Column chromatography: principle, procedure & applications

Column chromatography is the actual or the basic type of chromatography procedure which was developed during early stages of chromatography discovery. From it the principle of chromatography is described and these are still followed in the most advanced modes of chromatography. Other types of chromatography methods were developed with column chromatography as a module and with slight variations.

Column chromatography is basically a type of adsorption chromatography techniques. Here the separation of components depends upon the extent of adsorption to stationary phase. Here the stationary phase is a solid material packed in a vertical column made of glass or metal.

Principle: When a mixture of mobile phase and sample to be separated are introduced from top of the column, the individual components of mixture move with different rates. Those with lower affinity and adsorption to stationary phase move faster and eluted out first while those with greater adsorption affinity move or travel slower and get eluted out last.

The solute molecules adsorb to the column in a reversible manner. The rate of the movement of the components is given as follows

R= Rate of movement of a component / Rate of movement of mobile phase. i.e. it is the ratio of distance moved by solute to the distance moved by solvent.

Column chromatography animation

Procedure

Requirements:

The column chromatography requires a vertical column (preferably glass column) with a knob at the bottom end. This is preferably a burette shaped cylindrical column without graduations or readings.

Stationary phase or adsorbent, mostly fine sand will do or that recommended for the experiment. The particles of stationary phase should be of uniform size and shape without contamination.

Mobile phase preferably solvents of chromatography grade either a single solvent or a mixture of solvents as required for the separation.

Cotton wool or asbestos pad to plug the exit of column at the bottom and there by hold the column of stationary phase and let only escape of solvent and sample.

Procedure:

The stationary phase material is suitably moistened with mobile phase and packed sufficiently in the column with a cotton or asbestos pad at the bottom. The extract material or sample to be separated is placed on the top of packed stationary phase with a second cotton or asbestos pad in between.

The mobile phase is poured into the column over the sample. A collecting beaker is placed at the bottom of column near the end to collect the elute.

Precautions:

1. Keep the column in a clean and dust free place.

2. Do not disturb the column till the separation is complete.

3. Avoid gaps within the stationary phase packing.

Applications

♦ Column chromatography is best suited to separate active principle from plant materials.

♦ In separation of compounds after organic synthesis to obtain desired molecule.

♦ To separate or purify natural compound mixtures like alkaloids, glycosides.

For more details refer to column chromatography basics.

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Comments 56 comments

Phu Nguyen 4 years ago

wow great, thanks


Priya 4 years ago

Very informatical...


dera 4 years ago

very good and helpful


Sunita 4 years ago

nice


deepthi 4 years ago

good and an innovative idea


Iniya 4 years ago

very helpful


kanchan 3 years ago

helpful


honey 3 years ago

very good


himanshi 3 years ago

that's grt


rupesh 3 years ago

very good


sagar varma 3 years ago

very help full for exams


rehan 3 years ago

write some more about applications of column chromatography.


bheem profile image

bheem 3 years ago Author

Rehan! you are welcome to mention some more. It will be added here..thanks for stopping by.


Raja 3 years ago

Procedure is good and an innovative idea


bheem profile image

bheem 3 years ago Author

Raja! thanks for the appreciation but it is not innovative, it is an old established procedure to separate compounds..


Karthik 3 years ago

super,very useful for exams.


bheem profile image

bheem 3 years ago Author

Karthik! welcome again...


RUPA 2 years ago

super .............thanks


jatin 2 years ago

Its to gud column chromatography


a.s.sahu 2 years ago

must


lokesh j.h 2 years ago

very nice & it is important 1.


sharmila 2 years ago

nice...very usefull


soujanya 2 years ago

thank u..good info..


bheem profile image

bheem 2 years ago Author

@soujanya! You are welcome


roshna 2 years ago

nice....! very systematic.......


soni 2 years ago

Nice


chisom 2 years ago

its cool


bheem profile image

bheem 2 years ago Author

Hi Chisom! thnx for taking time to leave the comment


priyanka 2 years ago

There should be some more applications of column chromatography


sharanu.A.B 2 years ago

it is nice and very essential .


bheem profile image

bheem 2 years ago Author

There are many but due to more improvement in technology coming up, this method is less preferred for other applications.


Nita 2 years ago

Its very important information.........thanks


bheem profile image

bheem 2 years ago Author

Nita ! thanks for your complement..


preet 23 months ago

Its nice and easy to understand..... great work....


jot 22 months ago

very helpful thanksss :)


aniket 21 months ago

nice


Maneesh Guleria 18 months ago

Easy to understand..... @gud work


dhivya 18 months ago

Thankyou & good information


jyoti 17 months ago

very gud n excellent


plawan 17 months ago

very helpful....thnks


nilesh kinkar 16 months ago

Great & understendebal thanks


liya 13 months ago

good explaination


siva 12 months ago

Exalent information and thanks to beem sir


harini 12 months ago

good explaination

very helpful....thnks

--------------------------------------------------------------------------------


shilpa 12 months ago

good for assignments and

exams


Pranav 11 months ago

easy to understand....


naveen 11 months ago

Great


Teju 11 months ago

Too good


Sukhbir 10 months ago

Very good


zakia qurban 9 months ago

easy to understand and too great


Amol 8 months ago

Very important information thanx


Steve 7 months ago

Awesome


collins 6 months ago

good


Hemant 6 months ago

Thanx for the info. with best and convenient method


santosh 6 months ago

Useful


hamnah jabeen 2 months ago

thaaanks for this kind treatment gyues;

these app are very helpful for us for analytical chemistry'

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