Isolation of Caffeine

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Isolation of Caffeine from Soda

Pre-Lab Assignment:
Complete the pre-lab cover page and develop a procedure.
Develop an in-depth separations flowchart detailing the procedure in your pre-lab.
Learning Goals:
Understand the concepts and applications of natural product isolation
Reinforce liquid/liquid extraction techniques
Understand the pH dependent equilibrium of caffeine
Introduction:
Caffeine belongs to a family of heteroaromatic compounds known as xanthines. Other common
compounds found in this group are theophylline and theobromine. Theophylline is used in the
treatment of asthma and theobromine is found in chocolate. Xanthine, caffeine, and theobromine
are shown below. Note that the ring system and position of the nitrogen atoms are the same in
all three molecules.
O
H

H
N

H3C

N
H

Xanthine

CH3

CH3
N

N
CH3

Caffeine

CH3

Theobromine

A single cup of coffee can contain up to 200 mg of caffeine. Commercially, coffee is


decaffeinated by removing 97% or more of the naturally existing caffeine from the green beans
before roasting. Three methods for extracting caffeine from coffee are:

Direct Contact Method. After softening the beans by steam, the green beans are extracted
with methylene chloride, which dissolves the caffeine. The beans are steamed a second
time, heated and blown dry to remove almost all of the methylene chloride (US laws
allows no more than 10 ppm- 0.001% wt/wt- residual methylene chloride.).

Indirect Contact Method. The green beans are soaked in hot water, which extracts the
caffeine. The water is separated from the beans and extracted with either methylene
chloride or ethyl acetate. The extracted water, which is now caffeine-free, is put back
with the beans to restore some of the natural oils and flavor.

Water Filtration Method. After the beans soak for several hours in water, the water is
removed and passed through charcoal filters to remove the caffeine. The water, still
containing other flavor elements, is added back to the beans. This is often called the
"Swiss Water Process

In this lab you will isolate the caffeine found in common soft drinks, using extraction techniques
learned in earlier labs. Soft drinks are mainly carbonated water containing phosphoric acid,
caffeine, citric acid and other flavorings. Caffeine is an organic compound that in its neutral
state is weakly polar. Caffeine is also an organic base that when reacted with an aqueous acid
will become protonated to give an acid with a pKa of 3.6. Protonated caffeine is ionic and thus
polar, making it water soluble. Caffeines acid/base equilibrium is exploited in this lab to enable
separation from a complex mixture. The soft drinks used in this lab have been allowed to de-gas
over several days to facilitate handling. You will be provided with one can of de-gassed soft
drink that must be a shared with two other students. Do not forget to consider the starting pH
state of the soda, which will affect the initial protonation state of the caffeine.
Procedure:
Obtain a 250 mL separatory funnel and a hotplate from the stockroom. Once your separatory
funnel is secured in the iron ring (making sure the valve is closed), use a graduated cylinder to
add 125 mL of soda to the funnel. Using tweezers, add one pellet of sodium hydroxide (be
careful as sodium hydroxide is very basic: wash skin with water if contact is made with skin). Do
not pull out more than 1 or 2 pellets at a time, as they will absorb moisture from the air and you
wont be able to put them back. Cap and shake the funnel with frequent venting to dissolve the
sodium hydroxide. Remember to vent in your hood. Tilt the separation funnel upside down and
slowly open the drain valve to release any gas pressure.
Using litmus paper, determine the pH of your solution. Continue adding sodium hydroxide, one
pellet at a time, until the target pH of 8 is obtained. Why is a pH of 8 desired? Next, add 20 mL
of methylene chloride to the separatory funnel and shake vigorously, remembering to vent the
funnel into the hood frequently. Replace your separatory funnel in the ring, remove the cap and
allow the phases to separate (which layer is the methylene chloride?). You will notice a foam
layer between the organic and aqueous phases, this is called an emulsion. Carefully drain the
organic and the emulsion layer into a beaker. Add another 20 mL of methylene chloride and
repeat the extraction. Again, drain the organic layer into the same beaker. Pour your aqueous
layer into the sink while the cold water is running. Thoroughly clean your separatory funnel and
secure in the ring. Add the organic solution (your methylene chloride solution in the beaker)
back into the separatory funnel (again, making sure the valve is closed). Use an additional 10
mL of methylene chloride to rinse your beaker and add this rinse to the separatory funnel. To
this solution, add approximately 20 mL of saturated aqueous sodium chloride (brine), being
careful not to transfer any of the solids. Shake vigorously with venting, place back into the iron
ring and allow the phases to separate. (Brine makes the water layer more polar, decreases
emulsions, and helps remove polar compounds that are slightly soluble in methylene chloride.)
When the phases have separated, carefully drain the methylene chloride solution into a dry tared
125 mL Erlenmeyer flask.
Carefully boil off the methylene chloride using your heating plate and a boiling stick. At what
temperature does methylene chloride boil? Be careful to not pass this temperature by more than
20 degrees, to make sure the caffeine is not burned or sublimed. Why is a boiling stick used in
place of a boiling stone in this experiment? When all the solvent has evaporated, you will see an

off-white solid on the bottom of the flask: impure caffeine. After the flask cools, weigh the flask
and determine the amount of crude caffeine obtained.
Place a small amount of your crude caffeine into a disposable culture tube and obtain a small
sample of pure caffeine in a second tube. Add 1 mL of methylene chloride to each tube and
using disposable spotting pipets spot a TLC plate with your solutions (crude, pure, and co-spot).
What is the function of the co-spot? Develop the plate using ethyl acetate with 5% acetic acid as
the developing solution. Examine your plate under the UV light and mark the location of the
solvent front along with any spots that you see.

Post Lab:
Include what is required in the Lab Expectations handout.
Tape a properly labeled TLC plate to the observations section of your lab report, and
describe what it illustrates.
Answer the following questions separate from your conclusions.
1. Draw the equilibrium mechanism between caffeine and aqueous acid.
2. Using resonance structures, explain why one nitrogen is more basic than the others.
(Hint: Only one nitrogen can resonate a positive charge around the ring once protonated.)
3. Determine the total amount of crude caffeine in one can of soda pop using your
experimental yield in milligrams.
4. What does your TLC plate tell you about the purity of the crude caffeine you extracted?
5. Sublimation is a technique that can be used to purify your crude caffeine. What is
sublimation and how would it purify a compound?

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