What is Paper Chromatography: Principle, Types & Uses

What Is Paper Chromatography?

Paper chromatography is one of the types of chromatography procedures which runs on a piece of specialized paper. It is a planar chromatography system wherein a cellulose filter paper acts as a stationary phase on which the separation of compounds occurs.

Principle of paper chromatography: The principle involved is partition chromatography wherein the substances are distributed or partitioned between liquid phases. One phase is the water, which is held in the pores of the filter paper used; and other is the mobile phase which moves over the paper. The compounds in the mixture get separated due to differences in their affinity towards water (in stationary phase) and mobile phase solvents during the movement of mobile phase under the capillary action of pores in the paper.

The principle can also be adsorption chromatography between solid and liquid phases, wherein the stationary phase is the solid surface of paper and the liquid phase is of mobile phase. But most of the applications of paper chromatography work on the principle of partition chromatography, i.e. partitioned between to liquid phases.

Uses and Applications of Paper Chromatography

Paper chromatography is specially used for the separation of a mixture having polar and non-polar compounds.

For separation of amino acids.

It is used to determine organic compounds, biochemicals in urine, etc.

In the pharma sector it is used for the determination of hormones, drugs, etc.

Sometimes it is used for evaluation of inorganic compounds like salts and complexes.

Ascending Paper chromatography

Ascending type
Ascending type

Radial paper chromatography

Types or Modes of Paper Chromatography

Based on the way the development of chromatogram on paper is done in procedures, we have, broadly, five types of chromatography.

1. Ascending chromatography: As the name indicates, the chromatogram ascends. Here, the development of paper occurs due the solvent movement or upward travel on the paper.

The solvent reservoir is at the bottom of the beaker. The paper tip with sample spots just dips into the solvent at the bottom so that spots remain well above the solvent.

2. Descending chromatography: Here the development of paper occurs due to solvent travel downwards on the paper.

The solvent reservoir is at the top. The movement of solvent is assisted by gravity besides the capillary action.

3. Ascending- descending mode: Here solvent first travels upwards and then downwards on the paper.

4. Radial mode: Here the solvent travels from center (mid-point) towards the periphery of circular chromatography paper. The entire system is kept in a covered petridish for the development of the chromatogram.

The wick at the center of paper dips into mobile phase in a petri dish, by which the solvent drains on to the paper and moves the sample radially to form the sample spots of different compounds as concentric rings.

5. Two dimensional chromatography: Here the chromatogram development occurs in two directions at right angles.

In this mode, the samples are spotted to one corner of rectangular paper and allowed for first development. Then the paper is again immersed in mobile phase at a right angle to the previous development for second chromatogram.

Experiment Demo Video

Paper Chromatography Experiment Method

The experimental method involves:

a) Selection of suitable type of development: This depends on the complexity of the mixture, solvent, paper, etc. But in general ascending type or radial type chromatography are used as they are easy to perform, handle, less time-consuming and also give chromatogram faster.

b) Selection of suitable filter paper: Filter paper is selected based on pore size, the quality of the sample to be separated, and also mode of development.

c) Preparation of sample: Preparation of sample involves dissolution of sample in suitable solvent used in making mobile phase. The solvent used should be inert with the sample under analysis.

d) Spotting of sample on the paper: Samples are to be spotted at proper position on the paper preferably using a capillary tube.

d) Development of chromatogram: Sample spotted paper is subjected to development by immersing it in the mobile phase. The mobile phase moves over the sample on the paper under the capillary action of paper.

e) Drying of the paper and detection of the compounds: Once the development of chromatogram is over, the paper is held carefully at the borders so as to avoid touching the sample spots and dried using an air drier. Sometimes the detecting solution is sprayed in the developed paper and dried to identify the sample chromatogram spots.

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Comments 63 comments

israel okpari 4 years ago

what could say about this technology


houria 4 years ago

why do not you write references with this topic


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bheem 4 years ago Author

Hello Houria! this article including the figures is not copy pasted from some source or written by referring to some journal or book. It was written by teachings i received from my lecturers and experience with my seniors during my graduation. so I hardly remember the references.......

And the video you see is taken from YouTube and the source is found on the video itself when you click.

Thanks......


HARRISON 4 years ago

A the same u have done well


Roja 4 years ago

It's very useful for students


lhpk 4 years ago

Thank you .


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bheem 4 years ago Author

@HARRISON @Roja @lhpk! thanks for the complements and glad that you found the article helpful.......


Malik Ahmed Faheem 4 years ago

Usefull method for understanding


faima 4 years ago

its really help full for me...


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bheem 4 years ago Author

Hello Malik & Faima! happy to know this article on principle and types of paper chromatography was helpful to you...


Krupal 4 years ago

Nice not and very informative, thanks dear


mukesh 4 years ago

thank u


shalini 4 years ago

can u add some details about the mobile phases generally used in the detection of aminoacids by paper chromatography


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bheem 4 years ago Author

Hello shalini,

Amino acids are zwitter ions i.e they have both acidic and basic moieties on the same molecule. So amino acids separation rate varies based on pH of mobile phase used.

They are polar compounds so best mobile phases used are polar solvents like propanol, butanol and you may make their pH to basic by adding aqueous ammonia..

For more details do refer books on experimental organic, biochemical or phytochemistry with standard protocols for separation of set of amino acids by paper chromatography.


Sam 4 years ago

Thanks a lot. This really helped me with my homework :D


xswc.. 4 years ago

thanks .


Kakoli 4 years ago

sir !! its really helpful for me before few hours of my final exam....


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bheem 4 years ago Author

Sam, xswc, Kakoli! happy that you found the article useful...You are all welcome again...


rishabh 4 years ago

great work sir


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bheem 4 years ago Author

Thanks, rishabh...


Louisa 4 years ago

Thanx a lot for the article,was very useful in writing my practical report on P.chromatography


bheem profile image

bheem 4 years ago Author

Louisa! you are welcome...


azilah 4 years ago

i want ask a favour you..could you explain about the mobile phases generally used in the detection of lipid by paper chromatography


Amber 4 years ago

thx this rlly helped!


bheem profile image

bheem 4 years ago Author

Azilah! lipid detection in paper chromatography is done by reverse phase method where hydrocarbons form stationary layer and methanol, water, acetic acid like solvents are used as mobile phase.

Check for more Quantitative paper chromatography of lipids by Joanne L. Gellerman

Regards...


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bheem 4 years ago Author

Amber! welcome again...


MOON 4 years ago

Great work! We will now have the best group project


Vivek 4 years ago

Please reconfirm the principle as i have heard the principle is not coz of the water. Its due to difference in the affinity of the componenet between the stationary phase and mobile phase.


bheem profile image

bheem 4 years ago Author

Moon! happy to know the article was helpful to your group.


bheem profile image

bheem 4 years ago Author

Vivek! there is an update, as a clarification to your comment in the principle of the article....

Thanks for the point....


ayush 3 years ago

hello sir!

i am in class 12. i have to make a project on paper chromatography .please tell me the important headings related to it so that i can make the project upto 10 to 12 pages


bheem profile image

bheem 3 years ago Author

Hi ayush! try this as....

What is chromatography, its types, paper chromatography, principle, requirements, procedure or method, its advantages over other chromatographies, limitations, applications of .....

all the best...


Mbau 3 years ago

Very comprehensive and detailed thus i benefited abundantly; not only while doing tutorials but also in revision. Excellent work!


neelima 3 years ago

good sir


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bheem 3 years ago Author

Mbau & neelima! You guys are welcome again.. :-)


zurfishan 3 years ago

plz show me difference between mobile phase and stationary phase, with the help of diagram,please.


bheem profile image

bheem 3 years ago Author

Hi zurfishan! in the video you will notice a paper with sample at one end. This paper itself is stationary phase as it doesn't move at all. After putting the paper in the beaker having solvent, the solvent spreads over the stationary paper. So this solvent and solvent layer is called mobile phase......

hope this helps...


Neha 2 years ago

Nice description but i want to give referance to this so plz suggest that what should i write plz give some referance in detail... thanks


NEHA 2 years ago

nice description but i want a proper reference for this article please give me that.


bheem profile image

bheem 2 years ago Author

Neha!you can quote books on analytical techniques by chetwal and ravi shankar


pratue. 2 years ago

It is to good.


ashok 2 years ago

What is planer chromatography


sreedhar 2 years ago

ThankQ


asma 2 years ago

Plz write about comparative mobile phase pub of separating mixtures.

This article is very good


2 years ago

Nice and efficient


Rizmy 2 years ago

Excellent


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bheem 2 years ago Author

Thanks guys


reem 20 months ago

no doubt this article is very useful for students. suggestion for you is that you should also add lab and industrial applications of this technique.


sneha 18 months ago

its really helpful for me......... thank u


subbu 18 months ago

What is the correct principles of all chromatography? I need a best reason ?plzzz help me sir .......... I m B.Sc chemistry student , so, u tell me brief


vinay 18 months ago

Helped me a lot...thanks bheem sir


sweety 17 months ago

Thank you beem sir


frank. 15 months ago

great


Elumalai 15 months ago

Thank you beem sir


Dheeraj Raman 14 months ago

Sir this is very useful site and thanks


Melanie 14 months ago

It really helped me ... Good job


Rahul Patil 13 months ago

what is hplc chromatography


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bheem 13 months ago Author

Check related articles in the page you will find..


Reena 12 months ago

super sir i did nt understand


ali 8 months ago

it is nice


faraja frank 8 months ago

Thanks very much sir


Your Name 8 months ago

yea me too


shalini 7 weeks ago

This is useful to all thank you ....

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