Dandekar

Organic Chemistry Lab Manual

University of North Texas

Experiment 3: Liquid-liquid extraction of caffeine and derivative analysis

Objectives
a. Extract caffeine from a variety of common sources
b. Synthesize caffeine salicylate, a derivative of caffeine, for melting point analysis

Part A: Extract caffeine from a variety of common sources

Organic compounds are often extracted from plants and other natural sources for a variety of
purposes. Extraction methods can provide starting materials for drug synthesis or food additives
for preservation, flavoring, etc. Tea and coffee are familiar sources of caffeine, a drug that is
commonly used for its stimulant effects. When you drink tea or coffee, you are drinking an
aqueous extract of plant compounds! Perhaps you have taken a caffeine-containing supplement
to help you stay alert, such as a caffeine pill or an energy drink: the caffeine it contains was
probably first extracted from a plant in which it naturally occurs, such as tea or coffee, before
being used as an ingredient in the manufacture of the caffeine pill or energy drink. Naturally
brewed tea/coffee is essentially an aqueous extract, a complex mixture which contains, in
addition to caffeine, many other water-soluble components extracted from tea leaves and/or
ground coffee beans. Subsequently, by using an appropriate extraction method, one can separate
the caffeine from the remaining components of the initial extract. This example illustrates how a
variety of important natural products may be isolated from their natural sources.
There are several different methods to carry out extraction. Liquid-liquid extraction is a
commonly used technique that, as the term implies, allows for a substance to be extracted from a
solution in one liquid into another liquid in which it may be more soluble: the solute is
partitioned between the two liquids based on its relative solubility in the two liquids. You will
use this technique several times during the 2-semester laboratory sequence in organic chemistry.
Many organic substances are soluble in a range of solvents and some of these solvents are
mutually immiscible: this can be used to advantage in devising useful extraction methods. When
a substance is reasonably soluble in two immiscible solvents, it is possible to extract this
substance from its solution in one of these solvents into the second solvent. Thus, when the
solution of a substance X in solvent A is shaken with solvent B, X partitions between
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Organic Chemistry Lab Manual

University of North Texas

the two solvents in proportion to its solubility in the two solvents (partition coefficient). In this
experiment, you will use dichloromethane to selectively extract caffeine from various sources,
such as brewed tea, energy drinks, caffeine-containing pills, etc., by exploiting the greater
solubility of caffeine in dichloromethane, compared to its solubility in water.
Materials
Apparatus:
Ice bath, hot plate, filter paper, vacuum filtration apparatus (Fig. 1-9, p.28 Padas), transfer
funnel, gravity filtration apparatus, separatory funnel (125 mL), test tubes, test tube rack, Pasteur
pipette, pipette bulb, beakers (100 mL, 250 mL), melting point capillaries, Erlenmeyer flasks (25
mL, 100 mL, 250 mL) thermometer, watch glass
Chemicals:
Acetone, anhydrous calcium chloride pellets, dichloromethane, hexanes, petroleum ether,
salicylic acid, sodium bicarbonate
Caffeine sources: Tea bags, Excedrin tablets, Red Bull, 5-Hour Energy, No-Doz tablets
Procedure
Required reading: p.25-30, 128-130, 134-137 Padas
1. Obtaining an aqueous solution of caffeine from various sources:
Tea leaves:
To an Erlenmeyer flask, add 25 g tea leaves (tea bags are more convenient than loose-leaf
tea), followed by 20 g NaHCO3. To this mixture, add 200 mL boiling water, cover and
steep for 10 min: caffeine, along with other phytochemicals, will be extracted into the
water. Remove the teabags and use the cooled brew (25C) as your caffeine source.
Excedrin tablets:
Crush the tablet and add __________ mL water. Stir to dissolve the water-soluble
ingredients. If the solution is cloudy, filter (using gravity filtration) and use the clear
filtrate as your caffeine source.
No-Doz tablets:
Crush the tablet and add __________ mL water. Stir to dissolve the water-soluble
ingredients. If the solution is cloudy, filter (using gravity filtration) and use the clear
filtrate as your caffeine source.

Dandekar

Organic Chemistry Lab Manual

University of North Texas

Red Bull and 5-Hour Energy are already aqueous, and can be used directly as the caffeine
source.
2. Extraction:
Your TA will assign which caffeine source you are to use in your experiment. After preparing the
aqueous solution as described above, use a funnel to transfer it into a separatory funnel supported
on a ring stand: remember to close the stopcock of your funnel first and place a beaker under it to
catch any accidental leaks. Add 20 mL dichloromethane, close the top with the stopper, and
gently swirl the mixture: vent frequently, to release any built-up pressure. Remember that
dichloromethane is extremely volatile and its vapors can cause pressure to build up quickly in a
closed container, especially upon being shaken; not venting to release pressure can be disastrous:
it may cause the stopper to pop (violently!) and the contents to be splattered. Be careful to mix
just enough to allow the substances to come in contact with each other, but not to form an
emulsion: dichloromethane has a tendency to form stable emulsions, which can become a
nuisance during the separation of the two layers. As the two liquids get mixed, the caffeine gets
partitioned between the two solvents and most of it is extracted from the aqueous layer into the
organic layer. Support the separatory funnel on a ring stand, loosen the stopcock and allow the
mixture to sit undisturbed for a few minutes: the aqueous and organic layers will soon separate.
Despite your efforts to prevent the formation of an emulsion, it may still form: you will see a
cloudy zone in the region separating the two layers, and relatively clear liquid above and below.
A stirring rod, gently swirled through the emulsion sometimes helps to break it up. Alternatively,
you may try adding a few crystals of NaCl (stir to dissolve) or 1-2 mL of NaCl (aq, sat.) (stir to
mix and dissolve the crystals). Which layer settles on top and which settles on bottom? If you
cannot tell by looking at the mixture, refer to their densities. Drain the lower layer into an
Erlenmeyer flask by carefully opening the stopcock, being especially careful near the meniscus.
Repeat the process two more times, each time adding 5 mL dichloromethane to the aqueous
solution in the separatory funnel; combine the organic layers from the three extractions. Pour the
upper layer (through the top!)into a separate beaker: although this layer is essentially the
decaffeinated waste layer, it is generally advisable to not discard this layer until the extraction
is complete, just in case you have obtained the wrong layer or need to go back and extract more
of your desired product from the original mixture. Dry the combined organic extract with
anhydrous calcium chloride pellets. Remove the drying agent by decantation or filtration and
heat the solution gently (in the hood); the solvent evaporates, leaving behind the solid caffeine.
For larger amounts of solution, it may be advisable to remove the dichloromethane by
distillation; the collected dichloromethane must be discarded in the appropriate waste container.
Collect the crude caffeine product on filter paper and allow to air dry for a few minutes before
measuring and recording its mass. Also record its appearance: its color, whether it is powdery or
crystalline, etc. After filling a melting point capillary with the crude product, you may then
purify it further: dissolve it in 5 mL hot acetone and add hot hexanes, dropwise, until the solution
becomes cloudy. Cool slowly to room temperature and then place in an ice bath. Vacuum filter
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Dandekar

Organic Chemistry Lab Manual

University of North Texas

your product and wash the crystals with a small amount of ice-cold hexanes; allow the product to
dry, record its mass and observe its appearance. Does it look different from the crude product
first isolated? Determine the melting point of the crude and recrystallized caffeine and record
your observations in the table provided.
Part B: Prepare caffeine salicylate, a derivative of caffeine
Since extraction techniques are not perfect, undesired compounds that have a similar solubility in
the extraction solvent to your desired compound can sometimes find their way into your extract.
We commonly use melting point analysis to identify the products of an experiment, but if
multiple compounds happen to have similar melting points, how can you be sure that you have
obtained your desired compound at all?! In the previous experiment, you used mixed melting
point analysis to answer this question. Here, you will use an alternative method to confirm the
identity of the product: you will prepare a derivative, typically via a simple reaction, and
compare its melting point with published literature values: if the melting points of both, the
product as well as its derivative, match the published data, you can be pretty sure that have
isolated the desired product. This identification strategy is based on the following principle:
even if two substances have similar melting points, it is unlikely that they will both form
derivatives under the same reaction conditions; even if they do, it is unlikely that the melting
points of the two derivatives will also have similar melting points. Thus, caffeine salicylate, an
easy to prepare, well-known derivative of caffeine, can be used to confirm the identity of a given
substance suspected to be caffeine. The synthesis of caffeine salicylate simply requires the two
reagents to be mixed. However, in order to isolate the product, it must be crystallized from its
solution by adding another solvent in which its solubility is lower.

Materials
Apparatus:
Sand bath/hot plate, ice bath, vacuum filtration apparatus, filter paper, rubber tubing,
thermometer adapter, beaker, aspirator, 2-3 inch pieces of glass tubing, test tubes, test tube rack,
Pasteur pipette, pipette bulbs
Chemicals:
Caffeine (isolated in Part A), dichloromethane, petroleum ether, salicylic acid
Procedure
To a reaction tube (test tube) containing 10 mg caffeine, add 7.5 mg salicylic acid followed by
0.5 mL dichloromethane. Heat the solution in a sand bath. Add hot petroleum ether, drop-wise,
until the solution becomes slightly cloudy. If too much petroleum ether is accidentally added, add
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Dandekar

Organic Chemistry Lab Manual

University of North Texas

dichloromethane, drop-wise, until the solution partially clarifies: it must be a bit cloudy. Cool the
mixture slowly to room temperature and then place the reaction tube in an ice bath to crystallize
the caffeine salicylate. Leaving the reaction tube in the ice bath, carefully remove most of the
solvent with a Pasteur pipette taking care not to remove any crystals (Fig. 1-10, p. 29 Padas).
The last traces of solvent must be removed before measuring the melting point. This can be
accomplished by evaporating the residual solvent under vacuum. To connect the reaction tube to
the aspirator, use a thermometer adapter fitted with a short piece of glass tubing. Cover the
reaction tube with one side of the thermometer adapter. Use rubber tubing to connect the other
side of the glass tube to an aspirator. Start the vacuum and allow the last bit of solvent to
evaporate.
Use a spatula to transfer the crystals to a watch glass to collect a sample for melting point
analysis. Measure the melting point of the reaction product and record this value along with the
mass and the percent yield.
Pre-Lab Questions
1. Why is the extraction of the aqueous extract with dichloromethane to be repeated three
times: first with 20 mL, then 5 mL, then another 5 mL dichloromethane? Why not simply
use the entire 30 mL dichloromethane in a single extraction? Explain clearly why three
extractions are likely to be more efficient than a single extraction.
2. What is the purpose of using dichloromethane in the synthesis of caffeine salicylate?
3. The term petroleum ether is a misnomer: find out what the chemical constitution of
petroleum ether is, and comment on why the name can be misleading.
4. Use curved arrows to show what happens to caffeine after the addition of salicylic acid.
5. Chocolate contains some compounds very similar to caffeine. Find out what these are
and draw their structures.
6. Draw the structures of the purine bases found in RNA and DNA: adenine and guanine. In
what way are these structures similar to caffeine?
Post-Lab Questions
1.

2.

3.

When making the aqueous extract from tea leaves, you were instructed to add NaHCO3.
Why?
(Hint: Tea leaves also contain tannic acids: how might these react with NaHCO3 and how
might this reaction subsequently minimize the tannic acids being extracted into the
organic layer?)
If you were to use dichloromethane to carry out another liquid-liquid extraction in future,
how might you modify/refine your technique so as to minimize the formation of
emulsions?
Based on the melting range observed for your crude and recrystallized caffeine, how pure
was the crude caffeine you isolated?

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4.
5.
6.
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Organic Chemistry Lab Manual

University of North Texas

Crude caffeine isolated from tea leaves sometimes has a greenish tinge. Speculate on
what might account for this color.
Several caffeine sources were used in your class to isolate caffeine. Which of these had
the highest caffeine content? Explain how you reach your conclusion.
Did the observed melting points of caffeine and caffeine salicylate confirm that the
compound you isolated in Part A was indeed caffeine?
How pure do you think the caffeine salicylate was?