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Abstract
An extraction for caffeine from tea leaves by the use of solid-liquid and liquid-liquid
extractions was performed in order to determine whether if caffeine exist within tea leaves or
not, and from the extraction, is the extraction pure or not. The following techniques has been
liquid extraction. Moreover, the chemicals used within all of the process was identified and
described. At the end of the experiment, the existence of caffeine within tea leaves could not be
Objective
This experiment was conducted in order to isolate caffeine from the tea sample prepared
with water to determine whether if there is caffeine in tea leaves or not and whether if the crude
Introduction
This experiment includes the usage of various chemicals which are: dichloromethane,
ethanol, sodium sulfate, hexane, ethyl acetate, and iodine in order to extract caffeine from tea
leaves. The techniques used within this experiment are TLC (Thin Layer Chromatography),
have the melting point of 238 degree Celsius. Caffeine could be found in seeds, nuts, or leaves of
plants [)] . The appearance of caffeine is a soft, white crystal or powder. It is odorless and have a
slightly bitter taste. It is classified as a central nervous system stimulant. In 150 ml of tea, the
approximated amount of caffeine is 2-5 mg [+] . Caffeine is polar, hence, it is water soluble [,] .
commonly used as solvent. The density of dichloromethane is 1.33 g/3 .If exposed to high
temperature, dichloromethane could emit harmful, toxic fumes. It is also commonly used in the
process of decaffeinating coffee [0] due to the greater solubility of caffeine in dichloromethane
than in water. The solubility of caffeine in dichloromethane is 140 mg/ml when the solubility of
pungent, bitter taste commonly used as disinfectant. It is also used as solvent. The density of
used as drying agent. It dissociates itself in water in order to form sodium ions (5 ) and sulfate
ions (4 26 ). It usually come in the form of colorless crystals or fine white powder. It is odorless
and tastes bitter and is insoluble in alcohol and is hardly dissolved in water [7] .
extraction solvent. Hexane could be found in citrus, scallops, and some species of plants. It is a
clear, colorless liquid with petroleum-like odor. It is non polar and, hence, is insoluble in
water [8] .
Ethyl Acetate (, + , ) is a compound found in alcoholic beverages, radishes,
fruit juices, and cereal crops. It is commonly used as a solvent and typically used in the
manufacturing of decaffeinated coffee and tea. The physical characteristic of ethyl acetate is that
it is a clear, colorless liquid with fruity odor. It is a slightly polar compound due to the ester
Iodine (2 ) is a dark violet to black solid crystals with slightly metallic luster to it. The
odor of iodine is very sharp and pungent. It is very volatile [);] and could be used in thin layer
chromatography (TLC) to react with the spots to make the spots visible [))] .
the composing compound, helping with identifying the compound and the number of compounds
composing the mixture. TLC consists of three steps, spotting, development, and visualization.
The sample that would be analyzed would be dissolved in volatile solvent to produce dilute
solution to be spotted onto the TLC plate. The step of spotting consist of using a capillary to
transfer the diluted sample onto one end of the TLC plate. Then, the step of development
follows. The step of development includes soaking the TLC plate into the solvent and let the
solvent, with the sample to move along the TLC plate with capillary action. The outcome
depends on the polarity of each of the compound present. The compound with more polarity
would be closer to the starting line when compared to the compound with less polarity. Lastly,
visualization is the step that the difficulty depends on the substance. If the substance is colored,
then visualization is just looking at the plate. However, as some of the compound is clear and
colorless, visualization process is needed. The visualization process could be done by exposing
the TLC plate to iodine vapor and letting it react with the spots on the TLC plate. From the TLC
process, retention factor (= ) could be determined by dividing the distance travelled by the
solvent over the distance from the starting line to the center of the spot. By using the retention
Rotary evaporation technique is a technique used to separate volatile solvent and non-
volatile sample with most efficiency. It works by lowering the boiling point of the solvent
through the reduction of pressure. Then, the sample is rotated by the rotor vap in order to
increase the surface area of the sample, enabling the solvent to be evaporated while the sample is
heated. After that, the vapor of the solvent would be passing the coil filled with cold distilled
water in order to precipitate the solvent. After the process is done, the non-volatile sample would
be left within the rotating round bottom flask (evaporating flask) while the solvent would be
solid in a solvent at a temperature in which its almost boiling. Then, let the solvent cool down
and wait until the solution crystallizes. After the crystals developed, the crystals could be
collected, washed, and dried. This process do not involve any breakage of chemical bonds [),] .
laboratories. During this process, the solute separates into two immiscible (cannot be mixed)
1. Tea
2. Boiling water
3. Beakers
4. Graduated cylinder
6. Erlenmeyer flask
8. Watch glass
9. Dichloromethane (+ + )
11. Ethanol (, + )
16. Pump
22. Hexane (6 14 )
23. TLC chamber
24. Iodine (2 )
25. Thermometer
27. Forcep
Procedure
1. Prepare tea sample by letting the tea bags steep in boiling water. Leave the tea bag within
the water for a while in order to obtain the tea sample. This process is the solid- liquid
extraction. It is the extracting process of caffeine from tea leaves (solid) with hot water
(liquid).
2. Strain the tea leaves out of the tea solution by either picking out the tea bags or filter the
3. Retrieved 15 ml of the tea sample by using a graduated cylinder. Pour the obtained
sample into the separatory funnel. Making sure that the stopcock is closed before the
4. Obtain dichloromethane and pour it into a beaker. Cover the beaker with a watch glass in
dichloromethane into the separatory funnel. Close the beaker that contains
to release the gas that had been produced within the separatory funnel. Turn the stopcock
back to the horizontal position in order to close the funnel again and avoid spillage.
7. Slightly mix the solution in the flask by moving in C-shape. Then, carefully release the
gas created inside the flask (raise the end in order and point. Repeat this step two to three
times.
8. After letting the solution rest still for a while, the organic part of the mixture will be
sinken at the bottom of the flask due to the higher density of dichloromethane compared
to water. Separate the dichloromethane and caffeine mixture from the aqueous solution
by gradually open the stopcock of the separatory flask to let the solution flow and then
close the stopcock when it almost reach the layer of the tea solution.
9. Collect the dichloromethane and caffeine mixture (the organic part) in the Erlenmeyer
flask.
10. Repeat step 5 to 10 for three times in order to extract as much caffeine from the sample as
possible.
11. Dispose the leftover aqueous solution and clean the separatory funnel.
12. Pour the organic mixture back into the separatory funnel and add 15 ml of ethanol into
the funnel in order to get rid of any water residue within the mixture. Wait for the
separation of the layers. If there is any separations present, proceed to gradually open the
stopcock of the separatory flask in order to transfer the dichloromethane, which sinks due
more sodium sulfate to the dichloromethane until some of the sodium sulfate appears as
14. Prepare Buchner funnel with filter paper and place them on the suction flask which is
15. Open the pump, then carefully pour the solution into the Buchner funnel to filter out the
solid sodium sulfate from the mixture. Discard the filtered sodium sulfate.
16. Wash the funnel with dichloromethane, ensuring that all of the caffeine were transferred
into the flask and non were stuck onto the funnel and to recover some of the adsorbed
product.
17. Collect the organic solution, a mixture of caffeine and dichloromethane and transfer it
18. Perform rotary evaporation by using a rotary evaporator in order to obtain pure
dichloromethane in a beaker and the solid caffeine extract would be left within the round
bottom flask.
19. Transfer the caffeine extract from the round bottom flask onto a watch glass.
21. Collect some of the crude caffeine extract to be purified through the process of
recrystallization.
Part 2: Thin Layer Chromatography
2. Mark a line and two dots on the line, which is about one centimeter from the edge of the
plate, on one end of the TLC plate by using a pencil to indicate the starting line with the
points for the sample of crude caffeine obtained by the processes in the previous part and
the recrystallized caffeine. Be careful not to touch the matte side of the TLC plate.
3. Add some ethanol to the caffeine in order to make it easier to be transferred onto the TLC
plate.
4. Use a capillary tube to transfer the caffeine sample onto the TLC plate. Use separate
capillary tube for the crude caffeine and the purified caffeine.
5. Repeat step 4 for about 15-20 times, dotting the sample onto the TLC plate at the same
spot repeatedly in order to ensure that there is enough sample on the plate. Spot until a
6. Combine hexane and ethyl acetate by a 1:1 ratio to be used as the solvent (mobile phase)
7. Add the hexane and ethyl acetate mixture into the TLC chamber, trying to pour the
mixture into the chamber until it reaches about 1 cm over the bottom of the chamber so
that when the plate is placed, the solvent would be at the starting line.
8. Place the TLC plate into the chamber. Close the lid and let the process run until the
9. While TLC is running, prepare a petri dish with Iodine crystals. Close the lid of the petri
with the iodine crystals. Close the lid of the petri dish after the TLC plate was placed into
the dish.
12. Compare the distance of the dot from the starting line of both crude and purified caffeine.
2. Fill the crude caffeine sample into the melting capillary tube by using the open end of the
capillary tube to scoop the crystals into the tube. Tap the closed end of the capillary tube
against the table gently several times in order to force the sample to fall down into the
bottom of the tube. Repeat this step until there is 1-2 mm of packed sample within the
tube.
3. Repeat step 2 but replace the crude caffeine sample with the purified caffeine crystals
4. Insert both capillary tubes into the block of the melting point apparatus.
6. Reduce to 10@ /min once the thermometer reaches 60% of the expected melting point
7. Observe the melting of the substance through the eyepiece while recording the melting
Figure 2: Crude caffeine after dried overnight Figure 3: Purified caffeine acquired from recrystallization
Figure 4: The TLC plate with the crude sample on the right (the one with dots) and the purified caffeine sample on
the left (right plate) and the TLC plate with purified caffeine prepared by the instructor (left plate)
The resulting substance from the process of extraction appears to be a fine powder
(Figure 1). The coloring of the fine powder is yellow to white before being dried overnight. The
white powder is in the center of the watch glass while the powder that appears more yellow in
color is on the outer edge. After being dried overnight and mixed, the crude caffeine (Figure 2)
powder appears creamy in color. The color is not white nor yellow. On the other hand, the
sample that has been purified by the process of crystallization (Figure 3) was lessen in amount,
but appears white in color. The size of the recrystallized caffeine and the crude caffeine is
Through the TLC process, it could be observed that the crude caffeine is a mixture of at
least two to three substances due to the two to three spots showing on the TLC plate (Figure 4).
(Note: the spot is not very distinguishable, hence the approximated amount of spot is 2-3)
However, the substance within the recrystallized caffeine couldnt be distinguished since there is
no spot showing.
From the measurement of melting point by using the melting point apparatus, it could be
seen that the melting point of the crude caffeine measured is within the range of 225 227 .
However, the melting point of the recrystallized caffeine couldnt be measured since the crystal
didnt melt.
Discussion
From the process of TLC, it was illustrated that the crude caffeine obtained from the
extraction process wasnt pure and was composed with at least two to three substances. On
contrast, the information regarding the purified caffeine couldnt be obtained from the
experiment, and hence, the purity of the recrystallized caffeine couldnt be concluded through
In terms of melting point, while the melting point acquired from researches is238@ , the
melting point of the crude caffeine extracted is in the range of 225 227@ , only 9 13@
different from the reference, hence, there is a possibility that the crude caffeine extracted is a
mixture of caffeine and compounds with lower melting point. From the observation and the
comparison of the melting point of the samples, it was shown that neither the recrystallized
caffeine nor the crude caffeine extracted was pure due to the difference in melting point.
From the experiment, it could be determined that the strongest intermolecular force
present in caffeine is dipole-dipole. There are two intermolecular forces present in caffeine:
dipole-dipole and london force. Since caffeine is a polar compound due to the presence of
Oxygen and the absence of the overall cancelling of electronegativity within the compound, the
dipole-dipole intermolecular force is present. Moreover, since london force is present in every
compounds, london force which is weaker than dipole-dipole intermolecular force is also present
in caffeine
During the experiment, emulsion could be observed; especially during the liquid-liquid
extraction process. The emulsion could be a result of having a large amount of compounds with
low surface tension (which could be resulting from the weak intermolecular forces such as
london force), preventing the distinct separation of the two compounds. Another reason that
could be used to explain the presence of the emulsion is that the sample could contain
compound(s) with emulsifier-like properties and allow some of the polar and nonpolar
need to properly mix the two substances (in this case, the tea solution and the dichloromethane)
together. In order to break the emulsion, the separatory funnel could be gently shaken, tapped, or
swirled. Another easy way to break the emulsion is to just wait for the separation of the two
layers. Moreover, the addition of the drying agent into the organic solution could also help with
the breakage of the emulsion between the two layers. The drying agent, in this case the
anhydrous sodium sulfate, will pull the residue of the unwanted aqueous mixture out from the
wanted mixture of caffeine and dichloromethane. If the drying agent wasnt put into the
extracted organic mixture before evaporation, the evaporated product might be impure since
In this experiment, it couldnt be confirmed that caffeine is obtained from tea leaves due
to the error with the purified caffeine crystals obtained from recrystallizing the extraction.
However, it is likely that caffeine is obtained from the tea leaves due to its similar physical
characteristics and the similar boiling point. The crude sample obtained from extraction is
impure. The impurity of the sample could be confirmed by using TLC, where about two spots are
present on the TLC plate. Compared to the extract product with the others, it could be seen that
the extracted product is similar and both are impure. Two to three spots are present both the
The possible errors that lead to the inability to confirm whether if the crude sample
acquired through the process of extraction is caffeine or not are the lack of polarity of the
solvent. The reason that proves that the lack of polarity of the solvent could lead to the inability
of determining whether if the sample is caffeine or not is the clumping of the spots on the TLC
plate. This shows that the solvent (the mobile phase) doesnt have enough polarity to bring/repel
the components of the sample along the TLC plate. Moreover, the sample that was transferred
onto the TLC plate might not be enough for determining the components within the sample.
There are many possible errors that might lead to the impurity of the crude sample as
observed during the TLC process. Firstly, there might be other compound within tea leaves that
have similar properties as caffeine, meaning that it could also be extracted with dichloromethane
while the caffeine is being extracted. Secondly, there might be some impurities during the liquid-
liquid extraction, meaning that there might be some unwanted aqueous layer mixed with the
extracted organic layer. Also, during the liquid-liquid extraction process, the separation of layers
might not be completed due to the lack of time. During the TLC process, there might be some
contamination of the TLC plate due to touches and other compound that is preexisting on the
plate. Lastly, the waiting time during the process of transferring the sample onto the TLC plate
In order to avoid errors, a new solvent system that has more polarity might be used within
the lab and a new mobile phase that is more suitable to the sample might be used in the future.
Moreover, the time used in liquid-liquid extraction process might be added in order to let the
layers separate completely and the extraction could be done more carefully in order to lessen the
unwanted aqueous that is mixed with the wanted organic layer. Also, more sample could be used
during the TLC process in order to make sure that there would be some sample that could be
Conclusion
caffeine was extracted from tea leaves or not. However, the probability of the presence of
caffeine within the extraction from tea leaves is high due to the similar characteristic and boiling
point of the crude sample and the reference information on caffeine and it could be determined
that the extraction obtained from the tea leaves is impure due to the presence of more than one
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