused by ILs should be more studied since several studies
implied the necessary of such investigations (Bernot et al., 2005a,
2005b; Ventura et al., 2013). Bernot et al. (2005a, 2005b) showed
that exposure of D. magna to nonlethal IL concentrations during 21
days at constant food-resource levels resulted in production of
significantly fewer total neonates, first-brood neonates, and aver-
age neonates and therefore such reductions in the reproductive
output of Daphnia populations could cascade through natural fresh-
water ecosystems. Reproductive responses of D. magna following
chronic exposure to 1-methyl-1-propylpyrrolidinum bis(trifluoro-
methylsulfonyl)imide during 21 days were recently assessed by
Ventura et al. (2013) and chronic toxicity seemed to be slightly lower
than the acute toxicity.
5. Future directions – is it possible to design environmentally
acceptable ILs?
ILs present a group of outstanding chemicals with enormous
possible structural variations, and therefore a broad interdisciplin-
ary effort is already being made to understand the chemical and
environmental factors controlling IL behavior in the environment
in order to incorporate those factors into designing them as
environmentally friendly (Jastorff et al., 2005). The question that
arises is: can ILs fulfill environmental criteria while still retaining
their excellent application capacities?
As presented earlier, though the majority of tested ILs possess
a strong inhibitory effect on model organisms and are poorly
biodegradable, drastic changes in toxicity/degradability, depend-
ing on heterocyclic moiety, attached substituents and/or the
anionic part have been established, which leads us to believe
that it should be possible to manipulate their chemical architec-
ture in order to design “readily” or “ultimately” biodegradable ILs
with low toxicity. For example, the morpholinium head group
was shown to be the least toxicologically active head group in
several test systems (Stolte et al., 2007). Low toxicological activity
was also observed for ILs with short polar side-chains linked to
the head group (with toxicity over several orders of magnitude
lower compared to ILs with long hydrophobic side-chains) (Stolte
et al., 2007), while a promising design criterion towards achiev-
ing biodegradability is the use of the pyridinium head group and
the introduction of polar functional groups (e.g. ether, hydroxyl
or nitrile functions). Furthermore, different anions such as alkyl
sulfates, linear alkyl sulphonates, linear alkyl benzene sulpho-
nates and organic acid salts are recommended with respect to
their biodegradability and also from an (eco)toxicological point of
view, while the use of typical fluorine-containing anions should
be avoided (Docherty et al., 2007; Harjani et al., 2008; Garcia
et al., 2005; Gathergood et al., 2006). Therefore, in order to
improve old, or to create novel ILs, better understanding of the IL
structure – toxicity relationship is crucial and has already
intrigued scientist to develop various predictive models such as
Quantitative Structure Activity Relationship model (QSAR) in
order to estimate the toxicity or biodegradability of numerous
existing and hypothetic ILs, while avoiding the costs associated
with the production and testing of a new ILs (Alvarez-Guerra and
Irabien, 2011; Cho et al., 2013; Das and Roy, 2013; Fatemi and
Izadiyan, 2011; García-Lorenzo et al., 2008; Luis et al., 2010,
2007; Torrecilla et al., 2010). Aside from predictive ability, the
modeling of IL properties could also provide a better under-
standing of their environmental hazards by determining which
structural factors affect toxicity (Kulacki et al., 2008). Modeling of
toxicity endpoints exerted by ILs was studied for organisms such
as the marine bacterium V. fischeri, green algae S. vacuolatus and
8 M. Cvjetko Bubalo et al. / Ecotoxicology and Environmental Safety 99 (2014) 1–12
multicelular organism D. magna as well as for rat and human cell
lines. Similar to previously mentioned observations on the
influence of IL structure on the toxicity, these predictive models
found toxicity positively related to the length of alkyl side chains
of cations. Additionally, the presence of positively charged atoms
in anions and nitrogen in cationic aromatic rings, the presence of
polar head groups, molecular size and branching of side-chains
were established as important factors for toxic effects towards
tested organisms/cells (Das and Roy, 2013). Though predictive
value of certain trends is unquestionable, generalizations should
be taken cautiously, since the current understanding of the
modes of toxicity of ILs, their biodegradation pathways, behavior
concerning biosorption and bioaccumulation is limited (Petkovic
et al., 2010). Additionally, certain parameters such as sorption
potential, biological oxygen demand, pH and salinity should also
be kept in mind while developing new ILs since they interfere
with IL’ behavior. Studies on degradation and accumulation
coupled with QSAR modeling should also be anticipated in order
to gain a complete vision of ecotoxicological profile of ILs (Das
and Roy, 2013).
Even though some major guidelines for ILs design have been
established (Fig. 3), a matter for concern is that some readily
biodegradable ILs, such as those with long alkyl chains, are
Fig. 3. Some general guidelines for the synthesis of greener ILs.
simultaneously highly toxic due to their lipophilic character, while
ILs with a shorter alkyl side-chain are safer with respect to (eco)
toxicity issues, but pose a higher risk of persistency and mobility
due their lack of biodegradability and reduced sorption to organic
matter and clay minerals (Matzke et al., 2009; Mrozik et al., 2012;
Ranke et al., 2007; Stepnowski et al., 2007; Stolte et al., 2008).
These facts suggest that some novel substituents, or even com-
pletely novel cation and anion moieties that would fulfill all the
mentioned criteria, should be examined. To increase the number
of candidates for environmentally friendly ILs, scientists have
started to create previously mentioned natural ILs and the deep
eutectic solvents. The concept of deep eutectic solvents was first
described by Abbot et al. (2003) (referred to choline chloride/urea
mixtures), while Carter et al. (2004) were the first to employ
nontoxic precursors, saccharin and acesulfame (non-nutritive
sweeteners), to form environmentally benign ILs. Since then,
numerous reports on preparation and characterization of IL and
deep eutectic solvents from sources natural have been published
(Fukumoto et al., 2005; Gore et al., 2011; Gorke et al., 2008; Hou
et al., 2013a, 2013b; Tao et al., 2006; Wang et al., 2011). These
novel compounds consist of carefully selected naturally-derived
materials such as organic acids, amino acids, sugars, choline, or
urea and based on the properties of individual components are
 M. Cvjetko Bubalo et al. / Ecotoxicology and Environmental Safety 99 (2014) 1–12
claimed completely non-toxic and biodegradable. These solvents
have unique physicochemical properties very close to those of
common ILs (e.g. density, viscosity, refractive index, conductivity,
surface tension, chemical inertness) which make them applicable
in various industrial processes. Moreover, they consist of inexpen-
sive compounds that make them favorable candidates from the
economic perspective (Zhang et al., 2012). According to this
natural ILs strategy, cholinium is a promising candidate as the IL
cation, since cholinium-based ILs have been reported to exhibit
low toxicity and are readily biodegradable (Hou et al., 2013a,
2013b; Petkovic et al., 2010; Weaver et al., 2010). Hou et al.
(2013b) assessed the toxicity of cholinium amino acid ILs, demon-
strated as excellent solvents for selective extraction of lignin (Hou
et al., 2012), toward bacteria (E. coli, S. aureus, Salmonella enter-
itidis, and Listeria monocytogenes) and enzyme acetylcholine
esterase. These ILs displayed low toxicity toward the bacteria
tested, while inhibitory potentials to acetylcholinesterase were
weaker approximately an order of magnitude than the traditional
IL 1-butyl-3-methylimidazolium tetrafluoroborate. Furthermore,
ILs were classified as ‘readily biodegradable’ based on their high
levels of mineralization (62–87 percent), and what is quite
important, for most of the cholinium based ILs low toxicity
correlated with good biodegradability. Recently, toxicity of several
phosphonium- and choline chloride-based deep eutectic solvents
toward brine shrimp and Gram positive bacteria was studied
recently by Hayyan et al. (2013a, 2013b). It was shown that though
choline-based eutectic solvents were completely harmful for
tested organisms, phosphonium-based mixtures exhibited slight
antibacterial activity and toxicity toward brine shrimp. Interest-
ingly, the cytotoxicity of tested eutectic mixtures was much higher
than that of their individual components, indicating noticeable
synergistic effect. We have also studied in vitro toxicity and
phytotoxicity of choline–chloride based eutectic solvents and
observed that they were harmless, with EC50 values for growth
inhibition higher for several orders of magnitude compared to
traditional imidazolium based ILs (data not published). Initial
comparative hazard assessment of protic ILs that are derived from
aliphatic amines and organic acids was conducted by Peric et al.
(2013). It was shown that protic ILs are, in terms of toxicity as well
as in terms of biodegradability, much favorable then the classical
substituted imidazolium and piridinium chlorides.
Therefore, though recent publications stressed that is possible
to design inherently safe conventional ILs based on quaternary
ammonium salts by structural modifications, it is more likely that
the upcoming era of novel green solvents originating from liquid
salts or their mixtures belongs primarily to ILs from natural
sources and to deep eutectic solvents (Zhang et al., 2012; Dai
et al., 2013). However, common ILs based on imidazolium or
pyridinium cation will probably be used in certain processes
whereby special care regarding their handling and disposal should
be taken. It should be herein pointed out that though toxic nature
of certain ILs may be undesirable from the aspect of green
chemistry, tunable property of these compounds may be used
beneficially in the development of antimicrobials and other
pharmaceuticals (Coleman et al., 2012). For example, antimicrobial
activities of 1-alkylimidazolium and 1-alkoxymethyl imidazolium
lactate ILs against a range of antibiotic resistant strains, including
Methicillin-resistant S. aureus (MRSA), was reported by Pernak
et al. (2004), while Coleman et al. (2012) screened a series of chiral
imidazolium salts for toxicity to bacteria and fungi, including
clinical pathogen strains and identified MRSA selective antimicro-
bial compound. Studies of anti-cancer activity and cytotoxicity of
three different classes of ionic liquids (imidazolium, phosphonium
and ammonium) on National Cancer Institute's 60 human tumor
cell lines have demonstrated the potential of ILs to be developed as
therapeutic agents (Malhotra and Kumar, 2010).
9
6. Conclusion
Though the environmental impact of ILs and their preparation
methods and pathways have been well studied and point out the
potential risks of their use, there is much to learn about these
heterogeneous compounds, their structural possibilities and envir-
onmental hazards. Conventional ILs, though not benign, could
become part of sustainable products and processes due to their
excellent technological properties, but it is necessary to ensure the
safety of employees handling these solvents and to ensure IL
removal from processing effluents via the described regenerative
methods. However, novel ILs and deep eutectic solvents originat-
ing from natural products could possibly be the future direction in
IL design. Overall, creating a database of environmentally benign
structure moieties of ILs and deep eutectic solvents, based on their
toxicological and biodegradation data, would be of practical use as
a guideline for manufacturers and regulators to properly develop
and regulate the use of these novel, truly green solvents.
Acknowledgments
This work was supported by the Ministry of Science, Education
and Sports of the Republic of Croatia (Grants nos. 058-0582261-
2256 and 058-0582184-2414).
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