EXPERIMENT 1: ISOLATION OF CAFFEINE FROM A TEA BAG

OBJECTIVES: To determine the amount of caffeine in a tea bag.

INTRODUCTION:

Structure of caffeine
Caffeine belongs to the family of heterocyclic compounds known as purines. It has
the systematic name 3,7-dihydro-1,3,7-trimethyl-1H-purine-2,6-dione; it is also
known as 1,3,7-trimethylxanthine, and 1,3,7-trimethyl-2,6-dioxopurine. Caffeine can
be classified as analkaloid , a term used for substances produced as end products of
nitrogen metabolism in some plants. The chemical formula is C 8H

10

N 4 O 2 . Caffeine

has a molar mass of 194.19 grams (6.85 ounces). It is soluble in water and in many
organic solvents, and it appears in pure form as white crystals. Caffeine can be
prepared by extraction from natural sources or by synthesis from uric acid. More
than sixty plants, including those that give us coffee, tea, cola, and cacao, produce
caffeine from the purine xanthine. Whereas caffeine is a natural constituent in
coffee, tea, chocolate, and some cola drinks, it is added to consumer products such
as soft drinks, diet pills, and analgesics . Caffeine is said to be the most widely used
drug in the world, and more than 100 million people in the United States consume
caffeine each day. It has pharmacological uses: as a cardiac and respiratory
stimulant and as an agent that promotes kidney diuresis. A therapeutic dose of
caffeine is about the same as the amount found in an average cup of coffee,
between 100 and 200 milligrams (0.0071 ounces). Decaffeinated coffee can be

prepared through extraction with a solvent (such as methylene chloride), water

extraction, or steam extraction.
Caffeine enters the bloodstream about ten minutes after its ingestion and stays in
the body for up to twelve hours. Like other alkaloids, caffeine has powerful
physiological effects on humans and animals. It stimulates heart muscle and relaxes
certain structures that contain smooth muscle, including the coronary arteries and
the bronchi. It is a diuretic. Theophylline and theobromine, two other plant alkaloid
derivatives of xanthine, have physiological effects similar to those of caffeine.
Caffeine acts as a stimulant of the central nervous system (CNS) through several
proposed mechanisms. The most important seems to be its interference with the
ability of the neurotransmitter adenosine to bind to its nerve cell receptor . Also,
caffeine inhibits the enzyme cyclic nucleotide phosphodiesterase, which breaks
down intracellular cyclic adenosine monophosphate (cAMP), another messenger
involved in the transmission of nerve signals from hormones originating outside the
central nervous system
To separate caffeine from tea solution extraction technique was used. Liquid-liquid
extractions using a separatory funnel are essentially the only kind of extraction
performed in the organic teaching labs. Liquid-liquid means that two liquids are
used in the extraction procedure. The liquids must be immiscible: this means that
they will form two layers when added together, like oil and water. Some compounds
are more soluble in the organic layer and compounds are more soluble in the
aqueous layer.

APPARATUS: Beaker, small conical flask, separatory funnel, cotton, rotary

evaporator, dropper.
MATERIAL: distilled water, sodium carbonate, methylene chloride, anhydrous
sodium sulphate.
PROCEDURE:
1. A 100 mL beaker was used and 50 mL of water was added inside the beaker
to make the tea solution
2. Then, the hot tea solution was transferred into a small conical flask.

3.
4.
5.
6.

0.5 g of sodium bicarbonate (Na2CO3) was added into the tea solution.
The solution was filtered using cotton into small separatory funnel.
15 mL of methylene chloride was added into and the extraction begins.
The lower methylene chloride layer was drain off and 10 mL of fresh
methylene chloride was added.
7. The step of adding methylene chloride was repeated 2 times.
8. Combined all methylene chlorine solution and dried it with anhydrous sodium
sulphate.
9. Dried methylene chloride was transferred into a pre-weighted small round
bottom flask and the solution was evaporated using the rotary-evaporator.

RESULT:
Weight of empty beaker

= 49.33 g

Weight of Na2CO3

= 0.51 g

Weight beaker+anhydrous+ organic layer

Mass of teabag

= 63.46 g

= 2.17 g

Melting point

= 213.7 C

Mass of dry caffeine+beaker

= 49.43 g

Mass of caffeine

= 0.10 g

DISCUSSION:
Caffeine is a common substance and is found in coffee, tea, soft drinks and
chocolates. It can be isolated from natural sources like tea leaves, cola nuts and
cacao beans. Caffeine is an alkaloid, and more specifically, a member of the
methlxanthines. Caffeine has been found to act as receptor antagonists by
inhibiting the effect of adenosine. Adenosine is a modulator that regulates sleep and
other neural phenomena in human. This inhibitory effect of caffeine makes it a
central nervous system stimulant. High doses of caffeine in human adult can be
fatal. The acute lethal dose of caffeine in human adult has been estimated to be 10
g/person. To isolate caffeine from tea, extraction is used to separate the caffeine
from other components of tea. Extraction is a separation technique based on the
differences in the solubility of substances in two immiscible solvents.
For the purpose of this experiment, methylene chloride was used as the solvent
because caffeine is more soluble in methylene chloride than in water. Caffeine was
extracted by adding 15 ml of methylene chloride to the tea solution. 0.5g of sodium
carbonate was also added to the mixture. Sodium carbonate was added in order to
convert tannins and gallic acid to sodium salts which are soluble in water. When the
extraction was carried out, an emulsion was formed. An emulsion is dispersion of
liquid in another liquid. The emulsion was broken into two layers by centrifugation.
The organic layer which is denser than water settled on the bottom of the tube,
while the aqueous layer was on top. The high density of the organic layer is because

of the two chlorine atoms bonded to an alkyl group in methylene chloride. The
organic layer was separated from the aqueous layer and dried by adding sodium
sulfate. Drying is complete when the sodium sulfate clumps together. When this had
occurred, the organic layer is separated from the hydrated sulfate using a Pasteur
pipet.
The dry organic layer was then evaporated by heating using rotary-evaporator. After
the methylene chloride had evaporated, solid caffeine was collected which had
some greenish stains on the solid crystals. The melting point of the crude crystal
was found is 213.7 0C. This suggested that the crystals contained impurity. The
crude crystal weighed 0.10 g. This mass was calculated by subtracting the mass of
the beaker from the mass of the beaker when it contained crude caffeine crystal.
The crude crystal was purified by sublimation under reduced pressure. The pure
crystal recovered was whitish and had a powdery texture. The percent recovery was
calculated by dividing 0.10 g which is the amount recovered by 2.17 g, amount of
impure crystal and multiplying the result by 100.

Percentage of recovery (Caffeine)

0.10
= 2.17

X 100

= 4. 608 %
The melting point of the recovered crystal lied between 213.7 0C. This range differed
from the pure range of 235.30C-238.50C. The literature melting of caffeine is 236 0C.
The difference in melting point was as a result of operator error and the crystal
absorbing moisture from the ice cooled tube used in the set up and from
surrounding air.
CONCLUSION:
From this experiment, the amount caffeine required from the tea solution is 0.10 g.
The percentage of caffeine recovery was 4.608 %.

REFERENCES:
(1) Pavia, D.; Lampman, G et al. Organic Lab.Tech. 3rd ed.; Cengage: Belmont, 2011,
77.
(2) Isolation of caffeine from
tea, http://www.oxford.net/~mavarod/portal/school/caffeine.html(accessed
22/05/2015)
(3) Role and Reg. of
adenosine, http://www.cscb.northwestern.edu/jcpdfs/dunwiddie01.pdf.(accessed
25/03/2015)
(4) Nawrot, P.; Eastwood, J, e tal. Food Additives And Contaminant[online], 2003, Vol.
20, 2. http://ehis.ebscohost.com.db20.linccweb.org/ehost/pdfviewer/pdfviewer?
vid=3&sid=4415681e-3b1b-4d6a-bda9-f720bd9c776d%40sessionmgr10&hid=16 (a
ccessed 25/3/2015)
(5) Extraction, http://chemistry.csudh.edu/faculty/noel/CHE317L/Extraction
%20Experiment.htm (accessed 22/03/2015)

Question:
1. Why was the sodium carbonate added in experiment 11A and 11B?
Sodium carbonate was added in order to convert tannins and gallic acid to sodium
salts which are soluble in water.
2. The crude caffeine isolated from the tea has a green tinge. Why?

Because the tea leaves have chlorophyll and chlorophyll color is green based on
studies.
3. What are some possibilities explanations for why melting points of your
isolated caffeine may be lower than the true value (236 degree celcius)?

The difference in melting point was as a result of operator error and the crystal
absorbing moisture from the ice cooled tube used in the set up and from
surrounding air.
4. What would happen to the caffeine if the sublimation step was performed at
atmospheric pressure?
Caffeine would decomposed and produce less yield and more heat required.
5. Use

flow

weight a teabag
and prepare 50 mL
of tea solution

filter into
separatory funnel

add 0.5g sodium

carbonate

chart

to

outline

the

separation

repeat step above

using 10mL
methylene
chloridefor twice.

the drained layer

was added with
anhydrous sodium
sulphate.

drained the
bottom layer
(organic layer)

dry the solution

and then weight it.

add 15 mL
methylene
chloride, shake
and wait for
emulsion

technique