Column Chromatography
                     Column Chromatography
Introduction
Column chromatography is described as the useful technique in 
which the substances to be isolated are presented onto the 
highest point of a column loaded with an adsorbent (stationary 
phase), go through the column at various rates that rely upon 
the affinity of every substance for the adsorbent and the solvent 
or solvent mixture, and are typically gathered in solution as they 
pass from the column at various time. 
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Column Chromatography principle
The main principle involved in column chromatography is the 
adsorption of the solutes of the solution with the help of a 
stationary phase and afterward separates the mixture into 
independent components.
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Individual components 
At the point when the mobile phase together with the mixture 
that requires to be isolated is brought in from the top of the 
column, the movement of the individual components of the 
mixture is at various rates.
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Stationary phase 
The components with lower adsorption and affinity to the 
stationary phase head out quicker when contrasted with the 
greater adsorption and affinity with the stationary phase. The 
components that move rapidly are taken out first through the 
components that move slowly are eluted out last. 
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Mobile phase and delivery system
This phase is made up of solvents that complement the 
stationary phase.
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Column 
For liquid chromatography: 2-50cm long and 4mm internal 
diameter, fabricated with stainless steel
For gas chromatography: 1-3m long and 2-4mm internal 
diameter, fabricated either with glass or stainless steel
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Injector system 
Responsible for delivering test samples to the column’s top in a 
reproducible pattern.
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Detector and Chart Recorder 
This gives a continuous record of the presence of the analytes 
in the eluate as they come out from the column.
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Preparation of the column
● Mostly the column is comprised of a glass tube with an 
appropriate stationary phase
● The bottom end of the column is packed with a glass 
wool/cotton wool or an asbestos pad after which the 
stationary phase is packed.
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Major applications 
1. To isolate active constituents
2. To separate compound mixtures
3. To remove impurities or carry purification process
4. To isolate metabolites from biological fluids
5. To estimate drugs in drug formulations or crude extracts
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