Experiment No. 6 - ThinLayerChromatography

CENTRAL PHILIPPINE ChE 2101
UNIVERSITY COLLEGE OF ORGANIC

ENGINEERING CHEMISTRY
Jaro, Iloilo City, Philippines
Names: Cordova, Lorenz Priam Date Performed:
Lozada, Kristen Thea Experiment No. December 21, 2023
Maagma, Andrew Louie 6:
Relano, Shannelle Ivan Thin Layer
Sucgang, Karla Joy Chromatography
of Analgesic
Course BS ChE - 2 Drugs Date Submitted:
& Yr: December 21, 2023
I. Objective
1. To separate and analyze the components of common analgesic drugs, namely
aspirin, ibuprofen, and acetaminophen, using thin layer chromatography (TLC)
2. To determine their presence and assess the purity of the drug samples.
II. Theory
Thin layer chromatography (TLC) is a chromatographic technique that separates
and analyzes compounds in a mixture based on their differential migration on a thin layer
of adsorbent material. The stationary phase is a thin layer of material like silica gel on a
flat support. A mobile phase, typically a solvent or solvent mixture, moves up the plate
by capillary action, carrying sample components with it. The separation occurs as
components move at different rates depending on their affinity for the stationary and
mobile phases. Visualization and analysis of separated components are facilitated by
calculating Rf values, making TLC a quick and widely used method in various scientific
disciplines (Aryal, 2023).
Thin layer chromatography (TLC) operates on principles of adsorption and
partition chromatography. The stationary phase, typically silica gel or alumina, provides a
surface for adsorption, with polar compounds binding more strongly. As the mobile phase
ascends the TLC plate via capillary action, compounds with higher affinity for it move
more rapidly, leading to separation. The Rf value, calculated as the ratio of compound
movement to solvent movement, quantifies this behavior. Visualization techniques, such
as UV exposure or chemical staining, exploit the inherent properties of separated
components, making TLC a powerful tool for rapid qualitative analysis based on
fundamental principles of molecular interactions and partitioning (Saurabh, 2021).
III. Materials and Apparatus
Materials: Aspirin, Ibuprofen, and Acetaminophen tablets (as samples), Ethyl
acetate and methanol (solvents), Iodine solution
Apparatus: Silica gel-coated TLC plates, Capillary tubes, Developing chamber,
UV lamp, Ruler, Mortar and pestle, Filter paper, Watch glass
IV. Set-up Diagram
V. Procedure
1. Grind a tablet of each analgesic drug separately using a mortar and pestle.
2. Extract the drug components by adding each ground tablet to a separate
container with a mixture of ethyl acetate and methanol.
3. Filter the solutions to obtain clear extracts.
4. Apply a small volume of each extract to the baseline of a silica gel-coated TLC
plate using capillary tubes.
5. Develop the TLC plate in a closed chamber with a mobile phase consisting of
ethyl acetate and methanol.
6. After development, visualize the separated components using a UV lamp for
fluorescent compounds and iodine vapor for non-fluorescent compounds.
7. Measure the distance traveled by the solvent front and each spot.
8. Calculate the Rf values for each component using the formula Rf = distance
traveled by compound / distance traveled by solvent.
9. Compare the Rf values obtained with literature values or known standards to
identify the analgesic drug components.
VI. Data and Computations
Compound Distance Traveled Distance Traveled Rf Value

by Solvent Front by Spot (in mm)
(in mm)
Aspirin 100 75 0.75
Ibuprofen 100 60 0.60
Acetaminophen 100 45 0.45
Computation of Rf Values:
The Rf value is calculated using the formula:
𝐷𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑇𝑟𝑎𝑣𝑒𝑙𝑒𝑑 𝑏𝑦 𝑆𝑝𝑜𝑡
𝑅𝑓 = 𝐷𝑖𝑠𝑡𝑎𝑛𝑐𝑒 𝑇𝑟𝑎𝑣𝑒𝑙𝑒𝑑 𝑏𝑦 𝑆𝑜𝑙𝑣𝑒𝑛𝑡 𝐹𝑟𝑜𝑛𝑡
1. Aspirin
75 𝑚𝑚
𝑅𝑓 = 100 𝑚𝑚
= 0. 75
2. Ibuprofen
60 𝑚𝑚
𝑅𝑓 = 100 𝑚𝑚
= 0. 60
3. Acetaminophen
45 𝑚𝑚
𝑅𝑓 = 100 𝑚𝑚
= 0. 45
VII. Discussion and Interpretation of Results

The results of the thin layer chromatography (TLC) analysis for aspirin,
ibuprofen, and acetaminophen yielded valuable insights into the separation and
identification of these analgesic compounds. The Rf values obtained, 0.75 for aspirin,
0.60 for ibuprofen, and 0.45 for acetaminophen, closely aligned with literature values,
affirming the accuracy of the compound identification. Notably, the higher Rf value for
ibuprofen suggested its greater mobility in the mobile phase, indicating a weaker
interaction with the stationary phase compared to aspirin and acetaminophen.
Visualization techniques, including UV fluorescence and iodine staining, further
validated the presence of specific compounds. Ibuprofen exhibited fluorescence under
UV light, consistent with its known properties, while iodine vapor staining confirmed the
presence of aspirin and acetaminophen, which lack fluorescence. The results underscore
the potential utility of TLC in assessing the purity of drug samples, with deviations in Rf
values serving as indicators of impurities or variations. Overall, the comprehensive
approach employed in this study supports the reliability and applicability of TLC for the
analysis of analgesic drugs.
VIII. Conclusion
In summary, the thin layer chromatography (TLC) analysis effectively identified
aspirin, ibuprofen, and acetaminophen with Rf values that align with the results of other
reference experiments. The higher Rf value for ibuprofen indicates its increased mobility
in the mobile phase. UV fluorescence and iodine staining validated compound presence.
The results affirm the utility of TLC for rapid assessment of drug purity, supporting its
potential integration into pharmaceutical quality control. This study establishes TLC as a
reliable tool for the straightforward analysis of analgesic drugs, offering practical
applications in routine laboratory procedures and pharmaceutical research.
IX. Application
Thin layer chromatography (TLC) is a widely used technique with diverse
applications. It serves as a rapid and cost-effective method for qualitative analysis,
allowing the identification of components in mixtures based on their migration on a TLC
plate and their characteristic Rf values. TLC finds applications in pharmaceutical
industries for routine quality control, drug analysis, and monitoring chemical reactions. It
is also employed in environmental monitoring to detect pollutants, in forensic
investigations for substance identification, and in biochemical research for separating
biomolecules. Additionally, TLC plays a role in the analysis of food products, natural
extracts, dyes, and clinical samples, making it a versatile tool in various scientific fields.
Its simplicity and efficiency also render it useful in educational settings for teaching
chromatographic principles and techniques. Overall, TLC is an invaluable tool for both
routine analysis and research across chemistry, biology, and related disciplines (Santiago
& Strobel, 2013).
X. References
Aryal, S. (2023, September 11). Thin layer Chromatography: principle, parts, steps, uses.
Microbe Notes. https://microbenotes.com/thin-layer-chromatography/
Saurabh. (2021, January 11). Thin Layer Chromatography (TLC): Principle, Procedure &
Applications. Lab-Training.com. https://lab-training.com/thin-layer-chromatography-tlc/
Santiago, M., & Strobel, S. A. (2013). Thin layer chromatography. In Methods in Enzymology
(pp. 303–324). https://doi.org/10.1016/b978-0-12-420067-8.00024-6

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CENTRAL PHILIPPINE ChE 2101

UNIVERSITY COLLEGE OF ORGANIC

IV. Set-up Diagram

VI. Data and Computations

Compound Distance Traveled Distance Traveled Rf Value

Aspirin 100 75 0.75

Ibuprofen 100 60 0.60

Acetaminophen 100 45 0.45

VII. Discussion and Interpretation of Results

Microbe Notes. https://microbenotes.com/thin-layer-chromatography/

Applications. Lab-Training.com. https://lab-training.com/thin-layer-chromatography-tlc/

(pp. 303–324). https://doi.org/10.1016/b978-0-12-420067-8.00024-6

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