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What is Paper Chromatography: Principle, Types & Uses

Updated on December 16, 2015

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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    • profile image

      israel okpari 5 years ago

      what could say about this technology

    • profile image

      houria 5 years ago

      why do not you write references with this topic

    • bheem profile image
      Author

      bheem 5 years ago

      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......

    • profile image

      HARRISON 5 years ago

      A the same u have done well

    • profile image

      Roja 5 years ago

      It's very useful for students

    • profile image

      lhpk 5 years ago

      Thank you .

    • bheem profile image
      Author

      bheem 5 years ago

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

    • profile image

      Malik Ahmed Faheem 5 years ago

      Usefull method for understanding

    • profile image

      faima 5 years ago

      its really help full for me...

    • bheem profile image
      Author

      bheem 5 years ago

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

    • profile image

      Krupal 5 years ago

      Nice not and very informative, thanks dear

    • profile image

      mukesh 5 years ago

      thank u

    • profile image

      shalini 5 years ago

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

    • bheem profile image
      Author

      bheem 5 years ago

      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.

    • profile image

      Sam 5 years ago

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

    • profile image

      xswc.. 5 years ago

      thanks .

    • profile image

      Kakoli 5 years ago

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

    • bheem profile image
      Author

      bheem 5 years ago

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

    • profile image

      rishabh 5 years ago

      great work sir

    • bheem profile image
      Author

      bheem 5 years ago

      Thanks, rishabh...

    • profile image

      Louisa 5 years ago

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

    • bheem profile image
      Author

      bheem 5 years ago

      Louisa! you are welcome...

    • profile image

      azilah 5 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

    • profile image

      Amber 5 years ago

      thx this rlly helped!

    • bheem profile image
      Author

      bheem 5 years ago

      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...

    • bheem profile image
      Author

      bheem 5 years ago

      Amber! welcome again...

    • profile image

      MOON 5 years ago

      Great work! We will now have the best group project

    • profile image

      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
      Author

      bheem 4 years ago

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

    • bheem profile image
      Author

      bheem 4 years ago

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

      Thanks for the point....

    • profile image

      ayush 4 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
      Author

      bheem 4 years ago

      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...

    • profile image

      Mbau 4 years ago

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

    • profile image

      neelima 4 years ago

      good sir

    • bheem profile image
      Author

      bheem 4 years ago

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

    • profile image

      zurfishan 4 years ago

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

    • bheem profile image
      Author

      bheem 4 years ago

      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...

    • profile image

      Neha 3 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

    • profile image

      NEHA 3 years ago

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

    • bheem profile image
      Author

      bheem 3 years ago

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

    • profile image

      pratue. 3 years ago

      It is to good.

    • profile image

      ashok 3 years ago

      What is planer chromatography

    • profile image

      sreedhar 3 years ago

      ThankQ

    • profile image

      asma 3 years ago

      Plz write about comparative mobile phase pub of separating mixtures.

      This article is very good

    • profile image

      2 years ago

      Nice and efficient

    • profile image

      Rizmy 2 years ago

      Excellent

    • bheem profile image
      Author

      bheem 2 years ago

      Thanks guys

    • profile image

      reem 2 years 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.

    • profile image

      sneha 2 years ago

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

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      subbu 2 years 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

    • profile image

      vinay 2 years ago

      Helped me a lot...thanks bheem sir

    • profile image

      sweety 2 years ago

      Thank you beem sir

    • profile image

      frank. 2 years ago

      great

    • profile image

      Elumalai 2 years ago

      Thank you beem sir

    • profile image

      Dheeraj Raman 24 months ago

      Sir this is very useful site and thanks

    • profile image

      Melanie 23 months ago

      It really helped me ... Good job

    • profile image

      Rahul Patil 22 months ago

      what is hplc chromatography

    • bheem profile image
      Author

      bheem 22 months ago

      Check related articles in the page you will find..

    • profile image

      Reena 21 months ago

      super sir i did nt understand

    • profile image

      ali 17 months ago

      it is nice

    • profile image

      faraja frank 17 months ago

      Thanks very much sir

    • profile image

      Your Name 17 months ago

      yea me too

    • profile image

      shalini 10 months ago

      This is useful to all thank you ....

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