M.Arun Babu (4FSHA) .Toxi

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M. ARUN BABU
21BSR14091
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PC – PAPER CHROMATOGRAPHY
Chromatography technique that uses paper sheets or strips as

the adsorbent being the stationary phase through which a solution is
made to pass is called paper chromatography. It is an inexpensive
method of separating dissolved chemical substances by their different
migration rates across the sheets of paper. It is a powerful analytical
tool that uses very small quantities of material. Paper chromatography
was discovered by Synge and Martin in the year 1943.
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Paper Chromatography Procedure :
Below we have explained the procedure to conduct Paper
Chromatography Experiment for easy understanding of students.
1. Selecting a suitable type of development: It is decided based on
the complexity of the solvent, paper, mixture, etc. Usually
ascending type or radial paper chromatography is used as they are
easy to perform. Also, it is easy to handle, the chromatogram
obtained is faster and the process is less time-consuming.
2. Selecting a suitable filter paper: Selection of filter paper is done
based on the size of the pores and the sample quality.
3. Prepare the sample: Sample preparation includes the dissolution
of the sample in a suitable solvent (inert with the sample under
analysis) used in making the mobile phase.
4. Spot the sample on the paper: Samples should be spotted at a
proper position on the paper by using a capillary tube.
5. Chromatogram development: Chromatogram development is
spotted by immersing the paper in the mobile phase. Due to the
capillary action of paper, the mobile phase moves over the sample
on the paper.
6. Paper drying and compound detection: Once the chromatogram
is developed, the paper is dried using an air drier. Also,
detecting solution can be sprayed on the chromatogram developed
paper and dried to identify the sample chromatogram spots.
Paper Chromatography Applications :
There are various applications of paper chromatography. Some of the

uses of Paper Chromatography in different fields are discussed below:
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 To study the process of fermentation and ripening.
 To check the purity of pharmaceuticals.
 To inspect cosmetics.
 To detect the adulterants.
 To detect the contaminants in drinks and foods.
 To examine the reaction mixtures in biochemical laboratories.
To determine dopes and drugs in humans and animals
Forensic Significance of Paper Chromatography

 For separation of components of the ink.
 For separation of amino acids.
 To detect any kind of adulteration in foods & drinks.
 To determine drugs in humans and animals.
Paper chromatography uses are not confined to any particular field. A

number of the necessary areas include:
 Reaction monitoring – The progress of the reaction can be

estimated by developing the chromatogram over different time
intervals by spotting the reactants.
 Isolation & Purification – This technique is useful in the
purification and isolation of components of mixtures. Here, the
separated components on the paper are cut, dissolved in suitable
solvents and using spectroscopic methods, their absorption is
characterised at specific wavelengths.
 Foods – Analysis of food colours in synthetic drinks and beverages,
ice creams, sweets, etc. Only edible colours are permitted for use,
this is why identification and quantification are of utmost
importance.
 Forensics – Provides a basis for identification and comparison
against reference standards for drugs and their metabolites. Paper
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chromatography offers a vital role in the viable analysis of samples
that are available in milligrams or microlitre quantities.
 Pharmaceuticals – Provides information related to the development
of new drugs molecules, reaction completion and progress of
manufacturing processes. This process is cost-effective and hence
used as an alternative method in monitoring the active ingredients
present in the drug forms. Paper chromatography is also applicable in
colour identifications of pharmaceutical formulations.
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The technique of Paper Chromatography is being extensively used for the

last several years and still have preserved their ground associated with the
separation of different classes of compounds.
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Types of paper chromatography:
1. Ascending Paper Chromatography – The techniques goes with its
name as the solvent moves in an upward direction.
2. Descending Paper Chromatography – The movement of the flow of
solvent due to gravitational pull and capillary action is downwards,
hence the name descending paper chromatography.
3. Ascending – Descending Paper Chromatography – In this version
of paper chromatography, movement of solvent occurs in two
directions after a particular point. Initially, the solvent travels
upwards on the paper which is folded over a rod and after crossing
the rod it continues with its travel in the downward direction.
4. Radial or Circular Paper Chromatography – The sample is
deposited at the centre of the circular filter paper. Once the spot is
dried, the filter paper is tied horizontally on a Petri dish which
contains the solvent.
5. Two Dimensional Paper Chromatography – Substances which have
the same rf values can be resolved with the help of two-
dimensional paper chromatography.
FORENSIC APPLICATION: -
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Paper chromatography is a powerful analytical technique used in
forensic science for the separation and analysis of various chemical
substances. In forensic science, paper chromatography is used to
analyze trace amounts of compounds found in samples such as blood,
urine, and saliva.
The principle of paper chromatography is based on the separation of a

mixture of chemical compounds into their individual components. The
sample is placed on a paper strip, and the strip is then placed in a
solvent. As the solvent moves up the paper, it carries the various
components of the mixture with it. The components separate based on
their differing solubility in the solvent and the paper, resulting in
distinct bands or spots.
Forensic scientists use paper chromatography to identify the presence of

drugs, poisons, and other substances in bodily fluids or other materials
found at crime scenes. For example, if a suspected drug dealer is
arrested, paper chromatography can be used to analyze the drugs found
on their person or in their possession. Similarly, if a person is suspected
of poisoning someone, paper chromatography can be used to analyze the
contents of their stomach or other bodily fluids for traces of the poison.
Paper chromatography is a valuable tool in forensic science because it is

relatively simple, inexpensive, and can be used to analyze small sample
sizes. It can also be combined with other analytical techniques such as
mass spectrometry for even more precise identification and analysis of
substances.
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1

Chromatography technique that uses paper sheets or strips as

Paper Chromatography Applications :

There are various applications of paper chromatography. Some of the

 To detect the adulterants.

 To detect the contaminants in drinks and foods.

 To examine the reaction mixtures in biochemical laboratories.

To determine dopes and drugs in humans and animals

Forensic Significance of Paper Chromatography

Paper chromatography uses are not confined to any particular field. A

 Reaction monitoring – The progress of the reaction can be

The technique of Paper Chromatography is being extensively used for the

The principle of paper chromatography is based on the separation of a

Forensic scientists use paper chromatography to identify the presence of

Paper chromatography is a valuable tool in forensic science because it is

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