DISCUSSION

In this experiment, Thin Layer Chromatography (TLC) was performed for separating organic
compounds. The purpose of this experiment are to distinguish polar and non-polar solvent, to
perform analysis technique using TLC and to 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. First step is dissolving the sample with a
volatile solvent, ethanol solution. Then, spotting is using a capillary tube to transfer or make a
spot of dilute sample at the origin (0.7 cm from the bottom of TLC plate). Development consists
of placing the bottom plate into a beaker containing 5mL of chloroform. The beaker was closed
with petri dish as chloroform is evaporate quickly into surrounding air. 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 had travelled to the front line (0.5 cm from top of TLC plate). the plate was
removed. 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 coloured compounds.
TLC plate was placed under UV lamp and the spots was marked using pencil. TLC plate is
impregnated with a fluorescent material that glows under ultraviolet light.
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;

System Nisbah Mixture

A 0.5:4.5
B 2.5:2.5
C 2.5:2.5
D 4.7:0.3
E 4.7:0.3
hexane:chloroform
hexane:dichloromethane
hexane:diethyl ether
chloroform:methanol
dichloromethane:methanol

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, dichloromethane is polar, diethyl ether is non-polar, methanol is polar and
chloroform is non-polar. Hexane, chloroform and diethyl ether are non-polar, then system A,
B and D are non-polar. In system D, thought methanol is polar but due to presence of 4.5
chloroform in 0.3 methanol which decreasing the polarity of system D. Meanwhile,
dichloromethane and methanol are polar, then system B and E are polar.
Based on the result obtained, when hexane and chloroform was used no movement was
spotted on TLC plate. When dichloromethane and methanol was used not much movement was
observed on the TLC plate. When hexane and dichloromethane was used as the solvent system
more movement was observed in the spots. When hexane and diethyl ether was used a lot of
movement was observed. The solvent system that worked best was probably hexane:diethyl
ether and hexane:dichloromethane because more movement was observed on TLC plate for
both sample. And least suitable system solvent probably hexane:chloroform because less
movement was observed on TLC plate for both sample.
The marked spots were determined. The distance of the origin to the solvent at front
line (b) was measured. The distance of origin to the centre of the spot (a). The Rf value was
calculated by using formula below. For sample 27, the Rf value obtain is 0.82 and Rf value for
sample 28 is 0.79.
distance travelled by the compound (a)
𝑅𝑓 =
distance travelled by the solvents (b)
During experiment, there were some errors where it causes to 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 ensure
the TLC plate is placed in the beaker evenly and do not tilt the TLC plate.

CONCLUSION

The thin layer chromatography (TLC) was conducted successfully thought there were some
errors in the beginning of experiment. However, precaution was taken after the problems was
done. The Rf value for sample 27 and sample 28 are 0.81 and 0.79 respectively. In general,
there are three steps to perform TLC that is spotting, development and visualization.
TLC is widely used in analytical because of its simplicity, low cost, high sensitivity and
speed of separation. The purpose of this experiment has been achieved which is the Rf value
was determined for both samples, the analysis technique by using TLC was successfully
performed and polarity of the solvent was able to identify.
 There are many uses of TCL in our daily lives. Some of the these are: in food industry
TCL is used in separating and studying various aspects of food items like colours, sweetness
and preservations. in cosmetics industry TCL is used to separate and analyse different cosmetic
products and their components.

REFERENCES

[1] Lisa Mickey. Thin Layer and Column Chromatography. Retrieved from:
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[3] Thin Layer Chromatography. Retrieved from:
https://people.chem.umass.edu/samal/269/tlc.pdf.
[4] Thin Layer Chromatography. Retrieved from:
https://www.chemguide.co.uk/analysis/chromatography/thinlayer.html.
[5] Thin-layer Chromatography. Retrieved from:
https://www.chem.ucla.edu/~bacher/General/30BL/tips/TLC1.html.