QUANTITATIVE DETERMINATION OF

TOTAL ION CONCENTRATION BY ION

EXCHANGE CHROMATOGRAPHY
J. OLIVEROS1 and M. N. SALES1
1

INSTITUE OF BIOLOGY, COLLEGE OF SCIENCE

UNIVERSITY OF THE PHILIPPINES, DILIMAN, QUEZON CITY 1101, PHILIPPINES
DATE SUBMITTED: 6 MAY 2015
DATE PERFORMED: 29 APRIL 2015

ABSTRAC
T
The purpose of the study was to discuss the principles behind ion-exchange
chromatography and its use as a technique for separation. The study also asserts
the importance of the ion exchange chromatography, including its concept of
isocracy and electroneutrality, resin polymerization and sulfonation, stoichiometric
ratio, hydration and particulates, in chemical analysis and determination of total
ion concentration of Cu 2+ from the sample. This report discusses basic mechanisms
and materials of specifically cation exchange columnar chromatography. Dowex 50
cation exchange resin was the stationary phase and the solvent containing the
analyte ion was the mobile phase. Ion chromatography conditions were kept
paramount during the duration of the experiment. 10 mL of the analyte sample was
then eluted to the preconditioned column for three trials. Per trial, the eluate was
allowed to flow to a fresh Erlenmeyer flask until its pH = 6. This fresh pool was then
titrated with previously standardized sodium hydroxide NaOH against a primary
standard KHP. Cu 2+ concentration in ppm was then calculated using the titrimetric
data. The theoretical concentration was 2500 ppm Cu2+, the calculated value was
671 ppm Cu2+ of 73.16% error, yielding a % purity of 26.48% Cu2+.
INTRODUCTI
ON
Before, the only efcient and feasible
method known in the purifcation of
rare earth ions involved tens or hundreds
of tedious fractional precipitation, which
later on was revealed to still be relatively
impure. A method therefore, that can
separate one trace compound from another,
even in the presence of a large excess
adsorbable cation, is needed [1].
Ion exchange is probably the most
frequently used chromatographic technique
for the separation and purifcation of
proteins,
polypeptides,
nucleic
acids,
polynucleotides,
and
other
charged
biomolecules. The reasons for the success

of ion exchange are its widespread

applicability, its high resolving power,

its high capacity, and the simplicity and

controllability of the method [2].
Ion exchange refers to the separation
techniques in analytical chemistry that
allow diferent ionic materials to be
selectively retained on an ion exchange
resin.

Separation
in
ion
exchange
chromatography
depends
upon
the
reversible adsorption of charged solute
molecules to immobilized ion exchange
groups of opposite charge [2].
Separation is obtained since different
substances have diferent degrees of
interaction with the ion exchanger due to
diferences
in
their
charges, charge
densities and distribution of charge on their
surfaces.
These interactions
can be
controlled by varying conditions such as
ionic strength and pH. The diferences in
charged properties of biological compounds
are often considerable, and since ion
exchange chromatography is capable of
separating species
with
very
minor
diferences in properties, e.g. two proteins
difering by only one charged amino acid,
it is a very powerful separation technique
[2].
The classical titrimetric and gravimetric
methods have long been solely tantamount
to analytical chemistry until the early
Twentieth
century. Chemical analysis is
such a trending pursuit that analytical
methodologies
have
been
advancing
[1]
exponentially.
Hence,
instrumental
methods for analysis were developed. This
report
features
ion
exchange
chromatography as a chromatographic
instrumental
method
for
analytical
chemistry and as a separation science.
Elution chromatography is a columnar
chromatography
technique
involving
washing the solute through a stationary
phase by quantitative additions of the
mobile phase.
Table 1. List of Ion Exchange
Materials

Chromatography,
as
an
instrumental
method, separates, identifies, determines
and quantifies the chemical components of
a
mixture. Methodologically, it can be
planar using porous paper support or
columnar using narrow tube support. It is
composed of a stationary phase that is
fxed in the support and the mobile
phase that moves through the stationary
phase.
Table 1 provides diferent types of ion
exchange
materials
used
in
ion
chromatography. As shown, Dowex 50,
the resin used in the experiment, is a
type of a strong acid cation exchanger.
A cation resin has a stationary, functional
group with a negative charge. This means
that it will exchange positive ions with the
eluent solution.
In ion exchange chromatography one can
choose whether to bind the substances
of interest and allow the contaminants to
pass through the column, or to bind the
contaminants and allow the substance
of interest to pass through.
In general, chromatography can be planar;
a porous paper support is used, or
columnar; a narrow tube support is utilized.
Also, it has a stationary phase and a
mobile phase that are both essential for
the set-up. For the purpose of the
experiment, elution chromatography, a type
of columnar chromatography that involves
washing the solute through the stationary
phase by adding the mobile phase, was
used to achieve maximum output.

METHODOLO
GY
The experiment aimed to determine the
concentration in ppm of Cu2+ in a given
sample using the Ion chromatography as a
separation technique. In the experiment,
Dowex 50 cation exchange resin was the
stationary
phase and
the solvent
containing the analyte ion was the mobile
phase. A column previously inserted with a
wad of absorbent cotton for resin support
and packed with sulfonated resin was kept
hydrated with distilled water washings in
an improvised Ion chromatography burette

tube. A flow rate of 30 drops per minute

was maintained by precise manipulation of
the stopcock. The mobile phase was allowed
to flow until the pH of the eluate is equal to
the pH of distilled water, which was equal
to 6. 10 mL of the analyte sample was
then eluted to the preconditioned column

for three trials. Per trial, the eluate was

allowed to flow to a fresh Erlenmeyer flask
until its pH = 6. This fresh pool was then
titrated with previously standardized sodium
hydroxide NaOH against a primary standard
KHP. Cu2+ concentration in ppm was then
calculated using the titrimetric data.

The copper cation was separated from the

sample at par with the displacement of
hydrogen
protons from the column.
Separation is made possible by the
adsorption of charged analyte molecules to
immobilized resin ion exchange function
group of opposite charge. The displaced
ions
from
the column exist in a
stoichiometric ratio or factor with the
analyte ion. Hence, the displaced ion will be
titrated
and
the
calculated
amount
multiplied by the said propagated ratio or
factor will be the amount of analyte
constituents.
There is an exchange of
ions,
hence the name, ion exchange
chromatography.

thrice of an amount of H+ is equal to one

amount of Aluminum 3+. All of which can be
explained by the interplay of ionic charges
during the stoichiometric reaction.

RESULTS
DISCUSSION

AND

The set-up was concerned of separation of

various types of charged species in a
system for it is an ion exchange
chromatography, thus the system is
comprised of separate phases. A principal
stoichiometric reaction was then applied at
all analyses
to
maintain
the
ionic
integrity of the system.
To attest to the importance of
this
stoichiometric ionic reaction, look intently to
these chemical equations
I.
nrSO3 H+ + Mn+ (rSO3)nM
+
+ nH II.
2rSO3 H+ + Cu2+
(rSO3)2Cu + 2H+ III.
2H+ +
2OH- 2H2O
Equation (I) shows the general chemical
reaction in cationic exchange. nrSO3 H+
refers to the sulfonated cation exchanger
resin, Mn+refers to the metal / cation
analyte species, and H+ refers to the
displaced proton ion. Also, n refers to the
stoichiometric factor. [3]

Now grounded to the notion that the

calculations
are
dependent
on
that
stoichiometric ratio, this working equation
for determination of the amount of Cu2+
concentration in a given sample is derived:
(IV) Ppm Cu2+
=
M V

ppm Cu =

(1 mol H

/ 1 mol OH

) (1 mol Cu

2+

/ 2 mol H

OH OH

Volume of sample, L

) (63.55g / mol ) (1000mg / 1g )

pt

Equation (II) is the particular reaction in the

experiment. Dwelling on the stoichiometry
of the equation, for every 1 mole of copper
II species, 2 moles of the cation exchanger
reacts, therefore, 2 moles
of
H+ are
displaced. These displaced H+ are then
titrated with any strong base, NaOH for
example, and both species react with each
other in one-to-one correspondence. [3]
Therefore, twice the amount of OH- and
twice the amount of H+ is equal to one
amount of Cu 2+. Also,

The
calculated
ppm
from
the
experiment was
(mean);
671.4236668
ppm.
The
theoretical was
2500ppm. It can be observed that even
though there was a small deviation
between each trial for the ppm, there is
still a large diference between the
calculated values and the theoretical, this
can be explained by the errors and
inaccuracies of the techniques used during
the experiment
Principal Conditions: Before conducting
the Ion Chromatography, a
set of
parameters must be paramount and
maintained throughout the duration of the
experiment. That is why a pH of 6 (the pH
of distilled water must be observed. A pH of
6 means that the system is isocratic (not
Resins: The resin used was Dowex 50.
Cation exchanger contain sulfonic acid
groups nSO3 H+ attached / chelated to the
aromatic ring of insoluble inorganic
molecule nR.[5] Gel-based resins are
manufactured/formed by polymerizing with
usually
styrene-divinylbenzene
S-DVB
copolymer. The
cross linkages with the copolymer give the
resin physical strength to withstand
subsequent ion reactions. The spherical
shape narrows particle size allotment. The
polymerization yields inactive resins
and
must
be
activated. [5]
Sulfonation.:
Inactive
resins
have
hydrophobic
spheres
and
must
be
introduced to sulfonic acid nrSO3
to
expose the functional hydrophilic sites for
exchange. As the resins react with the
acid, water forms and swells the beads
making it a suitable medium for reaction.
Other acids like carboxylic acid and other
bases like quaternary ammonium can
functionalize resins. [5]
Notice that a
written with H+
H+). This shows
and the H+

resin functional group is

written posteriorly (nrSO3
that the resin is activated
are now ready to be

comparable)
and therefore is highly
deprotonated from H+ after the cation
exchanger has fully reacted with the cation
analyte (all H+ have been displaced and are
now ready for titration with strong base). [4]
Electro-neutrality:
The
stationary
adsorbent phase, the column of resins (site
of ion exchange), must be solvated with
deionized and distilled water. [4]
Flow rate should be 1.5 mL or 30 drops
per minute: This is because flow rate is
crucial for the ion exchange to take place.
Water is the medium for reaction. Rapid
flow or imprecise flow will never assure
the optimum completion of the reaction and
the total liberation of H+. A modest rate of
flow neutralizes the turbulence and high
fluid pressure at the narrow end of the tube.
[4]
cation exchanger than H+, therefore, H+
is eluted frst.
Table 1. Standardization of NaOH solution
Trial

10
Standard
weight,
g

.1008
1002

.1270

3.85
5.2
Net
3.9
volume
NaOH,
mL
exchanged with another cation species.
The resins
are
also
perforated
with
micropores for liquid pathways to increase
surface area of adsorption. These pores are
generated by poragens like toluene.
Water influence: The resin must be
hydrated at all times. Water level should not
fall below the resin level. Aside
from
maintaining the isocracy of the system,
water solvates the system and serves as a
suitable medium for reaction to take place. It
also displaces entrapped gases. [5] The air
pockets/bubbles
cause
pulsations/disturbances in the flow and may
react with the resins forming altered canals.

These canals cause apparent loss in column

capacity and adsorptive power.[4]
Liberation of H+: Since it was stated that
ion exchange explicitly depends on the
ionic interactions, therefore, the ionic
interactions are functions
of
ionic
strength. Separation is possible due to the
diferences in ionic strength. Because
transition metals like Cu2+ have greater
ionic affnity with the sulfonic acidic
functional group of the

M NaOH
.1256
Average M NaOH
1242

.1279

.1194
.

Titration: As mentioned, the liberated H+

are titrated with a strong base, NaOH to
determine the concentration of Cu 2+. Table
1 shows the standardization data of titrant
NaOH of all the groups.
It is grounded on the stoichiometric
ratio that 1
mole Cu2+ = 2 moles H+.
Therefore,
using the working equation (IV), the
concentration of Cu 2+ in ppm
was
determined per sample. See appendix
table 2 for detailed titration data of all
trials.
Disadvantages of IC: Like any other
instrumental method and as a still growing
chromatographic science, it is assailed
with a
few
limitations.
One notable
disadvantage is that it is costly. The class
improvised
a
burette
type
Ion
Chromatography tube. Amidst the greater
accuracy than that of classical methods,
like any other instrumental titration, it has
many demands and conditions to be
observed.
Errors and Inaccuracies: The setup
may
encounter
chromatographic
anomalies. An example is water dip,
otherwise known as carbonate water dip.
[5] There are also minor shifts in retention
and calibration curves. Temperature must
also be kept constant since minor
fluctuations can alter the ionic kinetic
energy. There are times that to maintain
the isocracy, a buffer is needed. However,
bufer may react with the resin or even the
solvent containing the analyte. The method
provides no direct information on events
occurring at the surface of the stationary
phase,
because
the
ion-exchange
equilibrium is always determined by the
balance

between
the
H+
and
analyte ion
interaction with active sites of a resin.
Impurities may also afect the data. These
flaws may have lead to statistical deviations

SUMMARY
CONCLUSIONS

AND

The results show that the unknown sample

has an average cation concentration of
671 ppm or
0.01056 M. The theoretical value for the
actual concentration was 2500 ppm. A large
deviation can be observed when comparing
the two values. To compare this to the
actual cation concentration in the sample,
the value obtained from the experiment has
a percent error of 73.16%. This error
could have been minimized if
the
experiment was done more carefully and
with more meticulous methods while doing
the techniques. Hence, the values obtained
could have been more accurate and
precise.

All of these factors plus additional ones

which were discussed before are main
elements that greatly afect the results
calculated from the experiment, thus errors
couldve been committed, may it be by
method,
instrument
or
even
by
indeterminate systems. All of these can be
accounted and should be accounted during
the experiment.

However, even if the feasibility of Ion

Exchange Chromatography can be affrmed
basing from the experiment results, there
were still errors that greatly afected the
veracity of this type of technique in
analytical separation. Although, it can be
concluded that the total ion concentration
can be accurately determined via Ion
Chromatography, and that the mechanism
of
isocracy
of
ionic
charges
and
stoichiometric ratio are of vital importance
for calculations involving the said technique.
The highly accurate value affirms the
feasibility
of
Ion
exchange
chromatography
as
a
means
of
determining total ion concentration.

[2] Wurts, William. "WATER HARDNESS

There are a lot of factors that can afect

the ion- exchange. These factors include
the surface area of the stationary phase
(resin bead size), density of exchange sites
on the stationary phase surface (crosslinkage), flow rate of the mobile phase (resin
bead size and column geometry; system
pressure in high-pressure chromatography,
and the chemistry of the mobile phase
(ionic strength of the sample solution,
concentration of mobile phase) [6].

REFERENC
ES
[1] Fischer, R., Peters, D. 1968. Basic
Theory and Practice of
quantitative chemical analysis.

-- CALCIUM & MAGNESIUM."

WATER HARDNESS -CALCIUM
& MAGNESIUM. Kentucky State
University CEP at UK Research and
Education Center, 1 Jan. 2008. Web.
1
Apr. 2015.
[3] Dyer A., Hudson MJ.,et al. 1993. Ion
Exchange Processes: Advances
and Applications. The Royal
Society of Chemistry.
[4] Skoog D., West D., et al. 1996.
Fundamentals of Analytical
Chemistry, 7th Edition. Saunders
College Publishing
[5] Walton, Harold. 1976. Ion Exchange
Chromatography. Dowen,
Hutchinson & Ross, Inc.
[6] Smith, Frank. 1983. The Practice of
Ion Chromatography. WileyInterscience Publications

APPENDIX
Table 1. The water hardness
scale. Water Hardness
ppm CaCO3
Soft

0 20

Moderately Soft

20 60

Moderately Hard

61 120

Hard

121 180

Very Hard

> 180

Table 2. Standardization of EDTA solution

Trial

Working
Ca(II)
standar
Net Vol of
EDTA
M of EDTA

10
mL

10
mL

10
mL

8
mL

8.5
mL

8.3
mL

9.878915223x10 M

9.297802562x10 M

9.521845998x10 M

Table 3. Hidden Spring Water Sample Analysis

Trial
Volume of water sample, mL
Net Volume of EDTA, mL
Total Hardness, ppm CaCO3
Averag
e
RSD
Confidence Interval

1
50
13
248.
7

2
50
13
248.
7
248.
7
0
ppt
24
8.7 +/- 0

3
50
13
248.
7

Calculations
Standardization:

(.1008 / 204.2g/mol x .998 )

= .1279601104
( 3.85mL/1000mL)
M
(.1270/ 204.2g/mol x .998 )
M of NaOH =
( 4.2mL/1000mL) = .1193644994
M
(.1002 / 204.2g/mol x .998 )
M of NaOH =
( 3.9mL/1000mL) = .125567938
M
.1279601104M + .1193644994 M+ .125567938 M
Average M of NaOH =
= .1242974345 M
3
M of NaOH =

ppm Cu2+:

.1242974345 M x 1.6 mL / 1000 x 1 x 1/2 x 63.55 x 1000

= 631.928157 ppm
( 10 mL /1000 mL)
ppm of Cu
.1242974345 M x 1.7 mL / 1000 x 1 x 1/2 x 63.55 x 1000
=
= 671.4236668 ppm
2+
( 10 mL /1000 mL)
ppm of Cu trial 2
.1242974345 M x 1.7 mL / 1000 x 1 x 1/2 x 63.55 x 1000
=
= 671.4236668 ppm
2+
(
10
mL
/1000
mL)
ppm of Cu
2+
trial 1

trial 3

2+

Average ppm of Cu trial 1. 2. 3 =

631.928157 + 671.4236668 + 671.4236668

= 658.2584969 ppm
3

New Mean: 671.4236668 ppm

i =1

SD =

X X

n 1

(671.4236668

671.4236668

) + (671.4236668
2

671.4236668

2 1

=0
RSD =

sd
X

Confidence Limit = X

x1000 ppt

0
x1000 ppt
671.4236668
=0
=

% purity =

tSD
n

= 671.4236668

(12.70) (0)
3

= 671.4236668 0

1 gram/liter [g/L] = 1000.000002 part/million [ppm]

671.4236668
g/L=
= .67142366655
1000.000002
.67142366655
M of Cu 2+ =
= .01056528191 M
63.55
.01056528191 M x 10/1000
% purity =
.1242974345 M x 1.6 mL x 1 x 1/2 x
2+
ppm of Cu trial 1
63.55
=
= 6319.28157 ppm
( 10 mL /1000
mL)
2+
ppm of Cu trial 2
.1242974345 M x 1.7 mL x 1 x 1/2 x 63.55
=
= 6714.236668 ppm
( 10 mL /1000
2+
ppm of Cu trial 3
mL)
.1242974345 M x 1.7 mL x 1 x 1/2 x
63.55
=
= 6714.236668 ppm
( 10 mL /1000
mL)
Average ppm of Cu

2+

6319.28157 + 6714.236668 + 6714.236668

= 6582.584969 ppm
3

trial 1. 2. 3

New mean: 6714.236668 ppm

n
2

(X
X)

SD =

i =1

n
1

(6714.236668
=

6714.236668

) + (6714.236668
2

6714.236668

2
1

=0

Confidence Limit = X

tSD
n

= 6714.236668

RSD =

(12.70) (0)
3

sd
X

x1000 ppt

0
x1000 ppt
6714.236668

= 6714.236668 0

=0