Technique of Ion Exchange Chromatography

Selection and preparation of the resin

A wide variety of resins are available commercially from which one must select an appropriate mesh -
size, cross linking and quality. Analytical grade resins are preferred because they have been more
carefully sized and washed to remove foreign organic and inorganic materials. It may be necessary to
convert the resin from one form to another, for example, the hydrogen form can be converted to the
sodium form by extensive washing in the column with strong sodium chloride solution until the effluent
is neutral. Used resins are regenerated in this way.

If it is important to know the weight of resin used, it must be dried or brought to known moisture
content in a hygrostat (a vessel with controlled, constant humidity). In any event, before packing it must
be equilibrated with water by prolonged soaking. After the resin has settled, the fins which float in the
water are poured off.

Packing the Column

Simple column are constructed from glass tubing with a reservoir at the top for the eluent and a friend a
fritted disk or glass wool plug at the bottom to support the resin bed. Usually the bottom of the column
is drawn to a narrower diameter and bent in a double U shape so that the outlet is higher than the top
of the resin bed. This prevents air bubbles from leaking into the column. The resin is packed as an
aqueous slurry and allowed to settle with occasional tapping. Once packed, the level of liquid should
never be allowed to drain lower than the top of the resin bed or air bubbles will be entrapped. The best
way to remove air bubbles or channeling is to back-flush the column with an upflow of water. A paper
or glass fiber disk placed on top of the bed will minimize disturbance of the resin when adding the
sample.

Total capacity of the Column

The total exchange capacity influences the maximum sample size and is used to check the long term
stability of the resin. The capacity of resin in milli-equivalent per gram of dry resin is normally marked on
the bottle by the manufacturer. Experimentally, it is most readily determined by converting the resin
entirely to the hydrogen form (if it is cationic), and then eluting with a sodium chloride solution until it is
completely converted to the equivalent to the capacity of the column-easily determined by titration
with sodium hydroxide. Common resins have a capacity of 1 to 5 meq/ml or roughly 1 to 5 N in acid or
base.

Detection Methods
The difficulty of detecting small amount of sample components in the presence of a large concentration
of eluting ion is one of the major disadvantages of ion exchange methods. Continuous recording is not
common, although in specific applications, light absorption, refractive index, pH, radioactivity, or
polarographic measurements have been utilized. The most common, practice is to collect numerous
small, equal, volume fractions and analyze each fraction for the species sought.