ANALYTICAL

CHEMISTRY
(Chromatography)
Dr.S.SURESH
Assistant Professor
Email:avitsureshindia@gm
ail.com

Adsorption
Chromatography

Separation based on their adsorption onto the surface

of solid (stationary phase).
A solid which is insoluble in the solvent chosen may
be used as the fixed phase.
The mode of interaction between the components of
the mixture and the fixed phase is adsorption, Hence
the method is called adsorption chromatography.
Ex; Column chromatography, TLC etc.,

Partition chromatography
Partition chromatography, is a chromatographic
technique in which the solute is separated based
on their partition between a liquid mobile phase
and a liquid stationary phase coated on a solid
support.
The support material used in partition
chromatography is usually silica

Thin-Layer Chromatography
Here the mobile phase is a liquid
Flowing past a thin layer of powder on a
solid support.
Substances that are less attracted to
the solid or are more soluble in the
liquid move faster
Stationary phase
glass or plastic plates coated with thin
layer of adsorbent
Silica gel, alumina, cellulose
Mobile phase
Solvent or mixture of solvents
Fall, 2008

Thin layer chromatography

It involves the separation of substances of a mixture
over a thin layer of an adsorbent.
A thin layer(about 0.2mm thick) of an adsorbent (silica
gel or alumina) is spread over a glass plate of suitable
size. The plate is known as thin layer chromatography
plate.
The solution of the mixture to be separated is applied as
a small spot about 2 cm above one end of the TLC plate.
The glass plate is then placed in closed jar containing
the solvent (Below 2cm height).
As the solvent in the jar moves up the plate, the
components of mixture move up the plate to different
distances, depending on this degree of adsorption
separation takes place.

The relative adsorption of each component of the

mixture is expressed in terms of its retention factor
i.e., Rf value

Rf value
Rf values are used in identification of each of the
component.
The retention factor, or Rf, is defined as the distance traveled by
the compound divided by the distance traveled by the solvent

For example, if a compound travels 2.1 cm and the solvent front

travels 2.8 cm, the Rf is 0.75:

Paper Chromatography
In this technique, the stationary phase is
considered to be the cellulose network of
the paper.
The
mobile
phase
known
as
the
developing solvent consists of either one
solvent or a mixture of different solvents.

Paper Chromatography
In this the mixture of compounds is
applied on the paper as a spot little
above the lower end and then this end
is dipped in the solvent. When the
solvent has risen more than two third
length of the paper, then it is removed
from the solvent. The paper is dried and
is known as chromatogram.
Now, the spots of different compounds
can be visualised using some suitable
chemicals.

Rf value
The ratio of the distance travelled by the compound in
a particular solvent to that the distance travelled by
the solvent is a constant and is known as retention
factor (Rf).

For example, if a compound travels 2.1 cm and the

solvent front travels 2.8 cm, the Rf is 0.75:

Ion Exchange
Chromatography

Ion Exchange Chromatography

Ion exchange chromatography is a special
name given to column chromatography
when the stationary phase is an ion
exchange resin.
Synthetic ion exchange resins are high
molecular
weight
polymeric
materials
containing large number of ionic functional
groups per molecule. Cation exchangers
contain sulphonic acid groups (RSO3 H+) or
carboxylic acid groups (RCOO H+). Anion
exchange resin contains amines attached
to the polymer molecule RN(CH2)3+OH).

Ion Exchange Chromatography

In ion exchange chromatography a column is packed with an
acid resin and treated first with hydrochloric acid to make sure
that all exchange points were occupied by hydrogen ions.
A mixture of rare earths as their chlorides is sent down the
column. This resulted in the displacement of hydrogen ions by
rare earth cations.
The rare earth ions could then be eluted one after another. Since
elution with water was very slow, a solution of citric acid was
used as eluting solvent.
The cations moved at different rates depending on the stability
of the corresponding complex with citric acid.

Gas Chromatography

Gas Chromatography (GC)

GC is currently one of the most popular methods for
separating and analyzing compounds.
This is due to its high resolution, low limits of
detection, speed, accuracy and reproducibility.
GC can be applied to the separation of any
compound that is either naturally volatile (i.e., readily
goes into the gas phase) or can be converted to a
volatile derivative.
A simple GC system consists of:
. G 1. Gas source
2. Injector or sample application system
3. Chromatographic column
4. Detector & computer or recorder

GC structure

Gas source
It provides all the necessary gas supplies.

The most widely used gases are H, He, N 2 and

air.

Injector
We use a syringe to inject sample onto the
column.
It is situated inside a thermostatically
controlled enclosure.

Column
It contains the column and an oven.
The column is the essential device to
achieve the necessary separation.
The oven is used to control the
column temperature.
The column has two kinds:
Packed column
Capillary column

Detector
There are wide range of detectors available each
having unique operating parameters and its own
performing characteristics.
The output of detector is electronically modified.

Types of
detectors
The two types of detectors are
TCD: Thermal Conductivity Detector
ECD: Electron Capture Detector

TCD Detector
A
TCD
detector
consists
of
an
electrically-heated
wire.
The
temperature of the sensing element
depends on the thermal conductivity of
the gas flowing around it.
Changes in thermal conductivity, such as
when organic molecules displace some of
the carrier gas, cause a temperature rise
in the element which is sensed as a
change in resistance.
The TCD is not as sensitive as other

ECD Detector
Uses a radiactive Beta
emitter (electrons) to
ionize
some
of
the
carrier
gas
and
produces
a
current
between a biased pair
of electrodes.
When
an
organic
molecule that contains
electronegative
functional groups, such
as
halogens,
phosphorous and nitro
groups, pass by the