DECLARATION

I hereby declare that the work being presented in the project

report entitled “Paper Chromatography” is an authentic record
of my own work carried out in the supervision of Naveen
Jaiswal, Chemistry Department, DELHI PUBLIC SCHOOL
KALYANPUR

SAMRIDHI SHREE
Roll. No. : 21
Class: 12 S2
 CERTIFICATE
This is to certify that the above statement made by the
candidate is correct to the best of my knowledge.

Naveen Jaiswal
PGT, Chemistry Department
DELHI PUBLIC SCHOOL KALYANPUR
 INDEX
1) Acknowledgement
2) Aim
3) Apparatus
4) Introduction
5) Types of chromatography
6) Paper chromatography
7) Theory
8) Developer
9) Experimental Procedure
10) Procedure
11) Observations
12) Result
13) Precautions 14) Bibliography
 ACKNOWLEDGEMENT

I express my sincere gratitude to my chemistry teacher Mr.

Naveen Jaiswal under whose guidance I was able to take up and
complete the project, “PAPER CHROMATOGRAPHY (SEPARATION
OF IONS). I am indebted to them for their invaluable support,
guidance and encouragement.
 AIM
To separate:

1) Cu(II) ions and Fe(III) ions.

2) Ni(II) ions and Fe(III) ions.
3) Ni(II) ions and Cu(III) ions.

Using paper chromatography technique.

 APPARATUS
1) Glass jar with cover or a wide mouthed test tube.
2) Capillary with fine tube.
3) Glass rod
4) 100 ml beakers
5) Whatman filter paper
6) Sharp pointed pencil
7) Scale
8) 10 ml graduated cylinder
9) Weight box
10) Watch glass
11) Sprayer

CHEMICALS:
1) Ethanol 4) Nickel(II) chloride
2) Distilled water 5) Potassium Ferrocyanide
3) Ferric(III) chloride 6) Copper(II) chloride

INTRODUCTION
Chromatography is an excellent and most extensively used
technique for separation and purification of compounds. It may
be defined as “the process of selective retardation of one or
more compounds of a liquid or gas uniformly migrating
through a stationary phase”.

In this technique, the mixture of components is put over a solid

or a liquid (mounted on a solid support) called stationary phase.
Another phase, liquid or a gas, known as mobile phase, is then
allowed to move slowly over it and selective retardation in their
rate of movement separates them. The reason of retardation of
the components i.e. separation is the process of adsorption or
both depending upon the nature of stationary phase.
TYPES OF CHROMATOGRAPHY

Name of Stationary Mobile Methods of Basic

Chromatography phase phase fixing the principle
stationary involved in
phase separation
Column Solid Liquid Held in Adsorption
tabular form
Thin Layer (TLC) Solid Liquid Finely divided Adsorption
solid on glass
plate
Gas-Solid (GSC) Solid Gas Held in Adsorption
tabular form and
volatility
Partition Liquid Liquid Adsorbed Partition
porous solid
held in a
tabular form
Paper Liquid Liquid Held in pores Partition
of a thick
paper
Gas-Liquid Liquid Gas Adsorbed on Partition
a porous solid
held in a tube
 PAPER CHROMATOGRAPHY
Paper chromatography technique was devised by Martin and
Synge in 1941.
It has been widely used for the separation and identification of
the components of mixture especially when it is available in
small amounts.
The basic principle of separation in this process is partition of
the components of mixture of two liquids. Stationary phase is
also a liquid (usually water) held in the pores of a thick paper
(filter paper).

THEORY

Rf Value

The separate components of a mixture are identified by their R f

values.
 Rf stands for “Retention Factor” or “Retardation Ratio” and can
be defined by the expression:

Rf

The Rf value of a substance depends on:

1) Nature of mobile phase
2) Quality of paper
3) Nature of substance
4) Temperature

DEVELOPER
It is a substance used to give color to different bands formed on
the fixed phase to make them more distinct. The developer
used i.e. gas forms a complex compound with the solute which
is thus responsible for the color.

Cations and anions are divided in various groups depending on

a particular developing solvent.
 EXPERIMENTAL PROCEDURE
Paper chromatography may be divided into following four steps
from practical point of view:

1. Preparation of filter paper:

Cut a strip of Whatman filter paper about the size of the
length of the jar or test tube. Then draw a straight line called
the original line or initial line horizontally along its width about
3 cm from one of its edge with a pencil. For radial
chromatography, make two parallel curs 2 mm part from the
edge upto centre of a circular Whataman filter paper having
diameter of about 11 cm. Bend this piece of perpendicular to
the circular paper so that it serves as a wick.

2. Spotting of filter paper:

Apply the test solution with a fine capillary in the middle of the
original line. While applying the test solution, it should be
ensured that the spot diameter does not exceed 3 mm.

3. Development:
The dry strip is positioned in a chromatographic jar in such a
way that its spotted end comes in contact with the mobile
phase. For example, the strip is suspended vertically in such a
way that its spotted end dips about 1 cm in the liquid phase
 The mobile phase can be single substance or a mixture of
substance or a mixture of substances. The composition of the
mobile phase (solvent developer) is decided by the nature of
the species to be separated. The selection of an appropriate
mobile phase for a new separation procedure has often been
determined by empirical experiments. However, the following
points are observed while selecting a suitable solvent (mobile
phase):
a) The components should be more soluble in the stationary
phase as compared to mobile phase.
b) The solvent should not react chemically with any of the
component of paper.
c) Rf values of any two components mist differ by at least 0.950 in
the chosen solvent
d) The composition of solvent doesn’t differ with time.
e) The solvent should not interfere with the detection of the
developed chromatogram

When the solvent has risen to a reasonable distance (say 15cm)

from the original line, the strip is removed from the jar and the
position of the solvent is marked by drawing a small line with a
pencil. This line is called solvent front. The paper strip is then
dried in air is known as chromatogram.
 4. Identification:
The components of mixture separate in the form of spots lying
between the original line and the solvent line. Number of
components are usually equal to number of spots appeared.
Visualization of the spots is done from their colors. But if the
sports are not colored, then the strip is prayed with a reagent
called visualization reagent or spraying reagent. This reagent
causes the components to form a derivative which is either
colored or fluoresces when exposed to UV light.

PROCEDURE
 Cut three strips of Whatman filter paper of length and width
almost equal to that of the jar.
 Draw a line of about 3 cm from one end of the strip with a
pencil.
 Prepare solution of Ferric Chloride, Copper Chloride and Nickel
Chloride of about 0.1M strength each.
 Apply the spot of (Ferric Chloride + Copper Sulphate), (Ferric
Chloride + Nickel Chloride) and (Nickel Chloride + Copper
Sulphate) solution on three different strips almost in the middle
 of the pencil line with the help of a fine capillary. Dry the spot.
Repeat the process 2-3 time putting every drop on the same
spot.
 Pour mobile phase, a mixture of ethanol and dil. HCl in the ratio
9:1, upto a depth of 2cm in the jar.
 Put the paper strips in the jar in such a way that the pencil line
just dips into the mobile phase.
 Keep the jar undisturbed and allow running the mobile phase at
least 10-12 cm above the pencil line.
 Remove the paper strips and mark the solvent front with the
pencil
 Dry the strips and identity them by spraying the respective
spraying agents over it.
 Identify each ion from the colors and calculate their R f values by
measuring the distances of the spot and solvent from the initial
spot.

OBSERVATIONS
 For Cu(II) and Fe(III) ions

1) Spot is applied with 0.1 M aqueous solution of

Copper Chloride and Ferric Chloride.
2) Mobile phase = Ethanol(90 ml) + 5M HCl(10ml) 3)
Spraying or visualization agent:

 Pottasium Ferrocyanide for detection of Fe(III) ions and

Cu(II) ions.

S. No. Coulor of Distance Distance Rf Value

the spot travelled by travelled
different by
components solvent
(cm) (cm)
1. (Cu2+) Chocolaty
brown 4 7.6 0.52
2. (Fe3+) Blue 5.7 7.6 0.75

For Ni(II) and Fe(III) ions

 1) Spot is applied with 0.1 M aqueous solution of Nickel
Chloride and Ferric Chloride.
2) Mobile phase = Ethanol(90 ml) + 5M HCl(10ml) 3)
Spraying or visualization agent:

 Pottasium Ferrocyanide for detection of Fe(III) ions and Cu(II)

ions.
 DMG (Dimethyle glyoxime) reagent for detection of
Ni(II) ions.

S.No. Color of Distance Distance Rf Value

the spot travelled by travelled
different by
components solvent
(cm) (cm)
1. (Ni2+) Rose red 3.1 6.9 0.44
2. (Fe3+) Blue 5.5 6.9 0.79

For Ni(II) and Cu(II) ions

1) Spot is applied with 0.1 M aqueous solution of Nickel
Chloride and Copper Chloride.
2) Mobile phase = Ethanol(90 ml) + 5M HCl(10ml) 3)
Spraying or visualization agent:

 Pottasium Ferrocyanide for detection of Cu(II) ions.

 DMG (Dimethyle glyoxime) reagent for detection of
Ni(II) ions.

S.No. Color of Distance Distance Rf Value

the spot travelled by travelled
different by
components solvent
(cm) (cm)
1. (Cu2+) Chocolaty
brown 6.5 10.1 0.64
2. (Ni2+) Rose red 4.4 10.1 0.43

RESULT
Mixture of ions is separated using paper chromatography
technique.

The Rf value for different ion is found to be:

1) Cu2+ : 0.52
Fe3+ : 0.75

2) Ni2+ : 0.44

Fe3+ : 0.79

3) Ni2+ : 0.64

Cu2+ : 0.43

Within experimental errors, Rf values of:

 Fe3+ ions is between: 0.75 to 0.79  Cu2+ ions is between: 0.43

to 0.52
 Ni2+ ions is between: 0.44 to 0.64

PRECAUTIONS
 Use a very fine capillary for applying the spot. Diameter of the
sport should not exceed 2 to 4 mm.
 The distance of the spot from the end should not be more than
3 cm.
 Allow the spot to dry before putting another drop on it.
 The spot must be dried before keeping the strip in the bottle or
jar containing the solvent.
 Do not put the strip in oven or above the flame for drying.
 The spot should not dip in the mobile phase.
 The strip should not touch the walls of the bottle or the jar.
 Do not disturb the jar or bottle after putting the strip.
 In order to get good results, saturate the jar with the vapors of
the mobile phase. This can be done by shaking the jar up and
down after pouring in the mobile phase.

BIBLIOGRAPHY

 https://en.wikipedia.org/wiki/Thinlayer_chromatography
 https://www.khanacademy.org/testprep/mcat/chemical-
processes/separationspurifications/a/principles-of-
chromatography
 https://en.wikipedia.org/wiki/Retardation_factor
 Comprehensive PRACTICAL PHYSICS class XII