ANALYTICAL SCIENCES JUNE 1996, VOL.

12 381

Original Papers

Retention Volume of the Second System Peak in

Ion-Exchange Chromatography with
Monobasic Acid Eluent Systems

Hideki WATANABE Yukio YOKOYAMA and Hisakuni SATOt

Laboratory of Analytical Chemistry, Faculty of Engineering, Yokohama National University

Hodogaya, Yokohama 240, Japan

A general, functional relationship between the retention volume of the 2nd system peak and the chromatographic
parameters (the acid dissociation constant of the eluent component, the eluent concentration, the acid/salt ratio in the
eluent, the acid-partition coefficient) in ion chromatography with monobasic acid eluent systems (acids, salts, and their
mixtures) have been derived. The relations precisely reproduced the results by computer simulation based on the plate
theory, and were confirmed with independent experiments. By using these relations, the partition isotherm of eluent
acids to the stationary phase can be obtained easily.

Keywords System peak, ion-exchange chromatography, computer simulation, monobasic acid eluent system

Computer simulations''2 based on the plate theory and

the acid partition model have revealed the origin and Theoretical Premise and Computer Simulation
fundamental behavior of the system peaks3-5 often
observed in ion-exchange chromatography with bulk- A monobasic acid eluent system contains HB, B-, H+
property detectors. By these simulations, however, we and A+ (some cation other than H+). The total concen-
can only obtain a chromatogram (concentration profile tration of B species in the mobile phase of a theoretical
vs. the volume of mobile phase) for a set of parameter plate is represented as CBand the concentration of A+is
values after calculations taking several tens of minutes, CA. A+ was assumed to be indifferent to the acid-base
and the calculation sometimes is terminated because the reaction.
equilibrium calculation in a certain plate does not The acid-base equilibrium in the mobile phase is
converge. In the calculation of 1000 theoretical plates, represented as:
even the use of quadruple-precision variables with a
mainframe computer was sometimes unsuccessful.2 HB~H+ + B-: Ka = [H+]m[B-]m/[HB]m (1)
Detailed evaluation of the retention characteristics of
the second system peak is thought to be impossible unless The ion-exchange equilibrium is represented as:
we know the functional relationships between the
retention volume and such chromatographic parameters X-+B-RIB-+X-R: Ke=[X-]S[B-]m/[X-]m[B-]5,
as the acid dissociation constant of the eluent, as well as
the eluent concentration, pH or acid/salt ratio in an (2)
eluent, acid-partition coefficient, and so on.
Although Yamamoto et al.6 tried to obtain such a where X- is the analyte ion and R is the skeleton of ion-
functional relationship, they considered the capacity exchanger. The suffixes, m and s represent mobile and
ratio of the 2nd system peak only for acid eluents. stationary phases, respectively.
Because the appearance of the 2nd system peak is not The acid partition equilibrium can be represented as:
limited to acid eluents, the equation is not utilized in
general. So, we have derived a general relationship (HB)m!(HB)S: Kd=[HB]S/[HB]m (3)
consistent with the results obtained by the computer
simulations) for monobasic acid eluent systems (acids, Another reaction equilibrium is the auto-protolysis of
salts, and their mixtures), and have confirmed the solvent water.
relation experimentally.
H2O~H++OH-: KW=[H+][OH-] (4)

t To whom correspondence should be addressed . Mass balances in a theoretical plate can be represented
382
as:
Ce= [B-]S+ [X-]S (5)
Tx = [X ]m+ [X ]s (6)
CB= [B-]m+ [HB]m (7)
CA= [A+]m= [B-]m+ [X-]m+ [OH-]m - [H+]m (8)
where Ce is the ion-exchange capacity of the stationary
phase. The phase ratio was assumed to be 1.0 for the
simplicity in the computer simulation. Values of Tx,
CB,and CAin ith plate at jth transfer of the mobile phase
are given from the initial conditions and the previous
processes (transfer 1-* j-1).
It was assumed in the present theoretical treatment and
computer simulation based on the plate theory (denoted
below by "PT calculation") that the above 8 equations
hold for every theoretical plate. Other than these
simplifications, we tried to treat the systems as rigorously
as possible.
The computer program for the simulation of mono-
basic acid eluent systems was written in MS-FORTRAN"
and run on an IBM personal computer (PS/V,
80486DX). The scheme of the programs was shown in
the previous paper.' The theoretical plate number was
set at 100 for almost all calculations for the convenience
of computer time. Source program lists are available on
request.
Experimental
Apparatus
The chromatographic system consisted of a Tosoh
(Tokyo, Japan) CCPM pump, a Rheodyne (Cotati, CA,
USA) 8125 sample injector with a 100 µl sample loop, a
Tosoh CM-8000 conductivity detector, and a Tosoh UV-
8000 absorbance detector. The separation column was
a Tosoh IC-anion PW (4.6X50 mm).
Reagents
All chemicals were of analytical grade and were
purchased from Wako (Osaka, Japan) or Tokyo Kasei
(Tokyo, Japan), and were used without further purifica-
tion. Water was purified by using a mixed-bed ion-
exchange column and a Milli-Q Labo water purification
system.
Chromatographic procedure
Water solutions of salicylic acid, sodium salicylate,
and salicylic acid+sodium salicylate were flowed with a
pump at 1 ml/min. The acidity of the eluent was
changed with the mixing ratio of the acid and the salt.
As the sample, water or an aqueous solution of sodium
chloride was injected. All experiments were performed
at about 22° C. Output from a detector was stored in the
disk memory of a personal computer (Fujitsu, FM-R50)
ANALYTICAL SCIENCES JUNE 1996, VOL. 12
and processed with homemade software written in
BASIC.
Measurement of partition isotherm
The quantities of acids adsorbed by a separation
column were measured for the salicylic acid-sodium
salicylate system by using the method described in a
previous paper.'
Results and Discussion
Derivation of thefunctional relationship
The second system peak is thought to be caused by the
perturbation of the adsorption equilibrium of the eluent
acid in the analytical column after the sample intro-
duction."6 Although this case is a sort of vacancy
chromatography', the retention volume of the second
system peak ( V2)is thought to be given by the well-known
relations for usual partition chromatography as,
V2- Vm+ Kd' VS (9)
where, Vmand VSrepresent the volumes of the mobile and
stationary phases, respectively, in a column. In Fig. 1,
some results by computer simulation are plotted. Each
group of the same Ka value and acidity shows a linear
relation, but a different slope. Therefore, Eq. (9) is
invalid for the present case.
When the partition isotherm is not linear at a certain
Fig. 1 Consistency between the results of PT calculations
(symbols) and Eq. (15) (lines). Parameters: CB=0.005, Vm=
V=0.5, Kex=1.0*, Ce=0.01, acid sample. 0, Ka=5.0X10-4
(acid eluent); ~, Ka 5.OX10-4 (salt eluent); Q, Ka 5.OX10-3
(acid eluent); (, Ka 5.OX10-3 (salt eluent); ~, Ka5.OX 10-2
(acid eluent); /, Ka5.OX 10-2 (salt eluent). * Selectivity
coefficient of ion-exchange reaction between B- and a sample
ion.
ANALYTICAL SCIENCES JUNE 1996, VOL. 12 383

temperature, the partition coefficient (Kd) is given9 as

Kda[HB]S
a[HB ] (10)
m

where, [HB]m and [HB]S are the concentrations of HB in

the mobile and stationary phases, respectively.
Because the acid molecule in the eluent dissociates
partly (weak electrolyte) or perfectly (strong electrolyte),
the retention behavior of the 2nd system peak becomes
complicated. The sample injection causes the concen-
tration change of the total concentration of the eluent ion
(CB). Therefore the coefficient to VSin Eq. (9) was
presumed to be given by the partial differential co-
efficient, a[HB]S/a CB.6'8 Thus,

8[HB]S
V2= Vm+ C
B • VS (11) a
The differential coefficient in Eq. (11) can be represented
as: Fig. 2 Acid-partition isotherm for the salicylic acid and
sodium salicylate system. 0, CB=0.8 mM (constant), CA!
a[HB]S a[HB]S a[HB]m CB=variable; S, CA/ CB=0.8 (constant), CB=variable.
aCB a[HB]m aCB

a[HB]m
= were drawn by using Eq. (15). If [H+]mois equal to
aCB •Kd (12)
[B-]moas in the case of acid eluent , Eq. (15) becomes
The differential coefficient to Kd in Eq. (12) may be
Kd
considered as a sort of correction term for Kd. V2 = Vm + VS (16)
From the basic relations, Eqs. (1), (7) and (8), we Ka
1+
obtain the next relation between [HB]m and CB, 2[H+]m0
neglecting [X-] and [OW] terms as small.
This is the same relation as that given in the report by
[HB]m2-[HB]m(2CB- CA +Ka)+ CB2- CACB=O Yamamoto et al.6

(13) Experimental confirmation

The general equation (Eq. (15)) for the retention
The partial differential coefficient in Eq. (12) can be volume of the 2nd system peak was confirmed
obtained by the differentiation of Eq. (13), and by the experimentally for the salicylic acid-salicylate eluent
substitution of the mass conservation equation and the system. The partition isotherm for this eluent system
electric neutrality rule. was first measured independently from the chromatog-
raphy.) Because the isotherm (Fig. 2) was not linear,
8[HB]m [H+]mo+ [Br]mo the measured values for [HB]S and [HB]m were adjusted
(14) to Freundlich's equation, [HB]S=k[HB]ml/n,by the least-
a CB [H+]mo+ [B-]mo+ Ka
squares method. Although Yamamoto et a1.10 used
where, [H+]moand [B-]moare the concentrations of H+and Langmuir's equation to the similar system, our measured
B-, respectively, in the eluent. data fitted better to the former equation than to the latter
Thus, we obtain the next general relation for the one in the present work. By using the k and n values
retention volume of the 2nd system peak. thus obtained (1.43 and 3.97, respectively), Kd values
were calculated for any values of [HB]m with Eq. (10).
The retention volume of the 2nd system peak could be
Kd
V2 = Vm + • VS
(15) estimated by using Eq. (15) and is shown by the solid line
Ka
in Fig. 3. The values of Vm and VSin Eq. (15) were
1+
[H+]m0+ [B-]mo obtained from the retention volume of the first system
peak (water dip) and the column size. Experimental
Equation (15) is consistent with all the simulation values (marks) well fit to the theoretical relation, even
results (PT calculation) for monobasic acid eluent though the partition isotherm is not linear. Thus,
systems (HB+AB) as shown in Fig. 1, where solid lines general relationships between the retention volume of the
384 ANALYTICAL SCIENCES JUNE 1996, VOL. 12

these values by measuring V2 in chromatographic ex-

periments.
The intensity of the 2nd system peak depends on the
amounts of the concentration change of eluent species at
the sample injection and on the method of detection.
This point will be presented separately.

References

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Fig. 3 Experimental cvalues (0) and the theoretical relation K. Hayakawa and M. Miyazaki, J. Chromatogr.,482,
(line) for the retention volume of the 2nd system peak in the 145(1989).
salicylic acid and sodium salicylate system. 7. C. N. Reilley, G. P. Hildebrand and J. W. Ashley Jr.,
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2nd system peak and chromatographic parameters have
10. A. Yamamoto, A. Matsunaga, E. Mizukami,
been obtained for monobasic acid eluent systems. Now K. Hayakawa and M. Miyazaki, J. Chromatogr.,644,
we can estimate V2 for any monovalent eluent systems 183(1993).
easily without performing PT calculations if we know Kd 11. Product of Microsoft Corporation.
and Ka. Although such basic data as the values of Kd are
not yet known sufficiently today, we can easily obtain (ReceivedJanuary 8, 1996)
(AcceptedFebruary 26, 1996)