Isolation of

Caffeine from Tea

Leaves

Advanced Chemistry 2017-2018

Mahidol University International
Demonstration School (MUIDS)
September 15,18 2017

Mui Roong ATOM pizza lilly

Pimchanok, Rawisara, Yanisa, Supichaya, Nitinan
 Abstract
The goal of the project was to perform a caffeine extraction as well
as Thin Layer Chromatography, also known as TLC technique. The
caffeine was extracted from tea, which was later used in TLC experiment
in order to examine the different types of compounds it contained.
Unfortunately, some errors were made, resulting in not as clear separation
of the polar and nonpolar compounds on the TLC plate.

Objective
- To observe the extraction of caffeine from tea.
- To learn how TLC technique can be applied to use within the experiment
of caffeine extraction.
 Introduction
Caffeine is an odorless white powder with bitter taste. It is a common compound
found in tea beverages, which can play an important role in stimulating humans brains
and central nervous system. At the same time, it can also prevent people from being tired
and sleepy (Petre, 2016). The caffeines chemical properties include both its melting and
boiling points. While its melting point is 460 F, it has a boiling point of 352 F at 760
mmHg (National Center for Biotechnology Information, 2017). For this experiment, the
caffeine will be extracted in order to see the elements that are contained in the tea
leaves.
Dichloromethane is a solvent used for separating caffeine from the tea solution. It
is a colorless liquid with a boiling point of 103.6 F at 760 mmHg and melting point of
-142.1 F(National Center for Biotechnology Information, 2017).
In order to run the caffeine up the TLC plate, ethanol is required to dissolve the
caffeine crudes and crystals into liquid before they are placed onto the TLC plate. Its
physical property is a clear colourless mobile liquid that has a mild smell like whiskey. The
point where it starts to boil is 173.3 F at 760 mmHg, yet it will start to melt at -173.4 F
(National Center for Biotechnology Information, 2017).

Another important solution used in this experiment is iodine. It acts as a crystal that
gives off its colour to the TLC plate, in which makes it easier to see the point of each
compound that are composed within the sample. The physical properties that can be
observed are violet-black crystals with a metallic luster and a sharp odor. The boiling point
at 760 mm Hg is 365 F, meanwhile its melting point stands at 236 F (National Center for
Biotechnology Information, 2017).
The experiment of caffeine extraction involves in conducting two main different
processes : extracting the caffeine from tea leaves and using TLC techniques to observe the
composition of caffeine itself, which is a technique used to separate nonvolatile mixtures.
The first process can be done by using the substance called Dichloromethane to help
pulling out the caffeine from the tea. The two substances, Dichloromethane and tea, were
put in the separatory funnel, which should be mixed together but not overmixed causing
the emulsion. To perform the process effectively, it was important to shake the separatory
funnel in a C shape slowly and carefully. Once the first part was done, giving the caffeine
crude and crystal, the TLC technique was conducted. The two forms of caffeine were mixed
with cold Ethanol, which were later put on the TLC plate by capillary. Normally, with the
second process, the different compounds should run up to the different spots on the plate,
which their properties could be identified from this part based on the type of types of their
structures : polar and nonpolar. The less polar the structure is, the longer in distanceit will
go due to the fact that it tends to move away farther from the water, which is a polar
molecule. However, there were some errors occurred in the experiment, resulting in an
unclear separation of the compounds on the plate.
 O-Ring Clamp

Watch Glass

Thermometer

Suction Flask

Glass Funnel
 Dropper

Round Bottom Flask

Chemical Used

1. Tea Leaves
6. Sodium Sulfate
2. Dichloro-
7. Iodine
methane Capillary
8. Silica Gel
3. Ethanol
9. Hexane
4. Ethyl Ace-

tate

Petri Dish Melting Point Machine

 Procedure
1. Remove the staple from the tea bags and place it
on the weighing scale. Records the weight of both
bag and tea leaves and place the tea leaves back
into the bag. Do not forget to stable the tea bag.
2. Place the tea bag into the bottom of the beaker - Caffeine Extraction -
and pour 30 ml of water into it. Place the beaker on
the hot plate and gently boil the substance in the
beaker for 10 minutes. Use the stirring rod to push
10. Open the stopper and wait for a few minutes to let
the tea bag down if the tea bag rise up.
the bubble inside the separatory funnel disappear. In
3. Pour hot liquid into a new beaker and add 20 ml the separatory funnel, there must be two separated
of water. Bring that beaker to the hot plate and layers of the liquid: brown layer (tea extraction) on the
heat up. Again pour the liquid into the beaker anoth- top and clear layer (dichloromethane) beneath it.
er beaker. Use the cork to press the tea bags in or-
11. Slowly open the stopcock to let the beneath layer
der to obtain the liquid as much as possible and
of the liquid (dichloromethane) flow down into the
transfer these liquid into the erlenmeyer flask. Leave
erlenmeyer flask. Prevent the brown layer to flow
the tea extract for a while to let it cool down and
down into the erlenmeyer flask by close the stopcock
remove the tea bag away
as the brown layer is getting near the neck of the
4. Pour the tea extract solution into the separatory separatory funnel.
funnel.
12. Repeat step 5-11 for 3 times.
5. Add 5 ml of dichloromethane into the same
13. If the bubble appears too much add a small drop
separatory funnel and put the stopper on the top to
of sodium chloride solution.
close the separatory funnel. Take the separatory
funnel out of the ring stand and hold it by grabbing 14. After complete the extraction discard the aqueous
the neck of the funnel with your right hand while the layer.
other hand need to control the stopcock to open 15. Slowly add sodium sulfate into the dichloromethane
and close. Holding the stopper firmly and invert the extraction and use your hand to swirl the erlenmeyer
funnel to the other side so that the liquid is gathered flask. Add the sodium sulfate until the sodium sulfate
at the stopper side. The liquid inside the funnel inside the solution does not clump with each other.
should not flow to stopcock side and point the stem
of the funnel away from the people. 16. Place the buncher funnel on the suction flask and
6. Shake the separatory funnel in C-shape but do connect the suction flask with the vacuum machine.
not shake it vigorously because the pressure may Place the filter paper inside the buchner funnel.
build up inside the funnel.
17. Pour all of the dichloromethane extraction that
7. Open the stopcock to release the pressure that
contain sodium sulfate inside into the buchner funnel.
build up inside the funnel after shaking.
All of the liquid will pass through the paper and sodi-
8. Repeat step 6-7 for 3-4 times. um sulfate will remain on the top of the filter paper.
9. Close the stopcock tightly, turn the separatory
funnel upside down and place it back to the ring 18. Discard the sodium sulfate and the filter paper.
stand. Place the erlenmeyer flask underneath the Disconnect the vacuum machine from the suction
separatory funnel. flask.
 Procedure
- Caffeine Extraction -

19. Pour all of the liquid extraction inside the suction flask into the round bottom flask by using glass funnel to
prevent from spill.
20. Pour the water into the hot bath. The water should be enough for the round bottom flask to sit in it.
21. Open the rota vapor machine and connect the round bottom flask that contain the liquid extraction to the
machine. Adjust all of the temperature and pressure inside the vacuum according to the instructor.
22. Wait until all of the liquid condense and flow down to round bottom flask at another side of the machine. In
order to determine whether the separation is successful or not the liquid which have the higher boiling point
will be left over in the original round bottom flask.
23. After finishing the process, obtain the caffeine extracted and divide in equal amount for 2 groups. One
group will go through the process of crystallization and another one will be kept as a crude. Then, both of
them will be used in the TLC process.

TLC Test Melting Point-

Boiling Point Test
1. Draw 1 c.m. lines on both end of the TLC plate by pencil.
2. Use the dropper to drop a small amount of the cold ethanol
to the crude of caffeine extracted and the crystal from crystal- 1. Put the same small amount of
lization. both samples in the test tubes
separately.
3. Use the capillary to tap the crude which already dissolved in
the ethanol on the line drawn on the TLC plate . The amount of 2. Put both samples in the melt-
the crude should be be enough to be seen easily. ing point machine.

4. Use the capillary to tap the crystal which already dissolved in 3. Put the thermometer inside the
the ethanol on same line, same TLC plate besides the crude. machine.
The amount of crystal should be enough to be seen easily. 4. During the experiment, use cell
5. Wait until the cold ethanol evaporates from the plate and on- phones camera to record the
ly the crude and crystal are left over, put the TLC plate inside changes occurred by connect the
the TLC chamber containing Hexane and Acetate (1:1) by let the phone with the machine and
line which contain the samples dips in the solvent. let the camera of the phone place
at the hole which used to ob-
6.Wait until the solvent (Hexane and Acetate) goes up the plate serve the changes.
to another line drawn, bring the plate up and wait until all of
the solvent evaporate. 5. Open the machine and ob-
serve both samples until they
7. Put the TLC plate in the petri dish containing Iodine (Iodine melt, then record the results.
Chamber) until the marks on the plate appear.
8. Obtain the TLC paper after the marks appear and analyze
 Results
For the first day, the result is caffeine extracted in the form of cream color
liked powder called crude as its not pure. And because half of the caffeine
extracted must be crystallized, there is just a small amount of the crude.

For the second day, from the TLC test, the result should be able to
obtained but due to the error occured, there is no result shown in the TLC
plate. None of the spot appeared on the TLC plate. However, during the
melting point and boiling point test, the result shows the difference of boiling
point/melting point between the crude and crystal which the crystal has 238
degree celsius while the crude has 225-227 degree celsius. The difference
between these two is 11 -13 degree celsius.
Discussion..

1. Explain why emulsion can be observed during liquid-liquid extraction?

How to avoid it? If it occurs, what is simple methods to destroy it?
Ans. The emulsion can be observed during the liquid-liquid extraction because of the process of
shaking the separatory funnel. By shaking it too fast, the two solvents will tend to produce an unclear
separation layer, which can cause the method of getting rid of water from the caffeine to be harder.
It can, however, be avoided by shaking the separatory funnel slowly and carefully. In the case that
emulsion actually occurs, there is a simple method of adding sodium chloride into the solution. The
sodium chloride will then act as a compound to trigger the two layers of the solvents to separate more
in which can helps to eliminate the emulsion.
2. Why do we need drying agent (Na 2SO4 ) after finishing liquid-liquid
extraction and before evaporation process?
Ans. After finished liquid-liquid extraction, Sodium Sulfate (Na2SO4) is added before evaporation
process to decrease the amount of water in the solution. Sodium Sulfate will catch water and create
clumps in the solution. Repeat this process until sodium sulfate the added become powder which
mean that it saturated. Then use the filter paper to filter the clumps and start evaporation process.
3. What are errors during the experiment and how to avoid them or
improve your technique?
Ans. During the first part of the experiment, the separatory funnel containing the tea and
Dichloromethane might have been shaken too much, which resulted in the huge amount of bubbles in
the substance. Besides that, the separatory funnel might have not been sitting long enough to let the
bubbles to disappear before releasing the excess Dichloromethane down the beaker. Because of the
bubbles, the caffeine was unable to be extracted as purely as it could be, causing the unclear
separation of the compounds during the TLC technique. To avoid the problems, the experiment should
be conducted from step by step more slowly and carefully. That way the bubbles would be seen less,
giving us a more pure caffeine extract. With the more pure extract, the different compounds would be
able to run up the TLC plate better, allowing us to identify the type of their structure more effectively,
whether they are nonpolar or polar molecules. Besides that, there were some contaminations occurred
during the experiment on the TLC plate. This is due to the fact that there might have been some
particles or molecules of other elements lie on the plate, when it was being transferred from one place
to another. For example, when it was brought to the chamber. This error decreased the purity of the
caffeine. Moreover, the caffeine crude and crystal might have not been put on the plate enough,
which limited the multiple compounds in itself from separating as clearly as it could have been.
4. How Do you know you get caffeine from tea leaves? Is it pure?
Compare your caffeine extract product with the others?
Ans. The cream powder which is the result from the experiment would be determined through the TLC
test and the boiling/melting point test to test whether its caffeine or not. First, TLC test is a process
where 2 or more samples will be identified the similarities or difference by testing their polarity. Since
the same compound will have the same polarity; if the samples are spotted at the same place and
have only one spot, they are the same compounds and pure. However, if they are not the same
compound, they will be in different spots. Moreover, even though they are the same compound but
different in purity, the one that is not pure will be considered as a mixture and leave more than 1 spot
on the plate which means there are many other components in the compound. However, due to the
errors occurred during the experiment, there is no result shown in the TLC plate. Therefore, the result
can not be conclude yet whether they are caffeine or not. But there is another process called melting
point and boiling point test. This test can also help to identify the compound since the pure compound
will have a certain melting/boiling point. However, the boiling/melting point test cant identify clearly
whether the samples are different compounds or just different in purity, because the purity and
different compounds can both affect to boiling point and melting point. So, this process is more
appropriate in determine the purity of the known compounds since the range of exploring is reduced
and more specific. Thus, the result cannot be concluded whether it is caffeine or not due to the error
occurred and the lack of reference. However, predicting that the compound is caffeine, the crude
can be determined its purity by comparing its melting/ boiling point with the pure caffeine which the
result shows that it is not pure because there is a difference in melting/boiling point for 11-13 degree
celsius compares to the pure caffeine. Finally, the caffeine extracted of our group cant be compared
to the other groups because there is no result shown in the TLC plate of our group and during the
melting point/ boiling point process, only a small sample that gathered from many groups is tested.
However, the result should be the same because during the rota vapor process, all of the solution
(caffeine and dichloromethane) is mixed together. Therefore, during TLC test, every groups use the
same sample to test.
5. Determine the strongest intermolecular force present in Caffeine.
The molecular formula of caffeine is C8H10N4O2. Caffeine is a polar molecule because its chemical
structure that contains four different chemicals and each chemical has different electronegativity.
Moreover, its molecular geometry allows the molecule become polar as well because the negative and
positive charges of each atom in the molecule does not cancel with each other. According to its
polarity, the intermolecular forces that can exist between each caffeine molecule are Dipole-Dipole
force and London force. However, the strongest intermolecular force that exists between each
caffeine molecule is the Dipole-Dipole force because London force is just a weak temporary force
that caused by the motion of the electrons of two atoms inside the molecule as they moving closer
near each other. In addition, the Dipole-Dipole force is the strongest intermolecular force because the
attraction of the negative and positive charges.
Conclusion
From this experiment, there is no result shown in TLC plate
because caffeine that extracted is not pure enough according
from the melting point which is 226 C instead of 235 C. There are
errors occurring in the experiment such as the contamination on
TLC plate, too less drops of crude and crystal on TLC plate, other
compounds in tea dissolve in dichloromethane and the period of
time that wait before the tea and dichloromethane separation
to decrease amount of bubbles.

Reference