Paul Henry Camacho

Mara Krista Coo

Ma. Kathleen Pearl Grande

Date Performed: February 1-4, 2016

Date Submitted: February 12, 2016

Experiment 1
Isolation of Caffeine From Tea
Introduction
Caffeine or 1,3,7 trimethylxanthine with the chemical
formula C8H10N4O2, is an organic substance commonly present in
coffee, teas, energy drinks and medicine. It is a crystalline solid of
white color and bitter taste. It can be found in nuts, seed and leaves of
some plants. It is widely used as a brain stimulant due to its effect on
the central nervous system which causes non-drowsiness, alertness
and energy boost in humans. It is also considered a weak diuretic or
can cause loss of little amounts of fluids in the body without dehydrating it. It was also proven
that caffeine can improve the metabolism by boosting the metabolic rate and increase the burning
of fats. Caffeine can be addictive too and can cause withdrawal symptoms if consumption is
stopped after addiction.
Amount of caffeine present in different substances can vary depending on the level or
significance of usage. Caffeine content in anti-fatigue and headache medicine is a lot higher than
those in coffee and teas. Caffeine is usually consumed by humans through infusion or extraction
in water. One common example of this is tea which uses caffeine-containing tea leaves that
contains a small amount of caffeine of about 3%.
Tea is a beverage made by steeping leaves and leaf buds of Camellia sinensis or the tea
plant. The active ingredients in tea are alkaloid groups with similar structures. Alkaloids are
mixed group of plant constituents that contain a nitrogen-bearing molecule which are
pharmacologically active like theobromine and theophylline. The most active alkaloid present in
tea is caffeine which belongs to purine alkaloid group.
In this experiment, caffeine was isolated from tea leaves using step-by-step and specific
methods to eliminate other organic compounds and components of the leaves which were also
water-soluble like flavonoids, cellulose, chlorophyll, theobromine and others. Extraction was
used in order to isolate the caffeine from other compounds present in the leaves. Several types of
extraction was used like solid/liquid extraction which was done in the form of brewing. Also,
Liquid/liquid extraction was done and distillation was also performed. Also, certain chemicals
were used in order to eliminate other components of the tea leaves which were specific to every
target compound/component to be eliminated.
The objectives of this experiment were to isolate the caffeine form tea by extraction and
find its mass and percent recovery. The determined mass and percent recovery was compared to
the amount claimed by the brand of tea. The tea brand used in this experiment was Lipton Tea.

The experiment was limited to tea leaves only since other substances also have a different series
and sets of steps and protocols used in separating organic compounds from them
Data and Results
Tea
Pre-Weighed Vial
Vial + Caffeine
Caffeine
g solute caffeine
%Caffeine
100
g tea sample
0.0115
%Caffeine
100
10.0523
0.1144%

Weight (g)
10.0523
20.6343
20.6458
0.0115

Observations

During the salting out of caffeine part, the following were observed:
After NaCl and Ca(OH)2 were added and the solution was heated for 15 mins., a brown,
chocolate-like liquid was formed. Then, after vacuum filtration using a Buchner Funnel, a
dark green to brown-colored filtrate was gathered.
Then, during the liquid-liquid extraction:
After the addition of DCM, small amount of air escaped through the stem of the separatory funnel
when allowed to vent. Then after mixing and gently shaking, two layers of liquid were observed.
One with a dark brown color and light brown color which was the aqueous layer and the organic
layer which was clear. Also, bubbles were formed at the aqueous layer. After the third addition of
DCM, more bubbles were formed and only a small amount of caffeine extract was separated and
most of the liquid in the funnel was already bubbly.
During distillation:
Only a small amount of caffeine was separated and deposited on the bottom of the flask. The
separated caffeine extract had an aromatic and fresh tea drink-like smell.
Lastly, during drying:
Small amount of white crystals were deposited at the side and bottom of the vial. The crystals still also
had an aromatic smell.

Discussion
In this experiment, one of the main objectives is to isolate the caffeine from tea through methods
of extraction and distillation. In the extraction process, caffeine was isolated through salting out with
water. In this process, after adding 100mL of boiling water to the pre-weighed tea bags and pressing out
as much water, sodium chloride and calcium hydroxide were added to the extracted solution. Sodium
chloride was added because water will be attracted to salt which will leave the less polar caffeine, thus,
isolating it.[1] Also, calcium hydroxide was added to react with the tannins present in tea to isolate these
components from caffeine. Adding calcium hydroxide also makes the solution basic. Undissolved solids
were separated using a Buchner funnel.[1]
Another procedure done in this experiment was the liquid-liquid extraction of caffeine. In this
process, dichloromethane (DCM) was added to the tea extract. This was done to dissolve the tea since it is
more soluble in DCM than in water. After the addition of DCM, the solution was mixed by gently rocking
it to avoid formation of emulsions. During the experiment, salt was added every time there is presence of
intolerable amounts of emulsions. The organic layer was then drained separating it from the aqueous
layer. This whole process was repeated three times.
The third part of the procedure was the washing and drying of caffeine extract. The caffeine
extract was washed with 10% sodium hydroxide solution. This was done to minimize caffeine lost along
the process. The extract solution was transferred into an Erlenmeyer flask and was dried by adding
anhydrous sodium sulfate. There was a formation of slightly solid compound which was decanted and
used for the distillation process.
In the distillation process, the extraction solvent was distilled from the caffeine extract using a
water bath as a heat source. The temperature was monitored until it reached 40 degrees Celsius, just in
time when the flask is dry and no more distillate is being collected. The crude caffeine residue was
transferred into a pre-weighed vial, rinsed with small amounts of acetone and was exposed to air to
evaporate the acetone. The vial was weighed again to determine the mass of caffeine.
In this experiment, the possible sources of error were the 1) presence of intolerable amounts of
emulsion during the liquid-liquid extraction, 2) contamination of extracts and solutions along the process
since this was a lengthy experiment and 3) impurities that might be present in the containers and
glasswares used in the experiment.
Conclusion and Recommendations
Extraction is a method for the separation of organic compound from a mixture.
In this experiment caffeine was extracted from tea. Different processes were
undergone in order to isolate caffeine from tea. Caffeine was first salted out by
adding NaCl and Ca(OH)2, liquid-liquid extraction followed, by adding
dichloromethane to separate the organic compound. It was then washed and dried
ready for distillation. Crude caffeine was the resulting residue.
The mass of the crude caffeine was 0.0115g and the computed percent caffeine
is 0.1144%. The claimed mass caffeine of Lipton Yellow Tea is 0.0055g per bag. In
the experiment where approximately 6 bags were used, the calculated gram
caffeine recovered was 0.0115g. The recovery of the caffeine was very small due to
some errors made during the experiment. Some reactions may have not gone into
completion or there was a loss of sample during transfer or during some certain
procedures.

To improve the reliability of the results of the experiment, it is recommended

that the experiment be conducted in 1 meeting to avoid contamination of extracts
and solutions along the process. Also, it recommended that the glasswares and
apparatuses used be properly washed and free from impurities that might cause
error.
Questions and Answers:
1
Discuss the role/s of the following in the isolation of caffeine:
a Dichloromethane- This is an organic solvent used in liquid-liquid extraction
because caffeine is more soluble in dichloromethane than in water.[1]
b NaOH aqueous solution- Sodium Hydroxide is a basic solution, it is used to
remove impurities and minimizes the caffeine lost. [1]
c Ca(OH)2 - Since Calcium hydroxide is a base and tea is consist of other
components including chlorophyll, tannins and flavonoids which are acidic,
Ca(OH)2 will react with tannins to form salt. This will isolate them from
caffeine. This also makes the solution basic. [1]
2
Explain why salt was added to the water before the liquid-liquid extraction of
caffeine. What is happening, why and how?
Salt is added before liquid-liquid extraction because water will be more attracted
to the salt which is very polar than that of the less polar caffeine. This will salt out
the caffeine from the solution. Adding salt also minimizes the formation of
emulsions. [1]
3
How efficient is the extraction of tea leaves containing 1.0g of caffeine with 3
20-mL portions of dichloromethane over that of a single step extraction?
It is efficient to extract caffeine from tea with 3 20mL portions of
dichloromethane over that of a single step extraction for it can obtain much of the
pure sample. It extracts the caffeine that where not extracted from the previous
one. [1]
4
What are emulsions and why do they form during extractions? How are they
minimized and removed?
Emulsions are mixture of two or more liquids in which one is present as droplets.
These are results of vigorous shaking of the separatory funnel. To avoid these
emulsions, mixtures should be gently mixed or add salt to aqueous layers. [1]
5
Why is it necessary to remove the stopper from the separatory funnel when
liquid is being drained from it through the stopcock?
Removing the stopper from the separatory funnel when liquid is being drained
will avoid the building up of vacuum above the liquid. This vacuum will reduce the
rate of draining and can get air in that will mix the solution again. [1]

References
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2016
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[2] Marsiano, M., Alkaloids. The Naturopathic Herbalist.

http://thenaturopathicherbalist.com/plant-constituents/alkaloids/.
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Retrieved From
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[3] Uzunalic, Amra, et. al., Extraction of active ingredients from green tea (Camellia
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from

http://www.drugs.com/caffeine.html

[5] Caffeine Content for Coffee, Tea, Soda and more. Nutrition and healthy eating.
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[6] Persad, Leeana, Energy drinks and the neurophysiological impact of caffeine.
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