Name: Aruzhan Temiresh Date:

Experiment No: 2
Title of Experiments: Isolation of caffeine from Tea
I. INTRODUCTION
The objectives of the experiment are: to extract caffeine from the tea based on solid-liquid
and liquid-liquid extraction technique; to determine the percentage yield of the extraction; to
calculate recovery percentage of the sublimination process; and to identify the melting point
range of extracted caffeine; by comparing experimental melting point with literature
temperature, to observe the purity of produced caffeine.
Caffeine is an alkaloid, structurally related to the purine bases as shown in a figure 1.

Figure 1. Structure of caffeine.

The following experiment regarding the extraction of the caffeine performs batch extraction
(the tea bags are mixed with water and heated) and liquid – liquid extraction of the tea
mixture with methylene chloride. The amount of the caffeine in tea leaves approximates 2-3.5
percent. While caffeine has medium solubility in water, it is more soluble in the organic
solvent methylene chloride since the partition coefficient for caffeine between chloroform
and water estimates 10 (ref. equation 1 ), indicating higher affinity of this alkaloid to the
methylene chloride (methylene chloride is similar to chloroform in its physical and chemical
properties but it is much less hazardous). The solubility of caffeine in water is 1.8 g in 100
ml, while in dichloromethane 18.2 g/100 ml:
g
18.2
100 ml
Kch2 cl 2/ h 2 o= ≈10 (1)
g
1.8
100 ml
Water and methylene chloride are used as the solvents in a liquid-liquid extraction stage due
to their physical properties: the solubility of CH2Cl2 (methylene chloride) in a water is 1.6,
the solubility of the water is also low; regarding their density, CH2Cl2 is heavier (denser)
than water, 1.326g/ml and 1g/ml respectively. That’s why the mixing of the solvents results
in two-layer system (ref. figure 2). The upper layer contains water (less-dense solvent), while
the lower contains dichloromethane.

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 figure 2. Two-layer system.
The caffeine is expected to be concentrated in methylene chloride solvent, which enables the
recovering of the caffeine through the evaporation of the organic layer.
The procedure for the following experiment includes:
1. Preparation of the brewed tea.
Contents of tea bags were taken from a packet and weighed, the average mass of the tea for
one bag was determined. 180 ml of water was placed into 500 ml Erlenmeyer flask with 6 tea
bags and heated on a hot plate for 20-25 minutes. 5 g of CaCO3 was put into the flask with
the beforehand measured water. Calcium carbonate is used to hydrolyze the ester bond in
tannin, breaking it into glucose and gallic acid. Tannins are compounds with medium polarity
in water, consequently, this organic compound diffuses to the organic layer. In order to lower
its affinity to the organic layer, ester bonds are broken by water in the presence of calcium
carbonate producing polar compound – glucose, and highly ionic gallic acid, which are
highly soluble in a water.
2. Filtration
After water penetration into tea leaves, tea bags were removed and squeezed out against the
flask wall to extract a tea solution as much as possible. Buchner funnel, filter paper are used
to perform final stage of solid-liquid extraction. The filter paper was dampened with tea
mixture, turning vacuum on. Then, filtrated solution was cooled in an ice water bath.
3. Two-layer system
5 g of NaCl and 25 ml of CH2CL2 were mixed with the cooled tea filter in 250 ml separatory
funnel by gently rotating to avoid emulsion formation. Sodium chloride is added to decrease
the miscibility of the aqueous and organic layers, which prevents from an emulsion. The
pressure in the funnel was eased by the opening of a stopcock, allowing organic solvent with
caffeine to be collected in a 125 ml Erlenmeyer flask. The secondary liquid-liquid extraction
was proceeded by ch2cl2 addition into separatory funnel. After layers separation, 1 g of
drying agent, MgSO4, was added to the flask. Magnesium sulfate removes water molecules
from organic layer that are trapped in it due to emulsion by forming hydrates with water, also
MgSO4 breaks emulsion. The drying agent was formed by filtration through the glass funnel
with cotton in its neck, organic layer was collected into 100 ml round bottom flask, which
mass were weighted beforehand. MgSO4 was rinsed with 2 ml of methylene chloride.
4. Evaporation of the solvent
Ice added to the cold trap of a rotatory evaporator to condense evaporating organic solvent.
Round flask was attached to the apparatus and lowered into the hot bath. Rotation and pump
are turned on until completed evaporation of the solvent. Produced caffeine at the bottom of
the round flask were weighted.
5. Purification of caffeine
Caffeine was scratched out from the bottom of the flask by using spatula onto the pre-tared
petri dish. Petri dish with caffeine was placed onto the hot plate, 3 filter were put to cover the
petri dish. The lowest filter paper (where caffeine will be collected) was weighted beforehand
the sublimation procedure. On top of the construction flask with cooled water was placed.

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The sublimation took place for 10 minutes by gradually increasing the temperature till 315℃.
Purified caffeine was weighed, and its melting point have been determined.

II. MATERIALS
The chemical equipment consisted of: 1x 500 ml Erlenmeyer flask, separatory funnel (1x 250
ml), Buchner funnel, Erlenmeyer flask (1x 125 ml), glass funnel, rounded flask (1x 100 ml),
graduated cylinder (1x 10ml), pipette (1x 50 ml), petri dish, hot plate, vacuum pump and
melting point apparatus.

III. CHEMICALS
Table 1. Physical-chemical properties of reagents
Compound MM, m.p., b.p., Density, Solubility Flamm Toxicity/
g/mol oC o
C g/mL ability hazards
calcium 100.09 825 - 2.7-2.9 Slightly - Causes eye
carbonate ℃ soluble in irritation, may
a water cause skin
irritation.
Ingestion of
large amounts
may cause
gastrointestinal
irritation.
[1]
Water 18.013 0 99.9 0.997 Poorly - -
4 8 liquid at soluble in [2]
25℃; aliphatic
0.917 and
solid aromatic
hydrocarb
ons
Methylene 84.93 -97 39 1.33 - 1 Causes skin and
chloride eye irritation,
may cause
drowsiness,
cancer, damage
to organs
through
prolonged
exposure
[3]
Sodium 58.44 801 1461 2.165 360 g/L 0 Causes skin and
chloride (20℃) eye irritation
[4]
Magnesium 120.37 1124 - 2.65 26 g/100 0 May cause skin
sulfate ml and eye
irritation
[5]

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IV. CALCULATIONS
The percentage yield of the extraction:
m(crude caffeine )
yield= (1)
m(tea leaves)
The recovery of the sublimation:
m( purified caffeine)
recovery = (2)
m(crude caffeine )
The mass of the recovered caffeine after liquid-liquid extraction can be calculated as:
Mass of impure caffeine = mass of round glass – mass of round glass = 63.21 – 63.09 = 0.12g
The percentage yield of the extraction:
% = 0.12/(6x1.79g) x100 = 1.1173 percent
The recovery of the sublimation:
The mass of the purified caffeine can be calculated as:
m = -(filter paper weight with caffeine - filter paper weight) = -(0.85-0.88) = 0.03g
The recovery of the sublimation:
Recovery = 0.03/0.12 x 100 = 25%
*the negative sign is assigned since filter paper during sublimation lost its mass due to water
evaporation from it, consequently, the observed mass of the filter paper weight with caffeine
will be lowered.
Results:
Table 2. Physical properties of impure and pure acetanilide
Impure Pure
Characteristics Solid, greenish crystals White tiny crystals
Melting point range, ℃ 224.7-244.3 233.5-235.1

Table 3. Experimental and theoretical recovery mass

Crude Pure
impure caffein
caffein e
e
Mass, g 0.12 0.03
Yield of 1.1173
extractio
n%
Recovery 25
%

V. DISCUSSION
The experiment is aimed to extract caffeine from the tea by applying solid-liquid and liquid-
liquid extraction methods based on solubility of the caffeine in used solvents.
It was determined that the melting point of purified (after sublimation) caffeine is in the range
of 233.5-235.1℃, while for crude one is 224.7-244.3℃. By comparing the melting point
ranges 1.6℃ and 19.6℃ for pure and impure caffeine respectively, it can be concluded that
impurity in a compound (such as tannins) decrease the actual melting point and expand the
melting temperature range. The literature temperature estimates that caffeine starts to melt at
233℃ and ends at 236℃ . The experimentally obtained caffeine fits that range ideally,
indicating high purity of the product.

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According to the recovery mass of the caffeine, it was determined that 25 percent of product
is recovered from its impure compound. However, the loss of paper filter weight due to the
water evaporation can lower the percentage even more, which means that usage of the filter
paper in a sublimation stage is inappropriate. Instead beforehand dried papers can be used to
exclude water loss. The low recovery percentage can be explained by loss of the product
during scratching out of the round-bottom flask, and inefficiency of the liquid-liquid
extraction of this protocol. The efficiency of the liquid-liquid extraction depends on not only
partition coefficient, but also on the volume of the solvent used and the number of
extractions. The increase in a volume of the solvent and further repeating of the extraction
could result in a higher recovery.
By calculation, the amount of caffeine per 11 grams (6 bags) of tea was determined to be
1.1173%. According to the theory, 2-3.5 percent of the caffeine is expected to be
concentrated in a tea. The obtained data is lower due to during the extraction of caffeine,
emulsion between immiscible layers. Emulsion occurs because of the formation of
suspension of one liquid in the other one. Consequently, during phase separation, the
amount of caffeine that have been separated depends how gentle the mixing of solvents has
proceeded.
VI. CONCLUSION
In this laboratory work, the solid-liquid and liquid-liquid extraction have been performed to
produce pure caffeine from tea. The melting point range of pure caffeine corresponds to
expected value (tabled temperature, 233-236℃ ), indicating high purity of the product.
Consequently, it is concluded that the main goal of the experiment to extract pure caffeine
was succeeded. Nevertheless, the recovery percentage and yield of extraction are low, 25 and
1.1173 per cent respectively. Even when lacking absolute accuracy, the procedure is useful
for practical purposes, the solvents with higher partition coefficient can be used and
extraction can be repeated several times. The objectives of the experiment have been
achieved; hence, it can be claimed the experiment proceeded successfully.

Pre-lab question answers:

1) During the caffeine extraction carbohydrates, tannins, amino acids, saponins and
pigments are extracted too.
2) Ester bonds of tannins are easily broken by water in the presence of caco3, producing
gallic acid and glucose. CaCO3 will react with tannic acid resulting in calcium tannate,
which is insoluble and precipitates. CaCO3 and calcium tannate are removed easily via
filtration process.
3) The addition of the sodium chloride decreases the miscibility of solvents, and helps in
avoiding emulsion formation: addition of salts increases the ionic strength of water
phase. Magnesium sulfate acts as drying agent, it removes water molecules by
producing hydrates from organic layer.
4) No, methanol is soluble in a water, the separation of two layers does not procced,
which makes impossible to do liquid-liquid extraction.
5) 1x 500 ml Erlenmeyer flask, separatory funnel (1x 250 ml), Erlenmeyer flask (1x 125
ml), glass funnel, rounded flask (1x 100 ml), graduated cylinder (1x 10ml), petri dish
7)
Solubility in water = 1*100/55 = 1.82 g/100ml
Solubility in chloroform = 1*100/5.5 = 18.2 g/100ml
Solubility in diethyl ether = 1*100/530 = 0.19 g/100ml
Solubility in benzene = 1*100/100 = 1g/100ml

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 Kchloro/water = 18.2/1.82 = 10
Kdiethyl/water = 0.19/1.82 = 0.1
Kbenzene/water = 1/1.82 = 0.55
The partition coefficient for the chloroform has largest value, indicating the higher affinity
of the caffeine to the chloroform. That’s why chloroform is appropriate solvent for
extraction.
8)
Methylene chloride and toluene are less toxic/hazardous than chloroform and benzene.

In-lab questions:
1) The objectives of the experiment are: to extract caffeine from the tea based on solid-
liquid and liquid-liquid extraction technique; to determine the percentage yield of the
extraction; to calculate recovery percentage of the sublimination process; and to
identify the melting point range of extracted caffeine; by comparing experimental
melting point with literature temperature, to observe the purity of produced caffeine.
The liquid-liquid and batch extraction have been introduced as the new techniques.
2) The percentage yield of the extraction:
% = 0.12/(6x1.79g) x100 = 1.1173 percent
According to the theory, 2-3.5 percent of the caffeine is expected to be concentrated in
a tea. The obtained data is lower due to during the extraction of caffeine, emulsion
between immiscible layers.
3) The recovery of the sublimation:
Recovery = 0.03/0.12 x 100 = 25%
The low recovery percentage can be explained by loss of the product during scratching
out of the round-bottom flask and insufficient number of repeated extractions.
4) The melting point range of pure caffeine (233.5-235.1℃) corresponds to expected
value (tabled temperature, 233-236℃ ), indicating high purity of the product.
7)
Due to the presence of polar keto group, acetone possesses a permanent dipole
moment. Petroleum ether due to the absence of polar carbonyl group is accounted as
nonpolar compound. So, acetone is more polar.
8)
Petroleum ether is insoluble in water which enhance the formation of immiscible layers.

Bibliography:
[1] – Material Safety Data Sheet for calcium carbonate. Retrieved from:
https://fscimage.fishersci.com/msds/03880.htm
[2] – Material Safety Data Sheet on water. Retrieved from:
https://fscimage.fishersci.com/msds/00199.htm
[3] – Material Safety Data Sheet on methylene chloride. Retrieved from:
https://www.fishersci.com/msdsproxy%3FproductName%3DD3720%26productDescr
[4] – Material Safety Data Sheet on Sodium chloride. Retrieved from:
https://fscimage.fishersci.com/msds/21105.htm
[5] – Material Safety Data Sheet on magnesium sulfate. Retrieved from:
https://fscimage.fishersci.com/msds/13510.htm

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