A Review of Ionic Liquids

in Chromatographic and Electromigration

Techniques

2008, 68, 1–10

Bogusław Buszewski&, Sylwia Studzińska

Chair of Environmental Chemistry and Bioanalytics, Faculty of Chemistry, Nicolaus Copernicus University, 7 Gagarin Str.,
87-100 Toruń, Poland; E-Mail: bbusz@chem.uni.torun.pl

Received: 17 January 2008 / Revised: 1 April 2008 / Accepted: 9 April 2008

Online publication: 27 May 2008

literally billions of different structures

that may form an IL [7]. Some of the
Abstract different cation structures and anion
pairs are shown in Fig. 1. The most
Although there have been numerous studies on the use of ionic liquids (IL) as solvents for commonly employed IL anions are
synthesis and catalysis, there are many potential new fields for their application. The number polyatomic inorganic species, such as
of studies dealing with the use of ILs as additives to the mobile phase in LC and CZE and as a PF6 and BF4 . Most prominent cations
stationary phases in GC is constantly increasing. The main goal of the present paper is to are pyridinium and imidazolium [1–10].
gather together studies concerning the use of ILs in chromatographic techniques. The The novelty of ILs is due to the pos-
application of these substances as stationary phases, mobile phase additives and electro- sibility of changing them by synthesis to
osmotic flow modifiers is discussed. Conversely, the application of separation methods in the give a range of distinctive properties.
analysis of ILs is also the subject of this review. These include high ionic conductivity,
low volatility and low vapor pressure,
non-flammability, high thermal stability
(compound specific), wide temperature
Keywords range as a liquid phase, good solvation
power for many organic and inorganic
Column liquid chromatography
materials and a wide range of electro-
Gas chromatography
chemical stabilities [4, 9–18]. Table 1 lists
Capillary electrophoresis
a few of the most popular ILs and their
Micellar electrokinetic chromatography
Isotachaphoresis physico-chemical characteristics [9–18].
Ionic liquids The properties of ILs make them inter-
esting compounds for utilization in many
different applications. These include
organic synthesis, catalysis, biocatalysis,
enzyme reactions, solvent extraction,
Introduction terparts of molten salts. The asymmetry electrochemistry, electrolytes in batteries,
lowers the lattice energy, and hence the electrolytes for dye-sensitized solar cells,
An ionic liquid (IL) is a salt in which the melting point of the resulting ionic elements of polymer gel electrolytes, in
ions are poorly coordinated. Conse- medium. In some cases, even the anions metal deposition, formation of surfac-
quently these compounds are liquid are relatively large and play a role in tant aggregates, high surface tension
below 100
C or even at room tempera- lowering the melting temperature [1–6]. materials and tissue preservation [1–10].
ture (RTIL’s) [1–6]. Their chemical The composition and associated prop- Since ILs have effectively no vapor
composition consists of large asymmet- erties of ILs depend on the specific pressure, they do not release harmful
ric organic cations and inorganic coun- cation and anion combinations: there are vapors to the environment. Over few last

Review Chromatographia 2008, 68, July (No. 1/2) 1

DOI: 10.1365/s10337-008-0662-y
0009-5893/08/07  2008 Vieweg+Teubner | GWV Fachverlage GmbH

Fig. 1. Most common ILs structures
decades it can be seen (by the range and
quality of contributions and the enor-
mously increased number of publica-
tions) that significant efforts are being
made to utilize ILs in green chemistry
[19, 20].
Apart from the environmental
friendly properties of ILs, most publi-
cations are dominated by reviews con-
cerning the application of RTILs as
solvents for synthesis and catalysis, their
use in electrochemical devices, finding
new fields for their application, and even
their history [2–6, 8, 9, 12, 16]. On the
other hand there is an increasing number
of papers dealing with the advantages of
ILs as separating agents especially in
chromatographic techniques are de-
scribed by, e.g. Koel [6], Pandey [2],
Stalcup and Cabovska [21] and Poole
[22]. ILs have intrinsically useful prop-
erties for the improvement of both GC
and LC. The salts are used as additives
to mobile phases in LC and CE and also
as stationary phases in GC. Conversely,
separation techniques are becoming
more popular in the analysis of ILs. The
role of ILs in chemical separation selec-
tivity may, in the future, be more
important than their basic function as a
solvent.
The main aim of this review is to
deal with the utilization of ILs in chro-
matographic techniques and also their
analysis by chromatographic methods
(Fig. 2). Chromatographic techniques
2 Chromatographia 2008, 68, July (No. 1/2)
increase our knowledge of their proper-
ties and also aid their detection in vari-
ous matrices. The application of
chromatographic methods in the analy-
sis of ILs is important because of their
possible migration to water and soil. As
the popularity of Ils in industry in-
creases, the probability is that they will
find their way into the environment.
Consequently there is a need to develop
methods for their determination in
environmental samples.
Ionic Liquids in
Chromatographic
Techniques
High Performance Liquid
Chromatography
Although LC is one of the most popular
separation techniques, it has still one
great disadvantage—poor results in the
analysis of polar compounds due to the
activity of residual silanol groups. These
lead to undesirable effects such as
asymmetric peaks, low efficiencies and/
or irreproducible retention [23, 24] There
are two ways to avoid the problems in
the analysis of polar or ionizable sub-
stances; working with stationary phases
synthesized on adsorbents other than
silica, or adding solutes such as trieth-
ylamine to the mobile phase which can
interact with residual silanols [25–27]. It
seems that ILs may also have this
potential.
Due to their properties IL are now
starting to be employed as promising
replacements for amines. The strong
proton-acceptor properties of these
analytes can be utilized to suppress del-
eterious effects of free silanols on LC
separations. The chaotropic character of
the anion may introduce ion-pairing
with cationic solutes and adsorption on
the stationary phase. The hydrophobic-
ity of the cation may further induce
stationary phase adsorption [28, 29].
Poole et al. [30, 31] first showed that
liquid alkylammonium nitrate and thio-
cyanate salts could be used as mobile
phases in RP-LC when mixed with a
second solvent of low viscosity. Results
of their experiments suggested that these
salts are good solvents for adjusting
eluent strength. Another study concern-
ing the utilization of alkylammonium
ILs as mobile phases was devoted to the
application of a concentrated solution of
ethylammonium acetate as a mobile
phase replacement for organic solvents
[32]. It was shown that the solution of IL
acts like an organic solvent with a
polarity similar to that of methanol.
Despite the relatively high viscosity of
ethylammonium acetate, plate count
values averaged only about 15% less as
compared to methanol. Moreover the
ILs utilized gave faster separations of
water-soluble vitamins on a reversed
phase column designed for a totally
aqueous mobile phase.
He et al. were the first to use 1-butyl-
3-methylimidazolium tetrafluoroborate
as an additive to the mobile phase in the
analysis of ephedrines [33], resulting in a
better separation of those compounds
than by the use of triethylamine. Similar
advantages in application of ILs as
residual silanol masking agents were also
obtained in their next study [34], where
the influence of those compounds on the
analysis of catecholamines was dis-
cussed. The separation of polar com-
pounds, amines and phthalic acids, with
1-alkyl-3-methylimidazolium ILs as mo-
bile phase additives was also studied [35].
More detailed study of the effects of free
silanol suppression in liquid chroma-
tography by imidazolium tetrafluorobo-
Review
Table 1. Physico-chemical constants for selected ionic liquids [3, 17, 18]

Ionic liquid Melting point Density Refractive Viscosity UV max Dielectric

(
C) (g mL 1) index (cP) (nm) constant

Ethylammonium nitrate 12.5 1.122 1.4537 32.1 292 13.4

n-Propylammonium nitrate 4.0 1.157 1.4561 66.6 294 18.1
Butylammonium thiocyanate 20.5 0.949 1.5264 97.1 275 33.9
Tetra-n-heptylammonium chloride 9 0.882 – 598 – –
Tetra-n-butylammonium 2-Hydroxy-4-morpholinopropanesulfonate – – 1.4778 4699 227 –
1-Butyl-3-ethylimidazolium bis(trifluoromethylsulfonyl)amide – 1.404 1.4285 48 – –
1-Butyl-3-ethylimidazolium perfluorobutanesulfonate 21 1.427 1.4025 323 – –
1-Butyl-3-methylimidazolium Hexafluorophosphate 10 1.373 – 450 – –
1-Butyl-3-methylimidazolium tetrafluoroborate 81 1.208 – 219 – –
1-iso-Butyl-3-methylimidazolium bis(trifluoromethylsulfonyl)amide – 1.428 1.4289 83 – –
1-n-Decyl-3-methylyimidazolium tetrafluoroborate 25 1.072 – 928 – –
1,3-Diethylimidazolium trifluoromethanesulfonate 23 1.330 1.4367 53 – –
1,3-Dimethylimidazolium bis(trifluoromethylsulfonyl)amide 22 1.559 1.4220 44 – –
1-Ethyl-3-methylimidazolium trifluoroacetate 14 1.285 1.4405 35 – –
1-n-Hexyl-3-methylimidazolium hexafluorophosphate 61 1.304 – 585 – –
1-n-Octyl-3-methylimidazolium tetrafluoroborate 79 1.110 – 439 – –

rate ILs was the subject of work by CE

Kaliszan et al. [36] with a mixture of
derivatives of phenothiazine, salicylic
acid and phenol. The silenol masking
potential of ILs was also investigated in
the analysis of a series of proton-accep-
tor basic drugs compounds [37]. Other
studies concerned the influence of addi-
tion of Ils to the mobile phase on the
separation of b-blockers [38], amino and
nucleic acids [39], amino benzoic acids
[40], guanine and hypoxanthine [41] and
mixtures of nucleotides [42]. ILs signifi-
cantly affected polar analyte retention in
all the systems tested [33–42].
The results from all these papers
showed that ILs are clearly superior to
the classical additives in terms of effi-
ciency, and peak shape enhancement.
The silanol-suppressing potency of the
Ils studied markedly exceeded that of the
standard mobile phase additives, like
triethylamine, dimethyloctylamine and
ammonia. The efficiencies and asymme-
try factors achieved for the group of
polar compounds were improved when
the cationic modifiers were added to the
aqueous-organic mobile phase, as com-
peting additives for the active silanol
sites. The improvement was most nota-
ble on the asymmetry factor.
When ILs are used as mobile phase
additives at millimolar concentration, all
their unique properties are gone. The
substances then just become dissociated
salts with hydrophobic cations and cha-
otropic anions [28]. Many effects of ILs
on the chromatographic process influ-
MEKC CZE
LC electroosmotic
flow modifiers
TLC
mobile phase
additives
improvement of selectivity in
chromatographic techniques
IONIC LIQUIDS
determination of ionic liquids and
estimation of their properties
LC
IEC ITP
GC
Fig. 2. Types of ILs application in chromatographic techniques
ence the final results—improving sepa-
ration, asymmetry factor or number of
theoretical plates, decreasing tailing,
reducing band broadening—are con-
nected with the retention mechnism,
which is not simple when ILs are present
in the mobile phase. The dual nature of
these salts is beneficent in the analysis of
polar substances since both anionic and
cationic parts of the IL additive can
participate in the retention mechanism.
IL cations coat the surface of the sta-
tionary phase on which they suppress
free silanols and at the same time they
MEKC
CCC LC
CE
stationary
phases
GC
CZE
become competing agents on the silica
surface for the solutes. Depending on the
IL’s properties, they partly move with
the mobile phase and interact with the
solutes. In the case of basic compounds
the increase of the length of the alkyl
subsitutent of the imidazolium cation
usually causes a decrease of retention
factors (Fig. 3a). A different effect
occurs in the separation of acidic sub-
stances due to the delocalization of the
charge (Fig. 3b). Although the IL
cations are mostly responsible for inter-
action with the stationary phase silanol

Review Chromatographia 2008, 68, July (No. 1/2) 3

groups they may also form ion-pairs
with acidic substances thereby increasing
their retention factors. The chaotropic
character of the anions constituting ILs
is responsible for possible ion-pairing
with the cationic solutes. These two
processes of silanol screening and ion-
pairing have a beneficial influence on the
chromatographic results [28, 33–38].
Figure 3c presents possible interactions
during the chromatographic process
when mixtures of basic, acidic and neu-
tral compounds are analyzed using ILs.
Two final advantages of the use of
ILs as mobile phase additives should be
noted. The first concerns the replace-
ment of the less effective and environ-
mentally harmful alkylamines with ILs,
to give improved separation efficiency
and greater handling safety [36, 37]. The
second advantage, that ILs have no
influence on the mobile phase pH unlike
TEA and amines, is much more impor-
tant [38].
The problems encountered with silica
supports in the analysis of polar basic
compounds may be omitted by not using
traditional stationary phases based on
silica. In countercurrent chromatogra-
phy (CCC), mobile and stationary phases
are both liquids. The chromatographic
retention in this case is based only on
liquid–liquid partitioning. During elu-
tion, solutes undergo a number of liquid-
phase exchanges. Berthod et al. [17] have
shown that the solvent properties of
1-butyl-3-methylimidazolium hexafluoro-
phosphate makes it a possible stationary
phase in CCC. The aim of their work was
to use an IL–water system with a CCC
chromatograph to measure the distribu-
tion coefficients of a set of aromatic
compounds containing acidic, basic, and
neutral substituents. Results obtained in
the study indicate that an appropriate
biphasic water/acetonitrile/IL system is
suitable for CCC. The use of 1-butyl-
3-methylimidazolium hexafluorophos-
phate or any other IL however causes
UV absorbance limitations [17].
One of newest directions in the
application of ILs in LC is their use as
stationary phases [43, 44]. New packing
materials were obtained with the 1-alkyl-
3-methylimidazolium cations [43] or
with the derivatives of 1-methyl-3-(tri
methoxysilylpropyl)imidazolium bromide
4 Chromatographia 2008, 68, July (No. 1/2)
and 1-butyl-3-(trimethoxysilylpropyl)
imidazolium bromide [44]. The first tests
of columns packed with these materials
were performed using aromatic carbox-
ylic acids as model compounds [44].
Separation appeared to involve multiple
interaction mechanisms including ion
exchange, hydrophobic interaction, and
other electrostatic interactions [43, 44].
However more detailed studies are re-
quired in this direction.
Gas Chromatography
ILs became a subject of interest in gas
chromatography thanks to their low
volatility, non-flammability, good sol-
vent properties and high viscosity.
However early work in this area gave
poor results. ILs-based stationary phases
exhibited selective interactions between
polar and hydrogen bond-forming sol-
utes, but showed no significant retention
of n-hydrocarbons [45, 46]. Other dis-
advantages were low liquid range, ther-
mal stability, and column efficiencies
[45–47]. In spite of these limitations ILs
have one great advantage for GC—their
dual nature, which was a subject of de-
tailed study by Armstrong et al. [48].
They used inverse GC with 1-butyl-
3-methylimidazolium hexafluorophos-
phate and the analogous chloride salt as
stationary phases. The Rohrschneider–
McReynolds constants were determined
for both ILs and showed considerable
differences. In particular for ILs proton-
donor and dipolar interactions appear to
be very strong, followed by proton
acceptor capabilities. It is apparent that
the chloride containing IL interacted
more strongly with proton-donor and
proton-acceptor molecules, while the
hexafluorophosphate tended to be
somewhat less polar and interacted more
strongly with non-polar solutes. The IL
anion influences the hydrogen bond ba-
sicity and dipolarity, while the cation
(depending on its structure) can influ-
ence the ability of interaction via non-
bonding and p-electrons. On the other
hand the nature of both the cations and
anions causes dispersion forces and
hydrogen bond interactions. During the
chromatographic process they behave as
if they were polar in the separation of
molecules with somewhat acidic or basic
functional groups or as non-polar phases
with hydrophobic compounds) [48].
During the last few years most of the
investigations have concerned the syn-
thesis of IL-based stationary phases,
which will confer unique IL properties
such as high thermal stability and effi-
ciency on the column [49–54].
Utilization of 1-benzyl-3-methylimi-
dazolium and 1-(4-methoxyphenyl)-3-
methylimidazolium trifluoromethane-
sulfonates gave good thermal stability up
to 260
C, symmetrical peak shapes, and
faster separations of complex mixtures
of molecules (e.g. alcohols, alkanes,
polycyclic aromatic hydrocarbons,
polychlorinated biphenyls and chlori-
nated pesticides) in comparison with a
commercial methylphenyl polysiloxane
[49]. However better thermal stability
and packing selectivity were gained,
when lightly cross-linking (by free radi-
cal reactions) of a new class of IL
monomers was applied. Those ILs-based
stationary phases gave high efficiency
separations up to temperatures of
350
C compared to commercial col-
umns [50]. One of the latest achieve-
ments in this area is the synthesis of a
geminal dicationic IL, 1,9-di(3-vinylimi-
dazolium)nonane bis[(trifluoromethyl)
sulfonyl]imidate stationary phase [54]. It
was found that this column exhibited a
much better selectivity for polar and
non-polar compounds than the poly-
siloxanes [54].
GC is one of the first separation
techniques in which ILs were used in
chiral separations. Extending these
compounds to chiral separations can be
done in two ways: a chiral selector can
be dissolved in an achiral IL, or the IL
itself can be chiral. Berthod et al. [51]
used the first approach. Permethylated
b-cyclodextrin and 2,6-di-methyl-b-
cyclodextrin were dissolved in 1-buthyl-
3-methylimidazolium chloride to prepare
GC stationary phases for chiral separa-
tions but the enantio-resolving capabili-
ties of those columns were much lower,
compared to commercial columns with
the same chiral selectors. The reason of
such an effect is probable blocking of the
cyclodextrin cavity by making an inclu-
sion complex with the imidazolium ion
pair. However ILs are used in chiral
Review
separations, mostly by the utilization of a)
chiral ILs such as N,N-dimethylephe-
drinium-based phases. Ding et al. were
the first to demonstrate enantiomeric = Si O

separations using chiral IL stationary

phases. The column packings developed = N N
ionic liquid
were effective for the separation of chiral cation

alcohols (including diols), chiral sulfox- = protonized solute

ides, some chiral epoxides, and acety-

F
lated amines. =
F
ionic liquid anion
B F
Chromatographic retention data give F

solute activity coefficient at infinite dilu-

tion, gas–liquid partition coefficients and
b)
other thermodynamic properties. Utili-
zation of these parameters and appropri-
ate models allows understanding of the
intermolecular interactions responsible
= Si O
for solvation in the stationary phase
[55, 56]. Liquid stationary phases for GC
based on ILs have recently been used to = N N
ionic liquid
determine activity coefficients at infinite cation
dilution of various substances in different = ionized solute
types of such columns. These investiga-
tions were performed with both polar c)
and non-polar substances for: 1-ethyl- N
3-methyl-imidazolium bis(trifluorometh-
N
ylsulfonyl) imidate [57], 1-butyl-3-meth-
ylimidazolium octyl sulfate [58],
1-methyl-3-octyl-imidazolium tetrafluo- = Si O

roborate [59], 1-methyl-3-ethyl-imidazo-

lium bis(trifluoromethyl-sulfonyl) amide, = N N
ionic liquid
1,2-dimethyl-3-ethyl-imidazolium bis(tri- F
cation

F ionic liquid
fluoromethyl-sulfonyl) amide [60], = F
anion
B
1-methyl-3-butyl-imidazolium bis(trifluo- N
F

romethyl-sulfonyl) imide [61], 1-hexyl-3- = protonized solute

methylimidazolium bis(trifluoromethyl- = ionized solute

sulfonyl) imide [62], 4-methyl-n-butylpy- N = neutral solute

ridinium tetrafluoroborate [63], 1-hexyl-
3-methylimidazolium tetrafluoroborate
[64], trimethyl-butylammonium bis(tri-
fluoromethylsulfonyl) imide [65], tri-
hexyl(tetradecyl)-phosphonium tris(penta-
fluoroethyl) trifluorophosphate [66] and
1-methyl-3-octyl-imidazolium chloride
[67]. One of the latest investigations in
this area is the work of [68]. The
measurements were carried out for 29
organic compounds in two RTILs,
1-butyl-3-methylimidazolium octyl sul-
fate and 1-ethyl-3-methylimidazolium
tosylate.
Electromigration Techniques
The chemical properties of ILs are often
more dependent on the anionic part of the
SILICA
Fig. 3. ILs participation in high performance liquid chromatography: a analysis of cationic
solutes with IL addition to mobile phase; b analysis of anionic solutes with IL addition to mobile
phase; c contribution of IL anions and cation in the analysis of different types of solutes
molecule, which makes them good can- fiers [69–71]. One of the first reports of
didates for electroosmotic flow (EOF) the use of ILs as buffer electrolytes in
modifiers in organic solvents. ILs have non-aqueous CE is the work of [72].
good electrical conductivity, are slightly They demonstrated the separation of
more viscous than organic solvents and water-insoluble dyes in acetonitrile,
therefore low concentrations may be re- which were previously not resolved using
quired for buffer modification to achieve traditional CE methodology. The addi-
better separations. In the field of capillary tion of 1-butyl-3-methylimidazolium
electrophoresis (CE), ILs can be used as fluoroacetate to the electrolyte provided
electrolytes, as additives to electrolytes change in the electrophoretic mobility of
and as dynamic or covalent coating re- the system. When using ILs as electrolyte
agents of the capillary. The first two additives or for dynamically coating the
directions seem to be the most popular. capillary wall several different types of
In some early CE studies, alkylam- analytes were separated—polyphenols
monium salts were used as EOF modi- [73–75], mixtures of phenols and

Review Chromatographia 2008, 68, July (No. 1/2) 5

a) interest. IL cations and anions influence
the migration behavior of analytes in
several different ways, mainly by associ-
ation with solutes (Fig. 4a, b). The
resolving power of the system is more
EOF often connected with activity of the
background electrolyte IL cation. It
plays two important roles, as an EOF
modifier and as an association agent for
compounds being analyzed (Fig. 4a).
The imidazolium ions coat the walls of
b)
the capillary. As a consequence analytes
can interact with the imidazolium
cations on the capillary wall or with the
free IL cations in the electrolyte solution,
EOF what leads to better separation [72–80].
This approach has also been applied
for the analysis of basic proteins
including lysozyme, cytochrome c, tryp-
sinoge, and a-chymotrypsinogen A [81].
In the analytical method described,
c) 1-alkyl-3-methylimidazolium-based ILs
were added to the running electrolyte to
OH dynamically coat the capillary for the
O
HO O
OHOHO
OH
O
S separation of proteins. Negatively
OH
HO

O
O
OH
HO
O
charged pure silica capillary material
S OH
EOF
HO
OH
O
OH
O
electrostatically attracts the positively
OH

O OH OH
OHO
OOH
HO
O
OH
d)
Fig. 4. The use of ILs in capillary electrophoresis: a analysis of cationic solutes with dynamically
coated capillary; b analysis of anionic solutes with dynamically coated capillary; c CE analysis of
substances with cyclodextrine and IL addition to running buffer; d MEKC analysis of solutes
with the addition of IL
aromatic acids [76], chlorophenoxy and
benzoic acid herbicides [77], monohalo-
genated phenols [78], carboxylates as
copper complexes [79] and pharmaceu-
tical analysis of nicotinic, isonicotinic
and picolinic acids [80]. All the CE
methods proposed appear to be simple
and provide good reproducibility in
terms of migration times. Experiments
O
OH
charged parts of proteins, however
addition of ILs to the buffer gives
improvements in resolution. The sepa-
ration mechanism was again suggested
to involve association between imidazo-
lium cations and proteins [81].
= ionized silanols A different application of ILs in CE
involves covalently coating the capillary
= ionic liquid cation
S wall. The EOF of a bare silica capillary
= ionic liquid anion
was reversed by the covalently bon-
S
ded 1-ethyl-3-methylimidazolium cation
EOF
S = solute [82]. The schematic capillary wall mod-
S ification process is shown in Fig. 5. The
= protonized solute utilization of ILs increases the working
pH range of the running buffer with
= ionized solute
good efficiency. Capillaries prepared in
this way were applied to the separation
of: NH4+ in human urine from the K+
ions and the separation of K+, Na+,
Li+, Ca2+, Mg2+ and Ba2+ ions in red
wine. The resolution of metal ions in
proved that resolution of solutes was the CE mode improved with the use of
better when the IL was used as an a freshly coated capillary. In other work
additive. Peaks in the electropherograms the authors reported utilization of cap-
were well separated and the analysis time illaries coated with Ils for DNA [83]
was often shorter in comparison with and positively charged drug separations
conventional buffers. [84]. The compatibility of those capil-
The separation mechanism of com- laries with mass spectrometry (because
pounds analyzed by CE, when ILs are of the low volatility of ILs) is their main
used as mobile phase additives is of advantage [82–84].

6 Chromatographia 2008, 68, July (No. 1/2) Review

 ILs were also used in CE chiral sep-
arations by the application of 1-butyl-3-
methylimizolium tetrafluoroborate as
running electrolyte with b-cyclodextrin
as modifier for the separation of
anthraquinone extracts of Chinese herbs
[85]. The utilization of ILs as the main
running electrolyte solution provided
separation of four solutes. In the case of
cyclodextrins in the running buffer, the
analyzed compounds may associate with
the imidazolium ions or with the
b-cyclodextrins (Fig. 4c). Analytes may
be entirely or partly embedded in the CD
cavity and as a consequention the asso-
ciation with the free imidazolium ions in
the bulk solution is weaker.
The application of ILs in electromi-
gration techniques also extends to
micellar electrokinetic chromatography
(MEKC). In this technique, a surfactant
is added to the buffer to separate either
chiral or achiral compounds. ILs
(1-butyl-3-methylimidazolium tetrafluo-
roborate, l-ethyl-3-methylimidazolium
tetrafluoroborate, l-ethyl-3-methylimida-
zolium hexafluorophosphate, 1-ethyl-3-
methylimidazolium trifluoromethanesul-
fonate and 1-ethyl-3-methylimidazolium
chloride) were used as mobile phase
modifiers along with polymeric surfac-
tants to separate achiral and chiral
compounds, such as alkyl aryl ketones,
phenols and a mixture of three chiral
binaphthyl derivatives [86]. The success-
ful separation of the analytes investi-
gated with the assistance of ILs was
reported. The final resolution depended
mainly on the interaction of the analytes
with the polymeric surfactants.
IL additives have been also used for
the quantification of the active compo-
nents of lignans found in medicinal herbs
[87]. It was reported that the addition of
1-butyl-3-methylimidazolium tetrafluo-
roborate to the micellar system resulted
in the complete separation of all the
compounds, which was earlier impossi-
ble with the use of pure SDS. The results
obtained showed that positively charged
imidazolium cations could be electro-
statically attracted to the negatively
charged sodium dodecyl sulfate (SDS)
micelle surface, which neutralized the
effective head group charge and reduced
electrostatic repulsion between the
charged hydrophilic headgroups of the
Review
Cl
Cl
Si OH
Si
Cl
Si OH
Cl
N Si
Cl
Si O
Si NH
Si O
Fig. 5. Schematic illustration of IL covalently bonding to capillary wall
surfactant molecules (Fig. 4d). ILs af-
fected the size and shape of the micelles
and thus altered the separations.
Determination of Ionic
Liquids by Chromatographic
Techniques
In the last few years ILs have become of
great interest in green chemistry. Many
methods, which should dramatically re-
duce the use of hazardous and polluting
organic solvents (especially in industry)
have been developed using ILs. However
the real green character of ILs still re-
quires enlargement of our knowledge
about their entire life cycle. This is the
main task for chromatographic tech-
niques in the analysis of ILs. Chro-
matographic techniques also allow the
study of the properties of ILs and their
interactions with other molecules. Their
determination is still quite a new subject
of study and only a few papers have been
published so far.
GC is a suitable tool for the deter-
mination of gas–liquid partition coeffi-
cients, activity coefficients and thermo-
dynamic constants. The solvation
parameter model provides a general tool
to define the contributions of cavity
formation and individual intermolecular
interactions to gas–liquid partition.
Furthermore application of chemometric
procedures to a database of system
constants for ILs allows improvement of
knowledge about their unique solvent
Chromatographia 2008, 68, July (No. 1/2)
Si O
Cl
Cl
Si
Si O
H2N NH
H
N
Cl
O
Br
Si NH
Si O
properties [88–91]. The use of liquid–li-
quid chromatography for the determi-
nation of partition coefficients for ILs
has also been reported, however this
approach still has many limitations [92].
On the other hand the method has a
number of advantages including speed,
accuracy and minimal consumption of
sample. Moreover liquid–liquid chro-
matography is effective for identifying
suitable systems for liquid–liquid
extraction.
Another application of separation
techniques in the prediction of the main
interactions in which ILs takes part is
the work of [93]. They used a group of
eight different ILs and LC with different
column types. This allowed the deter-
mination of the retention mechanisms of
the ILs and the main interactions
between stationary phase ligands and
analyzed chemical entities. Butyl, octyl,
octadecyl, phenyl, aryl, mixed (contain-
ing functional groups, such as octyl,
octadecyl, phenyl, aminopropyl and
cyanopropyl), alkylamide and cholester-
olic packing materials were chosen.
Retention of these salts increased with
the increase of stationary phase hydro-
phobic character. The lowest retention
factors were determined for packings
containing protonized ligands on their
surface as a consequence of electrostatic
repulsion forces. The main interactions
contributing to retention of the analyzed
compounds were dispersive and p…p
type [93]. In the case of aryl stationary
phases (phenyl, aryl) the predominant
7
role in the retention process is played by
p…p type interactions.
Buszewski et al. [94] studied four
different LC stationary phases for reso-
lution of IL cation mixtures: mixed,
cholesterolic, monolith octadecyl and
butyl. Significant differences were shown
in the results. Packings containing func-
tional groups bonded to the silica surface
that are able to undergo protonization
were found to be unsuitable for the
separation of the compounds investi-
gated. The best results were obtained for
two alkyl stationary phases: butyl and
octadecyl. The butyl phase allowed the
determination of IL compounds in
22 min. On the other hand the use of
monolithic silica modified with octadecyl
chains permitted complete resolution of
six IL cations in 13 min by the use of a
suitable gradient. The results clearly
indicate that good resolution of ILs may
be obtained by the change in mobile
phase and that choice of LC column
packing is also important [94].
The second work devoted to the
separation of 1-alkyl- and 1-aryl-3-
methylimidazolium-based ILs cations
describes routine LC with the use of an
octyl stationary phase [95]. Method
development was based on optimization
of mobile phase for peak resolution,
sensitivity and high reproducibility of
retention values. Using optimized con-
ditions, the authors were able to effi-
ciently separate a mixture of ILs, even
though resolution was still unsatisfac-
tory for the early eluting compounds
(short chain ILs). The applicability of
the method was tested by the determi-
nation of some examples of IL species
present in the medium used in cytotox-
icity studies. The compounds were well
isolated, identified and quantified [95].
All of the above studies on the utili-
zation of LC for the analysis of ILs were
performed using buffered mobile phases.
The use of mobile phases containing
only water and acetonitrile, without any
added salts was first attempted by Ruiz-
Angel and Berthod [96]. Eleven 1-alkyl-
3-methyl imidazolium ILs were analyzed
by LC with a commercial octadecyl col-
umn. When a buffer was added to the
mobile phase the analyzed compounds,
which differed only in their anions, could
not be separated. ILs with the same
8 Chromatographia 2008, 68, July (No. 1/2)
cation and different anions could be
successfully analyzed with the use of salt-
free mobile phases. According to the
authors, from a quantitative analysis
point of view, the figures of merit are
better when there is a salt present in the
mobile phase, because of the better peak
symmetry [96].
ILs have also been studied by ion
exchange chromatography. Stepnowski
and Mrozik [97] investigated the chro-
matographic behavior of eight different
ILs with a strong cation exchanger sta-
tionary-phase. During analysis several
interactions occurred with varying
strengths: cation exchange, non-specific
hydrophobic interactions, and adsorp-
tion, depending on the mobile phase
composition. The data obtained were
compared to reversed phase chromato-
graphic behavior of the selected ILs in
order to evaluate weaknesses and
strengths of both methods. The separa-
tion of ILs in the reversed phase mode
was less selective, especially in the range
of the highly hydrophilic solutes.
Appropriate modifications of the mobile
phase enabled even the most polar cat-
ions to be separated and analyzed, which
is the main difficulty in RP-LC separa-
tion. The authors suggest that the opti-
mized method may be directly applicable
to the analysis of IL cations in aqueous
environmental or industrial samples [97].
In a recent study special attention
was paid to short-chain hydrophilic
entities [98]. The results of LC analysis
with polar (phenyl) stationary phases
indicate the overall usefulness of p…p
interactions, which can be exploited to
reinforce the retention of highly hydro-
phobic IL cations. A very good resolu-
tion of salts identical in lipophilicity was
obtained. Additionally, the separation of
1-butyl-3-methylimidazolium chloride
and its degradation products demon-
strated that the method is readily appli-
cable to the control of various
experiments with ILs [98].
Besides GC and LC, CE also appears
to be a powerful tool in the analysis of
IL cations. Qin et al. [99] reported the
separation and the detection of 1-alkyl-
3-methylimidazolium, including isomers
(1-butyl-3-methylimidazolium—BMIM,
1-iso-butyl-3-methylimidazolium—iBMIM)
and related imidazole derivatives by
CZE. Cyclodextrin was chosen for the
separation iBMIM and BMIM. Its
cavity diameter is slightly larger than
that of the imidazole ring, so that both
ILs could be resolved, based on their
slight steric differences. The method was
also employed in the detection of
commercial chemical impurities and in
synthesis process control of ILs [99]. The
proposed method would, therefore,
appear to be a potential analytical tool
for analysis of industrial and environ-
mental samples, as in the case of RP-LC
or IEC.
CE was found to be a good method
for the routine separation of commonly
used ILs cations, without the addition of
cyclodextrins [100]. The best separation
of cationic constituents of ten imidazo-
lium ILs was achieved using citric buffer
as the electrolyte solution. The method
was found to be simple and reliable with
good quantitative results. Another
advantage of this method is the possi-
bility of theoretical prediction of migra-
tion times of compounds which were not
analyzed. This technique was applied
during photodegradation experiments
on ILs.
The electromigration techniques, as
with LC, offer the possibility of just IL
cation determination. One of the recent
studies in this area deals with the utili-
zation of isotachophoresis (ITP) in the
analysis of IL cations and anions [101].
The results indicated that this technique
offers a very promising alternative to the
chromatographic methods for analysis
of ILs. The main advantage is the
possibility of determining cationic and
anionic cores in a reasonable time.
Furthermore in the case of water or
wastewater samples no sample pre-
treatment is required before analysis, in
comparison with LC. Due to its low cost
and high speed, the method may be used
in qualitative routine analysis [101].
Due to the unusual characteristics of
ILs as well as their potential application
in industry as ‘‘green’’ solvents, the
analytical methods for their determina-
tion in environmental samples is the
subject of many investigations. There is
no doubt that, in the near future, ILs will
be used on an industrial scale and their
presence in the environment will be
monitored by chromatography.
Review
Concluding Remarks
Although ILs are not new they have
become very popular during last few
decades due to the discovery of their
interesting properties:
• by combining different anions and
cations, it is possible to generate a
huge number of different ILs, each
with their own specific solvation
properties,
• ILs are of very low volatility, hence
they may be used in high vacuum
systems and they eliminate many
contamination problems,
• they are good solvents for a wide
range of both inorganic and organic
materials and unusual combinations
of reagents can be brought into the
same phase.
ILs are useful additives for LC, GC
and CE. These kinds of compounds
might soon supplant the common mobile
phase additives and flow modifiers, as
they may be able to separate analytes
better than the ones previously used. ILs
have gained increasing interest within
the field of green chemistry, with respect
to their future potential industrial
applications. Therefore progress in ana-
lytical methods for ILs is observed. The
most popular technique in this area
seems to be LC.
Acknowledgments
This work was supported by the Ministry
of Science and Higher Education
(Warsaw, Poland) Grant No. 2P04G 083 29.
References
1. Welton T (1999) Chem Rev 99:2071–
2084
2. Pandey S (2006) Anal Chim Acta
556:38–45
3. Liu J, Jiang G, Liu J, Jönsson JA (2005)
Trends Anal Chem 24:20–27
4. Marsh KN, Boxall JA, Lichtenthaler R
(2004) Fluid Phase Equilib 219:93–98
5. Olivier-Bourbigou H, Magna L (2002)
J Mol Cat A Chem 3484:1–19
Review
6. Koel M (2005) Crit Rev Anal Chem
35:177–192
7. Visser AE, Swatloski RP, Reichert WM,
Mayton R, Sheff S, Wierzbicki A, Davis
JH, Rogers RD (2002) Environ Sci
Technol 36:2523–2529
8. Forsyth SA, Pringle JM, MacFarlane
DR (2004) Aust J Chem 57:113–119
9. Holbrey JD, Seddon KR (1999) Clean
Prod Proc 1:223–236
10. Wilkes JS (2002) Green Chem 4:73–80
11. Aki ANVK, Brennecke JF, Samanta A
(2001) Chem Commun 5:413–414
12. Chiappe C, Pieraccini D (2005) J Phys
Org Chem 18:275–297
13. Domanska U (2005) Pure Appl Chem
77:543–557
14. Handy ST (2005) Curr Org Chem 9:959–
998
15. Heintz A (2005) J Chem Thermodyn
37:525–529
16. Koel M (2000) Proc Estonian Acad Sci
Chem 49:145–155
17. Berthod A, Carda-Broch S (2004) Anal
Bioanal Chem 380:168–177
18. Poole CF (2004) J Chromatogr A
1037:49–82
19. Renner R (2001) Environ Sci Technol
35:410A–413A
20. Earle MJ, Seddon KR (2000) Pure Appl
Chem 72:1391–1398
21. Stalcup AM, Cabovska B (2004) J Liq
Chromatogr Relat Technol 45:89–113
22. Poole CF (2007) Adv Chromatogr
27:1443–1459
23. Krupczyńska K, Buszewski B, Jandera P
(2004) Anal Chem 76:226A–234A
24. Vervoort RJM, Maris FA, Hindriks H
(1992) J Chromatogr 623:207–220
25. Claessens HA (2001) Trends Anal Chem
20:563–583
26. Reta M, Carr PW (1999) J Chromatogr
A 855:121–127
27. Righetti PG, Gelfi C, Verzola B, Castel-
letti L (2001) Electrophoresis 22:603–611
28. Berthod A, Ruiz-Angel MJ, Huguet S
(2005) Anal Chem 77:4071–4080
29. Pan L, Lobrutto R, Kazakevich Y,
Thompson R (2004) J Chromatogr A
1049:63–73
30. Shetty PH, Youngberg PJ, Kersten BR,
Poole CF (1987) J Chromatogr 411:61–79
31. Poole CF, Kersten BR, Ho SSJ, Coddens
ME, Furton KG (1986) J Chromatogr
352:407–425
32. Waichigo MM, Riechel TL, Danielson
ND (2005) Chromatographia 61:17–23
33. Lijun H, Zhang W, Zhao L, Liu X, Jiang
S (2003) J Chromatogr A 1007:39–45
34. Zhang WZ, He LJ, Gu YL, Liu X, Jiang
SX (2003) Anal Lett 36:827–838
35. Xiaohua X, Liang Z, Xia L, Shengxiang
J (2004) Anal Chim Acta 519:207–211
36. Kaliszan R, Marszałł MP, Markuszewski
MJ, Baçzek T, Pernak J (2004) J Chro-
matogr A 1030:263–271
37. Marszałł MP, Baçzek T, Kaliszan R
(2005) Anal Chim Acta 547:172–178
38. Ruiz-Angel MJ, Carda-Broch S,
Berthod A (2006) J Chromatogr A
1119:202–208
Chromatographia 2008, 68, July (No. 1/2)
39. Polyakova Y, Jin Y, Zheng J, Row KH
(2006) J Liq Chromatogr Relat Technol
29:1687–1701
40. Zheng J, Polyakova Y, Row KH (2007)
J Chromatogr Sci 45:256–262
41. Zheng J, Row KH (2006) Am J Appl Sci
3:2160–2166
42. Hua JC, Polyakova Y, Row KH (2007)
Bull Korean Chem Soc 28:601–606
43. Sun Y, Cabovska B, Evans CE, Ridgway
TH, Stalcup AM (2005) Anal Bioanal
Chem 382:728–734
44. Wang Q, Baker GA, Baker SN, Colón
LA (2006) Analyst 131:1000–1005
45. Gordon JE, Selwyn JE, Thorne RL
(1966) J Org Chem 31:1925–1930
46. Pacholec F, Butler H, Poole CF (1982)
Anal Chem 54:1938–1941
47. Pacholec F, Poole CF (1983) Chroma-
tographia 17:370–374
48. Armstrong DW, He L, Liu Y-S (1999)
Anal Chem 71:3873–3876
49. Anderson JL, Armstrong DW (2003)
Anal Chem 75:4851–4858
50. Anderson JL, Armstrong DW (2005)
Anal Chem 77:6453–6462
51. Berthod A, He L, Armstrong DW (2001)
Chromatographia 53:63–68
52. Ding J, Welton T, Armstrong DW
(2004) Anal Chem 76:6819–6822
53. Andre M, Loidl J, Laus G, Schotten-
berger H, Bentivoglio G, Wurst K,
Ongania K-H (2005) Anal Chem 77:702–
705
54. Qi M, Armstrong DW (2007) Anal Bio-
anal Chem 388:889–899
55. Poole CF, Poole SK (2002) J Chroma-
togr A 965:263–299
56. Abraham MH, Poole CF, Poole SK
(1999) J Chromatogr A 842:79–114
57. Deenadayalu N, Letcher TM, Reddy P
(2005) J Chem Eng Data 50:105–108
58. Letcher TM, Marciniak A, Marciniak
M, Domańska U (2005) J Chem Eng
Data 50:1294–1298
59. Heintz A, Verevkin SP (2005) J Chem
Eng Data 50:1515–1519
60. Heintz A, Kulikov DV, Verevkin SP
(2002) J Chem Eng Data 47:894–899
61. Heintz A, Casás LM, Nesterov IA,
Emelyanenko VN, Verevkin SP (2005)
J Chem Eng Data 50:1510–1514
62. Heintz A, Verevkin SP, Ondo D (2006)
J Chem Eng Data 51:434–437
63. Heintz A, Kulikov DV, Verevkin SP
(2001) J Chem Eng Data 46:1526–1529
64. Letcher TM, Soko B, Reddy P, Deena-
dayalu N (2003) J Chem Eng Data
48:1587–1593
65. Heintz A, Vasiltsova TV, Safarov J, Bich
E, Verevkin SP (2006) J Chem Eng Data
51:648–655
66. Letcher TM, Reddy P (2005) Fluid Phase
Equilib 235:11–17
67. David W, Letcher TM, Ramjugernath
D, Raal JD (2003) J Chem Thermodyn
35:1335–1341
68. Mutelet F, Jaubert J-N (2006) J Chro-
matogr A 1102:256–267
69. Huang X, Luckey JA, Gordon MJ, Zare
RN (1989) Anal Chem 61:766–770
9
70. Harrold MP, Wojtusik MJ, Riviello J,
Henson PJ (1993) J Chromatogr
640:463–471
71. Quang C, Khaledi MG (1993) Anal
Chem 65:3354–3358
72. Vaher M, Koel M, Kaljurand M (2001)
Chromatographia 53:302–306
73. Yanes EG, Gratz SR, Stalcup AM
(2000) Analyst 125:1919–1923
74. Yanes EG, Gratz SR, Baldwin MJ,
Robison SE, Stalcup AM (2001) Anal
Chem 73:3838–3844
75. Vaher M, Koel M, Kaljurand M (2002)
J Chromatogr A 979:27–32
76. Vaher M, Koel M, Kaljurand M (2002)
Electrophoresis 23:426–430
77. Yu L, Qin W, Li SFY (2005) Anal Chim
Acta 547:165–171
78. Cabovska B, Kreishman GP, Wassell
DF, Stalcup AM (2003) J Chromatogr A
1007:179–187
79. Laamanen P-L, Busi S, Lahtinen M,
Matilainen R (2005) J Chromatogr A
1095:164–171
80. Marszałł MP, Markuszewski MJ, Kalis-
zan R (2006) J Pharm Biomed Anal
41:329–332
10
81. Jiang TF, Gu YL, Liang B, Li JB, Shi YP,
Ou QY (2003) Anal Chim Acta 479:249–254
82. Qin W, Wei H, Li SFY (2003) J Chro-
matogr A 985:447–454
83. Qin W, Li SFY (2003) Analyst 128:37–41
84. Qin W, Li SFY (2002) Electrophoresis
23:4110–4116
85. Qi S, Cui S, Chen X, Hu Z (2004)
J Chromatogr A 1059:191–198
86. Mwongela SM, Numan A, Gill NL,
Agbaria RA, Warner IM (2003) Anal
Chem 75:6089–6096
87. Tian K, Qi S, Cheng Y, Chen X, Hua Z
(2005) J Chromatogr A 1078:181–187
88. Martin SD, Poole CF, Abraham MH
(1998) J Chromatogr A 805:217–235
89. Kollie TO, Poole CF (1992) Chroma-
tographia 33:551–556
90. Poole SK, Poole CF (1995) Analyst
120:289–294
91. Anderson JL, Ding J, Welton T, Arm-
strong DW (2002) J Am Chem Soc
124:14247–14254
92. Shetty PH, Poole SK, Poole CF (1990)
Anal Chim Acta 236:51–61
93. Kowalska S, Stepnowski P, Buszewki B
(2006) J Sep Sci 29:2625–2634
Chromatographia 2008, 68, July (No. 1/2)
94. Buszewski B, Kowalska S, Stepnowski P
(2006) J Sep Sci 29:1116–1125
95. Stepnowski P, Muller A, Behrend P,
Ranke J, Hoffmann J, Jastorff B (2003)
J Chromatogr A 993:173–178
96. Ruiz-Angel MJ, Berthod A (2006)
J Chromatogr A 1113:101–108
97. Stepnowski P, Mrozik W (2005) J Sep
Sci 28:149–154
98. Stepnowski P, Nichthauser J, Mrozik W,
Buszewski B (2006) Anal Bioanal Chem
385:1483–1491
99. Qin W, Wei H, Li SFY (2002) Analyst
127:490–493
100. Markuszewski M, Stepnowski P, Mars-
załł MP (2004) Electrophoresis 25:3450–
3454
101. Kosobucki P, Buszewski B (2008)
Talanta 74:1670–1674
Review