simple

1. Presented by
Liji jesu

3. INTRODUCTION
 ION EXCHANGE CHROMATOGRAPHY
is a process by which mixture of similar charge ions
can be separated by using ion exchange resins.

4. principle
 The reversible exchange of ions in solution
with ions electro statically bound to inert
support medium.

5. Factors in ion exchange
 Electrostatic force of attraction
 Relative charge
 Radius of hydrated ions
 Degree of non bounding interactions

6. Ion exchangers
• It is carried out in columns packed with ion exchanger. they
are of two types
• Cation exchanger; covalently bound to -ve
charged ions.
Anion exchanger ; covalently bound to +ve charge
Counter ions: ions bound electro statically.
The exchanger pprd form of fully charged.
E-y + + x+ = E- x+ +y+ C E

8. ELUTING X+
Increasing conc.
Of y+
Increasing ph of
solvent
ie converting
x+ to uncharged
sps
Y+ conc to elute
x+ depend on
quantity of x+
charge

9. Macromolecules
Ptns and nucleic acid which can posses +ve
And –ve charge
It can bind ,anion and cation exchangers
Make them +ve by increasing ph
make –ve by reducing ph

10. Types of ion exchange
resins
 Poly styerne
 Cellulose
Properties of resins
 Ion accessibility
 Chemical stability
 Mechanical stability

11. type nature
strong cation Salphonated polystyrene
weak cation Sulphoproponyl cellulose
Condensed acrylic acid
Carboxy methylcellulose
Strong cation Dimethyl
Quadatory amino cellulose
weak anion Dithylaminoethyl cellulose
Dithylaminoethyl agarose

12. Pptn of exchange medium
 Swelling of medium
 Removal of very small particles
 Exchanger lies to equilibrium

13. PPTN OF EXC DETAIL
SWELLING: ALSO known as precycling
Burial of charge func.groups.
Charging with acid then wt base for ce reverse to
ae
Chealator EDTA -REMOVE METAL IONS
REMOVAL OF VERY SML PARTICLES : fines to
be remove by washing when suspended in water
polymers get settle down
 EQUILIBRIATED WITH COUNTER IONS: This by
washing resins with different reagent depending
upon desired counter ions.

14. Use of buffer
 To maintain ph of column.
 Anionic IEC :- cationic buffer
 Cationic IEC:-anionic buffer
 pk of buffer should be near as possible as which
the system is buffered.This result in high
capacity which with stand the local changers ph
column sample .

15. BUFFER PH range
Ammonium acetate 4-6
Ammonium formate
3-5
Pyridium acetate 4-6
ammonium carbonate 8-10

16. procedure
 Ptn mixture transferred into low ionic
strength .(mb)
 absorbent is packed into a column the
column is pre equilibrated with the buffer of
identical ph and similar ionic strength as
protein mixture (prefr the same buffer as ptn
mixture)
 Ptn mixture is applied into column .ptns
charged oppositely to IE media are
temporarily retained in column .

17.  All other ptns simply pass through the column
and are collected during this step..
 Retained ptns are eluted from the column by
applying a modified buffer ,elution is most
commonly achieved by gradually increasing
their ionic strength of buffer via salt gradient
,and ptn are eluted in order of increasing their
net charges, is specific cases the elution can be
accomplished.

18.  a)A ph change
 B)Affinity methods

20. APPLICATIONS
OF IE CHROMATOGRAPHY
 Use for amino acid analysis
 To determine the composition of nucleic acid.
 Uses for water purification
 Separations of vitamins ,biological amines
and bases
 Separation of ultra pure metals

22.  Thank you