NEW WAY SENIOR

SECONDARY SCHOOL
SESSION-2019-20

NAME – ABHINAV SINGH

CLASS – X11-A1
SUBJECT- CHEMISTRY
TOPIC- CHROMATOGRAPHY
BOARD ROLL NO.-
SUBMITTED TO:- MR. ANUJ GUPTA
INDEX
 Acknowledgement
 Bonafide certificate
 Introduction
 History
 Types of Chromatography
1. Column chromatography
2. Thin layer chromatography
3. Paper chromatography
 Application of chromatography
 Experiment
 Conclusion
 Bibliography
ACKNOWLEDGEMENT

I would like to express my special thanks of gratitude to my Chemistry

teacher “Mr. ANUJ GUPTA”for their able guidance and support in
completing my Project.

I woud like to extend my gratitude to the principal ma’am ”Miss Jyoti

Vanchoo” for providing me with all the facilities that was required.
Bonafide Certificate
This is to certify that ABHINAV SINGH of class X11 of NEW WAY
SENIOR SECONDARY SCHOOL has completed the project on the
topic CHROMATOGRAPHY as per the norms of CBSE curriculum in
academic session 2019-20.

Internal Examiner’s External Examiner’s

Signature signature

Principal’s signature:
INTRODUCTION
Chromatography is a laboratory technique for the separation of a
mixture. The mixture is dissolved in a fluid called the mobile
phase, which carries it through a structure holding another
material called the stationary phase. The various constituents of the
mixture travel at different speeds, causing them to separate. The
separation is based on differential partitioning between the mobile
and stationary phases. Subtle differences in a compound's partition
coefficient result in differential retention on the stationary phase
and thus affect the separation.

Chromatography may be preparative or analytical. The purpose of

preparative chromatography is to separate the components of a
mixture for later use, and is thus a form of purification. Analytical
chromatography is done normally with smaller amounts of material
and is for establishing the presence or measuring the relative
proportions of analytes in a mixture. The two are not mutually
exclusive.

Some materials appear homogenous, but are actually a combination

of substances. For example, green plants contain a mixture of
different pigments. In addition, the black ink in the pens that are
used in this experiment is a mixture of different colored materials.
In many instances, we can separate these materials by dissolving
them in an appropriate liquid and allowing them to move through
an absorbent matrix, like paper. Chromatography is a method used
by scientists for separating organic and inorganic compounds so
that they can be analyzed and studied. By analyzing a compound, a
scientist can figure out what makes up that compound.
Chromatography is a great physical method for observing mixtures
and solvents. The word chromatography means "color writing"
which is a way that a chemist can test liquid mixtures. While
studying the coloring materials in plant life, a Russian botanist
invented chromatography in 1903. His name was M.S. Tswett.
Chromatography is such an important technique that two nobel
prizes have been awarded to chromatographers. Over 60% of
chemical analysis worldwide is currently done with
chromatography or a variation thereon. Chromatography is used in
many different ways. Some people use chromatography to find out
what is in a solid or a liquid. It is also used to determine what
unknown substances are. The Police, F.B.I., and other detectives use
chromatography when trying to solve a crime. It is also used to
determine the presence of cocaine in urine, alcohol in blood, PCB's
in fish, and lead in water. Chromatography is used by many
different people in many different ways. Chromatography is based
on differential migration. The solutes in a mobile phase go through
a stationary phase. Solutes with a greater affinity for the mobile
phase will spend more time in this phase than the solutes that
prefer the stationary phase. As the solutes move through the
stationary phase they separate. This is called chromatographic
development.
HISTORY
Chromatography was first employed in Russia by the Italian-
born scientist Mikhail Tsvet in 1900. He continued to work with
chromatography in the first decade of the 20th century,
primarily for the separation of plant pigments such
as chlorophyll, carotenes, and xanthophylls. Since these
components have different colors (green, orange, and yellow,
respectively) they gave the technique its name. New types of
chromatography developed during the 1930s and 1940s made
the technique useful for many separation processes.
Chromatography technique developed substantially as a result
of the work of Archer John Porter Martin and Richard Laurence
Millington Synge during the 1940s and 1950s, for which they
won the 1952 Nobel Prize in Chemistry. They established the
principles and basic techniques of partition chromatography,
and their work encouraged the rapid development of several
chromatographic methods: paper chromatography, gas
chromatography, and what would become known as high-
performance liquid chromatography. Since then, the
technology has advanced rapidly. Researchers found that the
main principles of Tsvet's chromatography could be applied in
many different ways, resulting in the different varieties of
chromatography described below. Advances are continually
improving the technical performance of chromatography,
allowing the separation of increasingly similar molecules.
The earliest use of chromatography—passing a mixture
through an inert material to create separation of the solution
components based on differential adsorption—is sometimes
attributed to German chemist Friedlieb Ferdinand Runge, who
in 1855 described the use of paper to analyze dyes. Runge
dropped spots of different inorganic chemicals onto circles of
filter paper already impregnated with another chemical, and
reactions between the different chemicals created unique color
patterns.[2] According to historical analysis of L. S. Ettre,
however, Runge's work had "nothing to do with
chromatography" (and instead should be considered a
precursor of chemical spot tests such as the Schiff test).[3][4]

In the 1860s, Christian Friedrich Schönbein and his student

Friedrich Goppelsroeder published the first attempts to study
the different rates at which different substances move through
filter paper.[5][6] Schönbein, who thought capillary action
(rather than adsorption) was responsible for the movement,
called the technique capillary analysis, and Goppelsroeder
spent much of his career using capillary analysis to test the
movement rates of a wide variety of substances.
TYPES OF
CHROMATOGRAPHY
Column chromatography
Column chromatography is a separation technique in which the
stationary bed is within a tube. The particles of the solid
stationary phase or the support coated with a liquid stationary
phase may fill the whole inside volume of the tube (packed
column) or be concentrated on or along the inside tube wall
leaving an open, unrestricted path for the mobile phase in the
middle part of the tube (open tubular column). Differences in
rates of movement through the medium are calculated to
different retention times of the sample.
In 1978, W. Clark still introduced a modified version of column
chromatography called flash column chromatography (flash).
The technique is very similar to the traditional column
chromatography, except for that the solvent is driven through
the column by applying positive pressure. This allowed most
separations to be performed in less than 20 minutes, with
improved separations compared to the old method. Modern
flash chromatography systems are sold as pre-packed plastic
cartridges, and the solvent is pumped through the cartridge.
 Thin layer chromatography
Thin layer chromatography (TLC) is a widely employed
laboratory technique used to separate different biochemical’s
on the basis of their size and is similar to paper
chromatography. However, instead of using a stationary phase
of paper, it involves a stationary phase of a thin layer
of adsorbent like silica gel, alumina, or cellulose on a flat,
inert substrate. TLC is very versatile; multiple samples can be
separated simultaneously on the same layer, making it very
useful for screening applications such as testing drug levels and
water purity. Possibility of cross-contamination is low since
each separation is performed on a new layer. Compared to
paper, it has the advantage of faster runs, better separations,
better quantitative analysis, and the choice between different
adsorbents. For even better resolution and faster separation
that utilizes less solvent, high-performance TLC can be used. An
older popular use had been to differentiate chromosomes by
observing distance in gel (separation of was a separate step).
 Paper chromatography
Paper chromatography is a technique that involves placing a
small dot or line of sample solution onto a strip of
chromatography paper. The paper is placed in a container with
a shallow layer of solvent and sealed. As the solvent rises
through the paper, it meets the sample mixture, which starts to
travel up the paper with the solvent. This paper is made
of cellulose, a polar substance, and the compounds within the
mixture travel farther if they are non-polar. More polar
substances bond with the cellulose paper more quickly, and
therefore do not travel as far
APPLICATIONS OF
CHROMATOGRAPHY
Chromatography is a process used to separate mixtures. The
word chromatography is derived from the Greek words
"khroma" and "graphein" meaning "color" and "to write" or "to
represent". The chromatography technique is first discovered
by Russian Biologists, Dr Michael Tswett in 1906 for the
separation of colored plant pigment on a column of
alumina. Now-a-days various types of chromatography are used
to separate almost any given mixture whether colored or
colorless into its component. Chromatography may be regarded
as an analytical technique employed for the purification and
separation of organic and inorganic substances. It is also found
useful for the fractionation of complex mixture, separation of
closely related compounds, such as isomers and in the isolation
of unstable substances. Although there are several
different types of chromatography, in each case a substance
is placed onto or into a medium and a solvent is passed through
the test substance. Inchromatography science, the solvent is
called "the mobile phase" or "the carrier fluid" and the medium
is called "the stationary phase".
EXPERIMENT
Aim: Separation of component from a mixture of red and blue
inks.
Materials Required : Watman filter paper , capillary tube , pencil ,
distilled water , isopropyl alcohol, glass rod , red and blue ink,
chromatographic chamber with lid, measuring jar, scale.
Procedure:

 Take a Watman filter paper strip and using a pencil draw a

horizontal line 4cm from one end of the paper. Then draw
another line lengthwise (vertically) from the centre of the
paper. Name the point at which the two lines intersect as
P.
 Using a fine capillary tube, put a drop of the mixture of red
and blue inks at the point P. let it dry in air.
 Put another drop on the same spot and dry again, so that
the spot is rich in the mixture.
 Pore equal amounts of isopropyl alcohol and distilled
water into a chromatographic chamber and mix it well
using a glass rod. This is used as the solvent.
 Suspend the filter paper vertically in the chromatographic
chamber containing the solvent in such a way that the
pencil line remains about 2cm above the solvent level.
 Close the jar with its lid and keep it undistributed.
 Notice the rising solvent along with the red and blue inks.
After the solvent has risen about 15 cm you will notice two
different spots f blue and red colors on the filter paper.
  Take the filter paper out of the jar and using a pencil mark
the distance that the solvent has risen on the paper. This is
called the solvent front.
 Dry the filter paper and put pencil marks at the centre of
the red and blue ink spots.
 Measure the distance of the two spots from the original
line and the distance of the solvent from the original line.
 Calculate the Rf values of the red and blue inks using the
formula.

Rf Value = Distance from Baseline travelled by Solute

Distance from Baseline travelled by Solvent (Solvent Front)

Observation can be recorded as follows:-

Si no. Components Distance Distance Rf value
travelled by travelled by
the the solvent
component from the
from the original
original line(cm)
line(cm)
1. Red
2. Blue
 CONCLUSION
Chromatography is one of several separation techniques
defined as differential migration from a narrow initial
zone. Electrophoresis is another member of this group. In this
case, the driving force is an electric field, which exerts different
forces on solutes of different ionic charge. The resistive force is
the viscosity of the non flowing solvent. The combination of
these forces yields ion mobilities peculiar to each solute.
Chromatography has numerous applications
in biological and chemical fields. It is widely used
in biochemical research for the separation and identification
of chemical compounds of biological origin. In
the petroleum industry the technique is employed to analyze
complex mixtures of hydrocarbons.
As a separation method, chromatography has a number of
advantages over older techniques—crystallization, solvent
extraction, and distillation, for example. It is capable of
separating all the components of a multicomponent chemical
mixture without requiring an extensive foreknowledge of the
identity, number, or relative amounts of the substances
present. It is versatile in that it can deal with molecular species
ranging in size from viruses composed of millions of atoms to
the smallest of all molecules—hydrogen—which contains only
two; furthermore, it can be used with large or small amounts of
material.
BIBLIOGRAPHY
Help from internet
 www.chromatography.com

 www.wikipedia.com

 Class 12th Chemistry NCERT book

Help from Library.

Help from Teacher.