What Is Paper Chromatography: Principle, Types, & Uses

Updated on August 3, 2019

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 the paper and the liquid phase is of the 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 to 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 the 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 moves from the center (mid-point) towards the periphery of circular chromatography paper. The entire system is kept in a covered Petri dish for the development of the chromatogram.

The wick at the center of paper dips into the 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 the mobile phase at a right angle to the previous development for the 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 the mode of development.

c) Preparation of sample: Preparation of the sample involves the dissolution of the sample in a 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 the 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 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.

Questions & Answers

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        Wynona 

        3 weeks ago

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        Shivam 

        4 weeks ago

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        Monoshree Banerjee 

        4 weeks ago

        Thanks....

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        Hussaina Ibrahim Garba 

        2 months ago

        Thank you sir

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        Artistic girl 

        3 months ago

        Thank you soo much it is very helpful.Try to upload in easy english

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        Harien 

        4 months ago

        very authentic it was

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        Sneha M. N YADAV 

        4 months ago

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        kazye mukwiza 

        5 months ago

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        Yadhu 

        6 months ago

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        learner 

        8 months ago

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        manshi 

        8 months ago

        not soo effective give a short term..

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        Varusha gautam 

        9 months ago

        Very helpful and Thanku so much

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        chokkalingam.s 

        9 months ago

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        9 months ago

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        9 months ago

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        10 months ago

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        10 months ago

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        Gokul (GS) 

        10 months ago

        Helpful, thanks

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        Kapoor 

        11 months ago

        Amazing

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        Kinza Amjad 

        11 months ago

        Very informative

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        Binaya kisan 

        11 months ago

        Very commendable for me. And I easily get this topic Tq......

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        rishabhsoni 

        11 months ago

        Thanks

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        Kajal 

        11 months ago

        Easy to understand

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        neelima gayathri 

        11 months ago

        helpful thanks

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        Alisha 

        12 months ago

        Very informative

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        Dharanya 

        12 months ago

        Easy to understand. .....short and sweet. ...Thank you sir...

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        Pradeep 

        13 months ago

        Helpful

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        Prajakta 

        13 months ago

        Thank you Sir

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        Aishwarya patil 

        14 months ago

        No bad.. Somewhat useful

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        Abishek 

        14 months ago

        Very easy to understand and very simple manner great....

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        Athira P A 

        14 months ago

        This is very useful

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        Simfukwe Boyd 

        15 months ago

        I have learnt something

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        Harshita 

        19 months ago

        Please add about the HPLC and gas chromatography . And Rf factor .

      • profile image

        linh 

        2 years ago

        i have a question. what is the component of paper chromatography?

      • profile image

        shalini 

        3 years ago

        This is useful to all thank you ....

      • profile image

        Your Name 

        3 years ago

        yea me too

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        faraja frank 

        3 years ago

        Thanks very much sir

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        ali 

        3 years ago

        it is nice

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        Reena 

        4 years ago

        super sir i did nt understand

      • bheem profile imageAUTHOR

        bheem 

        4 years ago

        Check related articles in the page you will find..

      • profile image

        Rahul Patil 

        4 years ago

        what is hplc chromatography

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        Melanie 

        4 years ago

        It really helped me ... Good job

      • profile image

        Dheeraj Raman 

        4 years ago

        Sir this is very useful site and thanks

      • profile image

        Elumalai 

        4 years ago

        Thank you beem sir

      • profile image

        frank. 

        4 years ago

        great

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        sweety 

        4 years ago

        Thank you beem sir

      • profile image

        vinay 

        4 years ago

        Helped me a lot...thanks bheem sir

      • profile image

        subbu 

        4 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

        sneha 

        4 years ago

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

      • profile image

        reem 

        4 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.

      • bheem profile imageAUTHOR

        bheem 

        5 years ago

        Thanks guys

      • profile image

        Rizmy 

        5 years ago

        Excellent

      • profile image

        5 years ago

        Nice and efficient

      • profile image

        asma 

        5 years ago

        Plz write about comparative mobile phase pub of separating mixtures.

        This article is very good

      • profile image

        sreedhar 

        5 years ago

        ThankQ

      • profile image

        ashok 

        5 years ago

        What is planer chromatography

      • profile image

        pratue. 

        5 years ago

        It is to good.

      • bheem profile imageAUTHOR

        bheem 

        6 years ago

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

      • profile image

        NEHA 

        6 years ago

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

      • profile image

        Neha 

        6 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

      • bheem profile imageAUTHOR

        bheem 

        6 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

        zurfishan 

        6 years ago

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

      • bheem profile imageAUTHOR

        bheem 

        6 years ago

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

      • profile image

        neelima 

        6 years ago

        good sir

      • profile image

        Mbau 

        7 years ago

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

      • bheem profile imageAUTHOR

        bheem 

        7 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

        ayush 

        7 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 imageAUTHOR

        bheem 

        7 years ago

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

        Thanks for the point....

      • bheem profile imageAUTHOR

        bheem 

        7 years ago

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

      • profile image

        Vivek 

        7 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.

      • profile image

        MOON 

        7 years ago

        Great work! We will now have the best group project

      • bheem profile imageAUTHOR

        bheem 

        7 years ago

        Amber! welcome again...

      • bheem profile imageAUTHOR

        bheem 

        7 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...

      • profile image

        Amber 

        7 years ago

        thx this rlly helped!

      • profile image

        azilah 

        7 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

      • bheem profile imageAUTHOR

        bheem 

        7 years ago

        Louisa! you are welcome...

      • profile image

        Louisa 

        7 years ago

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

      • bheem profile imageAUTHOR

        bheem 

        7 years ago

        Thanks, rishabh...

      • profile image

        rishabh 

        7 years ago

        great work sir

      • bheem profile imageAUTHOR

        bheem 

        7 years ago

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

      • profile image

        Kakoli 

        7 years ago

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

      • profile image

        xswc.. 

        7 years ago

        thanks .

      • profile image

        Sam 

        7 years ago

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

      • bheem profile imageAUTHOR

        bheem 

        7 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

        shalini 

        7 years ago

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

      • profile image

        mukesh 

        7 years ago

        thank u

      • profile image

        Krupal 

        7 years ago

        Nice not and very informative, thanks dear

      • bheem profile imageAUTHOR

        bheem 

        7 years ago

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

      • profile image

        faima 

        7 years ago

        its really help full for me...

      • profile image

        Malik Ahmed Faheem 

        7 years ago

        Usefull method for understanding

      • bheem profile imageAUTHOR

        bheem 

        7 years ago

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

      • profile image

        lhpk 

        7 years ago

        Thank you .

      • profile image

        Roja 

        7 years ago

        It's very useful for students

      • profile image

        HARRISON 

        7 years ago

        A the same u have done well

      • bheem profile imageAUTHOR

        bheem 

        7 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

        houria 

        7 years ago

        why do not you write references with this topic

      • profile image

        israel okpari 

        7 years ago

        what could say about this technology

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