Aleks Glod

TA: Lauren Pepi

Chem 2211
February 25th, 2017
Using Thin Layer Chromatography to Identify Commercial Names of Mixtures
I. Introduction: Thin layer chromatography is a quick, efficient and inexpensive technique that
can be used in the organic chemistry laboratory to determine the compound purity, identify
various component in a multiple compound mixture, and to monitor the progress of a chemical
reaction. TLC is a solid-liquid chromatography that separates components of a mixture based on
their affinities for a mobile liquid phase over a solid stationary phase. TLC is an excellent
technique to utilize for mixture identification, as only a very small sample of the mixture is
required for TLC.
The rate of capillary action up a TLC plate depends on the polarity of the different
compounds within a mixture. The absorbent that will be utilized in out TLC experiment is silica
gel, which will glow fluorescent green when exposed to an ultraviolet lamp. The absorbent is an
important aspect of TLC, as different compounds stick to the absorbent to different degrees when
the TLC plate is exposed to an organic solvent. An important aspect of the experiment is to avoid
making contact with the absorbent gel during the entirety of the experiment, as interference with
the gel may cause blurring and ruin your results. After the plate has been spotted, a name for
the exposure of the plate to the reference chemicals or unknown chemicals, the student can begin
the developing process of TLC.
Solvent choice also plays an integral role in TLC. A polar element will be more prone to
the mobile liquid phase when a polar solvent is used and more prone to the stationary, solid
phase when a nonpolar solvent is used. The opposite is true for nonpolar solvents. Ideally, the
solvent should move the targeted compound between 1/3 to 2/3 the way up a plate during
separation. In this experiment, the organic solvent will be a 1:1 by volume mixture of methylene
chloride and ethanol. On a polarity scale of common organic solvents commonly used for TLC,
methylene chloride is considered only slightly polar, while ethanol is fairly polar. An important
aspect to gaining successful TLC results is to remove the plate from the developing chamber
(composed of a beaker with the organic solvent mixture, plate, and a watch glass on top) once the
solvent is about 1 cm from the top of the plate and to mark where the solvent reached with a light
pencil line immediately.
II. Procedure:
1. Obtain a small amount of each of the six unknowns from the bottles under the fume hood.
Only collect about a tip of the spatula of each compound. Each compound should be put in its
own test tube, carefully labeled with the corresponding unknown number.
2. Create a 1:1 by volume methylene chloride and ethanol mixture (will need at least 6 ml total)
3. Add 1 ml of the methylene chloride-ethanol solution to each test tube to dissolve the unknown
samples for spotting.
4. Using a dull pencil lightly draw an origin line about 1 cm from the edge on each of the three
plates provided by the TA. Do not press too hard or the plates will be ruined and your results will
be skewed.
5. Mark lanes for each of the known compound on the first plate.
6. On the second and third plate, mark lanes for each unknown mixture as presented in the figure
below.

7. Obtain the prepared and dissolved solutions containing the known compounds Ibuprofen,
acetaminophen, caffeine, and aspirin (acetylsalicylic acid).
8. Spot each of the knowns on the origin line of the first plate with the capillaries provided. The
spots should be evenly spaced at least 5 mm apart. Spotting samples too closely will lead to
smearing, which will lead to poor results.
9. Add enough ethyl acetate (containing 5% acetic acid) to a 100-150 ml beaker to barely cover
the bottom of the beaker. Build a developing chamber, as illustrated below, using a 100-150 ml
beaker and a watch glass.

10. Monitor the chamber. When the solvent travels up to within 1 cm of the top of the plate,
remove it and make a line across the solvent front. This line needs to be drawn immediately,
before the solvent begins to evaporate.
11. Repeat steps 8-10 with the other 2 plates and 6 unknowns. Run the plates separately, ensuring
you mark the solvent line each time after removing the plate from the developing chamber.
12. Once all 3 plates have been marked, look under an ultraviolet lamp. The components should
clearly stand out against the fluorescent green silica gel. Carefully circle all the dark spots seen
(there can be more than one spot per lane depending on the nature of the mixture).
III. Safety
General: All those working in an organic chemistry lab need to exhibit caution due to
boiling water, hot glassware, and potentially dangerous chemicals. Safety glasses and gloves
must be worn for the entire experiment. Handle spotting capillaries carefully as they are very
fragile and sharp. With corrosive chemicals, like acetic acid, be careful when dispensing. With
flammable chemicals such as ethyl acetate and methylene chloride, keep away from sparks, hot
surfaces, and fumes. With toxic chemicals such as methylene chloride, keep the snorkel on
throughout the lab. With any glassware, check the temperature prior to handling allow to cool
before disassembling.
Acetaminophen: Overexposure can lead to nausea and vomiting; It can damage the liver,
kidneys and central nervous system; Eye and skin contact causes irritation and redness;
also may cause respiratory irritation and hinder breathing.
Acetylsalicylic Acid: Harmful if ingested; causes serious eye irritation; may cause an
allergic reaction, induce asthma or cause trouble breathing; repeated exposure may cause
damage to organs.
Caffeine: Irritation of the skin, eyes, mucous membranes and respiratory tract; difficulty
breathing if inhaled; skin and eye contact may cause irritation and redness; toxic if
ingested in excess.
Ibuprofen: Causes serious eye irritation; skin contact causes potentially severe irritation
and redness; may inhibit breathing and cause respiratory problems; prolonged exposure
can target organs due to toxicity to blood, lungs, nervous system , and mucous
membranes.
Ethanol: Flammable and toxic; eye contact causes severe irritation and pain; ingestions
causes nausea, vomiting, dizziness, headache, drowsiness and unconsciousness.
Inhalation respiratory tract irritation and suffocation.
Methylene Chloride (dichloromethane): Can enter the body through means of
inhalation, skin absorption, ingestion, skin and/or eye contact, leading to an individual
experiencing irritation to eyes, skin; weakness or exhaustion, drowsiness, dizziness;
numbness, tingle limbs and nausea. It is considered a potential occupational carcinogen.
Ethyl Acetate: headache, irritation of respiratory passages and eyes, dizziness,
drowsiness and nausea, weakness, loss of consciousness; prolonged exposure may cause
organ damage.
Acetic Acid: Flammable liquid and vapor; harmful in contact with skin; causes severe
skin burns and eye damage; may cause respiratory irritation, coughing, and shortness of
breath.
Compound Structure Molecular Boiling point Density pKa
name weight g/mol C
Acetic Acid

Compound name Structure Molecular Weight Boiling Point C Density

g/mol
Methylene Chloride
(dicholormethane)

Ethanol

Ethyl Acetate

Compound name Structure Molecular Weight g/mol Melting Point C

Acetaminophen

Acetylsalicylic
Acid

Caffeine

Ibuprofen
 Citations
"Acetic acid | CH3COOH - PubChem." National Center for Biotechnology Information. U.S.
National Library of Medicine, n.d. Web. 25 Feb. 2017.
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"Acetaminophen | C8H9NO2 - PubChem." National Center for Biotechnology Information. U.S.
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