Che Chromatography

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KENDRIYA VIDYALAYA NO.1, AFS PUNE
Name : Arpit Maurya

CLASS : 12th C
SUBJECT : Chemistry Investigatory Project
TOPIC : Paper Chromatography
ROLL NO. :
SUBMITTED TO: SUBMITTED BY:

MRS. BINI MENON ARPIT MAURYA
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ACKNOWLEDGEMENT
I would like to express my gratitude my
special thanks of gratitude to my
chemistry teacher MRS. BINI MENON for
her guidance and support in completing
my project.
I would like to extend my

gratitude to our Principal Sir Mr. R.N
Wadalkar for providing me with all facility
required.
Date: Arpit Maurya

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CErTIfICATE
This is to certify that Arpit Maurya a

student of class 12th C has successfully
completed the project under the guidance
of Mrs. Bini Menon during the academic
session 2022-23 for the partial fulfillment
of Term 2 Chemistry practical conducted
by AISSCE.
INTERNAL’s SIGN:
SUBJECT TEACHER SIGN:
PRINCIPAL SIGN:
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Objectives
 Introduction
 Principal of chromatography
 Working of chromatographic
paper
 Types of paper chromatography
 Applications of Paper
Chromatography.
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INTrODUCTION
Paper Chromatography is a chromatography
technique used to separate mixture of chemical
substance mixture of chemical substances into
its individual compounds.
It is an expensive method of separating dissolved

chemical substances by their different migration
rates across the sheets of paper. It is powerful
analytical tool that uses very small quantities of
material. It is used to determine organic
compounds.
Paper chromatography was discovered by Syne

and Martin in the year.
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Principal of Paper Chromatography
The principle of separation in mainly “Partition”.

Cellulose layers in filter paper contains moisture
which acts as stationary and phase and organic
solvents/ buffers are used as mobile phase.
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Working of Chromatographic Paper

 A horizontal line is drawn near one end (about 1.5 cm
from the bottom edge) of the paper.
 The sample needs to be separated is placed as a small
drop or line on to the paper using capillary tube.
Labelling the drop by a pencil with an alphabet or
number help to identify the compound later. The
drops are then soaked on the paper and dried.
 The paper is then placed into a sealed container with
a swallow layer of suitable solvent (M.P). The solvent
level must be lower than the pencil line or drop on it.
The container needs to be covered to stop the solvent
to evaporate.
 The solvent rises up the paper chromatography
taking each component of the sample with it. The
components travel with the solvent depends on three
things:
✓ The polarity of the sample molecule. The
non-polar components travel faster than the polar
component.
The attraction between the sample molecule and the
solvent or solvent mixture. The attraction between the
sample and the silica.
 When the solvent rises near the end of the paper then
the paper should be taken out from sealed container
and air dried. The paper with separated bands of
components are then observed under UV-light.
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Types of Chromatography
• Paper adsorption chromatography: -

 It is based on the differential adsorption of the
components on the adsorbed (stationary phase).
 This means that different components are absorbed on
adsorbent at different degrees.
 Migration of the substance id due to difference in
"PARTITION Coefficient ".
✓ Column Chromatography: -
✓ Thin layer Chromatography (TLC)
✓ Gas Chromatography
 Paper partition chromatography: -
 The basic principal of partition chromatography is the
continuous differential partitioning of components of
mixture between stationary phase and mobile phase.
 Separation of similar substances by repeated extraction
by two immiscible liquids.
➤ Migration of the substance is due to "ADSORPTIVE" Power.
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Rf values
The retention factor (R) may be defined as the ratio of the distance
traveled by the solute to the distance traveled by the solvent. It is
used in chromatography to quantify the amount of retardation of a
sample in a stationary phase relative to a mobile phase. R, values
are usually expressed as a fraction of two decimal places.
⚫ If Rf value of a solution is zero, the solute remains in the
stationary phase and thus it is immobile.
⚫ The ratio of the distance moved up or travelled by the
components from the origin or point of application to the
distance moved up by the solvent from the same point.
Rf = Distance travelled by the solute from the original line

Distance travelled by the solvent from the original line
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SAMPLE OBSERVATION
Sr.no Substance Distance travelled by Distance Rf Value

different compounds travelled
by solvent
1 Red ink 6.4 cm 7.2 cm 0.86cm
2 Blue ink 6.8 cm 7.2cm 0.94cm
Calculation:-
Where Rf =
For red ink -- 6.4 = 0.86 cm
7.2
For Blue ink – 6.8 = 0.94 cm

7.2
P a g e | 11
Factors affecting Rf value

 The temperature.
 The purity of the solvents used.
 The quality of the paper, adsorbents & impurities.
 present n the adsorbents.
 Chamber saturation techniques, method of drying and
development.
 The distance travelled by the solute & solvent.
 Chemical reaction between the substances being
partitioned.
 pH of the solution.
 In many cases it has been observed that the solvent front is
run off the end of the paper. Rx value is thus used.
 It is the ratio of distance travelled by the sample and the
distance travelled by the standard. Rx value is always closer
to 1.
 Mobile phase composition.
 Stationary phase.
 Concentration of analyte.
 Particle size.
 In liquid chromatography, in general, the mobile phase pH is
an important parameter.
 In reversed-phase liquid chromatography (hydrophobic
retention chromatography) the solvent type and
concentration are important factors.
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SEpArATING Of COLOrED pIGMENTS
 Paper chromatography is an effective technique for

separating colored pigments from a mixture.
 A few drops of the mixture of colored pigments are placed on
the filter paper (stationary phase) and it is then slowly
submerged into a jar of solvent (mobile phase).
 That is Because of different polarity, molecules of each
pigment leave the solution at different places, as the solvent
continues to rise up the stationary phase.
 Thus, each pigment rises up to a particular level on the
chromatography paper, and gets separated in the process.

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AppLICATIONS Of ChrOMATOGrAphY
 Separation of mixtures of drugs

 To study the process of fermentation and ripening.
 To check the purity of pharmaceuticals.
 To inspect cosmetics.
 To detect the adulterant.
 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.
 Separation of carbohydrates, vitamins, antibiotics, proteins,
etc.
 Analysis of metabolites of drugs in blood, urine....
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BIBLIOGrAphY
 NCERT
 Wikipedia
 Chemistry class 12 lab manual

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THANK YOU

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KENDRIYA VIDYALAYA NO.1, AFS PUNE

Name : Arpit Maurya

SUBMITTED TO: SUBMITTED BY:

I would like to extend my

Date: Arpit Maurya

This is to certify that Arpit Maurya a

SUBJECT TEACHER SIGN:

It is an expensive method of separating dissolved

Paper chromatography was discovered by Syne

Principal of Paper Chromatography

The principle of separation in mainly “Partition”.

Working of Chromatographic Paper

• Paper adsorption chromatography: -

Rf = Distance travelled by the solute from the original line

Sr.no Substance Distance travelled by Distance Rf Value

2 Blue ink 6.8 cm 7.2cm 0.94cm

For Blue ink – 6.8 = 0.94 cm

Factors affecting Rf value

SEpArATING Of COLOrED pIGMENTS

 Paper chromatography is an effective technique for

 Separation of mixtures of drugs

 Chemistry class 12 lab manual

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