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CHEM 153

Lab Report (Experiment 2)

Lance Macellones July 1, 2019

Matthew Justin Nolasco Dr. Aileen Angcajas
Maria Angela Olinan BSN – 1 NC

VI. Analysis
1. Discuss the principle behind the thin layer chromatography. Indicate its limitation of use.

Thin layer chromatography is used to separate the components of a mixture using a thin layer of
adsorbent material. TLC is an analytical tool widely used because of its simplicity, relative low cost,
high sensitivity, and speed of separation. TLC functions on the same principle as all chromatography:
a compound will have different solubilities for the mobile (liquid) and stationary (solid) phases, and
this affects the speed at which it migrates. The goal of TLC is to obtain well defined, well separated
spots. However, the length of separation is limited compared to other chromatographic techniques.
TLC operates as an open system, so factors such as humidity and temperature can be consequences
to the results.

2. Compare the R values of your analgesic drug (if both polar and nonpolar solvents) with
your standards. Based on your data, determine the identity of your unknown?

nonpolar - hexane, polar - acetic acid

When comparing two different compounds run under identical chromatography conditions, the
compound with the larger Rf is less polar because it interacts less strongly with the polar adsorbent
on the TLC plate. Considering this and from the results, the Rf of the analgesic drug tends to be higher
than the standard throughout the polar (acetic acid) and nonpolar (hexane) solvents and with
differences between the two components close to 0.1 meaning that the advil is less polar and had
traveled more in terms of distance than the standard. The unknown is the ibuprofen which is the

3. Why do you use 2 different solvent systems?

Proper selection of solvent is important in TLC. The ideal solvent system is simply the system that
gives the best separation. Acids, bases, and strongly polar compounds often produce streaks rather
than spots in neutral solvents. Streaks make it difficult to calculate Rf and may occlude other spots.
Considering this fact, solvents' polarity and volatility affect the result of the whole process. Therefore,
using different solvents help in separating mixtures to obtain varied polarity values since we will use
it to calculate for the Rf.

4. Describe and explain the possible effect on your results of the following errors.
a) You developed your chromatogram too long and you can’t find the solvent front

If you allow your chromatogram to develop too long then the solute and the solvent being
used will travel too far until it will possibly run off the plate. Any components that are
especially soluble in the mobile phase could run off the top of the chromatogram and be lost.
CHEM 153
Lab Report (Experiment 2)

In which case, it will be difficult to calculate the Rf values of both the solute and solvents
without a measured front. In order to avoid this error, one should watch and to make sure
that the development of chromatogram will not run off the plate.

b) The lab assistant who prepared the developing solvent mistakenly used aqueous ammonia in
place of acetic acid.

Ammonia is considered to be basic, which means if aqueous ammonia is used, the OH group
may react with the TLC plate and the acidic samples which eventually form a solid. Since
ammonia is basic the spot will be deprotonated and it will not move. This may greatly affect
the results of your experiment or the experiment will not work accordingly.

c) You marked the starting line with a ballpoint

Using a pen to mark the starting line would contaminate the plate because the ink is
pigmented and may disturb the Rf value of the experiment. To further elaborate, the ink
would move with the mobile phase because it is soluble in most solvents. This would create
a mess of the chromatogram, preventing you from calculating accurate Rf values.

d) You did not replace the cap of the jar or the watch glass cover on the beaker while developing
the chromatogram.

If you did not cover the beaker while developing chromatogram, then it will show no accurate
results upon observing and gathering of your data. This affects the equilibration of the mobile
phase with the atmosphere. And if it develops too long, the mobile phase may evaporate
which affects the accuracy of your results and the Rf values.

5. Draw the structures of the standard analgesics.

CHEM 153
Lab Report (Experiment 2)

VII. Conclusion
After conducting the experiment our group has come to a conclusion that the Advil (sample)
traveled the furthest in terms of distance (Cm) in both solvents: Ethyl Acetate: Hexane and Ethyl
Acetate: acetic acid with a distance of 5.0 cm and 5.3 cm respectively. Furthermore, we can also
identify that the Rf values of Advil were also significantly greater than those of Ibuprofen in both
solvent systems. Leading us to the conclusion that Advil is more soluble than ibuprofen based on the
data gathered during the Thin Layer Chromatography experiment.