Prithvi Dev Awasthi

09/14/2016

TLC analysis of Analgesic Drugs

Objective
The purpose of this experiment was to find the Rf value.

Experiment Scheme
Initially, 10 micropipettes were used to spot the plates. Surface of the
plated should not be touched. It should be handled in edges. Then, the given
samples were spotted on the plates. It is important that the spots arent too
small but small enough so that the sample do not overlap each other after
development. Small amount of development solvent was filled in a screwcap
jar up to the depth of about 0.5-0.7 cm. Then, the spotted plates were placed
in the development chamber allowed to develop. The plates were placed
parallel and not tilted so that the developing happens correctly. The chamber
were not moved until the development are completed. When the solvent had
risen about 0.5 cm from the top, the plate was removed from the chamber
on a paper towel to dry.
After the plate was dry, it was observed under the UV light in a dark
space and the observed spots were outlined with pencil carefully. After the
UV analysis was done, the plated were put in a chamber of Iodine and the jar
was warmed gently with hand heat. The visible colors were noted and
change in color was observed. Finally, all the observations were recorded.

Given Chemicals
1. Acetaminophen (AC)

3. Caffiene (CF)

2. Aspirin (Asp)

4. Salicylamide

Prithvi Dev Awasthi

09/14/2016

Chemical Equation
Rf =

Distancetravelled by spot
Distance travelled by solvent

Chemical Table
S.
N

Chemical
Name

Molecul
ar
Formula

C8H9NO2

Acetaminop
hen
Aspirin

Caffeine

C8H10O2

Salicylamide

C7H7NO2

TLC reading 1

C9H8O4

Molar
Mass
(g/mo
l)
151.1
6
180.1
5
194.1
9
137.1
4

Stat Densi
e
ty
(g/cm
3
)
S
1.26

Meltin
g
Point
(C)
169

Boili
ng
Point
(C)
-

1.4

236

365

Liver
damage
Stroke

1.23

276.8

284

insomnia

1.8

140.1
4

181

Damage
d fertility

TLC reading 2

Health
Hazard

Prithvi Dev Awasthi

09/14/2016

Table 1

Table 2
54.5

3.8 3.7
4
2.9
2.4
2.3

2.6
3 2.52.5
Distance travelled by spot 2 1.4

Distance travelled by spot 2

0

1
0

Given samples

10

Given Samples

Calculations for reading 1

Calculations for reading 2

Rf of 1 (I) (Anacin): 2.3/5.4 = 0.43

Rf of 1 (Acetaminophen): 4.5/5 =
0.9

Rf of 2 (I) (Excedrin): 3.8/5.4 = 0.7

Rf of 3 (Tylenol): 2.9/5.4 = 0.54
Rf of 4 (I) (Bufferin): 3.7/5.4 = 0.69
Rf of 5 (Reference): 2.4/5.4 = 0.44

Rf of 2 (I) (Aspirin): 2.5/5 = 0.5

Rf of 3 (I) (Caffeine): 2.6/5 = 0.52
Rf of 4 (Salicylamide): 1.4/5 = 0.28
Rf of 5 (I) (Reference): 2.5/5 = 0.5

Discussion
Thin layer chromatography is a special of adsorption which is used to
separate mixtures. TLC technique was performed in on a sheet of plastic
covered with silica gel. The layer was known as stationery phase. Silica gel is
a form of silicon dioxide. At the surface silicon atoms are attached to the
hydroxyl group whereas silicon atoms are joined via oxygen atoms in a giant
covalent structure. Silica gel can form hydrogen bonds with suitable
compounds around it as well as van der walls dispersion forces and dipoledipole attraction which is why it is popular for its hydroxyl group.
Different compounds in the sample mixture travels in a different rate
due to their attraction and reaction to the given solvent. After the solvent
reaches its desired distance, the plates are observed under UV light. UV light
helps us determine the exact spots the samples have travelled to. Extreme
precautions that we should take is UV light should not be exposed in our
eyes directly. It can cause permanent damage. After that, Iodine also helps
us to find the color that appears in the plate.
While we are drawing the lines on the TLC plate, we should use pencil
instead of the pens because it can contaminate our TLC plate and the
solution point may move with the pens ink.

Questions:
Q1: What happens if the spots are made too large when preparing a
TLC plate for development?
A: if the spots are made too large, they might overlap each other
after development.

Q2: Is it possible to distinguish two spots that have the same Rf value,
but represent different compounds? Give two different methods.
A: yes it is possible to distinguish between two spots with same Rf. We
use method of UV illumination and iodine staining to distinguish different
compounds.
The Rf was determined and the sample on the tablet was found to be
Caffeine.
Reference:

P.; L.; kriz; E. Organic chemistry laboratory manual, 3rd ed.; Cengage learning.
Budavari, S. The Merck index: an encyclopedia of chemicals, drugs, and biologicals; Merck:
Rahway, NJ, U.S.A., 1989.
Lehman, J. W. The Students lab Companion, 2nd ed.; Pearson.