Ion Exchange Chromatography

  Ion Chromatography

The ion chromatography involves the separation of  polar molecules and  ions based on their  charge. This separation technique can be used to purify any kind of charged molecules

The ionic functional groups  in the  stationary phase  interact with the analytes of opposite charge

Ion exchange Chromatography is a Adsorption Chromatography


The earliest report of ion-exchange chromatography date back to 1850, Thompson studied the adsorption of ammonium ions to soils. Spedding and Powell published a series of papers describing practical methods for preparative separation of the rare earths by displacement ionexchange chromatography in 1947

Beginning in the 1950s, Kraus and Nelson reported numerous analytical methods which are used for metal ions based on separation of their chloride, fluoride, nitrate or sulfate complexes by anion chromatography

Type of Ion Chromatography

Based on the charge of analyte to  be exchanged, ion - exchange chromatography can be sub-divided as:

  • Cationic Exchanger Chromatography
  • Anion Exchanger Chromatography

The stationary phase of cation exchange chromatography contains negatively charged functional groups which  retain positively charged  ions, while  stationary phase of anion exchange contains positively charged functional groups and they  retain negatively charged analytes

Ion  exchange chromatography is a valuable method due to its mild separation conditions, low cost, high capacity, versatility, high resolving power, and wide range applicability

Basics of Ion Chromatography



  Phases of Ion Chromatography

The first stage is equilibration in which the ion exchanger is brought to a starting state, in terms of pH and ionic strength, which allows the binding of the desired solute molecules. The exchanger groups are associated at this time with exchangeable counter-ions (usually simple anions or cations, such as chloride or sodium).



The second stage is sample application and adsorption, in which solute molecules carrying the appropriate charge displace counter-ions and bind reversibly to the gel. Unbound substances can be washed out from the exchanger bed using starting buffer





In the third stage, substances are removed from the column by changing to elution conditions unfavourable for ionic bonding of the solute molecules. This normally involves increasing the ionic strength of the eluting buffer or changing its pH




Desorption is achieved by the introduction of an increasing salt concentration gradient and solute molecules are released from the column in the order of their strengths of binding, the most weakly bound substances being eluted first



The fourth and fifth stages are the removal from the column of substances not eluted under the previous experimental conditions and re-equilibration at the starting conditions for the next purification.



Separation Basis


Separation is obtained since different substances have different degrees of interaction with the ion exchanger due to differences in their charges, charge densities and distribution of charge on their surfaces


These interactions can be controlled by varying conditions such as ionic strength and pH



Factors Affecting Chromatographic Separation


Ion Exchange Resin: The swelling factor and cross linking is important for the effective separation. The cross linking should be controlled as its affects the exchanger’s capacity


Swelling helps in proper exposure of charged functional groups for exchange of ions


Sample: The concentration and charge of ions


Buffer: The pH of the buffer should impart the same charge to the sample ions as present in the Column. Anionic Exchange Chromatography should be carried out with cationic buffers and vice versa because buffer ion will indulge in ion exchange, which will be of no use






  Ion Exchangers


Stationary Phase

Polymer-based materials

The best known stationary phases are issued from copolymers of styrene and divinylbenzene, in order to obtain packings hard enough to resist pressure in the column. They are made of spherical particles with diameters of 5 to 15µm that are modified on the surface in order to introduce functional groups with acidic or basic properties

Silica-based materials

Porous silica particles can serve to support, through covalent bonding, alkylphenyl chains carrying sulfonated groups or quaternary ammonium groups

Resin films

A polymer called ‘latex’, prepared from a monomer that contains organic groups, is deposited as an array of tiny beads 0.1–0.2µm in diameter) on an waterproof support to form a continuous film-like layer about 1–2m thickness. The support is made of micro-spheres of silica or glass or polystyrene of about 25µm diameter

Common Exchangers


Preparation of Ion exchangers

Preparation of the exchange medium is essential for satisfactory performance of ion exchange chromatography. Apart from removing impurities, there are three major steps that are absolutely important in ion exchanger preparation.

Swelling: For anion Exchangers it is done by treating it first with an acid (0.5 N HCl) and then with base (0.5 N NaOH). Reverse is true for preparation of cation exchangers.

Removal of fine particles: Large number of such particle can result in decreased flow rate and improper resolution. For this the exchangers are repeatedly suspended in large volume of water

Addition of counter ions: Accomplished by washing the exchanger with suitable reagent depending upon which counter ion to be introduced. For e.g., NaOH (Na+ ), HCl (H+ ), etc,.

Advances in IC


Schematic of an ion  chromatograph instrument

Ion Suppressor

The mobile phase contains ions that create a background conductivity, making it difficult to measure the conductivity due only to the analyte ions as they exit the column

To improve the signal to noise ratio, when using a conductivity detector, a device called a suppressor, designed to selectively remove the mobile phase ions is placed after the analytical column and before the detector

The principle consists to convert the mobile phase ions to a neutral form or replacing them by others of higher conductivity. Suppressor-based detection is more useful for anion analysis than for cation analysis


Environment Application

Ion chromatography can be also used for the analysis of ions in natural brine waters, which include seawaters, subsurface brines, geothermal brines, and high salinity ground waters

Toxins – Cyanide and Chromium

Air Pollutants - HF, HCl, NOx and SOx on stationary emission

The most popular applications of ion chromatography are determination of common anions (F-, Cl-, Br-, NO2-, NO3-, PO43-, SO42-) and cations (Na+, K+, NH4+, Mg2+, Ca2+) in water and wastewater

Water purification

In this application, ion-exchange resins are used to remove poisonous (e.g. copper) and heavy-metal (e.g. lead or cadmium) ions from solution, replacing them with more innocuous ions, such as sodium and potassium

Few ion-exchange resins remove chlorine or organic contaminants from water – this is usually done by using an activated charcoal filter mixed in with the resin

There are some ion-exchange resins that do remove organic ions, such as MIEX (magnetic ion-exchange) resins. Domestic water purification resin is not usually recharged – the resin is discarded when it can no longer be used.