Extraction and Isolation of Caffeine from Tea Organic Chem Lab Manual

Activity No. 7
Extraction and Recrystallization of Caffeine from Tea

Caffeine is a naturally occurring alkaloid found in over 60 plant species. Caffeine belongs
to a family of naturally occurring compounds known as xanthines. The xanthines, which come from
plants, are possibly the oldest known stimulants. Caffeine is the most powerful xanthine in its ability
to increase alertness, put off sleep and to increase ones capacity for
thinking. Caffeine is a vasodilator (relaxes the blood vessels) as well as a
diuretic (increases urination). Caffeine does not exist alone in tea leaves;
the leaves are mainly cellulose, pigments and chlorophylls, and tannins.
Caffeine is the principle stimulant found in tea and coffee, and it can
account for up to approximately 5% of the mass of the tea leaves. Caffeine is soluble in water and
a variety of organic solvents, and both can be used to extract caffeine from tea leaves or coffee
beans. In fact, a wide variety of solvents including water, water in combination with organic
solvents, methylene chloride, ethyl acetate, and supercritical carbon dioxide are used commercially
to extract caffeine from coffee.

The basic N in caffeine can be used to increase or decrease its water solubility. Acidic
conditions will form the conjugate acid salt giving caffeine increased water solubility as a cation.
On the other hand if caffeine is in a basic environment it takes the neutral form and is only
somewhat polar. Tea leaves contain tannins, which are acidic, as well as a number of colored
compounds and a small amount of undecomposed chlorophyll (soluble in dichloromethane). To
ensure that the acidic substance remains water soluble and that the caffeine will be present as the
free base, sodium carbonate is added to the extraction medium. In order to successfully extract
any substance from one solvent into another, the differences in solubility must be maximized.
Adding base to the solution has a second important effect. The water solution contains much more
than just caffeine, and some of these compounds are also soluble in dichloromethane. Making the
solution basic forms insoluble tannin salts which removes them from the solution before the
caffeine is extracted. The dichloromethane extract will primarily contain caffeine with small
amounts of impurities. Caffeine is also water soluble, but by keeping the washing solution basic it
minimizes the caffeine lost, while maximizing the removal of impurities.

In this experiment, the interest is not in maintaining the flavor of the tea upon extraction
of caffeine. Taking advantage of caffeine’s water solubility, the first step of extraction involves
brewing of tea. Once the caffeine is extracted from the tea leaves it will be extracted from the
water using an organic solvent.

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 Extraction and Isolation of Caffeine from Tea Organic Chem Lab Manual

Materials
100-mL beaker (1) 50-mL grad cylinder (1)
15-mL screw-cap centrifuge tube (5) 5-mL grad. Cylinder (1)
watch glass Pasteur pipette (2)
25-mL beaker, pre-weighed (1) stirring rod
250-mL beaker (1) thermometer
wire gauze electric stove (1 for the whole class)
water bath specimen vial, pre weighed, 1
centrifuge machine (to be signed out by monitor)

Chemicals
sodium carbonate (s) methylene chloride
hydrochloric acid, 6M sodium sulfate (anhydrous) – drying agent
acetone hexane

Procedure
I. Extraction of Caffeine
1. Place 1 teabags in a small beaker and add 20 mL of water. Add 3 pcs of boiling chips. Boil the
tea for 20 minutes. Add water as needed to maintain the volume of the solution. Take out the
tea bag and place in a watch glass and squeeze out the liquid into the beaker by pressing
between two watch glasses. Cool a little bit in an ice bath.
2. Add 0.5 g of sodium carbonate to convert the tannins into water-soluble salts.
3. Once the sodium carbonate has dissolved and the tea has cooled to room temperature,
transfer equally to two 15-mL screw-cap centrifuge tubes. Check if the centrifuge tubes has no
leak.
4. Add 1.5mL of methylene chloride (also called dichloromethane) to each test tube. Cap
and shake gently by simply tipping over. Remember to vent the tube.
5. Spin the tea–methylene chloride suspension in a centrifuge for five minutes to break the
emulsion. Remember to balance your centrifuge tubes in the centrifuge machine.
6. Carefully transfer the organic layer from each centrifuge tube to a third 15-mL centrifuge
tube, using Pasteur pipette. Screw in the cover. See illustration at the two last pages of this
activity.
7. Repeat steps 4, 5, and 6. Collect all methylene chloride layers to the same 15 mL centrifuge
tube.
8. With a clean dry Pasteur pipette, transfer the methylene chloride solution to a fourth 15-mL
centrifuge tube.

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 Extraction and Isolation of Caffeine from Tea Organic Chem Lab Manual

10. Dry the organic extract by adding small portions of anhydrous sodium sulfate (drying agent),
with gentle shaking. The drying agent clumps up as it absorbs moisture. Add some more until
you see particles of the drying agent no longer clumping up, which means the solution is
already dry. Do not add too much drying agent, it may physically absorb all of the liquid.
11. Decant the dry methylene chloride–caffeine solution to a pre weighed, dry 25-mL beaker.
12. Cover loosely with a course filter paper. Label and let it stay overnight under the hood to
evaporate.
13. Note the appearance of the crude caffeine. Obtain the weight.

II. Recrystallization of the Crude Caffeine

14. Dissolve the crude crystasl by adding drop by drop hot absolute ethanol. Do not add too much
or it will remain dissolved and will no longer recrystallize. You can heat in a water bath to
dissolve completely.
15. Cool a little bit and then place in an ice bath to complete the crystallization. Cool also some
amount of ethanol.
16. Suction filter and wash the beaker with the cold ethanol.
17. Scrape off the purified caffeine crystals and place in a pre weighed specimen vial.
18. Note the appearance and obtain the weight.

References

Adapted from Introduction to Organic Laboratory Techniques: A Microscale Approach, 3rd edition.
Pavia, Donald L., Gary M. Lampman, George S. Kriz, and Randall G. Engel (1999).

Murray, D.S.; Hansen, P.J., J. Chem. Educ., 1995 (72) 851.

Hampp, A., J. Chem. Educ., 1996 (73) 1172.

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 Extraction and Isolation of Caffeine from Tea Organic Chem Lab Manual

Microscale Laboratory Techniques - Extractions

In microscale experiments, the conical reaction vial is the glassware item used for
extractions. The two immiscible liquid layers are placed in the vial, and the top is sealed with a cap
and a Teflon insert (with the Teflon side toward the inside of the vial). The vial is shaken to provide
thorough mixing between the two liquid phases. As the shaking continues, the vial is vented
periodically by loosening the cap and then tightening it again. After about 5-10 seconds of shaking,
the cap is loosened to vent the vial, retightened, and the vial is allowed to stand upright in a beaker
until the two liquid layers separate completely.

Two basic procedures are possible, depending on whether the solvent being used to extract
the desired product is heavier or lighter than water. Method A is employed for extractions where
the lower layer is a heavy solvent such as dichloromethane:

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 Extraction and Isolation of Caffeine from Tea Organic Chem Lab Manual

Method B is employed for extraction with a solvent which is lighter than water, such as
diethyl ether. Note that in this technique, one draws both phases into the pipet and then returns
the heavy (aqueous) phase to the conical vial. Ether is so volatile that it is often difficult to hold it
in the pipet. Use of a filter-tip pipet for this procedure will help prevent the volatile organic layer
from squirting out in an uncontrolled way.

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