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ORGANIC

CHEMISTRY

Organic Chemistry Laboratory CH 200 (2014-2015) Experiment 3

Extraction of Caffeine from Lipton Green Tea Using Multiple Extractions


Michael Baladad, Kenny Bautista, Alyssa Charisse Capuchino* and Charmaine Chan
Department of Biological Sciences, College of Science
University of Santo Tomas, Espaa Street, Manila 1008
Date submitted: September 4, 2014

Abstract:
The experiment was performed in order to obtain the percentage yield of caffeine, with a melting point of 238 oC[2], from the
Lipton Green Tea leaves as provided by the instructors using multiple extractions. From the weighed tea leaves, 11. 5129 g,
crude caffeine was obtained by extracting it with water and CH2Cl2 or Dichloromethane (DCM) for three times. With the
process, the group was able to get 0.59% of crude caffeine. Next, it was sublimated and recrystallized to obtain the pure
caffeine which yielded 0.19%. Then, the product was subjected to characterization using thin layer chromatography or TLC
wherein the retention factor or Rf value of the isolated and standard caffeine is 0.38 and 0.36, respectively.
Keywords: caffeine, tea leaves, solid-liquid extraction, liquid-liquid extraction, multiple extractions, distribution coefficient
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Introduction
In the early times, people have developed ways to extract substances from plants that are used for
different purposes such as flavouring, colouring, and even used in making beverages and medicines. One
common extract is tea powder from tea leaves of the tea plant, like Camellia sinensis, which contain a
number of compounds such as tannins, flavonoids, chlorophyll and xanthines which include caffeine.
Caffeine, 1,3,7 trimethylxanthine, is usually found in stimulating drinks or beverages such as tea,
coffee and carbonated drinks which students and professors usually resort to when the desire to be
awake for a longer period of time is needed. It can also be found in chocolates and certain medicines.
Caffeine is an alkaloid, a nitrogen-containing ring compound, which is usually bitter in taste. At room
temperature, it is a white solid. It has a stimulating property which stimulates the central nervous system
(CNS), heart, muscles and it can also increase ones blood pressure. It has a melting point of 238 oC and
its solubility in water is 22mg/ml at 25C, 180mg/ml at 80C, and 670mg/ml at 100C[2].
One of the ways used to separate a substance, such as caffeine from a mixture, like tea, is through
extraction. Extraction is the transfer of a target analyte from one phase to another (Tissue, 2013). It
selectively dissolves one or more compounds into a solvent. The dissolved compounds are called the
extracts. Different ways of extracting is used depending on the nature of the mixture. One way is using
the liquid-liquid extraction. Liquid-liquid extraction or solvent extraction involves two immiscible

liquids, which means two liquids that are not soluble to each other, having different densities which are
then separated. Two layers will be produced, the denser layer settling at the bottom where one contains
the extract and the other contains a small amount of the solute called the raffinate. Then, the pure form
of the solute is obtained from the extract phase and the solvent dissolved in the raffinate is recovered.
The separation maybe fast if the difference in their densities is small. A typical laboratory extraction,
such isolating organic compounds in aqueous solution, transfers the organic compounds out of the
aqueous phase and into an organic phase (Tissue,2013). Another way is the solid-liquid extraction which
is the process of separation of soluble constituents of a solid material using a suitable solvent (Dutta,
2013). This is also called leaching. This can be done by removing the unwanted soluble materials from
the solid using a solvent and the solid extracted is the desired product. Four steps are basically
performed in this type of extraction, first is the contact between the solid and the solvent, second is the
separation of the extract from the solid, third is the separation of the solved from the extract afterwards
purified, and fourth is the recovery of the solvent from the moist solid. A common example of this is the
preparation of the usual morning drink, coffee when dissolved in water, the flavours are extracted from
the powder. This extraction is further classified into diffusional extraction, washing extraction, leaching,
and chemical extraction. They merely differ in the characteristics of the solid involved and the solvent.
Another type of extraction is the multiple extraction which is more efficient than just a simple extraction
because a better percentage yield is obtained. This is a series of simple extraction but dividing the
solvent used in just one simple extraction into equal portions allowing for many extractions.
Aside from extraction, sublimation and re-crystallization is performed as well to obtain the desired solid
extract. Sublimation is the conversion of a solid into a gas skipping the liquid phase, while recrystallization is a method of purifying a compound.
In some experiments, the extracted compounds are also subjected to characterization which is usually
performed with thin layer chromatography or TLC. It is a type of chromatography that is used to
separate non-volatile mixtures. It used a TLC plate which is usually a sheet of glass, metal, or plastic
which with a thin layer of a solid adsorbent. A small amount of the mixture to be analysed is spotted
near the bottom of this plate. The TLC plate is then placed in a shallow pool of a solvent in a developing
chamber so that only the very bottom of the plate is in the liquid. This liquid, or the eluent, is the mobile
phase, and it slowly rises up the TLC plate by capillary action. With the help of the UV light, the spots
produced will be seen and need to be traced in order to compute for the retention factor value or Rf.
Methodology
For this experiment, the instructors have provided the students with 5 Lipton Green Tea bags and the
contents were weighed using an analytical balance that can read up to 4 decimal places. The weight was
then recorded on the log book. The 5 tea bags were then placed on a 250 mL beaker containing 150 mL
of water and heated to obtain the extract, using the set-up for heating which includes the iron stand,
tripod, Bunsen burner, rubber tubing, wire gauze and watch glass to help avoid burning of the tags of the
tea bags. The boiling was done for 5 minutes and afterwards the bags were further squeezed using a
glass rod to obtain the extract and were discarded. Two pieces of ice cubes obtained from the technician
were added to the tea extract to cool the liquid down and after melting, it was transferred to a separatory
funnel with the stopcock at the vertical position preventing the liquid to come out. 20 mL of
dichloromethane or DCM was added to the extract in the separatory funnel as the solvent for extraction
since caffeine is more soluble in DCM than water. Then the separatory funnel was covered with a

stopper and inverted then the stopcock was turned into a horizontal position to let the air out maintain an
open system as it was swirled to let the two layers separate. After swirling, the lower layer or the layer
residual water was carefully drained into a beaker while avoiding including a part of the upper layer.
This was repeated twice and then the upper layer was discarded. All of the lower layers obtained were
transferred into a clean separatory funnel and 20 mL of 6M sodium hydroxide or NaOH was added.
Again, it was swirled and the two layers were allowed to separate. The upper layer was discarded and
the lower layer was drained into a clean and dry 125 mL Erlenmeyer flask and a spatula of anhydrous
sodium sulfite or NaHSO4 was added in order to remove the water from the lower or organic layer since
NaHSO4 is an insoluble inorganic solid that could absorb water. Then, the flask was swirled and
decanted. The residue was discarded while the decantate was transferred in a pre-weighed evaporating
dish and was put on a set-up of a beaker with hot water bath on top of the hot plate under the fume hood.
It was then heated to dryness to evaporate the liquid. Then the evaporating dish with the crude caffeine
was weighed and the weight was subtracted to the weight of the pre-weighed empty evaporating dish
and then the percentage caffeine was computed. This was kept for a few days covered with a paper.
Another heating set-up for purification was prepared
including a filter tube with a fitted cold finger,
which provided a cool surface for sublimation, which
is simply a small test tube fitted against a hole made
on a cork that fits the filter tube, an empty 555 tuna
tin can with holes punctured at the bottom and an
aluminium foil. The tin can was put on the tripod
Figure 1. Extraction setup
above the Bunsen burner for heating. The crude
caffeine on the evaporating dish was scraped using a popsicle stick
and was transferred into the filter tube. This was placed on top of
the tin can with part of it inside the can and covered with
aluminium foil while a part of it was held by an iron clamp. The
gas was turned on and the Bunsen burner was lighted with a match
stick to produce fire. Then the cold finger was filled with cold
water. And before placing it on top of the filter tube was wiped dry
to prevent the condensed water outside from dripping onto the
filter tube. Sublimation was performed in order to obtain the pure
caffeine. After 15 minutes, the filter tube was checked whether
there was still left crude caffeine. The process was continued until
all the crude caffeine has been sublimated. After another 15
minutes, the fire was put off and the sublimate was scraped from
the bottom of the cold finger and from inside the filter tube while
the residue was discarded. The solid white crystals scraped were
transferred to a pre-weighed vial. The vial with the pure caffeine
was weighed on the analytical balance and was recorded on the
log book. Then the percentage yield of the pure caffeine was
Figure 2. Purification setup
solved.
This was followed by characterization using thin layer chromatography with the analgesics provided by
the instructors in vials. A 250 mL beaker lined with a filter paper was poured with 10 mL of developing
solvent ethyl acetate methanol acetic acid or EtOAc: HOAc: H2O (25:1:1) and was covered with a

watch glass. A 2 mL solution of methanol-toluene was added to the pure caffeine. Then a TLC plate
prepared by the instructor was obtained and the different compounds were spotted on it using a
microcapillary tube by dipping it onto the compound and touching it to the assigned point on the plate
three times. For point A is the isolated caffeine, point B standard caffeine, point C Acetaminophen, point
D Phenacetic, point E unknown, point F Aspirin, point G Ibuprofen and point H Mefenamic acid. After
spotting the plate was developed by placing it inside the chamber and was not removed until the solvent
reach the solvent front. The plate was then air-dried after reaching the front and was visualize under the
UV light. The dark spots were traced for the calculation of the retention factor or Rf values and were
recorded on the log book.
Results and Discussion
Table 1. Tabulation of Data for Extraction of
Caffeine
Weight of tea leaves (g)
Volume of water used (mL)
Volume of dichloromethane used (mL)
Volume of 6M NaOH used (mL)
Weight of empty evaporating dish (g)
Weight of evaporating dish and crude caffeine (g)
Weight of crude caffeine (g)
% crude caffeine (%)
Description of crude caffeine
Weight of empty vial (g)
Weight of vial and sublimate (pure caffeine) (g)
Weight of pure caffeine (g)
% pure caffeine (%)
Description of pure caffeine

from tea leaves. DCM was used as the


extraction solvent since it is much denser than
water having a density of 1.33 g/mL and
caffeine is found to be more soluble in DCM
thus the caffeine will easily move out from the
coffee and into the DCM. It can be seen that out
of 11.5129 g of tea leaves, only 0.022 g of pure
caffeine was extracted Using 20 mL of DCM
three times rather than using 60 mL under 1
extraction processes provided a much better
yield. In order to get the amount of caffeine in
the dichloromethane, the distribution coefficient
of water and dichloromethane was needed
which is KD = 10. Distribution coefficient is
defined as the ratio of the amounts of solute
dissolved in two immiscible liquids at
equilibrium.

s o
s a
KD=

The data gathered during the experiment was


presented on Table 1. The first part of the
experiment is the extraction of crude caffeine

, where KD distribution coefficient.

If KD is large, the solute will tend toward


quantitative distribution in organic solvent. With
this, the amount of caffeine in the DCM can
now be solve using,

KD caffeine (CH2Cl2/water) = 4.6 =


x
20 mL CH 2 CL2
,
0.035 gx
150 mL

where 20 mL is the volume of DCM used, 150


mL is the volume of water used and 0.035 g is
the accepted value of caffeine in Lipton Green
Tea leaves

x1 = 0.013 g
x2 = 0.008 g
x3 = 0.008 g
x1 + x2 + x3 = 0.013 g + 0.008 g + 0.008 g = 0.029 g caffeine
Doing this for the all the repetitions of the extraction process using DCM with the value of x, the amount
of extract in DCM, changing after each extraction, the total amount of theoretical caffeine is 0.029 g.
The experimental weight of crude caffeine is deviated from the actual weight of crude caffeine since
there were errors involved. Some of the errors are the amount of tea leaves at the beginning of the
experiment was not the same when it was boiled due to some tea leaves lost during weighing when
transferring from the tea

bag to the beaker and back again to the tea bag. Also, during the heating process, the extract was not
fully acquired because the tea bag was not properly squeezed to release the liquid extract left inside.
Followed by the DCM, NaOH was added to the extract. This is used to wash the caffeine in order to
remove any acids or polyphenols (tannins) extracted along the caffeine. The weight of crude caffeine
was obtained by subtracting the weight of the evaporating dish and crude caffeine to the weight of the
empty evaporating dish resulting to 0.0683 g and the percentage crude caffeine was obtained by dividing
the weight of the crude caffeine and the weight of tea leaves and multiplied to 100.
Weight of crude caffeine:
Wevaporating dish + crude caffeine Wevaporating dish = Wcrude caffeine
124. 9283 g 124. 86 g = 0.0683 g
Percentage of crude caffeine:
Wcrude caffeine
x 100
Wtea leaves
0.0683
x 100=0.59
11.5129
The crude caffeine is characterized by a yellow color and powder in form when scraped from the
evaporating dish.
For the purification process, the scraped crude caffeine produced 0.022 g of pure caffeine and 0.19%
pure caffeine. Sublimation was first performed and made possible with the use of the cold finger that
provided the cool surface for the caffeine to adhere. This process produced values deviated from the
accepted value due to the errors involved. The formation of the solid caffeine could have been affected
by the insufficient heat used in sublimation as well as the temperature of the cold finger. Also, the
scraping process had also affected the experimental yield because not all of the pure caffeine extracted
was collected in the vial with the interference of air and the crystals that might have remain stuck to the
scraper as well as those burned ones. The pure caffeine and its percentage were obtained by using the
same method of obtaining the weight and percentage of crude caffeine.
Weight of pure caffeine:
Wvial+ pure caffeineWempty vial=Wpure caffeine
10.1686 g10.1466 g=0.022 g

Percentage of pure caffeine:


Wpure caffeine
x 100
Wtealeaves
0.022 g
x 100=0.19
11.5129 g
Comparing the experimental weight of pure caffeine which is 0.022 g or 22 mg to the actual weight of
caffeine in Lipton Green Tea which is 35 mg [3], it can be deduced that we obtained 37% error which
were caused by the mentioned source of errors during the experiment.
Characterization was also performed and the Rf values of 0.38 and 0.36 for isolated and standard
caffeine were obtained based on the TLC chromatography performed. And based on the results obtained,
the isolated caffeine has a near Rf value with the standard caffeine which means that they are the same
compound.
Rf value:
Rf =

x
y , where x is the distance traveled by the spot and y is the distance traveled by the solvent

Rf isolated caffeine=

2.1cm
=0.38
5.6 cm

Rf standard caffeine=

2.0 cm
=0.36
5.6 cm

Conclusion
Extraction of caffeine from tea leaves involves different processes such as solid-liquid and liquid-liquid
extraction utilizing two different solvents, water and DCM, washing to remove unwanted materials
using NaOH, drying, and purification using sublimation. The process is a bit tedious since it involves a
lot of repetition in extraction or multiple extractions to ensure a more accurate result but unavoidable
errors still cause the result to deviate from the actual value. The percentage caffeine obtained from doing

the processes is 0.19% for the Lipton Green Tea leaves and using the TLC or thin layer chromatography,
the retention factor or Rf value of the isolated caffeine is 0.38 and the standard caffeine has 0.36.
Overall, the objectives were met though characterized by some inaccuracies which are always part of an
experiment.
References
[1] Dutta, B. (2007). Principles of mass transfer and separation processes. Delhi, India: PHI Learning
Private Limited
[2] Extraction of caffeine from tea. Retrieved from http://amrita.vlab.co.in/?
sub=3&brch=64&sim=169&cnt=1
[3] Pure green tea 40oct. Retrieved from http://www.caffeineinformer.com/caffeine-content/lipton-tea
[4] Tissue, B. (2013). Basics of analytical chemistry and chemical equilibria. Hoboken, NJ: John Wiley
&Sons, Inc.
[5] Zubrick, J. W. (2011). The organic chem lab survival manual: A students guide to techniques.
Hoboken, NJ: John Wiley & Sons, Inc.
Appendix

s o
s a
Distribution coefficient: KD=

, where KD distribution coefficient

Weight of crude caffeine: Wevaporating dish + crude caffeine Wevaporating dish = Wcrude caffeine
Percentage of crude caffeine:

Weight of pure caffeine:


Percentage of crude caffeine:
Wpure caffeine
x 100
Wtealeaves

Wcrude caffeine
x 100
Wtea leaves
Wvial+ pure caffeineWempty vial=Wpure caffeine

Retention factor:
by the solvent

Rf =

x
y , where x is the distance traveled by the spot and y is the distance traveled