NAME: Glendon Lewis

DATE: February 6, 2015

TITLE: THIN LAYER CHROMATOGRAPHY
AIM: To identify the components in a plant extract using thin layer chromatography
APPARATUS/MATERIALS: Silica TLC plate (plastic backing), Capillary tube, Beakers, Hexane,
Acetone, Hibiscus (plant extract)
METHOD:
1)
2)
3)
4)

A baseline was drawn approximately 1cm from the bottom of the TLC plate.
The extract was obtained and dissolved in 60% hexane and 40% acetone,
A clean capillary tube was used to introduce dissolved extract into TLC plate.
The spotted TLC plate was placed into the beaker consisting of the suitable solvent 60%

hexane, 40% acetone.

5) The plate was removed when the solvent front was approximately 0.5cm from the top of the
plate.
6) All visible spots were circled with a dull pencil on the chromatogram.
7) Two spots were identified and used to calculate the RF value.
OBSERVATION:

TLC PLATE SHOWING RESULTS OF THE SEPARATION OF COMPONENTS OF A SOLVENT

MIXTURE OF HEXANE AND ACETONE
CALCULATIONS:

Retention Factor = Distance moved by solute / distance moved by solvent

Distance moved by solvent = 5cm
Therefore RF value = distance moved by solute / 5
For Spot A, distance moved = 1.9cm
Therefore RF = 1.9cm/ 5 cm
= 0.38
For Spot D, distance moved = 3.6cm
Therefore RF = 3.6cm/5cm
= 0.72
Rf Value of B = 0.42
Rf Value of C = 0.44
DISCUSSION:
Chromatography is a technique used the separation and analysis of colored dyes and plant pigments.
There are many methods of chromatography. Four of these include, paper chromatography, thin layer
chromatography, gas liquid chromatography and column chromatography .All chromatography methods
use the same principle of mobile phase a liquid or a gas moving past a stationary phase. The stationary
phase may be a solid onto which the solutes are adsorbed. The solute molecules partition themselves
between the stationary phase and the moving phase. this partition is not a true equilibrium since the
mobile phase is constantly moving past the area on the stationary phase where a particular solute is
adsorbed or dissolved .Thus a fresh partition with pure mobile phase , is continually being setup. The
mobile phase should never become saturated with the solute.
In thin layer chromatography the stationary phase is a thin layer of a solid such as alumina or silica
supported on an inert base such as glass aluminum foil or insoluble plastic. This plate can be prepared in
the laboratory b using a slurry o the powder and then carefully oven drying. The practice is necessary to
make standard and uniform coverings of the plates. They can also be obtained from laboratory suppliers
which is the normal practice. The plate is relatively small of around 5 cm by 12 cm. The mixture to be
analyzed is spotted at the bottom of the TLC Plate and allowed to dry. This area is labelled as the base
line or the lie of origin. The plate is placed in a closed vessel such as a covered container containing the

mobile phase so that the liquid level is below the spot. TLC Is very efficient and the solvent ascends the
plate by capillary action. During TLC the liquid is filling spaces between solid particles. This technique is
usually done in a closed vessel to ensure that the atmosphere is saturated with solvent vapor and that
evaporation from the plate is minimized before the run is complete. The partition here is between the
solute adsorbed onto the silica or alumina particles via polar and hydrogen bonded interactions and to a
lesser extent acid base attractions due to the slight acidity of the silica or alumina plate. The plate is
removed when the solvent front approaches the top of the plate and the position of the solvent front is
recorded before it is dried. This allows the calculation of the retention factor.TLC has many uses such as
determining the progress of reactions and in determining the components of products.
SOURCE OF ERROR:
1. Contaminations may have occurred while using the capillary tube.
2. Enough solution may not have been spotted to ensure that all the components would be produced.
PRECAUTIONS:
1. Use a very dull pencil instead of a sharp pencil to make markings on the TLC plate as the layer of
silica powder is very fragile and thus will break easily.
2. Ensure TLC plate does not settle on the side of the container as it will cause non-uniform
movement of solvent.
CONCLUSION:
Components were separated and identified using thin layer chromatography. Various Spots were
observed. The retention factors for spots A, B, C and D respectively are 0.38, 0.42, 0.44 and 0.72.