Chromatography

How Does It Work? Like Magic!

A mixture of various components enters a chromatography process,
and the different components are flushed through the system at different
rates. These differential rates of migration as the mixture moves over
adsorptive materials provide separation. Repeated sorption/desorption acts
that take place during the movement of the sample over the stationary bed
determine the rates. The smaller the affinity a molecule has for the
stationary phase, the shorter the time spent in a column.
In any chemical or bioprocessing industry, the need to separate and
purify a product from a complex mixture is a necessary and important step in
the production line. Today, there exists a wide market of methods in which
industries can accomplish these goals. Chromatography is a very special
separation process for a multitude of reasons! First of all, it can separate
complex mixtures with great precision. Even very similar components, such
as proteins that may only vary by a single amino acid, can be separated with
chromatography. In fact, chromatography can purify basically any soluble or
volatile substance if the right adsorbent material, carrier fluid, and operating
conditions are employed. Second, chromatography can be used to separate
delicate products since the conditions under which it is performed are not
typically severe. For these reasons, chromatography is quite well suited to a
variety of uses in the field of biotechnology, such as separating mixtures of
proteins.

The figure below will help you to follow along in our discussion.

The Column
Although there are other types of chromatography (e.g. paper
and thin layer), most modern applications of chromatography
employ a column. The column is where the actual separation takes
place. It is usually a glass or metal tube of sufficient strength to
withstand the pressures that may be applied across it. The column
contains the stationary phase. The mobile phase runs through the
column and is adsorbed onto the stationary phase. The column can
either be a packed bed or open tubular column.