DISCUSSION

Chromatography is used to separate mixtures of substances into their components. All forms of
chromatography work on the same principle. They all have a stationary phase (solid, or liquid
supported on solid) and a mobile phase (liquid or gas). The mobile phase flows through the stationary
phase and carries along the components of the mixture with it. Different components will travel at
different rates.
Thin Layer Chromatography (TLC) plate is a sheet of glass, metal, or plastic which is coated
with a thin layer of a solid adsorbent (usually silica or alumina). To separate the sample wanted to
analyse, small amount of the sample is spotted near the bottom of this plate. The plate is then placed in
a shallow pool of a solvent in a developing chamber so that only the very bottom of the plate is in the
liquid. This liquid is at the mobile phase, and it slowly rises up the TLC plate by capillary action.
As the solvent moves past the spot that was applied, equilibrium is established for each
component of the mixture between the molecules of that component which are absorbed by the solid
and the molecules in the solution. In principle, the components will differ in solubility and in the
strength of their absorption to the absorbent and some components will be carried farther up the plate
than others. When the solvent has reached the top of the plate, the plate is removed from the
developing chamber, dried, and the separated components of the mixture are visualized. If the
compounds are coloured, visualization is straightforward. Usually the compounds are not coloured, so
a UV lamp is used to visualize the plates.
In this experiment, since we are using oils, thus, our compounds are in yellow in colour, thus
our spots are in yellowish-brown. The plates are immersed in three different mixtures of compounds,
producing different rates of separation for each of the mixture. From the results, the first solvent is
more suitable for the soybean oil. This is because the separation occurs more than in other solvents.
For the second solvent, the spotted shown that there are very less separation while there are half of the

separation occurs in solvent 3. Five separations occur in the mixture of hexane, diethyl ether and acetic
acid for 90:30:1 ratio. Thus, this solvent is called the idea solvent. The organic solvent are needed to
mix to make the solvent either more polar or non-polarity.

The strength with which an organic compound binds to an adsorbent depends on the strength of
the following types of interactions: ion-dipole, dipole-dipole, hydrogen bonding, dipole induced
dipole, and van der Waals forces. The highly polar molecules will interact fairly strongly with the
polar SiOH groups at the surface of these absorbents, and will tend to stick or adsorb onto the fine
particles of the adsorbent while weakly polar molecules are held less tightly. Weakly polar molecules
generally tend to move through the absorbent more rapidly than the polar species. Thus, the
compounds needed to follow the elution order given above.

Retention Factor or Rf, is defined as the distance travelled by the compound divided by the
distance travelled by the solvent.

The larger an Rf of a compound, the larger the distance it travels on the TLC plate. The compound
with the larger Rf is less polar than the compound with small Rf because it interacts less strongly with
the polar adsorbent on the TLC plate. The Rf can provide the evidence to identity the compound in the
sample. If two substances have the same Rf value, they are likely to be the same compound. If they
have different Rf values, they are definitely different compounds.
From the experiment, the safety precautions need to take in this experiment is to make sure the
solvent fully absorb to the top line of the plate. If we take out the plate before the solvent absorb until
the top, the spot will be not fully travel or separated. Secondly, we need to make sure the developing
tanks are closed to make sure the solvent not evaporated into the air. Thirdly, we need to make sure not
to place the spot of the sample to little or too much. Because if the spotted is too low, the separation
would not be able to be absorbed or if it is too much or too concentrated, the spots will be streak or run
together.