Bheem is a writer and scientist who has a graduate degree in pharmacology. He enjoys health, fitness, and sharing pharmacology insights.
What Is Paper Chromatography?
Paper chromatography is one of the types of chromatography procedures that runs on a piece of specialized paper. It is a planar chromatography system wherein a cellulose filter paper acts as a stationary phase where compounds are separated.
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 held in the pores of the filter paper used, and the 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 the stationary phase) and mobile phase solvents during the movement of the mobile phase. This mobile phase moves 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 the mobile phase. But most of the applications of paper chromatography work on the principle of partition chromatography, i.e., partitioned between two liquid phases.
Uses and Applications of Paper Chromatography
- Paper chromatography is specially used to separate 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 to determine hormones, drugs, etc.
Sometimes it is also used to evaluate inorganic compounds like salts and complexes.
Ascending Paper Chromatography
Radial Paper Chromatography
Types or Modes of Paper Chromatography
Based on the way the development of chromatogram on paper, it is done in different modes. 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
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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, the solvent travels upwards and then downwards on the paper.
4. Radial mode
Here, the solvent moves from the center (mid-point) toward the periphery of the 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 the paper dips into the mobile phase in a petri dish, by which the solvent drains onto the paper and moves the sample radially to form the sample spots of different compounds as concentric rings.
5. Two-dimensional chromatography
Here, 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 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 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 the 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 the 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 off the paper and detecting the compounds
Once the chromatogram development 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.
© 2011 bheem
Mutinta Muchindu on March 22, 2020:
Love the notes, easy to understand.
Aditya on January 20, 2020:
Jaideep singh shekhawat on January 05, 2020:
It's realy very good for all person infact it's very esay to understand for those person whose english is not so good
Ku. Anjali Bhagat on November 29, 2019:
Thank you for this. It's make my doubts all clear.. Great job
Alekhya on November 20, 2019:
Delton Shikanda on November 15, 2019:
Thanks too it aware some
Wynona on October 20, 2019:
Thanks much its awesome
Shivam on October 14, 2019:
Very worst slide
Monoshree Banerjee on October 13, 2019:
Hussaina Ibrahim Garba on August 19, 2019:
Thank you sir
Artistic girl on July 19, 2019:
Thank you soo much it is very helpful.Try to upload in easy english
Harien on July 16, 2019:
very authentic it was
Sneha M. N YADAV on July 04, 2019:
Thanks for your great corporation
kazye mukwiza on June 16, 2019:
Super be sir
Yadhu on May 02, 2019:
learner on March 18, 2019:
manshi on February 20, 2019:
not soo effective give a short term..
Varusha gautam on February 12, 2019:
Very helpful and Thanku so much
chokkalingam.s on February 06, 2019:
Gnana deepa pasupuleti on February 02, 2019:
prabsi on January 25, 2019:
love this this fully helped me excel in my education and real life
Nandini on January 10, 2019:
Sush on December 28, 2018:
Gokul (GS) on December 20, 2018:
Kapoor on December 17, 2018:
Kinza Amjad on December 14, 2018:
Binaya kisan on December 10, 2018:
Very commendable for me. And I easily get this topic Tq......
rishabhsoni on December 06, 2018:
Kajal on December 03, 2018:
Easy to understand
neelima gayathri on November 22, 2018:
Alisha on November 14, 2018:
Dharanya on November 10, 2018:
Easy to understand. .....short and sweet. ...Thank you sir...
Pradeep on October 17, 2018:
Prajakta on October 08, 2018:
Thank you Sir
Aishwarya patil on September 16, 2018:
No bad.. Somewhat useful
Abishek on September 05, 2018:
Very easy to understand and very simple manner great....
Athira P A on August 31, 2018:
This is very useful
Simfukwe Boyd on July 28, 2018:
I have learnt something
Harshita on March 24, 2018:
Please add about the HPLC and gas chromatography . And Rf factor .
linh on September 03, 2017:
i have a question. what is the component of paper chromatography?
shalini on August 08, 2016:
This is useful to all thank you ....
Your Name on January 12, 2016:
yea me too
faraja frank on January 06, 2016:
Thanks very much sir
ali on January 05, 2016:
it is nice
Reena on September 12, 2015:
super sir i did nt understand
bheem (author) on August 30, 2015:
Check related articles in the page you will find..
Rahul Patil on August 08, 2015:
what is hplc chromatography
Melanie on July 14, 2015:
It really helped me ... Good job
Dheeraj Raman on July 05, 2015:
Sir this is very useful site and thanks
Elumalai on June 26, 2015:
Thank you beem sir
frank. on June 18, 2015:
sweety on April 14, 2015:
Thank you beem sir
vinay on March 30, 2015:
Helped me a lot...thanks bheem sir
subbu on March 15, 2015:
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
sneha on March 07, 2015:
its really helpful for me......... thank u
reem on January 23, 2015:
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 (author) on August 12, 2014:
Rizmy on July 31, 2014:
s on July 30, 2014:
Nice and efficient
asma on May 21, 2014:
Plz write about comparative mobile phase pub of separating mixtures.
This article is very good
sreedhar on December 19, 2013:
ashok on December 08, 2013:
What is planer chromatography
pratue. on November 22, 2013:
It is to good.
bheem (author) on October 29, 2013:
Neha!you can quote books on analytical techniques by chetwal and ravi shankar
NEHA on October 23, 2013:
nice description but i want a proper reference for this article please give me that.
Neha on October 22, 2013:
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 (author) on January 22, 2013:
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...
zurfishan on January 19, 2013:
plz show me difference between mobile phase and stationary phase, with the help of diagram,please.
bheem (author) on November 26, 2012:
Mbau & neelima! You guys are welcome again.. :-)
neelima on November 25, 2012:
Mbau on November 09, 2012:
Very comprehensive and detailed thus i benefited abundantly; not only while doing tutorials but also in revision. Excellent work!
bheem (author) on October 21, 2012:
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...
ayush on October 20, 2012:
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 (author) on July 26, 2012:
Vivek! there is an update, as a clarification to your comment in the principle of the article....
Thanks for the point....
bheem (author) on July 26, 2012:
Moon! happy to know the article was helpful to your group.
Vivek on July 17, 2012:
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.
MOON on May 24, 2012:
Great work! We will now have the best group project
bheem (author) on April 23, 2012:
Amber! welcome again...
bheem (author) on April 23, 2012:
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
Amber on April 22, 2012:
thx this rlly helped!
azilah on April 17, 2012:
i want ask a favour you..could you explain about the mobile phases generally used in the detection of lipid by paper chromatography
bheem (author) on March 18, 2012:
Louisa! you are welcome...
Louisa on March 18, 2012:
Thanx a lot for the article,was very useful in writing my practical report on P.chromatography
bheem (author) on March 18, 2012:
rishabh on March 17, 2012:
great work sir
bheem (author) on March 12, 2012:
Sam, xswc, Kakoli! happy that you found the article useful...You are all welcome again...
Kakoli on March 11, 2012:
sir !! its really helpful for me before few hours of my final exam....
xswc.. on March 11, 2012:
Sam on February 26, 2012:
Thanks a lot. This really helped me with my homework :D
bheem (author) on February 21, 2012:
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.
shalini on February 20, 2012:
can u add some details about the mobile phases generally used in the detection of aminoacids by paper chromatography
mukesh on February 12, 2012:
Krupal on February 07, 2012:
Nice not and very informative, thanks dear
bheem (author) on February 05, 2012:
Hello Malik & Faima! happy to know this article on principle and types of paper chromatography was helpful to you...
faima on February 04, 2012:
its really help full for me...
Malik Ahmed Faheem on February 03, 2012:
Usefull method for understanding
bheem (author) on January 22, 2012:
@HARRISON @Roja @lhpk! thanks for the complements and glad that you found the article helpful.......
lhpk on January 22, 2012:
Thank you .
Roja on January 22, 2012:
It's very useful for students
HARRISON on December 09, 2011:
A the same u have done well