Lab Partners: Date Performed: February 4, 2015

Bliss Capidos Joshua Emmanuel H. Pagulong Date Submitted: February 12, 2015
Kaye Antonette Chua Eli John Novela

I. Introduction
Tea is one of the most commonly used caffeinated beverages in the world. The caffeine (C8H10N4O2)
found in tea is a bitter, white, crystalline methylxanthine and a member of a class of compounds known as
alkaloids (Wang, 2011). Alkaloids are basic nitrogen containing compounds present in plants. The structure of
caffeine affects the functions it performs.
Alkaloids, such as caffeine, are often physiologically active in humans and are known central nervous
system stimulants and diuretics (Wang, 2011). Caffeine also causes an increase in respiration and heart rate, as
well as nervousness and insomnia. Though caffeine has demonstrated to have physical dependence, it is also
capable of improving alertness, learning capacity, and exercise performance (NCBI, 2013). Tea leaves, in which
caffeine is found, also contain acidic tannins, undecomposed chlorophyll, cellulose, and pigments.
In order to extract caffeine from tea leaves, caffeine must be present as the free base (Amrita, 2013). In
order to do so, the above-mentioned acidic substances must remain water-soluble . In order to extract caffeine
from tea, several methods are used. First, a solid/liquid extraction must take place in order to get the solid
natural product into the liquid solvent. This can be done simply brewing a cup of tea. In order to isolate the
desired reaction compounds from the natural product, liquid/liquid extractions are used. Neutral and acid/base
are two forms of liquid/liquid extractions (Williamson, 2011) . Caffeine extraction from tea leaves involves an
acid/base liquid/liquid extraction (Oneota, 2003) . The reaction involves a homogenous mixture of an organic
and aqueous layer. The ideal solvent in the extraction should have a low boiling point, not react with the solute
or other solvents, not be toxic or highly flammable, not miscible with water, be inexpensive, and should readily
dissolve caffeine at room temperature. A common liquid/liquid solvent pair for the extraction of caffeine is
water-dichloromethane (Williamson, 2011). Because water is present in the pairing, it possible to separate
inorganic compounds from organic compounds due to the fact that organic substances are immiscible in water
(Amrita, 2013).
When mixing the liquid pairs, the density of the both solvents predict which solvent is the top and which
is the bottom layer. Caffeine, which was present in the organic layer, was located below the aqueous layer
(Williamson, 2011). The product that is collected after extraction still has many impurities. Sublimation is one
way to purify the sample, because caffeine has the ability to pass directly from the solid to vapor and reverse to
form a solid all without undergoing the liquid phase. Caffeine has the ability to undergo sublimation under
different conditions than the impurities, and can thus be isolated (Tello, 2011). A series of techniques were used
to extract pure caffeine from tea leaves. The percent error and percent recovery were calculated to assess how
much pure caffeine was obtained, and to account for errors that may have occurred that led to a loss of product.
This experiment illustrates the isolation of a naturally occurring product from plant material -- caffeine
from tea leaves. The experiment will provide experience in handling relatively small amounts of material and at
the same time you will be exposed to several techniques and procedures which are fundamental for survival in
an organic chemistry lab.
Caffeine is an alkaloid, a class of naturally occurring compounds containing nitrogen and having the
properties of an organic amine base [alkaline, hence, alkaloid]. Typical alkaloids are caffeine, nicotine, morphine,
codeine, and cocaine.
One can develop both a tolerance and a dependence on caffeine. The dependence is real, and a heavy [>
5 cups of coffee per day] user will experience lethargy, headache, and perhaps nausea after about 18 hours
abstinence. An excessive intake of caffeine may lead to restlessness, irritability, insomnia, and muscular tremor.
Caffeine can be toxic, but it has been estimated that to achieve a lethal dose of caffeine one would have to drink
about 100 cups of coffee over a relatively short time.
Because of the central nervous system effects that caffeine causes, many persons prefer to use
decaffeinated coffee. The caffeine is removed from coffee by extracting the whole beans with trichloroethylene
 at 71 0C. Following this, the solvent is drained off, and the beans are steamed to remove any residual solvent.
Then, the beans are dried and roasted to bring out the flavor. Decaffeination reduces the caffeine content of
coffee from a range of 2% to 5% to the range of 0.03% to 1.2% caffeine
The isolation of caffeine from tea leaves presents the chemist with a major problem: caffeine does not
occur alone in tea leaves, but is accompanied by other natural substances from which it must be separated. The
major components of tea leaves are:
1. Cellulose − the major structural material of all plant cells. Since cellulose is virtually insoluble in water
it presents no problems in the isolation procedure.
2. Caffeine − one of the major water soluble substances present in tea leaves. Caffeine comprises as
much as 5% by weight of the leaf material in tea plants.
3. Tannins − high molecular weight, water soluble compounds that are responsible for the color of tea.
The term "tannin" does not refer to a single compound or even to substances having similar chemical structure.
Rather, "tannin" refers to a class of compounds that have certain properties in common. They contain
phenol groups, are acidic and are used to convert animal hides to leather [tanning].
4. Flavonoid Pigments − water soluble colored compounds that are widely distributed in plant life.
5. Chlorophylls − water soluble green plant pigments that enable plants to convert carbon dioxide and
water to carbohydrates and oxygen [photosynthesis].
The caffeine isolated from tea leaves can be purified by sublimation.
Caffeine melts and sublimes at 2380C. But during this experiment you will identify caffeine not from its
melting point but from the melting point of its salicylate. One way to identify an organic compound is to prepare
a derivative of it. Caffeine is an organic base and can therefore accept a proton from an acid to form a salt. The
salt formed from salicylic acid, even though ioic has a sharp melting point and can thus be used to help
characterize caffeine.

II. Data and Observations

Table 1. Measured Masses and Volumes
Tea brand: Lipton Tea (Yellow)
Theoretical Mass of Caffeine (6 tea bags) : 330 mg
Mass (g)

Tea Leaves 11.7692

Vial 16.5922 g

Vial + Caffeine 16.6244 g

Caffeine (Actual Yield) 0.0322 g

Volume (mL)

added DCM 60.0 ml

recovered DCM 2 ml

Table 2. Error and Percent Recovery

% Caffeine Recovery 0.2736 %

% Error 90.24 %
Observations:
The predetermined masses provided by the Lipton Tea manufacturers are accepted as the experimental
measurements of both tea and caffeine. The approximate weight of an individual Lipton tea bag is 2.00 ± 0.05 g,
containing 55 mg of caffeine per bag. In an experiment using 6 tea bags, 330 mg is the expected yield of caffeine
to obtain. To extract caffeine, the heated tea solution, along with DCM, was poured into an isolated separatory
 funnel and inverted to mix the solution completely. Due to the reaction, pressure built up inside the funnel,
requiring the stopcock to be opened to release excess gas following each inversion. Once settled, the solution
separated into 2 layers: the polar, brown tea solution on the top and the nonpolar, clear methylene chloride on
the bottom. The tea separated above the methylene chloride because while the density of water is 0.997 g/mL,
the density of methylene chloride is 1.32 g/mL. Between the two layers sat small bubbles or possible emulsions
that restricted the amount of methylene chloride that could be extracted. Despite this, the methylene chloride/
caffeine layer was effectively drained into a flask and the process was repeated 2 additional times to ensure that
all of the caffeine was reacted with. In doing so, the volatile methylene chloride began to evaporate into the
surroundings due to the increased room temperature. The remaining solution was heated briefly through
distillation until the volatile solvent evaporated, leaving behind solid caffeine. When weighed, the vial and
caffeine rendered a total mass of 16.6244 g. The difference in mass between the initial weight of the vial and
the final product was then the actual yield of caffeine, 0.0322 g. Basing from the total mass of tea used in the
experiment, the results only provided 0.2736 % yield of caffeine.
This very small percent yield and very high percent error from the theoretical expected value of mass of
caffeine that can be extracted is highly affected by the formation of emulsion in the solution in the separatory
funnel. Formation of emulsions may be brought by the mixing method of the solution and the dichloromethane.
The two solutions must be mixed slowly. Also, emulsions may also form because it can be that not all of the
interfering substances are eliminated in the previous processes. Also impurities in the substances used may also
affect the yield because it can be that not all interfering substances are not all reacted and eliminated
throughout the processes.
III. Discussions of Results
Caffeine is a commonly used addictive drug which is present in varying amounts in beverages
like coffee and tea. Together with some compounds like nicotine, morphine, strychnine and cocaine, it
is also classified as an alkaloid which are nitrogen-containing compounds that are found in plants.
Having nitrogen atoms, most of them tend to accept a proton and act as a base or alkaline substance.
Generally, these compounds taste bitter and are often physiologically active in humans. Caffeine is also
found to act as a stimulant, treat some types of headache and is diuretic in nature.

Figure 1. Molecular Structure of Caffeine

In this experiment, tea leaves (Lipton Tea Yellow) were used as the sample for the extraction of
caffeine. Since tea leaves are not completely composed of caffeine alone, several methods were
undertaken in order to isolate the obtained crude caffeine from other components like cellulose,
tannins, flavonoid pigments and chlorophyll. At the beginning, six bags of tea leaves amounting to
11.7692 g where weighed, poured with 100 mL of boiling water and steeped for about eight minutes in
order for caffeine to be released from the tea leaves. The muddy looking tea solution was then poured
into the cheesecloth in which removal of cellulose (as the solid matrix) was involved due to its
insolubility in water thereby getting a tea extract in the beaker. Addition of sodium chloride (NaCl) was
part of the salting out process wherein water separated from the rest since salt is immiscible in water
for both are polar compounds following the rule like dissolves like. Also, this decreased the solubility of
caffeine for it is only slightly polar in nature. The addition of calcium hydroxide (CaOH2) on one hand
which acts as a base was used to precipitate out the tannic acid as calcium tannate. Being converted
into salt, tannins were no longer extracted together with the solvent in the procedure that followed
and remained in the aqueous layer. After heating and gradual stirring for 15 minutes, the extract was
 vacuum filtered rather than gravity filtered to remove undissolved solids and save time since it is
faster.
In the liquid-liquid extraction of caffeine part, the cooled tea extract was put into the
separatory funnel and added with 20 mL of dichloromethane (CH2Cl2). The separatory funnel was
inverted and vented to the fumehood to release pressure upon addition of DCM. It was then mixed
gently to avoid formation of emulsions which may cause difficulty in separation of the layers. An
emulsion is signified by small liquid droplets which become dispersed in another immiscible liquid that
were found in our case. Addition of salt removed the emulsions since it made the emulsion more polar
allowing it to be less soluble with other substances. The aqueous layer formed is composed of
chlorophyll, tannins and flavonoid pigments while organic layer is comprised of DCM, caffeine and
impurities. When the layers were left to separate, aqueous layer (clear) is on top and organic layer
(brown) is at the bottom part since DCM is denser. After the organic layer is drained to a flask, the
procedures were repeated twice more for matters of accuracy.

Figure 2. Molecular Structure of Dichloromethane

The caffeine extract from the flask was put back into the separatory funnel and washed with
10% sodium hydroxide (NaOH) to remove acids and remaining impurities which may have been
extracted along with the caffeine. After it was transferred back to a flask, anhydrous sodium sulfate
(Na2SO4) was added in order to remove traces of water until the solution is clear with powdery
compositions. Decanting into the round bottom flask, the extract was then prepared for distillation
process. The extract was heated to a temperature that allowed the dichloromethane to boil which is
about 38 degrees close enough to its actual boiling point at 39.6 degrees however it was heated to a
final temperature of 42 degrees to assure that what remained in the round bottom flask was caffeine.
The distilled solvent was measured to be about 2 mL. The crude caffeine residue was rinsed with an
acetone and transferred to a vial where it was left to evaporate. The mass of the crude caffeine
obtained was 0.0322 g which signifies 0.273% caffeine in the sample tea.
As compared to the theoretical percentage of caffeine claimed by the manufacturer which is
about 3% in six tea bags, the obtained caffeine is only 0.3% which is far lesser than the actual. The large
margin of error may be due to the emulsions that were formed in the process. The emulsions may have
been partly remedied but there is still a large possibility that these may have restricted the amount of
caffeine drained from the separatory funnel having lesser yield. Another reason may be in the
decantation wherein some substance were spilled in the process. Filtration instead may cause a slight
increase in the caffeine yield.
IV. Conclusion and Recommendation
It was estimated based upon the theoretical values that the tea bags provided had a potential of 330 mg of
caffeine available for extraction. But we were only able to extract a total mass of 32.2 mg which is very small from
the expected value. There were several factors that could have affected the yield.
 During the liquid/liquid phase of extraction, the separating funnel was shaken very vigorously causing a thick
layer of emulsion to form between aqueous and organic layers. Even after 2nd extraction with dichloromethane,
there was still a heavy layer of emulsions, indicating that there was still a large amount of caffeine that would
not be available for recovery. This is why proper mixing of the solutions is highly recommended in order to avoid
formations of emulsions.
 The leaves are composed of many other compounds other that caffeine. Considering that all the mass of the
leaves is available for extraction of caffeine is a misinterpretation of the true amount. However exact
information regarding the mass or percentages of caffeine contained in a single tea bag was not available for use
for the percent yield calculations.
 Another important factors which affected the yield is the standard human error.
 This laboratory experiment successfully demonstrated the visuals for separation of two liquids in a mixture and
teaching the fundamental concepts like densities, polarities, boiling points, acid/base interactions, neutralization
reactions, etc. which are essential for complex chemical reactions.
In this experiment there are a lot of sources of error. In order to reduce sources of error in the future, it is
recommended to conduct many trials and to compare the results that have been acquired. This experiment is only
limited in getting or isolating caffeine from tea, so it is recommended to try looking for other products with caffeine
besides tea and to compare with commercial products. A big factor to the error in this experiment is may be due to
equipment due to equipment defect and many personal errors of the experimenter like in the evaporation of
dichloromethane which we need to be careful and we need to be careful of spilling of these chemicals because
caffeine will be wasted and the concentration will be lessen. So in future experiments experimenters should be
careful and should perform the experiment carefully.
Dichloromethane and other chemicals in the experiments are highly hazardous and some are carcinogenic and is
very harmful to the body if inhaled or comes in contact with it. When Dichloromethane is breathe it is metabolically
converted to carbon monoxide after systemic absorption, which yields increased concentrations of
carboxyhemoglobin in the blood. It may cause damage to some of our internal organs and may cause a lot of effects
in the body including dizziness and headache. It is highly recommended to be careful and be cautious of the
chemicals being handled in this experiment. It is also recommended to wear proper attire with mask and gloves to
protect self from these chemicals.
V. Answers to Questions
1. Discuss the role/s of the following in the isolation of caffeine:
a. Dichloromethane
The organic solvent dichloromethane is used to extract caffeine from an aqueous
extract of tea leaves because caffeine is more soluble in dichloromethane than it is in water.
The dichloromethane - caffeine mixture can then be separated on the basis of the
different densities of dichloromethane and water because dichloromethane is much denser
than water and insoluble in it. Residual water is separated from dichloromethane by drain out
the dichloromethane through separating funnel, thus dichloromethane passed through the
funnel while polar solvents such as water is still remains in the funnel. Water and
dichloromethane is slightly soluble in each other. So, after separating the solvents, residual
water will remain the organic layer.
b. NaOH aqueous solution
Washing 10% sodium hydroxide help remove any acids or polyphenols that were
extracted along with the caffeine. This solution contains the conjugate acid of caffeine. Since the
majority of powders contain a small portion of caffeine, isolation will not be performed.
c. Ca (OH)2
Ca(OH)2 was used to make the caffeine solution basic, so that it puts caffeine in its least
polar form and so more readily solvated in organic solvents and less attracted to water. Since
the water solution contains much more than just caffeine, and some of these compounds are
also soluble in organic solvents, Ca(OH)2 was added so that it would react with tannic acids in
the solution to form insoluble tannin salts which precipitate and so can be removed from the
solution before the caffeine is extracted.
2. Explain why salt was added to the water before the liquid-liquid extraction of caffeine. What is
happening, why and how?
When NaCl is added to the water layer, it decreases the solubility of caffeine. This is due
to the fact that caffeine is only slightly polar in nature.
3. How efficient is the extraction of tea leaves containing 1.0 g of caffeine with 3 20-mL portions of
dichloromethane over that of a single step extraction?
It is efficient in order to obtain purity of the product – the recovered caffeine residue.
4. What are emulsions and why do they form during extractions? How are they minimized and removed?
An emulsion is a mixture of two immiscible liquids. Chemically, they are colloids where
both phases are liquids. They are typically milky in appearance and the suspended material may
be colloidal in nature. When tannins are converted to their salts, they become an ionic
surfactant. These causes the materials that don’t dissolve in water to form an emulsion with
 water. This emulsion can be minimized reducing the energy input through slow and careful
shaking and swirling of the separatory funnel.
5. Why is it necessary to remove the stopper from the separatory funnel when liquid is being drained
from it through the stopcock?
It is necessary to remove the stopper from a separatory funnel in order for the liquid to
continually flow out. Removing the stopper would release the pressure so the liquid will flow
continuously. When stopper is not removed, the liquid will not drain out no matter how you
manipulate the stopcock.

VI. Calculations

m = 16.5922 g (vial)
m = 16.6244 g (vial + caffeine)
m = 0.0322 g (caffeine)

g crude caffeine 0.0322 g

% caffeine = X 100 = 11.7692 g x 100
g tea sample
% caffeine = 0.2736 %

g theoretical total mass of caffeine−g crude caffeine

% error = X 100
g theoretical total mass of caffeine
.330 g − 0.0322 g
= .330 g
x 100
% error = 90. 24%

VII. References
 McMurry, John, Organic Chemistry 7th edition, Thompson Learning Inc. Books/Cole 2008.
 Silberman, Robert. “Isolating Caffeine from Tea,” page 65-73, Signature Lab Series, CHE 2403, Cengage Learning.
Mason, Ohio: 2008.
 National Center for Biotechnology Information, U.S. National Library of Medicine. CaffeineCompound Summary.
http://pubchem.ncbi.nlm.nih.gov/summary/summary.cgi?cid=2519 (accessed Oct 7, 2013) (NCBI, 2013)
 Oneota. Isolation of Caffeine from Tea Leaves. [Online] 2003. http://employees.oneonta.edu/
knauerbr/chem226/226expts/226_expt06_pro.pdf (accessed Oct 7, 2013)
 Weaver, S.L. October 26, 2014. Extraction: Isolation of Caffeine from Tea Leaves Notes.
 May 21, 2013. Material Safety Data Sheet Dichloromethane MSDS
Links:
 http://vlab.amrita.edu/?sub=3&brch=64&sim=169&cnt=1
 http://www2.volstate.edu/CHEM/2010/Labs/Caffeine_Extraction.htm
 http://www.webassign.net/sample/ncsumeorgchem1/lab_3/manual.html
 https://www.coursehero.com/file/p3l0tpu/Isolation-of-caffeine-from-tea-2-3-Experiment-In-this-experiment-
caffeine-will/
 https://www.coursehero.com/file/p3692t/Conclusion-It-was-estimated-based-upon-the-theoretical-values-that-the-
tea-bags/
 http://www2.volstate.edu/CHEM/2010/Labs/Caffeine_Extraction.htm.
 http://wcam.engr.wisc.edu/Public/Safety/MSDS/Dichloromethane.pdf