A. Arma, J. A. Ines/Chemistry 26.

1 (2018) P a g e |1

Quantitative Determination of Total Ion Concentration by Ion-Exchange

Chromatography
A. Arma1, J.A. Ines2
1 College of Education, University of the Philippines, Diliman, Quezon City, 1101
2 National Institute of Geological Sciences, College of Science, University of the Philippines, Diliman, Quezon City, 1101

Performed: April 18, 2018 Submitted: April 25, 2018

ABSTRACT

Ion-exchange chromatography is a technique that separates ions based on their charge difference/affinity
with respect to the ion exchanger. It is widely used as a separation and purification technique for
biomolecules such proteins and deoxyribonucleic acid (DNA). This experiment aims to apply the
principles of ion-exchange chromatography in determining the unknown concentration of a Cu2+ solution.
In this experiment, Dowex 50 resin, a strong acid cation exchanger was used as the stationary phase while
the mobile phases were the concentrated HCl solution and Cu2+ solution. A burette was used as a
substitute for the ion exchange column. The eluate was obtained and was titrated with 0.0945 M NaOH
and phenolphthalein as indicator. It was determined from three trials that the unknown Cu 2+ solution has
an average concentration of 2452.24 ± 86.08 ppm (95% confidence) with a relative standard deviation of
14.14 ppt, indicating that the measurements are precise.

Introduction their affinity to the ion exchanger. Ion

Chromatography is a separation technique that
operates based on the differential partitioning of
mixture components between a mobile and a
stationary phase. The mobile phase travels
through the stationary phase in a defined
direction. The distribution of components
between the two phases depends on adsorption,
ionic interactions, diffusion, solubility, or
specific interactions. Depending on the
experimental design, the separation in a liquid
mobile phase may be carried out via column or
planar chromatography, on analytical or
preparative scales [1].
Since the introduction of this technique by M.
Tswett in 1906, chromatography has undergone
tremendous modifications. Nowadays, various
types of chromatography are in use to separate
almost any mixture, whether colored or
colorless, into its constituents [2].
One of its modifications is the ion-exchange
chromatography. This technique is used in
water purification, water analysis and quality
control. In fact, the phenomenon of ion exchange
was first reported by two British agricultural
chemists in 1850 who proved that soil can
remove potassium or ammonium salts from
water with release of an equivalent amount of
the calcium salt [2].
exchangers composed the stationary phase.
They are insoluble solid which carry
exchangeable cations or anions. They are
capable of reversibly removing ions from a
solution while at the same time replacing them
with ions of equivalent charge [3]. They are
divided into two: cation and anion exchangers.
Cation exchangers possess negatively charged
groups which will attract positively charged
molecules. They are also called acidic ion
exchange materials since their negative charges
result from the ionization of the acidic group.
Anion exchangers, on the other hand, have
positively charged groups which will attract
negatively charged molecules. They are called
basic ion exchange materials as their positive
charges usually result from the association of
protons with basic groups [4].
In ionic compounds consisting of cationic
species M+ and anionic species B-, cationic ion
exchange chromatography will retain the
cations since the stationary phase has a negative
charged functional group (1) while anions will
be retained by the anionic ion exchange
chromatography using positively charged
functional group (2).
R-X-C+ + M+B- → R-X-M+ + C+ + B- (1)
R-X+A- + M+B- → R-X+B- + M+ + A- (2)

Ion-exchange chromatography involves the The most important class of ion exchangers are
separation of ions and polar molecules based on
 A. Arma, J. A. Ines/Chemistry 26.1 (2018)
the organic ion-exchange resins. Their main
advantages are their high chemical and
mechanical stability, high-ion-exchange
capacity, and high exchange rates (Helfferich,
1995). Cation exchange resins contain acidic
functional groups, usually sulfonic acid groups,
attached to the aromatic ring of the insoluble
organic molecule. Dowex 50 is the most
common example of this. The hydrogen ion in
the ionizable RSO3H+ formed on the resin after
HCl treatment exchanges with other cations (3)
while in an anion exchange resin, the hydroxyl
ions exchanging with anions (4).
N RSO3-H+ + Mn+ → (RSO3)nM + n H+ (3)
N RNH3+OH- + An- → (RNH3)nA + n OH- (4)
Methodology
To prepare the column, a wad of absorbent
cotton was placed at the bottom of a clean ion-
exchange column. It was made sure that the
cotton was not too thick for a more controllable
flow rate of liquid and easier removal of the
resin. A quarter of the ion-exchange column was
then filled up with Dowex 50 cation-exchange
resin by gently pouring the resin as it was
suspended in concentrated HCl. The resins that
adhered to the sides were washed off with
distilled water. Then, the column stopper was
opened to allow some of the liquid to flow out
from the column. The excess HCl was washed
out with distilled water until the pH of the
eluate was equal to the pH of the distilled water
used. The drops of the latest eluate were caught
on a watch glass to test the pH using pH paper.
When the pH of the latest eluate was finally
equal to the pH of the distilled water, the column
was ready to be used.
In the determination of the total cation
concentration, a copper (II) solution with an
unknown concentration was obtained from the
instructor. A 10.00 mL of the solution was
poured into the prepared column with a 250-mL
Erlenmeyer flask under to receive the eluate.
The stopcock was opened until the rate flow was
about 30 drops per minute. The remaining
solution was poured into the column and then,
the column was washed again with distilled
water until the drop of the pH of the latest
eluate was equal to the pH of the distilled water
used. When the desired pH was attained, the
Erlenmeyer flask with the eluate and washings
was removed and titrated with a standard
0.1000 M NaOH solution using phenolphthalein
as indicator. The titration was performed in
triplicate.
P a g e |2
After the titration, to store the resin, 3.0 M HCl
solution was poured into the column until the
yellowish-brown color of the resin reverted
back. To transfer it back to the original
container, the burette was inverted to pour the
resin with acid to the original container. The
process was repeated until the resin has been
transferred.
Results and Discussion
For this experiment, ion-exchange
chromatography was used to determine the
unknown concentration of the Cu2+ solution.
This experiment used a cation ion-exchanger,
specifically Dowex 50 Resin. Cation ion-
exchanger has negatively charged covalently
bonding sites that attracts the cations in the
solution being analyzed [5], which is for this
experiment is the Cu2+.
The resin used for this experiment was soaked
in water hours before being used in the ion-
exchange column/burette in order to remove
the interferences of water soluble impurities
that may block the exchange sites [6]. Soaking
the resin in the water also makes it swell hence,
the surface of the resin being exposed to the
cations increases which also increases the
efficiency of the resin to adsorb cations. The
water level in the burette was always
maintained at a level higher to the resin to
ensure that air bubbles and packets will not
enter and fill the exchange sites of the resin [7].
When air bubbles are present, they block off the
exchange site, decreasing the efficiency of the
resin for ion exchange. This is also the reason
why a dry resin is not an ideal ion-exchanger.
Hydrochloric Acid was also added in the resin in
order to increase the strength of the resin and to
remove the impurities that are still bounded to
it [7]. After the addition of HCl in the resin, it
was assumed that the H+ ions in the resin is
greater than in amount compared to the Cu2+
that will replace it during the sample analysis.
HCl was again used during the storage of the
resin in order to remove the Cu2+ ions that were
bounded to it so that it can be used again for
future experiments. It should be noted that a
higher concentration of HCl compared to the HCl
used in preparing the resin must be used in
order to completely dissociate the Cu2+ ions in
the resin.
After the addition of the HCl to the resin, it was
then washed with distilled water until the pH of
the eluate is equal to the pH of the distilled
water being used. This was done in order to
remove the excess H+ ions that are not bounded
 A. Arma, J. A. Ines/Chemistry 26.1 (2018)
in the resin and to assure that the H+ that would
be titrated in the sample analysis will only come
from the H+ previously attached to the resin
which are replaced by the Cu2+ from the sample
[7]. Not washing the burette with distilled water
and correcting the pH will result to a higher H+
concentration reading since it accounts the
excess H +. Relating H+ and Cu2+
stoichiometrically, the sample analysis will
result to a higher Cu2+ concentration reading
than the true value.
During sample analysis, the burette was also
washed with distilled water and the washing
continued until the pH of the eluate is equal to
the pH of the distilled water being used, similar
to what is done during the preparation of the
column. This was done to ensure that the ion
exchange proceed into completion and all of the
H+ that was replaced by the Cu2+ flowed out of
the burette.
The rate flow of the sample was maintained at a
rate of 30 drops per minute or 1 drop in every
two seconds [8]. The rate was intentionally to be
slow so that there will be an ample time for the
Cu2+ to replace the H+ in the resin. Increasing the
flow may lead to an incomplete exchange
between the Cu2+ and H+, hence the sample
analysis will lead to a lower H+ and ultimately a
lower Cu2+ concentration reading than the true
value.
The exchange of Cu2+ and H+ in the cation-ion
exchanger can be physically observed by the
change of color of the resin from brownish
orange into dark green. This reaction is
explained by the following chemical equation
[5].
2RSO3-H+ + Cu2+ → (RSO3)2Cu + 2H+ (5)
The concentration of the H+ replaced in the ion-
exchanger is stoichiometrically related to the
Cu2+ that replaces it hence, titrating the eluate
will determine the H+ concentration and
ultimately, the Cu2+ solution.
Three trials of sample analysis were performed
with 0.0945 M standardized NaOH and
phenolphthalein as indicator. The NaOH titrant
was standardized with KHP (99.8% purity) (See
Appendix for computations). For each trial, the
combined solution of eluate and washing was
titrated with the standardized titrant. The
titration gives the concentration of replaced H+
and using Equation 5, the Cu2+ concentration
was computed.
P a g e |3
Table 1. Computed Concentration of the Cu2+
Solution, in parts per million.
Trial VNaOH, mL Cu2+ Concentration,
ppm
1 8.1 2432.22
2 8.3 2492.27
3 8.1 2432.22
It was determined from the three trials that the
Cu2+ solution contains an average of 2452.24 ±
86.08 ppm of Cu2+ (95% confidence interval)
with a relative standard deviation of 14.14 ppt,
indicating that the measurement was precise
and successful despite of using a burette instead
of an ion-exchange column. There is a 95%
probability that the true mean of the sample lies
within the range of 2366.16 ppm – 2538.32
ppm.
Possible sources of error included the
inconsistent and fast rate of flow of the eluate.
Fast rate of flow the eluate may lead to an
incomplete ion exchange which will result to a
lower Cu2+ measurement compared to the true
value. This error may be reduced by performing
a calibration trial before proceeding to the three
trials of sample analysis. Another source of error
is when the eluate and washing was titrated
even the pH equality is not yet achieved.
Inequality in pH signifies that there are still
replaced H+ ions inside the burette. Titrating the
said solution will result to a lower H+ and Cu2+
solution since not all of the H+ replaced by the
Cu2+ are accounted in the titration. Lastly, pH
paper reading may also lead to an error since pH
paper only gives us the approximate pH of the
eluate and distilled water. Also, color perception
is subjective per person. This error may be
reduced by assigning one person to observe the
color equality of the pH paper to reduce
personal bias.
Conclusion
Ion-exchange chromatography is the
appropriate method to be employed when
determining the total amount of desired ions in
the sample since the concentration of ions are
directly related to H+ which its concentration
can be determined using the principle of acid-
base titration.
The resulting concentration of Cu2+ from three
trials of the titration is 2452.24 ± 86.08 ppm
with a relative standard deviation 0f 14.14 ppt
which implies that the obtained data points are
close to each other. The experiment is a success
even if a burette was used instead of an ion-
exchange column since the obtained
 A. Arma, J. A. Ines/Chemistry 26.1 (2018) P a g e |4

measurements has a relatively low standard [8] Dudas, F. O. (2011).Analytical Technique for
deviation implying that the data precise and a Studying Environmental and Geologic Samples
narrow confidence interval which implies that
the measurements has a high accuracy.

It is recommended to use a more accurate

device in measuring the pH of the eluate and
distilled water for high accuracy of the
measurements. It is also recommended that
when using a pH paper, only one person should
observe the color change to reduce personal
bias. Ion exchange chromatography can be also
used to determine the concentration of other
types of ions such as the Ca2+ and Mg2+ which
determines the water hardness of a sample.

References

[1] Kovacs, M. & Hegyi, G. (2013).

Chromatographic Methods: Ion Exchange
Chromatography. In Introduction to Practical
Biochemistry (pp. 77-80). Eotvos Lorand
University.

[2] Singh, R. (2002). Ion Exchange

Chromatography and Electrophoretic
Techniques. In Chromatography (pp. 123-126).
New Delhi: Mittal Publications.

[3] Helfferich, F. (1995). Structure and

Properties of Ion Exchangers. In Ion Exchange
(pp. 14-15, 20). New York: Dover Publications,
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https://books.google.com.ph/books?id=F9OQM
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[eBook]

[4] Rathore, K. (2012). An Overview on Ion

Exchange Chromatography. Accessed at
https://www.academia.edu/3690641/Ion_Exch
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[5] Harris, D. (2010) Quantitative Chemical

Analysis 8th ed. Madison Avenue, NY: W.H.
Freeman and Company.

[6] Harvey, D. (2009). Modern Analytical

Chemistry; McGraw-Hill Companies

[7] Khopkar, S. M. (2015). Environmental

Pollution Monitoring and Control; New Age
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identified.