DISCUSSION

In this experiment, Thin Layer Chromatography (TLC) was performed for separating
organic compounds. The purpose of this experiment is to distinguish polar and non-polar
solvent, to perform analysis technique using TLC and differentiate the retention factor, Rf
for different compounds.
TLC is a chromatography technique that being used in separating organic
compounds. The principle of TLC is similar to other chromatographic technique. TLC consists
of three steps that is spotting, development and visualization. TLC is a method to analysis
the mixture by separating compounds in the mixture. There are two unknown samples that
was run on the TLC plate. There are five types of mobile phase (solvent system) used in the
experiment, that is

Label Eluent Composition Ratio of combination of the

components
A Hexane-acetone 1:1
B Hexane-acetone 9:1
C Acetone-water 9:1
D Methanol-acetone-hexane 1:1:2

First step is dissolving the eluent compositions according to the ratio of combination
of the components. Then, placed the green apple food coloring sample and yellow egg
coloring sample using a dropper at the origin (1.5cm from the bottom of the TLC plate and
paper plate). Development consists of placing the bottom plates into the labeled beaker
which were containing 20ml of the mixture of solvent. When the solvent travels up to the
plate, it is noticed that the original spot also moves over (as shown in results). When the
solvent travelled to the front line (1.5cm from bottom of TLC plate). The plate was removed
after 5 minutes. The plate was let to evaporate before marked the spot with pencil. The last
step is visualization which is the spots was observed after development with a colored
compound.
If the solvent system is very non-polar, the non-polar components of the sample will
travel further up the TLC plate than the polar components. If the solvent system is very
polar then the polar component will travel further up the TLC plate than the non-polar
components. Hexane is non-polar; acetone and methanol are polar and water is polar
covalent. Hexane and acetone are non-polar and polar respectively, then system A is
immiscible. In system B, thought acetone is polar but due to presence of 18ml of hexane in
2ml of acetone which is decreasing the polarity of the system D. Meanwhile, acetone and
water are polar, then system C is polar. In system D 5ml of methanol (polar), 5ml of acetone
polar) and 10 ml of hexane (non-polar) so it also considered immiscible.
Based on the result obtained, when hexane and acetone (9:1) was used no
movement was spotted on TLC plate and paper plate. When methanol, acetone and hexane
was used not much movement was observed on the TLC plate and paper plate. Acetone and
water were used as the solvent system a lot of movement was observed in the spots. The
solvent system that worked best was probably acetone and water because more movement
was observed on TLC plate and paper plate for both samples. Meanwhile, least suitable
system solvent probably hexane and acetone (9:1) because less movement was observed on
TLC plate and paper plate for both samples.
The marked spots were determined. The distance of the origin to the solvent at front
line (y) was measured. The distance of origin to the centre of the spot (x). The Rf value was
calculated by using formula below. For sample A, TLC plate the Rf value obtain is 0.89
(green) and 0.92 (yellow) while the paper plate Rf value is 0.87 (green) and 0.96 (yellow).

Rf = [Distance travelled by the compound (y)]

[Distance travelled by the solvents (x)]
The system B, C and D Rf value also calculate by the formula. The table A and table B shown
the result for TLC plate and paper plate.
During experiment, there were some errors where it causes to some incorrect data.
These errors were over-large spots that cause the spot difficult to be identified. The
problems could be avoided by spotting the sample around 1-2 mm in diameter and also
avoid overlapping of other components spots. Some precautions should be taken are the
sample should be above the solvent level, there should be enough solvents to travel up the
length of TLC plate and paper plate. Moreover, ensure the TLC plate and paper plate are
placed in the beaker evenly and do not tilt the TLC plate.