THIN LAYER

CHROMATOGRAP
HY

PRESENTED BY : AVIRAL UPADHYAY SECTION : D

SCHOLAR NO: 191113269 ROLL NO: 19D104
BG Image: https://images.app.goo.gl/GeaZhMEtpgxwaYY67
INTRODUCTION
• It is a chromatographic technique used to
separate non-volatile mixtures.
• It is a solid-liquid technique in which the
stationary phase is a solid and the moving phase
is a liquid.
• It works on the principle that different organic
compounds will have different polarities and
adsorption to the two phases between which
they are to be partitioned.
THEORY AND OVER VIEW

Image: https://en.wikipedia.org/wiki/File%3ATlc_sequence.svg

TLC Plate dipped The position of

TLC Plate Capillary action takes place; organic
in the liquid compounds are
with compounds which are more polar travel
mobile located via
added small distances on the plate (as the
phase(solvent) visualization
analytes. stationary phase is more polar) and thus
by edge. techniques to
get separated from the mixture of the less
calculate
polar compounds which travel large
retention factor
distances.
- The stationary phase is sprayed on an unreactive plate and dried,
the resultant known as TLC plate.

- The TLC plate is then prepared by adding analytes (sample

solutions of mixture of compounds to be separated and pure
compounds for reference) and dried.

- It is then dipped in the developing chamber which contains a

solvent as mobile phase.

- Solvent rises vertically up on the TLC plate due to capillary action.

- Due to difference in polarity of different organic compounds, they

are attracted differently to the rising solvent and stationary
adsorbent, and thus travel different heights on the plate.

- The plate is finally visualized and compounds separated from the

mixture are spotted and identified with the help of reference.
BG Image: https://youtu.be/qdmKGskCyh8
HOW IT WORKS: EXAMPLE OF SILICA
GEL
• It is a network solid of silicon dioxide with Image : https://youtu.be/I4u_1ST7Ezk

alternating silicon and oxygen atoms in every

direction.
• At the surface of this material there are many polar
hydroxyl groups which participate in dipole-dipole
interactions with other polar substances.
• When this plate with added analytes is put in the
solvent chamber, the solvent begins to rise and it hits
the added samples and drag the components up with
it.
• The two components A and B interact with the polar
hydroxyl groups differently.
• The more polar component A interacts strongly with
the hydroxyl groups and gets strongly adsorbed on
the plate, thus shows less vertical movement.
• The less polar component B interacts weakly with
the hydroxyl groups and thus gets weakly adsorbed
on the plate, thus shows more vertical movement.
RETENTION FACTOR VALUE Image : https://youtu.be/I4u_1ST7Ezk

• Retention factor value or Rf value of a particular

compound tells us how far a compound traveled
relative to the solvent front.

Distance of spotted compound from the origin

Rf value = Distance of the solvent front from the origin
_____________________________________________________________________________________________________________

• Every spot gets its own Rf value between 0 and 1.

• Relative Rf value of a compound is calculated Image : https://youtu.be/I4u_1ST7Ezk

with respect to another as the ratio of their

absolute Rf values.
• It can be used to identify compounds due to its
uniqueness to each compound at a given
temperature, layer thickness, sample size and https://youtu.be/I4u_1ST7Ezk

solvent parameters.
 PRECAUTIONARY MEASURES
DURING PREPARATION
Lightly touch capillary tube to
TLC plate.
Change capillary tube every
time while adding a new
analyte to avoid cross
contamintion.
Use pencil to avoid ink
contamination.
Don’t put spots too close to
each other on the origin.
DURING DEVELOPMENT DURING VISUALISATION
Hold the plate with tweezers Hold UV lamp so that light is
by the edge to avoid pointing straight down onto
contamination. table.
Make sure solvent is below Dry TLC plate properly in a
the origin. hood before visualization.
The solvent chamber should Again, use pencil for spotting
be properly capped to avoid the compounds. Do it lightly.
leakage of solvent vapours.
Make sure the TLC plate is not
touching sides of the filter
paper.
Refrain from moving or
bumping the chamber while
mixture is eluting.
Solvent front should not
touch end of the plate.
 Purity of sample:
If any impurity is Biochemical analysis: the
present, the constituents of body
analyte shows fluids can be separated.
extra spots.

Pharmaceutical
Identification: use: For
Natural products detection of
like volatile oil APPLICATIONS impurities in
and steroids can pharmacopoeial
be identified. chemicals.

Food and
Reaction cosmetics: To
Examination: The separate and
test may tell us if identify colors
the reaction is and
complete. preservatives
 ADVANTAGES AND DISADVANTAGES
ADVANTAGES
Very simple and Inexpensive technique
Once the best solvent is found, it can be
applied to other techniques such as high
performance liquid chromatography.
DISADVANTAGES
TLC plates do not have long stationary phases,
hence the length of separation is limited.
The detection limit is a lot higher compared to
other chromatographic techniques.

More than one compound can be separated on TLC plates operates as an open system, hence
a TLC plate. humidity and temperature affects the results
of the chromatogram.
It is possible to use several different solvents
depending on our desired result.
The chromatographic conditions can be easily
modified for resolution of a specific
components.

The identification of most compounds can be

done simply by checking Rf values.
 BIBLIOGRAPHY
Engineering Chemistry, Jain and Jain

Chemistry LibreTexts libraries, Supported by California State University

https://chem.libretexts.org

Wikipedia
https://www.wikipedia.org

Khan Academy
https://www.youtube.com/user/khanacademy