TYPES OF CHROMATOGRAPHY

1. Types of
Chromatography
Presented by : Prajapati Dimple B.
M.sc sem -2
Department of Life sciences ,Patan

2. Content
Types of Chromatography
Paper Chromatography
Gas chromatography
Thin layer Chromatography
High performance thin layer Chromatography

3. What is Paper Chromatography?
Paper Chromatography is an analytical
method used to separate colored
chemicals or substances.

4. Principle of Paper Chromatography
The principle involved is partition
Chromatography wherein the substances are
distributed or partitioned between liquid phases.
One phase is the water , which is held in the pores
of the filter paper used ; and other is the Mobile
phase which moves over the paper.

5. Paper chromatography

6. Application of Paper Chromatography
Paper Chromatography is specially used for the
separation of a mixture having polar and non polar
compounds.
For separation of amino acids.
It is used to determine organic compounds ,
biochemicals in urine etc.
In the pharma sector it is used for the
determination of hormones ,drugs etc.
Sometimes it is used for evaluation of inorganic
compounds like salts and complex.

7. What is gas chromatography?
Gas chromatography (GC) is a common type of
Chromatography used in analytical chemistry for
separating and analyzing compounds that can be
vaporized without decomposition.
In gas chromatography , the Mobile phase ( or ''
moving phase'') is a carrier gas , usually an inert gas
such as helium or an unreactive gas such as nitrogen .
The stationary phase is microscopic layer of liquid or
polymer on an inert solid support , inside a piece of
glass or metal tubing called a column.

8. Principle of gas chromatography
 The sample solution injected into the instrument
enteres a gas stream which transport the sample
into a separation tube known as the " column" (
Helium or nitrogen is used as the so called carrier
gas) .
The various components are separated inside the
column.

9. Gas chromatography

10. Application of gas chromatography
Gas chromatography ( GC) is an instrumental
technique used forensically in drug analysis ,arson
, toxicology analyses of other organic compounds.
Gas chromatography is a physical separation
method in where volatile mixtures are separated.

11. What is TLC ?
Chromatography in which compounds are
separated on a thin layer of absorbent material ,
typically a coating of silica gel on a glass plate or
plastic sheet.
 Thin layer of absorbent is known as the
stationary phase .
After the sample has been applied on the plate ,
a solvent or solvent mixture ( known as the
Mobile phase) is drawn up the plate via capillary
action.

12. Principle of TLC
TLC works on the same Principles.
 In thin layer Chromatography , the stationary
phase is a polar absorbent , usually finely ground
alumina or silica particles.
This absorbent is coated on a glass slide or plastic
sheet creating a thin layer of the particular
stationary phase.

14. Application of TLC
Purity of sample:
• Purity of sample can be carried out with TLC.
Identification of compounds :
• Thin layer Chromatography can be employed in
purification , isolation and identification of
natural products like volatile oil or essential oil,
fixed oil, waxes, terpenes ,alkaloids
,glycosides,steroid etc.
Examination of reaction

15. What is HPTLC?
High performance thin layer Chromatography
(HPTLC) is an enhanced form of thin layer
Chromatography (TLC).
A number of enhancement can be made to the
basic method of thin layer Chromatography to
automate the different steps, to increase the
resolution achieved and to follow more accurate
quantitative measurements.

16. Principle of HPTLC
HPTLC have similar approach and employ the same
physical principles of TLC ( adsorption Chromatography )
i.e., the principle of separation adsorption.
 The Mobile phase solvent flows through because of
capillary action .the components move according to
their affinities towards the absorbent .
The components with more affinity towards the
stationary phase travels slower . the components with
lesser affinity towards the stationary phase travels faster
.Thus the components are separated on a
Chromatographic plate.

18. Application of HPTLC
Application of TLC and HPTLC in the analysis of
semipermanent hair dyes.
Application of HPTLC for the determination of
active ingredients in herbal and pharmaceutical
formulation.
Cosmetic and environmental analysis.
Metallurgy , electroplating.
Toxicology , forensic analysis.

19. References
Principles & technique of Biochemistry &Molecular
Biology edited by Keith Wilson & Jhon walker
 https: // en . m. Wikipedia. Org
https:// www.slideshare.net
www.Biologydiscussion.com

20. Thank You