Isolation of Caffeine from Tea Solution

Abstract
The purpose of this lab is to extract the pure caffeine from tea sample and to know
more some methods that can use to identify the substance.​ ​We use four procedures
including extraction, filtration, TLC (by comparing with the pure one that we already know),
and melting point to identify that our extraction is pure or not. There was an error during the
experiment as we do not know the exact point that it is start to melt. However, the result of
this experiment is our extraction is not pure, it can be noticed because our substance begins
to melt before the pure caffeine sample, so we can assume that it is contaminated.

Introduction
People might not known that most beverages taken in present contains Caffeine
inside. This type of compound can be found in things people ingested in their everyday life
such as coffee, tea, cocoa, milk chocolate, baking chocolate, coca-cola, excedrin,and or
even in no-doz. Caffeine (​C8​ ​H1​ 0​N4​ ​O2​ ​) is a methylxanthine considered as a member of the
class called Alkaloids in organic compounds (Pubchem.ncbi.nlm.nih.gov, 2018) [1].
Naturally, containing basic compounds such as Nitrogen that can be found in plants. The
taste of caffeine is commonly bitter. Caffeine is mostly used as a pharmacological agent
affected in human’s nervous system helping in increase in alertness and agitation. Some of
these compounds included: nicotine, cocaine, morphine, and strychnine. A diagram of
caffeine’s structure is given below.

Caffeine is helpful in relaxing smooth muscles, stimulating cardiac muscles and

diuresis and also treating type of headache. However, it is physically addictive. People with
caffeine addiction and tend to stop taking it may possibly experiences insomnia, headache,
and nausea.
However, in this lab, we will mainly use the extraction method. This method will
separate the substances when they are mixed together in immiscible phrase such as oil and
water in this lab (Chemicool.com, 2018) [2]. In addition, the substance that is less soluble will
move to the substance that is more soluble, so we can see the separation of two layers in
this method and use the stopcock to control the volume that we want. However, the
substance that has more density will present in lower part and the substance that has less
density will present in the upper part. For example, when extract between water and
trichloromethane, trichloromethane is more dense, so it will present in the lower part in the
separatory funnel and water is less dense, so it will present in the upper part in the
separatory funnel. After that, we will use TLC (Thin Layer Chromatography), it is the method
that used to separate mixtures of substances into their components. All forms of
chromatography work on the same principle (Chemguide.co.uk, 2018) [3]. In this method, it
has 2 phases which are stationary phase and mobile phase. In addition, stationary phase is
the phase that support the mobile phase. The mobile phase will flow through the stationary
phase and carry the components and mixtures with it. Also, different components will flow in
the different rate (Chemistry LibreTexts, 2018) [4].

Purpose
To extract caffeine from tea solution and to learn more about the methods that are
used in this experiment such as extraction, filtration, TLC, and melting point.

Material
- tea bags
- 50 ml Erlenmeyer flask
- 125 ml separatory funnel
- Ring stand
- dichloromethane
- glass rod
- Capillary tube
- TLC plate
- rota evaporator
- vacuum
- TLC camber

Method
1. Transfer the tea extract from the ​50 ml Erlenmeyer flask to a ​125 ml separatory
funnel​ that is supported by a ring on ​a ring stand.

2. Add 20 milliliters of ​dichloromethane to the separatory funnel. Stopper the funnel and
grab the neck of the funnel with one hand, holding the stopper into the neck of the
funnel. With your other hand grasp the stopcock in such a way that you can turn the
plug in the barrel to open and close the stopcock. While holding the stopper tightly
into the neck of the funnel, invert the funnel so the liquid no longer is in contact with
the stopcock. Pointing the stem of the funnel away from everybody, open the
stopcock to release any pressure that may have built up inside the funnel. Close the
stopcock and agitate the material in the funnel without shaking it vigorously – you
want the contents to mix, but you do not want to generate an emulsion. Point the
stem of the funnel upward, and making sure no liquid is in contact with the stopcock
 open it to again relieve any pressure build-up. Close the stopcock. Agitate again as
before and reclamp the separatory funnel to the ring stand.

3. Allow the contents of the separatory funnel to settle. There should be two distinct
mostly clear layers. If there is an emulsion (cloudy) layer between two clear layers it
is sometimes possible to break the emulsion by swirling the contents of the funnel or
stirring the contents using a ​glass rod​. If the emulsion persists seek your instructors
help

4. Carefully drain the lower (dichloromethane) layer into a ​25 ml Erlenmeyer flask​. Try
to not include any of the aqueous (upper) layer. If there is a lot of emulsion, include it
in the Erlenmeyer flask.

5. Repeat steps 2 through 4 using a second 20 ml portion of dichloromethane.

6. Combine organic layers from step 2 - 4, then wash it with 20 mL of 6M NaOH
7. Carefully drain the lower (dichloromethane) layer into a ​25 ml Erlenmeyer flask​. Try
to not include any of the aqueous (upper) layer.
8. Wash the organic layer from step 7 with 20 mL of H​2​O, then rotovap the substance to
dryness
9. rinse the residue, and weight the dried substance
10. Prepare TLC plate by cutting it into rectangular shape, draw finish line (1 cm from the
upper edge) and start line (1.5 cm from the bottom edge). On start line, assign two
dot which are for the extraction we made and the pure caffeine sample.
11. Dissolve some part of dried caffeine extraction with distilled water, and use sharp-end
of a capillary tube to transfer the solution from petri dish, drop some of it on prepared
TLC plate (on the dot that is provided for extraction).
12. Dissolve the pure caffeine, then use sharp-end of a capillary tube to transfer the
solution from petri dish, drop some of it on prepared TLC plate (on the dot that is
provided for pure caffeine sample).
13. Place the TLC plate in to a chamber that filled with 80 mL ethyl acetate, 20 mL
hexane, and 1 mL of acetic acid. (about 1 cm high)
14. Observe the result
15. After getting results from TLC test. Carefully put the crudes and pure caffeine
substances in separate capillary tubes. So, we will get two capillary tubes to run in
the next melting point test.
16. Put the thermometer inside the melting point to check for temperature range along
the test.
17. Then put both of capillary tubes containing our crudes and pure substances beside
together to compare their melting point.
18. Wait for the temperature to hit around 200°C then looks through the melting point
machine to check for changed.
19. Record the range of temperature if one of the substances melt.

Result
 At first, we extracted caffeine from tea by using dichloromethane and get our caffeine
crudes. Then after getting caffeine crudes from extraction, we test the purity of our crudes by
using TLC paper and chamber. The results come out that we got caffeine from the tea
extraction because the spot is in the same position as our reference or pure caffeine
substance. ​However, our crudes were not pure substances because it also contain another
spot above which mean that there are contaminated in crudes. Next, we put our crudes into
capillary tube and another pure caffeine substance into another tube and then run them in a
melting point machine for test. From determining melting point of our caffeine crude and pure
substances of caffeine, our group have found out that ​our crudes were the first one to
dissolve and transform into liquid substances and then following by melting of pure
substances of caffeine. However, we have an error on doing this step because we cannot
catch up the range of Celsius that our substances melted up. According to reported in
literature, the melting point of caffeine should be 238°C. So, our range of temperature should
be around 200°C but not exceed 260°C.

Discuss

Based on theory, we already know the properties of substance consisting of melting point
and Thin Layer Chromatography. Our substance has almost the exact same height as the
pure one, but the pure one is a bit higher than our substance. Therefore, Our substance
might be contaminated since the other substances that were mixed with extracted caffeine
probably have stronger attraction to pull caffeine down, so it did not go as high as the
original one. Regarding melting point, our caffeine melted before the pure caffeine, hence it
is obvious to assume that our caffeine is not pure and contaminated.

Both of the substances including caffeine and dichloromethane, are polar, hence
emulsion can be happened. The way to avoid emulsion is not swirl the solution too much, so
that the substances would be mixed, but if there is emulsion, it can simply remove using
glass rod. We need drying agent (Na2SO4) after finishing liquid-liquid extraction and before
evaporation process because Na2SO4 can absorb water, so our extraction will loss amount
of excess water and remain pure caffeine.
According to the experiment, the pure caffeine is provided to clarify the expected
results for the test consisting of melting point and chromatography, so the extraction can be
compared to it. The result is that the extraction is not pure. As comparing to the initial result,
the melting point and result of chromatography are not the same.

4. What is the Rf value of caffeine?

The errors that appeared were misunderstanding the method and could not get the
exact temperature of melting point. Although the extraction was not contaminated, mixing
wrong substances could be very dangerous. Therefore, in the next experiment, the
experimenters should spend more time on procedure to make sure that they are clearly
understand how to perform the experiment. Other than that, we should be more careful for
observing temperature or even other reaction that could be reported. So, the result of the
experiment is more accurate and reliable. The strongest intermolecular force which presents
in Caffeine is dipole-dipole force because each molecule of caffeine has a dipole-dipole
force with different EN, so they will have dipole-dipole force with each other when they are
bound.

Conclusion
After doing this lab, the extraction of caffeine from tea solution, we have got crudes of
caffeine as a product. Also, after running TLC test we known that our crudes are caffeine but
also mixture and contaminated with other substances. Lastly, we run our crudes in melting
point machine and got a result that our crudes melt up first so its purity is not pure as pure
substance which confirmed the point that our crudes are contaminated.

Reference (APA)

[1] Pubchem.ncbi.nlm.nih.gov. (2018). ​Caffeine.​ [online] Available at:

https://pubchem.ncbi.nlm.nih.gov/compound/caffeine#section=Top [Accessed 13 Sep. 2018].

[2] Chemicool.com. (2018). ​Definition of extraction - Chemistry Dictionary.​ [online] Available

at: https://www.chemicool.com/definition/extraction.html [Accessed 13 Sep. 2018].

[3] Chemguide.co.uk. (2018). ​thin layer chromatography.​ [online] Available at:

https://www.chemguide.co.uk/analysis/chromatography/thinlayer.html [Accessed 17 Sep.
2018].

[4] Chemistry LibreTexts. (2018). ​Thin Layer Chromatography​. [online] Available at:
https://chem.libretexts.org/Demos%2C_Techniques%2C_and_Experiments/General_Lab_T
echniques/Thin_Layer_Chromatography [Accessed 17 Sep. 2018].