Ion exchange chromatography is a process that separates ions based on their charge. It works by reversibly exchanging ions in solution with ions bound to an inert resin. There are two main types of ion exchange resins: cation exchangers that bind positively charged ions and anion exchangers that bind negatively charged ions. Samples containing mixtures of ions are passed through a column packed with the resin, and ions of opposite charge bind to the resin while others pass through. The bound ions can then be selectively eluted by altering conditions like increasing salt concentration or pH. Ion exchange chromatography has applications in separating amino acids, nucleic acids, metals, vitamins and other biological molecules.
9. 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
10. 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
11. Types of ion exchange resins
• Poly styerne
• Cellulose
Properties of resins
• Ion accessibility
• Chemical stability
• Mechanical stability
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.
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