ION EXCHANGE

CHROMATOGRAPHY

By
Dr.Bhagure G.R.
DNYANASADHANA COLLEGE, THANE
 ION EXCHANGE

CHROMATOGRAPHY

ADSORPTION

Stationary
Liquid
phase
Mobile phase
Ion exchange Resin
 ION EXCHANGE

CHROMATOGRAPHY

Poly Nucleic Poly Charged

Proteins,
Peptides Acids Nucleotides Molecules
 Ion Exchange

Ion exchange is an adsorption phenomenon where the mechanism of

adsorption is electrostatic. Electrostatic forces hold ions to charged functional

groups on the surface of the ion exchange resin. The adsorbed ions replace ions that

are on the resin surface on a 1:1 charge basis. For example:

 + +
H Na
 The first ion exchangers were synthetic resins designed for Applications such as

demineralization, water treatment, and recovery of ions from wastes. The first ion exchangers

designed for use with biological substances were the cellulose ion exchangers developed by

Peterson and Sober.

Ion exchangers based on dextrin (Sephadex), followed by those based on agarose (Sepharose

CL-6B) and cross-linked cellulose (DEAE Sephacel) were the first ion exchange matrices to

combine a spherical form with high porosity, leading to improved flow properties and high

capacities for macromolecules

 THE ION EXCHANGE RESINS

Polymeric resins are made in 3-D networks by cross-linking hydrocarbon chains. The

resulting resin is insoluble, inert and relatively rigid. Ionic functional groups are

attached to this framework.

INTEGRAL PART
Ion Exchange Resin:
CHARACTERISTICS OF THE ION EXCHANGE RESINS

Ion exchange resin should have following characteristics.

It should be insoluble in aqueous medium.

 Should be denser than water
 Should have lose porous polymeric structure.
 It should be inert in nature

 Should have large exchangeable sites

 Should have high degree of cross linking.
 CLASSIFICATON

OF

ION EXCHANGE RESINS

CATION ANION

EXCHANGE EXCHANGE

RESINS RESINS

STRONGLY
WEAKLY STRONGLY
STRONGLY BASIC ANION
ACIDIC CATION BASIC ANION
ACIDIC CATION EXCHANGE RESIN
EXCHANGE EXCHANGE RESIN
EXCHANGE RESIN
RESIN
 STRONGLY

ACIDIC CATION

EXCHANGE RESIN

R SO3H
 WEALKLY

ACIDIC CATION

EXCHANGE RESIN

R COOH
 STRONGLY

BASIC ANION

EXCHANGE RESIN

R
-
OH
+ --
QAS Ex. R-(NCH3)3 Cl

(Integral part) (Labile ion)

 WEAKLY

BASIC ANION

EXCHANGE RESIN

-
R OH

R- Primary amino group

or secondary amino group

The charge on the ion.
 The size on the ion .
 The concentration of the ion

 Use of complexing agent and

ability to form complexes

 Nature of the resin.

1) The charge on the ion:

Attraction of the resin for an ion increases

with increase in the charge carried by the ion.

The preference of the resin can be follow the

order.

Ex. Th4+ >Al3+, >Ca2+>Na+

The size on the ion :

The size of the bare ion is not considered here because ions get hydrated when they are in the

aqueous medium.

The ions having small size get hydrate more and have less affinity with resin and vice versa i.e.

The ions having large size get hydrate less and have more affinity with the resin.

Ex. Li+ <H+ <Na+< NH4+ < K+<Rb+

Less affinity

Small
Resin LargeSi

Size
ze

More affinity
3)The concentration of the ion:

Higher the concentration of the ion (same

ion) in the solution higher is the affinity of

the ion with the resin.

4)Use of complexing agent and ability to form complexes :

The ion having ability to form complexes its size will increases and its

affinity with the resin is decreases and vice versa.

5) Nature of the resin.

Resin having spherical shape, high degree of cross linking, strength of

functional group decides its ability for the exchange of ions.

6) Effect of pH :

The pH can affect on separation by several ways, The ionization of the

resin in solution depends on the pH especially if the functional group is

weakly acidic or basic. The strength of acids or bases, the hydrolysis of the
of resin. The number of mill equivalent ion exchange by one gram of dry

resin is called as ion exchange capacity.

One gram of cation exchange resin is soaked in HCl acid to convert the resin in

H+ form. It is then placed in water. The glass tube like that of burette is

packed with the resin. 10 ml of 0.5N solution of Na2SO4 is placed on the resin

column. The elution is carried out using distilled water. The eluate is collected

in conical flask and then titrated with 0.1N NaoH solution using

phenolphthalein indicator.

Volume of NaoH required for the titration is found out

and cation exchange capacity is

determined by using following formula.

VxN

Cation exchange Capacity: ----------

One gram of anion exchange resin is soaked in HCl acid to convert

the resin in cl- form. It is then placed in water.

The glass tube like that of burette is packed with the resin. 10 ml of

0.5N solution of Na2SO4 is placed on the resin column. The elution

is carried out using distilled water. The eluate is collected in conical

flask and then titrated with 0.1N AgNo3 solution using Potassium

chromate indicator. Volume of AgN03 required for the titration is

found out and anion exchange capacity is determined by using

following formula. VxN

Cation exchange Capacity: ----------

V: - Volume of Burette reading.

EXPERIMENTAL TECHNIQUE

Water level

Ion Exchange Resin

Cotton Plug Or glass wool plug

 Terms involved

Elution:-The process of removing adsorbed

ions is known as elution.

Eluent :-The solution used for elution is called

as eluent .

Eluate.:- The solution resulting from the elution

is called as eluate.
APPLICATION OF ION EXCHANGE CHROMATOGRAPHY.


PREPARATION OF DEMINERLISED WATER OR DEIONISED WATER.

 Separation of Lanthanides.
 Separation of amino acids.

 Determination of concentration of trace constituent.

 Separation of similar ions.

 Separation of interfering cations and anions.

 Water from which all cations and anions are removed is called as

Deminerlised water or deionised water.

The process involves two steps.

a) In the first step water is passed through the cation exchange resin

column (H+ form). While passing through the column all cations such as

Na+, Mg2+ are exchanged for H+ ion .

3- +
R-SO3‑H+ + M+ --------- RSO M

-----------

b) In second step, water coming out from first column is allowed to pass

through an anion exchanger in basic form (OH-) In this anions like

Cl- ,SO42- are exchanged for OH-.

R- OH- + Cl-+ --------- R--Cl- + OH-.

-----------
containing mixture of lanthanides is poured on the top of the hen following

equilibrium takes place,

3+
3R-H + Ln3+ ---------------- R3 ____Ln 3H+

<--------------

In case of lanthanides the ionic radii of tripositive ions decreases with

increase in atomic number i.e. from lanthanum to lutecium.

Hydration of ion increases with decrease in ionic size and therefore hydrated

size of lanthnone ion increases with increase in atomic number.

Lanthnone ion having small hydrated size( La) will strongly adsorb on the

resin and vise versa (Lu). Hence lanthnone ion held by resin decreases from

La to Lu.

When HCL solution is passed as mobile phase separation of lanthanides

takes place in reverse order of atomic number. Lutecium ion will separate out
3) Separation of amino acids:-

Chromatographic column is packed with cation exchange resin. Solution of

metal ions like Cu2+,Cd2+ are exchanged on the column. When amino acid

mixture passed on the column it forms complexes with varying stability. Most stable

complex will elutes first and least stable will elutes at last.

Determination of concentration of trace constituent:-

Trace amount substance present in large volume can be found out.

Ex. If trace amount of Ag +ion is present in large volume of water .In this Column is

packed with Cation exchange resin. Then solution is passed.

Ag +ion exchange with cation. Silver is then eluted by using small amount of eluent.
Ex. Similar ions like Cl-,Br- and I- can be separated using ion exchange

chromatography. Column is packed with anion exchange resin like R-OH- .When

solution containing halides passed over the column these anions will adsorb at

various extent. When elution is carried out using 0.3M NaNo3 Solution Cl-will elutes

out first. When concentration NaNo3 increases to 0.6M After elution Br - ion eluted

similarly I- ion eluted at last using 0.9M NaNo3.

6) Separation of interfering cations and anion:-

Interfering ion can be removed by ion exchange chromatography. Ex.In the

estimation of Ca2+ and Mg2+ion by by sulphate method, Phosphate ion interfere in

the estimation. This Phosphate ion can be removed by passing solution through

cation exchanger, Ca2+ and Mg2+ions get exchanged for H+ ion. While phosphate

ion s pass through exchanger. Ca2+ and Mg2+ions held by resin are eluted and
 Thank You
For Kind Attention