BIOORGANIC

CHEMISTRY
BioOrganic Chemistry Laboratory - CH 205 (2015 2016) Experiment 3

Active Ingredients in Analgesic Tablets by TLC

Katrinna Delfin, Christian Paolo Eslava, Marielle Dominique Eusebio, and Ronell Angelo Esteban*
Department of Sports Science, College of Rehabilitation Sciences
University of Santo Tomas, Espana Street, Manila 1008
Date Submitted: September 15, 2015
Abstract:
This experiment is all about determining the active ingredients and its RF value residing in the analgesic tablets
(Aspirin, Acetaminophen, Ibuprofen, Mefenamic Acid, and Caffeine). In for us to determine the active ingredients, we used
the method of Thin Layer Chromatography (TLC). TLC can be used to determine the purity of a compound, to analyze the
composition of a mixture or to follow the progress of a reaction. Based on the results, the analgesic is the most active
ingredient that travels fastest to the mobile phase and the acetaminophen is the one that travels slowest and closest to the
stationary phase.

Introduction:
One of the purpose of this lab/experiment is to introduce to the technique of thin-layer
chromatography (TLC). TLC is an extremely effective method for separating a mixture of different
components. The benefits of this technique are that it: 1) It is relatively fast (<10min per run). 2) Itis
inexpensive.

is non-destructive (the sample can be recovered unchanged) 4) It requires very little

sample. 5) And it gives reproducible results. Because of these benefits organic chemists use TLC on a
daily basis for: Monitoring a reaction - the disappearance of starting material and appearance of product
on the TLC plate can be used as a method to check the progress of a reaction. Determining the purity of
a compound - TLC allows you to separate and visualize the components of a mixture. Identifying a
compound. - if a spot on a TLC plate lines up precisely with a known compound it is a good indication
(but not assured) that the mixture contains the known compound. The use of Thin Layer
Chromatography in this experiment is to determine the active ingredients of the Analgesic Tablets.
Another scope of this experiment is to determine the Retaliation Factor of the active ingredients of the
analgesic tablets in order to make the experiment more scientific rather than a mere interpretation by
sight. The Retaliation Factor or the Rf value is defined as the ratio of the distance moved by the solute
and the distance moved by the solvent (known as the Solvent front) along the paper, where both
distances are measured from the common Origin or Application Baseline, that is the point where the
sample is initially spotted on the paper. And at the end of the experiment, through the use of the
techniques mentioned above, the relation of the structure of the compounds with their chromatographic
behavior will be known.
Methodology:
A 250 mL beaker was lined inside with a filter paper. 10 mL of developing solvent was poured
inside and it was covered with an aluminum foil. Standard solutions was prepared containing 2%
solutions of acetaminophen, aspirin, ibuprofen, and mefenamic acid in an extraction solvent. Half of the
tablet was pulverized using a mortar and pestle and was placed in a test tube with an addition of 2mL

extraction solvent. The test tube was shaked to dissolve the tablet. The residue settled at the bottom of
the tube while the clear liquid was decanted in a clean and dry vial. A TLC plate pre-coated with the
silica gel was obtained from the professor. With a pencil, the ORIGIN was marked. A spot of the
standards and unknown analgesics was applied to the dots that was drawn on the bottom of the TLC
plate with the use of a capillary tube. The application was repeated and then the TLC plate was placed
into the developing chamber/beaker. It was covered with aluminum foil and the chromatogram was
allowed to develop undisturbed. The plate was remove from the chamber when the level of the solvent
was near the top of the plate. And the Solvent Front was marked with a pencil. The plate was allowed
to dry and it was visualized in the UV chamber. All the dark spots was measured and the chromatogram
was sketched in the logbook. And then the RF value of the standard analgesics and each components of
the tablet was calculated.
Results:

Legends
1. Acetaminophen
2. Aspirin
3. Analgesics
1
4

4. Ibuprofen
25. Mefenamic
3
Acid

Figure 1: TLC Plate with the Dark spots and its Legends

Solvent Front
Acetaminophen
Aspirin
Analgesics
Ibuprofen
Mefenamic Acid

Distance from the Origin

5 cm
2 cm
3.3 cm
3.7 cm
3.4 cm
3.5 cm

Figure 2: Distance from the Origin and RF value of the Analgesic Tablets

RF Value
.40
.66
.74
.68
.70

Discussion:
In this experiment, the RF Value was computed by dividing the distance from the origin to the
center of the dark spot, to the distance from the origin to the solvent front. The result of it was that the
analgesics was the most active ingredient and is the one that is closest to the mobile phase. Followed by
the Mefenamic Acid, Ibuprofen, and Aspirin. The first four analgesic tablets that I mentioned are close
together and they are closer to the mobile phase than the stationary phase. This means that they are
likely to be less polar. While Acetaminophen was the one that traveled slowest and it is more attracted to
the stationary, which means it is more polar than the rest. It is probable that our experiment did not meet
the ideal results due to a chance that the measurement was a little bit big or smaller, the calibration of
was solvent was not accurate, or that the solvent has an impurity. It is all because of human error. And
from this experiment we are able to identify the active ingredients in analgesic tablets, but what do we
really know about the active ingredient? The active ingredient is the metabolite that elicits the effect of
the drug in the body.
Conclusion:
The most active ingredients in the experiment are the Analgesics, Mefenamic Acid, Ibuprofen,
and Aspirin, with Analgesics as the one that travels farthest. While the Acetaminophen is the least active
ingredient. We are also able to compute the RF Value of each Analgesic Tablets: Acetaminophen - .40,
Aspirin - .66, Ibuprofen, .68, Mefenamic Acid - .70, and Analgesics - .74. And the relationship of their
structure in the TLC plate with their chromatogram behavior is that Analgesics, Mefenamic Acid,
Ibuprofen, and Aspirin are closer to the mobile phase than the stationary, which means that they are
likely to be less polar. And Acetaminophen as being closer to the stationary phase, it is more polar.
Reference:
Buschmann, H. & Christoph, T. (2005). Analgesics: from chemistry and pharmacology to clinical
application. USA: Wiley-VCH.
Komsta, L., Hajnos, M., & Sherma, J. (2014). Thin layer chromatography in drug analysis. USA: CRC
Press.
Case, M. & Corin, S. (2000). Active ingredients in analgesics by thin layer chromatography.
http://en.wikipedia.org/wiki/Thin_layer_chromatography Write-up for this lab to be completed on 9/22
and 9/24
Sinatra, R. (2010). The essence of analgesia and analgesics. England: Cambridge University Press.