Ion exchange chromatography

Ion exchange chromatography

• IEC , is a subdivision of liquid-solid

chromatography.
• Stationary phase : Resin or gel
Mobile phase : contains the inorganic
salt dissolved in a suitable solvent, is
applied to the column.
 Ion exchange chromatography
• Ion exchange chromatography (or ion
chromatography) is a process that allows
the separation of ions and polar
molecules based on their affinity to
the ion exchanger. It can be used for
almost any kind of charged
molecule including large proteins,
small
nucleotides and amino acids.
The separation occurs by reversible
exchange of ions between the ions present
in the solution ( mobile phase )
and those present in the ion exchanger (
stationary phase)
We can say it is an
adsorption phenomenon .
The adsorption is
electrostatic.
• Principle:
• Ion Exchange chromatography principle,
Exchange of ions is the basic principle in this type
of Chromatography. In this process two types of
exchangers i.e., cationic and anionic exchangers can
be used.
• Cationic exchangers possess negatively charged
group, and these will attract positively charged
cations . These exchangers are also called “Acidic ion
exchange” materials, because their negative charges
result from the ionization of acidic group.

• Anionic exchangers have positively charged groups

that will attract negatively charged anions. These are
also called “Basic ion exchange” materials.
• Applications of Ion Exchange Chromatography:
• It is extremely used in the analysis of amino acids. The
amino acid “Autoanalyzer” is based on in exchange
principle.
• To determine the base composition of nucleic
acids. Chargaff used this technique for established the
equivalence of Adenine and Thymine; Guanine and
Cytosine.
• This is most effective method for water purification.
Complete deionization of water (or) a non-electrolyte
solution is performed by exchanging solute cations for
hydrogen ions and solute anions for hydroxyl ions. This is
usually achieved by method is used for softening of
drinking water.
• Proteins are also successfully separated by this
technique.
• It is also used for the separation of many vitamins, other
biological amines, and organic acids and bases.
One of the main disadvantages of ion exchange
chromatography is its buffer requirement:
because binding to IEX resins is dependent on
electrostatic interactions between proteins of
interest and the stationary phase, IEX columns
must be loaded in low-salt buffers.
For some applications, this restriction may
require a buffer exchange step prior to ion
exchange chromatography.