Experiment # 2: Extraction

GONONG, D.A. G.

GUTIERREZ, H.A. G.*

HAGIWARA, K. L.

JAVIER, C.B. DLR.

LABAJO, A.M. F.

2C-MT, Faculty of Pharmacy, UST

Abstract

The experiment was performed so as to differentiate single from multiple extraction and to be
able to calculate the percentage yield of caffeine for both extraction procedures. Before the actual
experiment was performed, the principle of extraction and a brief run through of what to do in the
experiment was discussed. At first, the group hypothesized that there will be caffeine in the dried tea
leaves, but it would be insignificant since it is obvious that tea does not produce adrenergic responses
similar to coffee. Like all laboratory activities, this experiment may be dangerous if proper cautions
aren’t taken into consideration. For faster facilitation of the experiment and also to minimize confusion
and spoilage, division of labor was observed by the group. Each member performed their delegated
task with maximum efficiency and accuracy. After the solution of sodium carbonate was prepared and
the weighed tea bags were immersed, it was allowed to boil. While waiting for the time required to
boil the solution, so to be productive, some of the members were tasked to observe while the rest
weighed the evaporating dish and prepared the anhydrous sodium sulfate. The group was tasked to
perform single extraction and after the two layers was clearly visible, the organic layer or the lower
layer which contain DCM and the extracted caffeine was drained and was transferred in to Erlenmeyer
flask with anhydrous sodium sulfate and was decant to the weighed evaporating dish which was
evaporated to dryness under the hood. The residue was weighed and percentage yield of 4.0% was
calculated. This percentage yield is necessary to show the amount of caffeine present in the tea
sample. At the end of experiment, every member helped one another in cleaning the work place.

Introduction solvent extraction and partitioning which

The technique used to separate an
organic compound from a mixture of
compounds is called Extraction. It is the
process of obtaining something from a
mixture or compound using chemical,
physical, or mechanical means. It uses
two immiscible phases to separate a
solute from one phase into the other.
There are two types of extractions
performed in the laboratory: single
extraction and multiple extractions. Single
extraction introduced dichloromethane to
the tea solution only once, while multiple
extractions introduced dichloromethane to
the solution three times. In this
experiment, we used particularly Liquid-
liquid single extraction, also known as
uses a separatory funnel. Extraction
process selectively dissolves one or more
of the mixture compounds into an
appropriate solvent.
One of the objective of the
experiment is to separate the caffeine
content from the tea. Caffeine is a
naturally occurring alkaloid, a class of
naturally occurring compounds containing
nitrogen and having the properties of an
organic amine base, found in coffee, tea,
Cocoa, Guarana (Paullinia cupana) and
kola nuts. Caffeine belongs to a family of
naturally occurring compounds known as
xanthines. Caffeine is chemically 1,3,7-
trimethylxanthine (C8H10N4O2). Caffeine
acts as a stimulant. It stimulates the
heart, respiration, the central nervous
system, and is a vasodilator meaning
relaxes the blood vessels, as well as a
diuretic. It is a beverage produced by
steeping in freshly boiled water the young
leaves and leaf buds of the tea plant,
Camellia sinensis.
Fig.1 Structure of Caffeine
Fig.2 Structure of Xanthines
Extractions of certain solids can be
performed by utilizing the different
chemical properties of different solvents.
The solvent used in the extraction of
caffeine is water. Caffeine is soluble in
water at normal temperatures but it is
highly soluble at boiling water. The
solubility of caffeine to water is necessary
to create a solution of water and caffeine
which will be mixed with dichloromethane.
First, caffeine will be extracted from tea
leaves by boiling them in water. This
solution must be cooled before further
procedures. Although caffeine is soluble
with water, the caffeine will remain in the
solution together with other compounds in
particular, the tannins that are slightly
soluble in the dichloromethane which can
be eliminated by converting it to their
salts (phenolic anions) by adding sodium
carbonate, which remain in the water.
Fig.3 Structure of Tannin
Caffeine’s solubility in
dichloromethane is high even in room
temperature. So, when dichloromethane
is added to the previous water-caffeine
solution, the caffeine is immediately
transferred to dichloromethane. The
addition of dichloromethane to the
previous caffeine solution produces two
layers. Dichloromethane can be seen on
the bottom layer of the separatory funnel
since it is denser compare to water. This
dense lower layer contains the
chloroform-caffeine mixture. Upper layer
is separated from the lower layer by the
process of decantation, wherein the lower
layer is transferred to the evaporating
dish. After decantation, the solution is
exposed to heat for the purpose of further
separating caffeine from dichloromethane.
After heating, it is definite that caffeine
will be the only substance left in the dish
because dichloromethane is a highly
volatile substance.
Methodology
The experiment required a Bunsen
burner, separatory funnel, Erlenmeyer
flask, glass rod, evaporating dish, iron
stand, iron ring, iron clamp, unflavored
tea bags, sodium carbonate,
dichloromethane and sodium sulfate.
Single extraction was assigned to the
group. The first step in the experiment
was to weigh 4.4 grams of sodium
carbonate in a small Erlenmeyer flask
which was dissolved in a pre-heated
100ml of distilled water. Simultaneously,
10.00 g of the tea leaves contained in a
tea bag was weighed in the triple beam
balance and was immersed to the said Result and Discussion
mixture. The flask was covered and was
boiled for 10 minutes on a low flame. Single Extraction was assigned to the
group. Single and Multiple extraction is
After 10 minutes of boiling, the tea almost the same except that in multiple
bags were removed and were squeezed by extraction, there is repeated extraction
pressing it against the side of the flask process. It is usually more efficient than
using the glass rod so that all liquid will be single extraction because it allows more of
drained from the tea bags. Be careful so the caffeine in the mixture to be
as to not destroy the bag containing the extracted.
tea leaves or else might prolong the
experiment since it will require doing Notice in figure 4 that there are two layers
another purification technique which is the upper and the lower layer. The upper
filtration. layer in the experiment, the less dense is
the aqueous layer while the one in the
Before the mixture was transferred lower layer, the denser, is called the
in the separatory funnel, it was cooled organic layer. The organic layer
down for about 3 minutes. After cooling, constitutes of the dichloromethane and
the mixture was transferred in the the caffeine extracted.
separatory funnel containing 20mL of
dichloromethane. The separatory funnel
was turned upside down and the stop cock
was opened so to release the pressure
inside. To mix, the solution was swirled
and was turned upside down to release
again the pressure inside. This procedure
was repeated for two minutes. After
mixing, the separatory funnel was placed
in a steady position until the separation of
the two layers is clearly visible.

While waiting for the mixture to

settle down, using the triple beam
balance, the evaporating dish was
weighed and half spatula of anhydrous
sodium sulfate was prepared and was put
in the Erlenmeyer flask.
Fig.4 Extraction Set-Up
When the two layers were already
evident, the lower layer or the organic Caffeine can be extracted from tea
layer was drained into the Erlenmeyer leaves by boiling them in water yet,
flask containing the prepared anhydrous although caffeine is soluble with water,
sodium sulfate. The solution (sodium the caffeine will remain in the solution
sulfate + DCM + caffeine) was decanted together with other compounds in
onto a pre-weighed evaporating dish. It particular, the tannins which can be
was evaporated to dryness under the eliminated by converting it to their salts
fume hood until the residue is completely (phenolic anions) by adding sodium
in powder form and once again the carbonate
evaporating dish together with the residue
is weighed using the triple beam. Results
were gathered and percentage yield was
calculated.
 The purpose of the anhydrous
sodium sulfate on the other hand serves
as a drying agent that will remove any
water that may be present before starting
to evaporate the solvent off.
http://firstyear.chem.usyd.edu.au/prelab/
e28.shtml
http://itech.pjc.edu/tgrow/2210L/chm221
0LCafext.pdf

Data Gathered: http://www.orgchem.colorado.edu

Weight of tea 10.00 grams http://www.pharmainfo.net/reviews/extra

leaves used ction-caffeine-tea-leaves
Weight of empty 103.30 grams
http://www.scribd.com/doc/24382592/Ext
evaporating dish
raction-of-Caffeine-From-Tea-Leaves
Weight of 103.70 grams
evaporating dish http://www.scribd.com/doc/26317714/De
with extracted termination-of-Caffeine-in-Tea-Samples
caffeine
Weight of Caffeine 0.40 grams http://www.scienceteacherprogram.org/ch
emistry/Paul00-2.html

Using the formula:

wt. of crude caffeine
% yield = x 100%

wt. of tea leaves used

0.20 grams
% yield = x 100%

10.00 grams

= 4.0 %

A percentage yield of 4.0% was

computed after getting the ratio of the
weight of caffeine and weight of tea
leaves used and was multiplied by 100.
This means that for every 10.00 g. of tea
leaves, an amount of 0.04 grams of
caffeine is contained.

References:

Books:

Bayquen, A.V., Cruz, C.T., et al (2009).

Laboratory Manual in Organic Chemistry.
Quezon City: C&E Publishing Inc

Internet:

http://www.chemenv.titech.ac.jp

http://www.chemistryabout.com