ZION

INTERNATIONAL PUBLIC SCHOOL

Mappedu, Chennai -600126

CHEMISTRY INVESTIGATORY PROJECT

AISSCE (2022-2023)

NAME: A. REVANTH
STD: XII SEC: A
TOPIC: Paper Chromatography
 CERTIFICATION
Certified to

be bonafide investigatory project done

by …………………………… of …………...........

during the academic year …………….

Reg.No.

Teacher-in-charge Principal

Submitted for the Practical Examination held on

……………………….
at Zion International Public School, Chennai-600 126.

Internal Examiner External Examiner

 ACKNOWLEDGEMENTS

I have taken efforts in this project. However, it would not

have been possible without the Kind support and help of many
individuals.

I would like to thank my chemistry teacher Mr. P.ALBIN

Over the invaluable guidance which has sustained my Efforts
in all the stages of this project work.

I would also like to thank my parents and friends for their

continuous Support and encouragement for completing the
project.

I would thank Almighty God for being able to complete this

project with success.
 Index
S.NO TOPICS
1 Introduction
2 Principle of paper chromatography
3 Paper Chromatography procedure
4 Working if chromatographic paper
5 Types of paper chromatography.
6 Other Types of paper chromatography:
7 Rf Values
8 Factors affecting Rf Value
9 Sources of error
10 Separating colored pigments
11 Applications of paper chromatography
12 Bibliography
 Introduction

Paper chromatography is a chromatography technique used to

separate mixture of chemical substances into its individual
compounds.

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
quantiles of material. It is used to determine organic
compounds.

In the beginning of 1860, a scientific concept with the wok of

FRIEDRICH GOPPELSROEDER who was a pioneer of
paper chromatography he developed the theory of capillary
analysis by using paper strips while examining wine, milk,
alkaloid, dyes and oils among other.

Paper chromatography was discovered by Synge and Martin

in the year 1943.
Principle of paper chromatography
The principle of separation is mainly “Partition” rather than
adsorption. Cellulose layers in filter paper contains acts as a
stationary phase and organic solvents are used as mobile
phase.
Paper Chromatography procedure
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.
Working if chromatographic paper

 A horizontal line is drawn near one end (about 1.5 cm

from bottom age) of the paper.

 The sample needs to be separated is placed as a small

drop or line onto the paper using capillary tube. labelling
the drop by a pencil with an alphabet or number helps to
identify the compound later. the drops are then served on
the paper and dried.

 The paper is then placed into a sealed container with a

shallow layer of suitable solvent. The solvent level must
be lower than the pencil line or drop of 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 we get. The components
travel with the solvent depending on three things:
o 1.the polarity of the sample molecule. The nonpolar
compounds travel faster than the polar component.
o 2. the attraction between the sample molecule and
the solvent of solvent mixture.
o 3 that reaction 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
there observed under UV light.

 This is what causes the different liquids within our

original liquid mixture to spread out on the solid.

 The key thing to remember is your chromatography is a

surface effect.
 Types of paper chromatography.

Paper chromatography

Paper adsorption Paper partition

chromatograph 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 is due to difference in
“PARTITION COEFFICIENT”.
 Column chromatography
 Thin layer chromatography
 Gas chromatography

 Paper partition chromatography: -

 The basic principle 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 “ABSORPTIVE”
power.
 Other Types of paper
chromatography:

1. Ascending Paper Chromatography – The techniques go

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 center 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.
 Rf Values

The retention factor (Rf) may be defined as the ratio of the

distance travelled by the salute to the distance travelled by the
solvent. It is used in chromatography to quantify the amount
of retardation of a sample in a stationary face K mobile face.
RF value 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.
For example, if a compound travels 9.9 cm in the solvent front
travels 12.7 cm, and Rf value = (9.9/12.7) =0.779 or 0.78.
Rf value depends on temperature and the solvent used in
experiment, so several solvents offer several RF values for the
same mixture of compound a solvent in chromatography is the
liquid the paper is placed in, and the salute is the ink which is
being separated.
 Factors affecting Rf Value

 The temperature
 The purity of the solvents used
 The quality of the paper, absorbents and impurities
present in the absorbents
 Ph of the solution
 Particle size
 The distance travelled by the solute and solvent
 In many cases it has been observed that the solvent front
is run off the end of the paper.
 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.
 Sources of error

 Error during the application of the spots: -

 Apply minimum volume of the concentrated solution in
order to avoid diffusion through the paper which leads to
poor separation.
 Spots should be approximately of the same diameter.

 Development: -
 Improper adjustment of the paper in the tank leads to this
error so the paper should be held vertically.
 Do chamber saturation.

 Detection: -
 The spraying methods affect the final result.
 Separating 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 and it is then slowly submerged
into a jar of solvent.
 As the solvent rises of the paper, it dissolves the
molecules present in the mixture, the solubility
depending on the polarity.
 This is because of different polarity molecules of each
pigment leave the solution a different place, as the
solvent continuous to rise up the stationary phase.
 Thus, each pigment rises up to a particular level on the
chromatography paper and get separated in the process.
 Applications of paper
chromatography

 Separation of mixtures of drugs

 To study the processes 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…
 BIBLIOGRAPHY

 N.C.E.R.T text book

 https://www.youtube.com/watch?v=mz_xcNrTK_U
 https://www.slideshare.net/SandeshWalke1/paper-
chromatography-project-class-12?from_action=save
 https://byjus.com/chemistry/paper-chromatography/
#Paper-Chromatography-Diagram
 https://byjus.com/chemistry/rf-value/
 https://lab-training.com/paper-chromatography/
 https://microbenotes.com/paper-chromatography/