CHROMATOGRAPHY

Group B February 26th, 2018

‘Chromatography’ is an analytical technique As depicted above (Figure 2), the analyte

commonly used for separating a mixture of chemical is loaded over the silica bed (packed in the
substances into its individual components, so that the column) and allowed to adhere to the silica. Here,
individual components can be thoroughly analyzed. silica acts as the stationary phase. Solvent (mobile
There are many types of chromatography e.g., liquid phase) is then made to flow through the silica bed
chromatography, gas chromatography, ion-exchange (under gravity or pressure). The different
chromatography, affinity chromatography, but all of components of the analyte exhibit varying degrees
these employ the same basic principles. of adhesion to the silica (see later), and as a result
Let’s first familiarize ourselves with some terms they travel at different speeds through the
that are commonly used in the context of stationary phase as the solvent flows through it,
chromatography (Table 1; Figure 1) and the principle of indicated by the separation of the different bands.
chromatography (Figure 2). The components that adhere more strongly to the
stationary phase travel more slowly compared to
Table 1. The terms in chromatography
those with a weaker adhesion. Analytical
Term Definition
Mobile phase or solvent moving through the column chromatography can be used to purify compounds
carrier ranging from milligram to gram scale.
Stationary phase substance that stays fixed inside the column The commonly used chromatographic
or adsorbent
Eluent fluid entering the column techniques are mentioned below:
Eluate fluid exiting the column (that is collected in flasks) 1. Paper chromatography
Elution the process of washing out a compound through a 2. Thin layer chromatography (TLC)
column using a suitable solvent
3. Liquid column chromatography
Analyte mixture whose individual components have to be
separated and analyzed 4. Size exclusion chromatography
5. Ion exchange chromatography
6. Affinity chromatography
7. Gas chromatography

Discuss and answer the following questions with

your group. Find the answers in these
references, Boyer (2012) pages 115-159; Wilson
& Walker (2010) pages 433-476.

1. What is the principle of chromatography?

2. What are the factors affecting molecular
mobility on chromatography?
3. Compare the types of chromatography,
Figure 1. The illustration of column chromatography including their applications?

Figure 2. The representation of a column chromatographic separation set up

Chromatography