992 Electrophoresis 2006, 27, 992–1005

1*
Abolghasem Jouyban Review
Ernst Kenndler2

1
Faculty of Pharmacy, Theoretical and empirical approaches to
Tabriz University of Medical
Sciences, express the mobility of small ions in capillary
Tabriz, Iran
2
Department for Analytical electrophoresis
Chemistry,
University of Vienna,
Vienna, Austria A discussion is given about the concepts of the ion mobility, the analyte property which
governs migration and thus separation selectivity in CE. It deals with small organic and
inorganic ions, not with charged polymers or large particles like colloids. The discus-
Received September 17, 2005 sion is directed to two main concepts. (i) The first is based on physico-chemistry of ion
Revised November 4, 2005
conductance in solution, and distinguishes three types of mobility. The absolute mo-
Accepted November 7, 2005
bility is the limiting mobility at zero ionic strength; it depends on the solvent and the
temperature. It is obtained by extrapolation of the actual mobilities, those of the fully
charged particles at finite ion concentration. The observed reduction of the absolute
mobility with ionic concentration is related to an ion cloud, and is formulated by the
established theories of ion conductance. It explains the actual mobility as function of
(beside other factors) the ionic strength, the viscosity and relative permittivity of the
solvent, the temperature, the relaxation time of solvent polarisation and the distance of
closest approach between ion and counterion. The effective mobility, finally, is the
mobility when association and dissociation equilibria play a role. Most important are
acid–base reactions, but complexation, ion pairing and homo- and heteroconjugation
were considered as well. (ii) The second approach treats mobility data with different
mathematical methods, and formulates their dependence on variables like solvent
composition with appropriate algorithms. These empirical methods mainly include
least squares and neural network-based methods. The least square methods ranges
from the simplest model, which uses only the molecular weight of the analyte, to more
complicated model requiring three-dimensional structural descriptors of the solutes.
Neural networks have been applied to model the mobility using different input variables
and various architectures. Work comparing the accuracy of least squares and neural
network methods was discussed; the results showed that the neural network method
leads to a more accurate mobility calculation. However, the least squares methods
could give some information to the factors affecting the mobility of the analytes. The
resulting methods allow the prediction of mobilities under different experimental con-
ditions with certain accuracy. It has been shown that using such models, it is possible
to predict mobility of analytes after training the models by a minimum number of data
to speed up the method development stage.

Keywords: Capillary zone electrophoresis / Ionic strength dependence / Mobility /

Organic solvent / Review DOI 10.1002/elps.200500696

Correspondence: Professor Ernst Kenndler, Department for Analyti-

cal Chemistry, Faculty of Chemistry, University of Vienna, Währinger-
strasse 38, A-1090 Vienna, Austria
E-mail: ernst.kenndler@univie.ac.at
Fax: 143-1-4277-9523
Abbreviations: ANN, artificial neural network; CODESSA, compre-
hensive descriptors for structural and statistical analysis; DHO,
Debye-Hückel-Onsager; EtOH, ethanol; FA, formamide; MeCN, ace-
tonitrile; MeOH, methanol; MLR, multiple linear regression; PC, pro-
pylene carbonate; 1-PrOH, 1-propanol; RBFNN, radial basis func-
tion neural network
1 Introduction: The mobility of ions
It is a prerequisite for separation in CZE that the analyte
zones migrate with different velocity, in other words, that
certain separation selectivity is achieved. Whether or not
sufficient resolution of the sample zones is finally reached
* Second corresponding author: Dr. Abolghasem Jouyban,
Faculty of Pharmacy, Tabriz University of Medical Sciences,
Tabriz 51664, Iran
E-mail: ajouyban@hotmail.com

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Electrophoresis 2006, 27, 992–1005
depends then on the extent of peak broadening that takes
place during migration (and on the extra-column peak
effects). Peak dispersion, expressed by the plate height
or the plate number, and determining the separation effi-
ciency, is not the topic of this review. This subject was
discussed in previous reviews [1–5]. The present work is
directed to the mobility of the analyte, which govern the
migration velocity in the electrophoretic system. The
migration velocity, ui, of an ion, i, is directly proportional to
the applied electric field strength, E (Eq. 1a); the factor of
proportionality, mi, is named the mobility (Eq. 1b)
(a) ui = miE (b) mi = ui / E (1)
The mobility has the dimension m2V21s21. It is a single ion
property, which depends on external variables like the
temperature and the solvent, and on ion-specific proper-
ties like the particle size and shape, and its charge; it is
dependent on the ionic strength of the solution as well
(see e.g. [6]). In classical chemistry of ions in solution the
mobility is a parameter derived from the single ion con-
ductance, li, by
m1 = li / ziF (2)
where F is the Faraday constant, and zi is the valency of
the ion. li/zi is the single ion equivalent conductance; it is
derived from the electrolyte conductance and the trans-
ference numbers of the ions. With the increased distribu-
tion of CZE mobilities are measured from the migration
time, tm,i, the migration distance, Ld and the field strength
according to mi = Ld / tm,iE. It should, however, not to be
overlooked that mobilities or proportional quantities have
been measured also with other CE methods like ITP,
decades before CZE was commercially available. In the
textbook published by Everaerts, Verheggen and Beckers
[7] in 1976, the concept of mobility in CE was discussed in
detail, and mobilities in different solvents were tabulated.
Kiso, Hirokawa et al. [8, 9], e.g. presented mobility-related
values in the 1980s for nearly 300 anions in water, and of
60 anions in MeOH (methanol); in mixed solvents with
MeOH and DMSO as constituents mobilities were derived
from isotachophoretic step heights [10, 11].
It is obvious from Eq. (1) that the analyte migration behav-
iour in CZE can be forecasted given that its mobility is
known. Therefore, it would be an enormous advantage if
methods exist by which the mobilities could be predicted.
In this case any structured design to develop a CE method
for analysing a certain analyte and/or set of analytes would
speed up the process at the method development stage.
This contribution gives an overview about the theoretical
models of mobilities in connection with the theory of ion
conductance in solutions on one hand. On the other hand,
the published mathematical models representing the
 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
CE and CEC 993
electrophoretic mobility in CE with monosolvent and
multisolvent BGEs are discussed. We limit the review on
small inorganic and organic ions, which means that we
will not consider polymers or colloids. However, before
discussing such methods we must distinguish different
mobilities which describe ion migration, depending on the
chemical environment of the ion, and on its own ionization
properties. These are the absolute, the actual and the
effective mobilities. The absolute mobility is the limiting
mobility at zero ionic strength, whereas the actual mobility
is that of a fully charged ion at finite concentration. The
effective mobility takes into account that the ions are not
fully ionized. It has to be mentioned that if there is an EOF
present in the electrophoretic system, the overall, or total,
or apparent mobility is relevant for separation. However,
this is simply the sum of the individual ionic mobility and
that of the EOF. The mobility of the EOF will not be dis-
cussed in the present review. Note that we consider the
mobilities as signed quantities; those of cations have
positive and of anions negative signs.
2 Models for ion mobility
2.1 Absolute mobility
The absolute electrophoretic mobility, m0,i, is the average
velocity of a charged species, i, per unit electric field
strength at zero ionic strength. It is obvious that it cannot
be measured directly; it can be obtained by extrapolation
from single ion conductances or actual mobilities at dif-
ferent ionic strengths. In the most simplified model the
medium is considered as a continuum with a certain
macroscopic dynamic viscosity, Z, and the ion as a rigid
sphere with the hydrodynamic or Stokes radius, rSt,i, on
which the electric force is acting (see e.g. [6, 12]). The ion
is retained by hydrodynamic friction expressed by
Stokes’ law, whereby the friction coefficient, fhydr,i is
fhydr,i = 6pZrSt,i (3)
for ‘sticking’ conditions, that is when the solvent wets the
sphere. Due to the balance between the electric force,
zieE, and the frictional force, 6pZrSt,iui, the resulting
absolute mobility (at infinite dilution or zero ionic strength,
when no counterions are present) is then
zi e
m0;i ¼ (4)
6pZr St;i
where e is the elementary charge. The hydrodynamic
radius differs in many cases from the crystal radius; this is
because in solution the ion exists in a solvated form, and
the solvation shell contributes to its size. Equation (4)
would allow forecasting the mobility in different solvents
www.electrophoresis-journal.com
994 A. Jouyban and E. Kenndler
with different dynamic viscosity. It could also express the
change of mobility with temperature, because viscosity is
a temperature-dependent variable.
It is clear that this approach is an oversimplification due to
the following reasons: (i) the solvent is considered as
continuum, but the solvent molecules have about the
same size as the ions. (ii) The ions are taken as spheres,
but most ions are not spherical. (iii) The hydrodynamic
radius is considered as constant, but different solvents
can solvate the ions to a different extent, thus leading to
varying hydrodynamic radii; moreover, even for the same
solvation number the radii can differ due to the different
size of the solvent molecules. (iv) Only hydrodynamic
friction is considered to retain the moving ion, but dielec-
tric friction might play a role as well. Equation (4) leads to
the so-called Walden’s rule, stating that the product of
absolute mobility and solvent viscosity is constant and,
e.g., independent of solvent and temperature
m0,iZ = const. (5)
Due to the oversimplifications, the rule is often not well
obeyed; especially water as solvent gives normally very
different products for a certain ion when related to organic
solvents. The reason is that water is an excellent solvator
for anions and cations as well, in contrast to many organic
solvents. Indeed, the more pronounced the solvation
ability of a solvent, the less pronounced Walden’s rule is
followed when comparison is carried out with other sol-
vents. Constancy of the product in water is observed, on
the other hand, for a particular ion at varying temperature,
T; note that viscosity is a nearly linear function of T over a
not too wide temperature range. However, for organic
solvents, particularly those with low solvation ability, the
product deviates from constancy in several cases sur-
prisingly weakly, especially for large ions with low charge
density. To give an example: the absolute mobilities of an
individual ion differ by 700% in so different solvents like
acetonitrile (MeCN) and propylene carbonate (PC), but
the products stray by only several ten percent [13, 14].
Modifying the exponent of the viscosity from 1 to 0.9 led
to a variation of the average Walden products of only
610% for 17 aromatic acids in mixed aqueous–organic
solvents [15]. The mixed solvents consisted of aqueous
phosphate buffer with MeOH, ethanol (EtOH), 1-propanol
(1-PrOH) or MeCN as organic constituents at concentra-
tions up to 60% [15–18]. The actual mobilities at 20 mM
ionic strength were considered (see below).
An approach that takes into account that the solvent (still
considered as continuum) does not wet the spherical ions
(slipping conditions) expresses the friction factor by
fhydr,i =4pZrSt,i instead of fhydr,i =6pZrSt,i, and thus leads to the
modification of Eq. (4) by replacing factor 6 by factor 4 [19].
 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Electrophoresis 2006, 27, 992–1005
In another additional modification it is tried to consider the
deviation of the ions from spherical shapes, and the cor-
rection factor, f/f0 [20] is introduced. f is the frictional force
on an ion of hydrodynamic radius rSt, and (f/f0) is the ‘fric-
tional ratio’ to correct for the asymmetry of the ion. The
modified Eq. (4) reads
zi e
m0;i ¼ (6)
5pZr St;i ðf=f 0 Þ
It should be mentioned that for Z normally the macro-
scopic dynamic solvent viscosity is taken. However, in
solutions with polymeric additives it is rather the micro-
scopic viscosity that plays the relevant role. The former
parameter is that determined in the usual way, e.g., with
viscometers. On the molecular scale, on the other hand,
the microscopic viscosity can have values even for solu-
tion with high macroscopic viscosity, which do not differ
from those for the pure, polymer-free solvent. This was
shown by CE in [21] not only for mobilities but also for
diffusion coefficients. In extreme cases the macroscopic
viscosity of an aqueous solution of a long-chain PEG (with
average molecular mass 26106) was nearly ten times
larger than a polymer-free aqueous buffer solution, but
the mobility (and the diffusion coefficient) of the reference
ion remained approximately constant and independent of
the viscosity.
An extended model not only considers the hydrodynamic
friction but takes into account the so-called dielectric
friction as well. This contribution occurs because in each
infinitesimal volume element of the solution polarisation of
the solvent is established, the results of the orientation of
the solvent molecules under the influence of the electric
field of the ion. This polarization is characterised by the
dielectric relaxation time, t; this is the time after which the
solvent molecules are randomly oriented again after the
passage of the ion. An electrostatic drag force acts on the
moving ion within the relaxation time, leading to retarda-
tion additional to the hydrodynamic friction. According to
the theory of Hubbard-Onsager [22–24] (modifying the
approach introduced by Fuoss [25], Boyd [26, 27] and
Zwanzig [28, 29], see. e.g. [30]) the coefficient of dielectric
friction for sticking condition, when the solvent wets the
ion, is
tz2 e2
e
e1 
f diel;i ¼ 0:0607 i 3 (7)
ri e2
e is the absolute static permittivity of the medium e = er.e0,
er is the relative permittivity, e0 is the permittivity of
vacuum (8.854610212 F/m), e? is the permittivity at high
frequency. The resulting absolute mobility is then
zi e
m0;i ¼ (8)
tz2i e2
6pZr i þ 0:0607
r 3i e
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Electrophoresis 2006, 27, 992–1005
It can be seen that it depends two-fold on the solvent: via
the dynamic viscosity and the relaxation time. Lucy and
co-workers [31–38] have demonstrated in a series of
striking papers dealing with MeOH and MeCN that the
dielectric friction indeed plays a role especially for higher
charged ions. For those with low charge (e.g. zi = 1) the
effect is, on the other hand, negligible.
Roy and Lucy [37] rearranged Eq. (8) and used
1 6pr St;i tzi e
¼ þ 0:0607 3 (9)
Zm0;i zi e r i Ze
for correlating the anions mobility with charges of 21 to
24 in water–MeOH mixed-solvent buffers and reported
R2 values varying from 0.079 to 0.999. Plots of 1/Zm0,i
versus t/Ze given in Fig. 1 indeed show that the slope of
the fitted lines are dependent on the charge of the ion. It is
increasing with increasing charge number of the analyte.
This justifies the superiority of dielectric friction for higher
charged ions in comparison with ions with unit charge.
Following earlier contributions [39–41], Kiso and Hiro-
kawa [42] showed that the absolute mobility can be
expressed as function of the ion mass. They found that
the absolute mobilities were inversely proportional to the
square roots of formula weights for a number of n-alkyl-
ammonium, mono- and dicarboxylate ions analytes.
Another empirical attempt was used [31] to compute the
mobility of analytes taking their size and shape into
account, and used the following equation to test the
concept on the mobility of monoamines
6:3  10
3
m0;i ¼ (10)
0:620
ðM þ 0:22H
0:175SÞ
Figure 1. Correlation between change in ion mobility and
the solvent properties t, e and Z for H2O/MeCN media,
according to the rearranged Hubbard-Onsager equation
(Eq. 9). Solutes: benzenesulfonate, 2,6-naphthalenedi-
sulfonate, 1,3,6-naphthalenetrisulfonate. From [37] with
permission.
 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
CE and CEC 995
where M is the molecular mass, and H is the mean of
waters of hydration of the ion (the sum of the waters
hydrating the functional groups of the analyte calculated
using McGowan’s additive method). S is a shape factor,
which is a function of the number of carbon atoms in the
longest chain, the number of side chains on the amines (0
for primary amines, 1, 2 and 3 for secondary, tertiary and
quaternary amines, respectively) and the average number
of carbon atoms in the side chains. The percentage error
of Eq. (10) for representing the mobility of monoamines
was 3.7%.
C0
m0;i ¼ (11)
V C1 þ C2 pK
was also proposed by Lucy and co-workers [32] to cal-
culate the absolute mobility of analytes using size and
pKa (or pKb) of the analytes. V is the van der Waals mo-
lecular volume, and C0–C2 are the model constants. The
numerical values of C0 varies between 6.861023 and
7.861023 for acids and bases, C1 is nearly constant for
acid and bases (0.62) and C2 is ,0.66. V0.62 could be
considered as effective surface area as it is confirmed in
Offord’s model (which expresses the actual mobility as
function of the molecule mass, M, see below). The model
of Lucy and co-workers [32] combines the hydrodynamic
and dielectric frictions on the electrophoretic mobility
and has been derived based on Born’s model. Its accu-
racy has been checked using acidic and basic analytes
and produced errors less than 5% [32]. The model
should be trained for acidic and basic analytes sepa-
rately. In a multilayer neural network using an extended
delta-bar-delta algorithm proposed by Li and co-workers
[43] employing molecular weight, molecular volume, a
code for the charge of the ion (21 for anions and 11 for
cations) and the pKa of the analytes as input variables
using the absolute mobility of 56 mono-charged analytes
collected from the literature [32] more accurate results
have been reported [43]. The authors divided 56 mobility
data into three subsets, namely training, validation and
test sets. The artificial neural network (ANN) model has
been trained by 40 mobility data of the training set and
the trained model was validated using ten validation data
sets. Then, six data points of the test set have been
predicted using the trained and validated model. The
percentage error for training, validation and test sets
were 2.2, 3.7 and 2.8%, respectively, where the overall
percentage error was 2.5% which is significantly less
than 5% of Eq. (11). This equation was, by the way,
derived from theory and its constants have physical
meaning, whereas the ANN method uses weights with-
out any physical meaning. It should be mentioned that
many equations are unit dependent. In the present
review we do not give the appropriate units for the
www.electrophoresis-journal.com
996 A. Jouyban and E. Kenndler
quantities the equations consist of. These details are
certainly indispensable for book chapters or textbooks.
Here the readers are referred to the original papers.
In a modelling work on absolute mobility data, Xue and
co-workers [44] used a radial basis function neural net-
work (RBFNN) and seven descriptors of analytes studied,
which were calculated from the chemical structures of the
analytes using comprehensive descriptors for structural
and statistical analysis (CODESSA) software. Their ana-
lyte set include 115 carboxylic and sulphonic acids and
the RBFNN method gave an overall average relative
deviation of 3.8% for all data points [44].
2.2 Actual mobility
The actual mobility, the mobility of a fully charged ion in a
solution of finite ionic strength, is expressed mainly by
two approaches. One approach is based on the theory of
conductance in solutions; in the most established theory
– developed by Onsager, based on the work of Debye and
Hückel – it is tried to relate the deviation of the actual from
the absolute mobility to the presence of an ion cloud or
atmosphere formed by the counterions around the ion.
The other approaches use mathematical algorithm with
empirical variables to express the mobility as function of
e.g. the solvent composition. Both approaches are dis-
cussed as follows.
2.2.1 From theory of ion conductance in
solution
It was observed since the beginning of research on ion
conductance in solution that the relationship between
conductance and concentration of a strong electrolyte
decreases nonlinear with increasing concentration. The
theory to explain the influence of the ion concentration on
the chemical activity of an electrolyte was introduced by
Debye and Hückel and extended to ion conductance by
Onsager (see e.g. [12]). The according Debye-Hückel-
Onsager (DHO) limiting law of the conductance, L, for a
binary electrolyte as a function of the ionic strength, I, is
( )
2:801  106 jzc za jqL0 41:25ðjzc j þ jza jÞ pffi
L ¼ L0
pffiffiffi þ I
ðeTÞ3=2 ð1 þ qÞ ZðeTÞ1=2
(12)
Suffix 0 indicates zero ion strength, I, suffix c stands for
cations, a for anions. The ionic strength for a certain ion
concentration, c, is I ¼ 1=2ðz2c cc þ z2a ca Þ or generally
when X ion species, i, are present in solution
I ¼ 1=2 z2i ci . q in Eq. (12) is
 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Electrophoresis 2006, 27, 992–1005
jzc za j l0;c þ l0;a
q¼ (13)
jzc j þ jza j jza jl0;c þ jzc jl0;a
or, written for mobilities instead of single ion limiting con-
ductances, l0
jzc za j m0;c þ m0;a
q¼ (14)
jzc j þ jza j jza jm0;c þ jzc jm0;a
q has the value of 1/2 for symmetrical electrolytes
(jzc j ¼ jza j). The actual mobility for monovalent ions can
be derived from Eq. (12) by transformation of con-
ductance into mobility and reads
" #
8:204  105 4:275 pffi
mact;i ¼ m0;i
m0;i þ I (15)
ðeTÞ3=2 ZðeTÞ1=2
A simplified form was used in [45] to match mobilities as a
function of the ionic strength according to
pffi pffi
mact;i ¼ m0;i
Am0;i I
B I (16)
A and B are functions of relative permittivity and dynamic
viscosity, and depend thus on the solvent (the data for A
and B in many solvents are tabulatedpffiin [2]). As a con-
sequence, the slope of the mact versus I curve is strongly
solvent-dependent as well, in other words, the effect of
the ionic strength on the mobility varies with the solvent.
This can be seen from the plot in Fig. 2 (for hypothetical
ions with absolute mobility of 4061029 m2V21s21 in all
solvents) constructed from the data for er and Z for differ-
ent solvents. Whereas the reduction in formamide (FA) is
even less pronounced than in water, it is much stronger in
the common organic solvents MeOH and MeCN. This
was indeed found [46] and its consequence for separation
efficiency in CE clearly demonstrated in [45].
Deviations from Eq. (12) are found at concentrations
higher than 1023 M. Attempts were made to extend the
validity of Eq. (12) by adding other concentration-
pffi
Figure 2. Mobility vs. I in FA, PC, water, DMF, MeOH
and MeCN according to the DHO limiting law (Eq. 15) at
257C. Mobilities are in 1029 m2V21s21, ionic strength in
mol/L.
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Electrophoresis 2006, 27, 992–1005 CE and CEC 997

dependent terms. One semiempirical equation for the

conductance was introduced by Hsia and Fuoss [47];
adopted to mobilities it would read

mact;i ¼ m0;i
Sc1=2 þ Eclnc þ A0 c þ B0 c3=2 (17)

Here the factors S and E describe the effect of the ion at-
mosphere on the mobility; for low ionic strengths, S
approaches the Onsager limiting slope. A’ and B’ depend
on the same parameters as S and E, and on the distance
of closest approach between ion and counterion. This
means that this and further approaches to widen the
validity of the DHO limiting law to higher concentrations
take into account that the ions are not point charges, but
have finite radii. In this case an ionic diameter, a, is intro-
duced, which is – in a more strict sense – the distance of Figure 3. Actual mobilities, mact,i, of tetramethyl-
closest approach of ion and counterion. This extended ammonium (TMA), tetraethylammonium (TEA) and tetra-
theory for electric conductance was developed among butylammonium measured as a function of the ionic
others by Falkenhagen, Pitts and their co-workers strength in nitromethane as solvent. Temperature, 257C.
[48–51]. The according equation, again transformed for Full lines are fitted curves according to Eq. (18). From [53]
the ionic mobility of a monovalent ion in a symmetrical with permission.
electrolyte, is

mact;i ¼ m0;i
Equation (21) was also proposed based on classical so-
" # pffi
8:204  105 4:275 I lution theory of electrolytes [33]

m0;i þ pffi (18) !
ðeTÞ3=2 Zðer TÞ1=2 1 þ 50:29aðer TÞ
1=2 I pffi
I
mact;i ¼ m0;i
Az pffi (21)
Also in this equation the mobility is solvent-dependent, as 1 þ 2:4 I
it contains the specific parameters er and Z. Indeed not where A is a constant. The accuracy of Eq. (21) was tes-
only for common solvents like MeOH or MeCN, but also ted on mobility data of carboxylates, phenols and sul-
for unusual solvents in CE like PC, FA, DMF, N,N-di- phonates. It has been shown that pthe
ffi Onsager slope (A in
methylacetamide or nitromethane [14, 52–54] this equation most simplified mact;i ¼ m0;i
A I) deviates from con-
is well obeyed, as can be seen from Fig. 3. Equation (18) stancy for higher zi values as demonstrated in Fig. 4 [33].
can be simplified as follows, with B, B1 and B2 being de- Note that all the models could be used to determine the
pendent on the solvent properties just mentioned. In this absolute mobility of analytes by extrapolation.
form it was used to calculate in turn the limiting mobility of
the analytes in MeOH from the actual mobilities [55] The following equation, similar to Eq. (12), was used by
pffi pffi Roy and Lucy [35] in accordance with basic physico-
mact;i ¼ m0;i
½B1 m0;i þ B2   ð I=ð1 þ aB IÞÞ (19) chemistry
These limiting mobilities served as input data to recalcu- 82:5
late the actual mobilities by the same equation for mact;i ¼ m0;i
zi þ
Zðer TÞ1=2 F
depicting the electropherogram by computer simulation  pffi (22)
with the program PeakMaster (http:///www.natur.cuni.cz/ 1:40  106 2q I
þ jzþ z
j pffiffiffi m0;i pffi
,gas) adopted for methanolic solutions [55]. ðer TÞ1=2 1þ q 1 þ Ba I

All these equations stand for mono-charged ions. For The authors investigated the effects of ionic strength on the
symmetrical electrolytes with higher valency than unity electrophoretic mobility of anions with 21 to 24 charges at
the modified equation different concentrations of MeOH in binary mixed-solvent
  pffi buffers containing 0.05 M NaOH. They assumed that the
q I
mact;i ¼ m0;i
A1 m0;i pffiffiffi þ zi A2 pffi (20) ion size parameter, a, remains unchanged in the mixed-
1þ q 1 þ 1:5 I
solvent buffer. In Eq. (22), q is the electrolyte parameter
was used [56]; A1 and A2 are constants, and q is the same equal to 0.5 for a 1:1 electrolyte. In parameter B
factor as defined in Eqs. (13), (14). For unsymmetrical sffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffiffi
electrolytes it is necessary to calculate the limiting mobil- 8pNA e2
B¼ (23)
ity from mact by iteration. 1000er k B T

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998 A. Jouyban and E. Kenndler
er is the only varying term for the water-MeOH mixture (NA
and kB are the Avogadro and the Boltzmann constants,
respectively). It was shown that the ions with higher
charges are strongly influenced by dielectric frictions in
these mixed-solvent buffers.
All these equations do not take into account chemical
equilibria in which permanent ions are involved (we do not
consider at this stage protolysis of weak acids and bases).
These equilibria can lead to phenomena that hardly occur
in aqueous solutions, but are quite frequent in organic sol-
vents, especially in those with low dielectric constant: ion
pairing and homo- and heteroconjugation. The modified
Fuoss-Onsager equation for the actual mobility extends
the Hsia-Fuoss equation by a term for ion pair formation,
the last term of the following equation (see e.g. [19])
pffiffiffiffiffiffiffiffiffi
mact;i ¼ m0;i
S ðacÞ þ EðacÞlnðacÞ þ A0ðacÞþ
(24)
þ B0 ðacÞ3=2
K A mi g2 ðacÞ
where c is the initial total ion concentration, KA is the ion-
pair formation constant, a is the degree of dissociation of
the ion-pair and g6 is the mean activity coefficient
according to the Debye-Hückel limiting law. For solvents
with high dielectric constant and for low electrolyte con-
centrations the Fuoss-Onsager equation approaches the
Onsager limiting law. Barthel et al. [57–60] used this
equation to derive ion-pair formation constants for 1–1
electrolytes in several organic solvents (MeCN, MeOH,
EtOH, 1-PrOH, mixtures of PC and MeCN). In a very
recent work, Dai et al. [61] used the deviations from
Eq. (22) to derive ion-pair formation constants by CE in
mixed solvents consisting of MeCN and water.
 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Electrophoresis 2006, 27, 992–1005
Figure 4. Effect of the solute
charge on the Onsager pffi slope
(A in mact;i ¼ m0;i
A I) for
carboxylates (d) and sulfonates
(u) with charge number be-
tween 1 and 6. From [33] with
permission.
2.2.2 From empirical algorithms
A purely empirical equation for actual mobilities as a
function of ionic strength and analyte charge number, zi,
was derived by curve fitting of data measured by CE for a
set of 21 organic anions (sulfonates) with charge numbers
between 1 and 6 [62]. The model was adopted from a
previous model successfully implemented in HPCE
simulation program for CE [63]. The concentration range
was up to 100 mM. The resulting equation
pffiffiffiffiffi
mact;i ¼ m0;i expð
0:77 zi IÞ (25)
enabled to express the actual mobility for each ionic
strength with less than 5% deviation.
An empirical equation was used to calculate the actual
mobility of peptides in paper electrophoresis [64]
Kzi
mact;i ¼ (26)
M2=3
where K is a constant, M is the molecular mass and M2/3
could be considered as a function of the effective surface
area [64]. It was successfully used for modelling the mo-
bility of small ions of vinca alkaloids in aqueous and
aqueous–organic buffers [65] and also the mobility of
sulphonamides [66]. The only required data are the mo-
lecular weights of the analytes as size indicator. However,
there the dependence on ionic strength is missing.
In order to overcome the limitations from the assumption
of spherical geometry of the analytes, and also to intro-
duce a discriminative tool between different isomers of
the analytes, McKillop and co-workers used three-dimen-
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Electrophoresis 2006, 27, 992–1005
sional structural features of the analytes, which has been
ignored above. It is a fact that a number of isomeric
compounds having similar shape and identical charges
could be separated in CE and the models representing
the mobility should cover these observations as well.
McKillop et al. assumed that three contributing factors
could be considered as: (i) the overall size of the ion, (ii)
the effective radius of the ion and (iii) the resistance of the
ion due to its natural tumbling motion. The authors pre-
sented these factors: rvdw, the van der Waals radius; rrot,
the rotational radius and ri, the radius of inertia to tumbling
distance between centre of mass and charge. The fol-
lowing equation for the actual mobility was proposed
 
3:22 0:570 0:0114
mact;i ¼ þ

0:604  10
3 (27)
r vdw r rot ri
based on the data of a series of alkylpyridines [67]. As
evident from the coefficients of the independent vari-
ables, the size factor (rvdw) is the most important factor on
the mobility, followed by rrot and ri which are responsible
for the observed selectivity of the positional isomers. A
good predictability for the mobility of the analytes has
been shown and the model was successful in representing
the mobility of alkylpyridines using the independent vari-
ables computed using a molecular modelling package
called SYBYL [67]. The extension to other pyridine deriva-
tives gives a slight modification of the constants in Eq. (27),
and still a high correlation between the measured and the
predicted behaviour is demonstrated. In a work by Jalali-
Heravi and Garkani-Nejad [68], employing a data set of
pyridines, the mobilities were modelled using an ANN
method with architecture 3–6-1 with the reciprocal of van
der Waals radius of the molecules, the principal moment
of inertia of the molecule around the x-axis and the dipole
moment of the molecules as input variables. The ANN
model was trained by a back-propagation method.
Standard errors of training and prediction sets were 1.0
and 1.2%, respectively [68]. It has to be mentioned that
these models do not consider the ion strength depend-
ence of the actual mobility. In an earlier work of Jalali-
Heravi and Garkani-Nejad’s group, an ANN model with
the architecture of 3–4-2 was proposed to correlate the
actual mobility of the analytes. The accuracy of the ANN
method has been compared with that of multiple linear
regression (MLR) models. Their proposed MLR models
were
mact;i ¼ J0 þ J1 DHf þ J2 pK þ J3 SA (28)
for cationic sulfonamides and
mact;i ¼ J4 þ J5 DHf þ J6 PPCH þ J3 SA (29)
for anionic sulfonamides. J0–J6 are the model constants,
DHf is the heat of formation, PPCH is the most positive
partial charge (the most positive atom of the analyte), and
 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
CE and CEC 999
SA is the surface area. The superiority of ANN on MLR
models has been shown [69]. As an example, the reported
MLR model for the cationic form of sulfonamides is
mact;i ¼
1:091 þ 0:001DHf þ 1:335pK
0:002SA (30)
The intercept represents the effects of other affecting
variables which are not considered in the model, factor
0.001 reflects the effect of heat of formation, factor 1.335
means that the higher the pKa values the higher is the
mobility observed and factor - 0.002 means that the wider
the surface area of the ions, the slower is the mobility of
the ions. However, this is not the case for the ANN mod-
els, and one cannot provide such information from ANN
curve fitting parameters; they just provide better correla-
tions between input and output variables as shown by the
authors [69].
In another MLR model, the logarithm of the electropho-
retic mobility as dependent variable and computational
descriptors calculated using HyperChem as independ-
ent variables have been employed as
lnmact;i ¼ K 0 þ K 1 PQ þ K 2 V 2=3 þ K 3 TE þ K 4 DHf þ K 5 MR (31)
PQ is the net charge of the most positive (for cations) or
the most negative (for anions) atom on the analyte, V2/3 is
the surface area of the molecule with volume V, TE is the
total energy, a computational term calculated using
HyperChem software, MR is the molar refractivity and K0
to K5 are the model constants. The accuracy of Eq. (31)
was tested using five sets of various analytes where the
overall percentage errors for correlative and predictive
analyses were 1.4 and 5.6%, respectively [70]. The main
disadvantage of the mathematical equations representing
the mobility of the analytes using computational descrip-
tors, so-called quantitative structure–property relation-
ships (QSPRs), is that they calculate the mobility solely
from chemical structures of the analytes. The main lim-
itation of QSPRs is that they should be trained for each
analytical condition and there is no possibility to predict
the mobility under various conditions.
From cosolvency models representing the solute solubil-
ity in binary solvent mixtures an approach adopted to
calculate the electrophoretic mobility of analytes in
mixed-solvent buffers gave
lnmact;i ¼ f 1 lnm1 þ f 2 lnm2 þ K 0 f 1 f 2 þ K 1 f 1 f 2 ðf 1
f 2 Þ (32)
where f1 and f2 represent the fractions of the solvent 1
and 2 in the mixed-solvent buffer, m1 the mobility of the
analyte in the mono-solvent buffer 1 and m2 the mobility of
the analyte in the monosolvent buffer 2; K0–K1 are the
model constants [71]. The accuracy of Eq. (32) has been
assessed using various acidic and basic analytes in
aqueous and nonaqueous binary solvents and compared
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1000 A. Jouyban and E. Kenndler
with other algorithms [72]. The model provides better
correlations and more accurate predictions among similar
algorithms and is capable of predicting the electropho-
retic mobility of structurally related drugs in a given binary
buffer [73]. The model could also be extended to

lnm ¼ f 1 lnm1 þ f 2 lnm2 þ f 3 lnm3 þ f 1 f 2 M0 þ M1 ðf 1
f 2 Þþ
  0 0
þ M2 ðf 1
f 2 Þ2 þ f 1 f 3 M0 þ M1 ðf 1
f 3 Þþ
0   00 00 00
þ M2 ðf 1
f 3 Þ2 þ f 2 f 3 M0 þ M1 ðf 2
f 3 Þ þ M2 ðf 2
f 3 Þ2 þ
 000
000 000
þ f 1 f 2 f 3 M 0 þ M 1 ð f 1
f 2
f 3 Þ þ M2 ð f 1
f 2
f 3 Þ 2
for correlating the mobility in ternary solvent buffers and
provided accurate results using mobility of beta-blockers
in water–MeOH–EtOH mixtures [74]. In Eq. (33), f3 is the
fraction of the third solvent in the mixed-solvent buffer, m3
the mobility of the analyte in the mono-solvent buffer 3;
M0–M2, M0 0-M0 2, M00 0–M00 2 and M000 0–M000 2 are the model
constants. It has been shown that by collecting experi-
mental mobility data of an analyte in the mono-solvent
buffers and also three data points from each binary and
ternary solvent buffers (total required data points are 15),
and by computing the model constants of Eq. (33), one is
able to predict the mobility in all solvent compositions of
binary and ternary solvent buffers. In practice, employing
such predictive models, the process of method develop-
ment could be facilitated [74]. Recent investigations
showed that the ANN methods produced more accurate
predictions for mobility of analytes in mixed buffers in
comparison with Eqs. (32), (33) [75, 76].
Liu et al. [77] calculated more than 130 descriptors of 26
aromatic acids using CODESSA software and employed
five of the descriptors to model the mobility data of anions
in mixed-solvent buffers. Their proposed MLR model is
mact;i ¼ H1 TEIP
H2 MIA þ H3 Qmin


H4 HDSA1 =TMSA
H5 VOL þ H6
where TEIP is the topographic electronic index (all pairs),
MIA is the moment of inertia A, Qmin is the minimum partial
charge, HDSA1 is the hydrogen acceptor-dependent
hydrogen-donor surface area, TMSA is the total molecu-
lar surface area, VOL is the molecular volume and H1–H6
the model constants. Equation (34) should be trained for
each solvent mixture of the buffer. To provide a general
model for correlating mobility data of aromatic acids in
water–MeOH and water–EtOH binary mixtures, the
authors added dielectric constants of solvent mixtures
and the energy of the highest occupied molecular orbital
(EHOMO) of MeOH and EtOH to Eq. (34) and reported
R = 0.835 for 182 mobility data of the analytes studied. To
provide more accurate results, the authors also employed
 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Electrophoresis 2006, 27, 992–1005
two other numerical methods, namely RBFNN and sup-
port vector machine (SVM), and showed that the SVM
provides more accurate results in comparison with MLR
and RBFNN methods [77].
Ab initio predictions (prediction without using any experi-
mental data) of the mobility of analytes using computa-
tional methods are not available at the present time. As
discussed above, the present models possess a number
 of model constants and require experimental data points
 as training set. However, it is possible to train the available
mathematical models using a minimum number of
(33) experimental data points and predict mobilities under the
same analytical conditions. In an earlier work [73] experi-
mental mobility data of atenolol, alprenolol, labetalol and
metoprolol in aqueous buffer under a given analytical
condition, their molecular weight and pKb values were
employed to train the model of Fu, Li and Lucy [32]; and
the trained model was
394:600
mact;i ¼ (35)
MW 0:639
2:935pK b
To show the capability of this model, mobilities of three
beta-blocker drugs, i.e. propranolol, timolol and acebu-
talol, have been predicted with the overall percentage
error of 2.8% [73]. Since a large number of chemically or
biologically related compounds are synthesised and/or
extracted from natural sources to assess their biological
activities and finding an analytical method has its own
priority in such studies, one can estimate the importance
of such trained models in pharmaceutical and chemical
industries [73]. It is obvious that any predictive method
has some prediction errors. So, the question is what error
is acceptable in practice. It could be agreed that this error
range is less than or equal to the RSD of repeated
experiments; this is say 1% for mobility data [31]. Under
this precondition the accuracy of prediction is too low. If
we accept the threefold of the SD as acceptable, then
prediction is within this tolerable range.
(34)
2.3 Effective mobility
The effective mobility meff of the compound consisting of n
ionic forms is
P
n
ck mact;k zk
meff ¼ k¼1 (36)
c
where ck, mact,k and zk are concentration, actual ionic mo-
bility and charge number of the kth ionic species,
respectively, and c is the analytical concentration of the
compound. Again the mobilities are signed quantities in
this context.
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Electrophoresis 2006, 27, 992–1005 CE and CEC 1001

It is clear that all equilibria in which the analytes are
involved can influence the effective mobility; this holds,
e.g., for the ion-pair formation or conjugation as men-
tioned above. However, the most abundant type of reac-
tion that is relevant in the context of electrophoresis of
weak acids and bases is certainly protolysis. In this case
the application of the Henderson-Hasselbalch equation
transformed to express the degree of ionization, a, for
example of a monobasic acid HA to the anion A2
according to HA = H1 1 A2 gives
1 tions are mentioned. Homo- and heteroconjugation is a
a¼
1 þ 10pK a
pH
and consequently the expression for the effective mobility
as a function of the pH of the buffer
mact;A

meff;A
¼
1 þ 10pK a
pH
It is important that this equation is followed well not only in
aqueous solutions but also in organic solvents like MeOH
and MeCN [46, 78–80], even in such unusual solvents like
nitromethane [53], FA [54] or DMF [81] (see for example
Fig. 5).
If not only one group is ionised by protolysis, the appro-
priate equations are obtained accordingly. So is the
effective mobility for an acidic amino acids like aspartic or
glutamic acid as a function of the pH
mact;Aþ 10pK a;1
pH
mact;A
10pH
pK a;2
meff;A ¼
10pK a;1
pH þ 10pH
pK a;2 þ 1
with Ka,1 and Ka,2 being the dissociation constants of the
equilibria A1 = A7 1 H1 and A7 = A2 1 H1, respectively.
For a more detailed formal treatment of the mobility see
e.g. [82]; this formalism was used for the simulation pro-
gram PeakMaster, which is freely available on the Internet
(http:///www.natur.cuni.cz/,gas). Expressions for the
mobility dependence of zwitterions on the pH in mixed
solvents were used based on the mass action law for
quinoline derivatives by Barron et al. [83] in order to pre-
dict favourable separation conditions.
In principle the effective mobility is more adequate than
the actual mobility also when other than acid–base equi-
libria are present. In the first view, complexation reac-
tions, ion pairing or homo- and heteroconjugation reac-
(37)
phenomenon which can occur in solvents with low solva-
tion ability for the ion. Let us consider an acid HA, which
dissociates into H1 and A2. In water, with its excellent
solvation ability towards both cations and anions, the ions
are stabilized by hydration. If the solvent has a low solva-
(38)
tion ability, e.g. MeCN, the anion is not stabilized by the
solvent molecule (stabilisation of H1 is reflected by the
solvent’s basicity). The anion might be stabilized instead
by association with its parent molecule, the neutral parti-
cle HA, under formation of the ion AHA2. This reaction is
called homoconjugation. It is clear that the resulting ion has
a mobility different than A2. If reaction takes place with a
molecule HB, e.g. a buffer constituent present in the so-
lution, heteroconjugation can take place under formation
of AHB2. Also in this case the mobility of A2 is influenced,
and it is a matter of the position of the conjugation equi-
librium to what extent the mobility varies [84, 85]. In prin-
(39) ciple we could formally treat these equilibria like the acid–
base equilibria, introduce a kind of the degree of associ-
ation (related to the conjugation constants and the con-
centrations of the reactants) and discuss the resulting
mobility as effective mobility rather than as actual mobili-
ty. It is the same formalism as we apply it for complex
formation reactions, e.g. with CDs or other ligands. In

Figure 5. Measured effective mobilities, meff,i, vs. the pH of the BGE in organic solvents. Solid lines are
fitted according to Eq. (38). Left: nitromethane as solvent with BGEs of different pH. Right: FA as
solvent; mobility of 3,5-dimethylbenzoate vs. pH of the BGE. Temperature, 25.07C. pH was calculated
by means of the Henderson-Hasselbalch equation, and corrected for the activity of the buffer anion.
From [53, 82] with permission.

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1002 A. Jouyban and E. Kenndler
case of biological or pharmaceutical agents we name the
method affinity electrophoresis. In all cases the same
principle lies in the reaction, in the simplest case with 1:1
stoichiometry, according to
A 1 X = AX
In the case of protolysis, X is the proton; in other applica-
tions X can be CD, a protein in case of drug–receptor
interactions etc. We consider the kinetics of complex for-
mation and dissociation being fast enough. In practice the
ligand X is added to the BGE at a varying concentration (in
case of the acid–base reaction it is adjusted by the pH of
the buffer); the analyte is injected as usual in CZE. The
effective mobility of the analyte, meff,A, is composed from
its mobility, mA-, as free species A2, weighted by the frac-
tion of free analyte, f, present in the solution, and the
mobility of the complex, mAB, also weighted by the com-
plex fraction, which is (1 2 f). Fraction f is given by
½A
f¼
½A þ ½AX
which upon application of the mass action law gives
1 the mass action law but on an empirical approach using a
f¼
1 þ K c ½X
where Kc is the complexation constant. The effective
mobility as function of the ligand concentration is
accordingly
1 K c ½X m
þ mAB K c ½X
meff;A ¼ mA
þ mAB ¼ A
1 þ K c ½X 1 þ K c ½X 1 þ K c ½X
This equation expresses the effective mobility for reac-
tions we summarize under the terms complexation, ion
pairing, conjugation and affinity. It is obvious that for the
noncharged complex, e.g. in case of 1:1 ion pairing or
 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim
Electrophoresis 2006, 27, 992–1005
complexation of two ions with opposite charge Eq. (43)
reduces to
1
meff;A ¼ mA
(44)
1 þ K c ½X
(40) because the mobility of the complex is zero. In Fig. 6, an
example is given in which the deviation of the depend-
ence of mobility on ionic strength according to Eq. (22)
was related to ion pairing in buffered MeCN–water media.
From this deviations the authors derived ion-pair forma-
tion constants [61]. It is worth mentioning that Eq. (43)
also holds for acid–base equilibria. In this case the effec-
tive mobility of the anion A2 formed by dissociation of the
acid HA (its mobility is zero) reads after transformation of
Eq. (43)
mA
mA

meff;A ¼ ¼ (45)
1 þ K c ½X 1 þ 10pK a
pH
whereas [X] is [H1] and Ka, the ionisation constant, is the
reciprocal of the formation constant Kc. It can thus be
(41) seen that Eq. (44) is identical to Eq. (38).
The expressions for the effective mobility were derived so
far by simple thermodynamics. An equation not based on
(42)
MLR model describing the effective mobility as function
of pH and of the concentration of organic modifiers [86] is
lnmact;i ¼ A0 þ A1 lnf 1 þ A2 lnpH þ A3 ðlnf 1 Þ2 þ
þ A4 ðlnpHÞ2 þ A5 lnf 1 lnpH (46)
(43)
where Ai is the model constants and, f1 is the fraction of
organic solvent in the buffer. The model has been tested
on 12 benzoate derivatives at various f1 and pH values of
MeCN and phosphate buffer mixture where the correla-
tion coefficient was 0.999 [86].
Figure 6. Deviation of the mobility as for-
mulated by Eq. (20). Conditions: 35/65 MeCN/
buffer, 5.0 mM hydrochloric acid at pH 2.3, and
0–55 mM NaCl. Plot legends: (s) nortriptyline;
(u) amitriptyline; (n) perphenazine; dotted
lines, theoretical predictions based on Eq. (20).
It was assumed that perphenazine is doubly
charged. From [62] with permission.
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Electrophoresis 2006, 27, 992–1005
3 Conclusion
Advanced theories exist which formulate the mobilities as
function of many experimental parameters and variables.
Fundamental physicochemical theories based on ion
conductance in solution with increasing complexity have
been established. They start with the most simple concept
of the electrical movement of a spherical ion in a con-
tinuum with application of Stokes’ law, and try to explain
the deviations from this model by more sophisticated
concepts like introducing parameters for nonspherical
size, the presence of an ion atmosphere, the considera-
tion of the ions and counterions approaching each other
to a certain distance and the polarization of the solvent in
the vicinity of the ion by its electric field (so-called dielec-
tric friction). Association and dissociation reactions have
the effect that only a part of the analyte species is present
in ionic forms. The application of basic thermodynamics
and the mass action law allows expressing the resulting
effective mobility as a function of the ligand concentration
for equilibria in acid–base, complexation, ion pairing,
homo- and heteroconjugation or affinity reactions. In
many cases, however, the systems are too complex, and
then the mobility changes cannot be related to physico-
chemical quantities. In these cases mathematical algo-
rithm can be favourably applied to predict mobilities as a
function of some experimental variables. The presented
algorithms could be categorised in MLR and ANN meth-
ods. Usually ANN methods provided better correlation in
comparison with MLR. On the other hand, MLR models
are able to provide slightly more significant information on
the factors affecting the mobility of the ion. These MLR
and ANN models describing the mobility with respect to
ionic strength, solvent composition, pH of the buffer and
structural features of the analytes were presented.
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5 Addendum c Concentration
E Electric field strength
List of symbols F Faraday constant
H Mean of waters of hydration
a Ion size parameter, distance of closest approach
DHf Heat of formation
E Elementary charge, charge of an electron
I Ionic strength
f/f0 Shape correction factor, frictional ratio
KA Ion-pair formation constant
f1 Volume fraction of solvent 1
MR Molecular refractivity
f2 Volume fraction of solvent 2
M Molecular mass
f3 Volume fraction of solvent 3
NA Avagadro’s number
fdiel,i Dielectric friction coefficient PPCH Most positive partial charge
fhydr,i Hydrodynamic friction coefficient PQ Net charge of the most positive/negative atom of
kB Boltzmann constant the analyte
q Electrolyte parameter (1/2 for 1:1 electrolyte) S Shape factor, Onsager slope in different equations
rcryst,i Crystal radius SA Surface area
ri Inertial radius TE Total energy
rrot Effective radius V Molar volume
rSt,i Stokes (hydrodynamic) radius of the ion a Degree of dissociation
rvdw van der Waals radius e Absolute static permittivity
zi Charge on the analyte, valency of the ion e0 Permittivity of vaccum

 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim www.electrophoresis-journal.com

Electrophoresis 2006, 27, 992–1005 CE and CEC 1005

er Relative permittivity mI Mobility of an ion

g6 Mean activity coefficient m0,i Absolute electrophoretic mobility of an ion
Z Dynamic viscosity mact,i Actual electrophoretic mobility of an ion
l Single ion conductance meff,i Effective electrophoretic mobility of an ion
m1–m3 Electrophoretic mobility of analyte in mono- t Dielectric relaxation time
solvent buffer 1–3 L Conductance

 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim www.electrophoresis-journal.com