NATIONAL AUTONOMOUS UNIVERSITY OF

MEXICO

FACULTY OF HIGHER STUDIES

ZARAGOZA

BIOLOGICAL PHARMACEUTICAL CHEMISTRY CAREER

Report on the practice: Extraction of caffeine from

an energy drink (Red Bull)

Prepared by: Bravo Rangel Edgar Gabriel

Durán Aguirre Antonio Yair
Gómez Carbajal Lizeth

Advisor: López Ortiz Manuel

Group: 2355

05/28/2018
Summary:
Through the extraction method, the separation of caffeine was carried out from a
energy drink which in our case was Red Bull and later it
sublime and caffeine identification tests were conducted through a
murexide reaction and by thin-layer chromatography.
To the milliliters of Red Bull to be used, sodium carbonate was added to
neutralize the reaction that could occur when adding solvent.

A liquid-liquid extraction system was set up (see scheme 1 page), it

500 mL of Red Bull was placed in a separation funnel to separate the
aqueous phase of the organic phase. subsequently, 50 mL of was added
chloroform to perform 5 extractions (until separation is no longer observed)
of two phases) then brine was added to the phase that remained in
the funnel and a final extraction was made, finally sulfate was added to it
anhydrous sodium as a drying agent. Finally, it was placed in a flask
ball.

A sublimation system was set up using a cold finger (see diagram 2 p.).
With the help of a Fisher lighter, the solution was evaporated while on the finger.
cold sublimation of caffeine was carried out. The grams obtained were weighed.
caffeine obtaining ___.

A system was set up to carry out the murexide reaction to identify

caffeine (see diagram 3 page), a small sample was added to a crucible
of caffeine and 2 mL of concentrated nitric acid was added, subsequently with
help from a Fisher lighter, the solution evaporated, leaving a residue of
yellow-brown color, then add 2 drops of hydroxide of
ammonium, which turns purple.
Finally, chromatographic plates were used, comparing our crystals.
of caffeine with a substance provided to us by the teacher, in the sample
It was observed from the professor that there was only one point slightly off, whereas in
In our sample, two more points are observed, which shows that it has
impurities.

Introduction:

Extraction is the most used technique to separate a product from a

reaction mixture or to isolate it from its natural sources. It can be defined
as the separation of a component from a mixture by means of a
solvent.

Liquid-liquid extraction is a process widely used in laboratories.

organic, is carried out through two phases, an aqueous (water, solution
saturated aqueous sodium chloride, acidic aqueous solutions,
basic aqueous solutions, aqueous solutions of sodium bisulfite) and

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an organic compound, usually the desired compound is extracted into the phase
organic leaving many impurities in the aqueous phase. The organic compound
It is not usually obtained pure, therefore to eliminate the highest number of impurities
Washes are applied with a solvent that is usually immiscible with water.

Theoretical foundation of liquid-liquid extraction:

Liquid-liquid extraction is a technique used to separate a solute.

of 2 immiscible liquids with each other. Therefore, the aim of this technique is the
separation of a compound and its concentration in a specific phase

Generally, this technique is based on the distribution of the desired compound and
of the impurities between the organic medium and the aqueous medium. It is defined
distribution coefficient (K) as the solubility of the compound in the medium
organic divided by its solubility in the aqueous medium. For the process to
for extraction to be effective, the distribution coefficient must be high, since
it is the one that will indicate the number of washes that will be necessary to apply for
that the extraction of the desired compound is optimized. The distribution of
solution between two phases takes place until it reaches a dynamic equilibrium, due to
thus, the distribution coefficient can also be defined as the ratio of
concentrations of the solute between two immiscible liquids.

The affinity of a solute for a solvent is due to the interactions between the
molecules of the solute and the solvent, in such a way that a polar solute
it will show greater affinity in a polar solvent and little in a non-polar solvent
polar

Sublimation is a process in which substances in a solid state transition to

gaseous state without passing through the liquid state. This property is used for the
separation and purification of organic substances. Caffeine is a
alkaloid that can be separated by this process and is determined
qualitatively by the murexide reaction

objectives
General
Extract caffeine from an energy drink.
Specific
Carry out the corresponding number of washes with the help of the coefficient of
distribution
Conduct tests for caffeine identification by the murexide reaction and
thin layer chromatography
Hypothesis: 0.1679 g of caffeine will be extracted from 900mL of Red Bull (drink
energetic) taking as a reference its solubility in chloroform 1.47Kg/L

Variables:
Independent variables: Pressure
Dependent variables: Temperature, concentration of substances

Experimental part:
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List of materials:
Material Capacity Quantity
Test tube 1000 mL 1
Separation funnel 1L 1
Fisher Lighter 1
Bowl flask 1L 1
Three-finger clamp 1
Iron ring 1
Object holder 5
Plastic funnel 1
Pasteur pipettes 5
Asbestos cloth 1
Capillaries 1
Universal support 2
Precipitated vase 1000 mL 2
Cold finger 1
Crucible 1
Triplet 1
Crucible tongs 1
Porcelain triangle 1

Team:
Analytical balance
Recirculator

Reagents:
Reagent Formula
(condensed/semi-developed)
Water H2O
Chloroform Chlorine trifluoride3
Anhydrous sodium sulfate Na2SO4
Sodium carbonate Na2CO3
Ethyl acetate C4H8O2, CH3COOCH2CH3
Silica gel SiO2
Concentrated nitric acid HNO3
Ammonium hydroxide NH3

Technique
Measure 900mL of the energy drink using a graduated cylinder and transfer it to
a 1 L beaker. Approximately 6.75g of Na was added.2CO3
slowly until the Red Bull stopped fizzing. Added
450 mL of the substance into a separation funnel, then it is
50 mL of chloroform was added using a graduated cylinder, the funnel was stirred.

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separation and waited for it to separate into 2 phases to carry out the first
extraction, 4 washes were performed on each of the samples.
Finally, the aqueous phases were combined and a final extraction was carried out.
but in this case brine was added instead of chloroform. Next
sodium sulfate was added to the phase that remained above and was transferred to a
ball flask.
The system is set up as shown in diagram 2 (page). With the help of a
Fisher lighter evaporated the entire sample, leaving the caffeine crystals.
on the cold finger, he scraped the cold finger with a spatula and the crystals
They were placed on a glass of a clock and weighed.

Identification tests were conducted using the murexide reaction. It is set up.
system as observed in scheme 3 (p. ). A ...
small sample of the obtained crystals and 2 mL of nitric acid was added
concentrate, with the help of the Fisher lighter the mixture evaporated and remained a
yellow residue, to this was added a drop of ammonium hydroxide
and finally a purple color was observed

Preparation of plates.
Silica gel is added to a 50 ml beaker and it is added little by little.
little ethyl acetate until a thick appearance is achieved.
Three plates will be prepared, quickly adding the stationary phase in a
slide until a smooth and thin layer is found, leaving a
thickness of 0.5 cm of the plate without adding. Let it rest for 1 min
approximately to work with the plates.
Once the plates are made, a sample will be deposited (2 points) in the part
inferior where the stationary phase plate is totally covered,
measure approximately 0.5 cm from the bottom to the top and place
2 points (pure caffeine and caffeine obtained in the laboratory) with a distance
among them 0.5 cm. the samples will be taken with the help of capillaries.
Outside the laboratory, fire will be applied with a lighter in the middle of the
capillary, start to separate them so that a thinner capillary remains and apply a
show on the plate in a finer and more precise way.
In a 50 ml beaker, hexane and ethyl acetate will be added.
proportions 1-1. To a volume in such a way that at the moment of placing the
chromatography plate do not exceed the sample placement points.
Place the chromatography plate with the 2 sample spots in a glass.
precipitate with the pair of solvents (hexane, ethyl acetate) place a
watch glass in such a way that it covers the beaker. Wait for the
eluent rises, the solvent front should not reach the other end of the
plate. remove the plate and make a mark where the eluent has reached. With a
UV lamp observe the distribution of the spots, measure the Rf.

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 Flowchart

Caffeine extraction

In a funnel
Calcium carbonate is added 50 mL of is added of addition
a 900mL of Red Bull chloroform putfor450 mL
make extractions from the drink already
(4 extractions from each)
with the
sample carbonate of
sodium

System is installed If the add A final one is carried out.

from scheme 2 for sodium sulfate to extraction with brine
sublime the caffeine the phase that
contains caffeine
and is deposited in
a round-bottom flask

In a crucible add a
The grams of are weighed They are carried out sample of crystals and
caffeine obtained for add 2 mL of acid
tests of
calculate performance nitric concentrated
identification
evaporate y
with the reaction
later add 1
of murexide
drop of hydroxide
ammonium. It must be observed
a purple color.

Samples apply
of pure caffeine and the It is poured into a glass
obtained in the acetate precipitate 3 are being prepared

laboratory of style plates

chromatographies

The course of the

samples with a light lamp
ultraviolet. And the Rf is calculated.

Diagrams of the devices

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Scheme 1.

Illustration 1. system, liquid-liquid extraction.

Scheme 2.

Illustration 2. Sublimation of caffeine.

Scheme 3.
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 Illustration 3. Murexide reaction

Diagram 4

Illustration 4. chromatographic plates.

Cost of reagents
Name Formula Purity Amount Cost (MXN)
Chloroform CHCl3 99% 100 mL $848.00
Sodium carbonate2CO3 90% 500 g $1588.00
sodium
Acetate from C4H8O2 99.8% 1L $1648.00
style
Sulfate of Na2SO4 99% 500g $1,112.00
sodium
Silica gel SiO2 size pore 1Kg $8049.00

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 150 Å, 200-
425 mesh
Nitric acid HNO3 70% 500 mL $1494.00
concentrated
NH hydroxide3 99% 100 mL $1488.00
ammonium

Results.

Performance obtained.

Initial grams of cloves 100 g

Oil with impurities (water) 400 mL

Chloroform oil 180 mL

Pure Eugenol 7.43 mL

Percentage of performance

real yield
%performance= x100
theoretical yield

3.889g
%yield= x100=88.62%
4.388g

% Performance 88.62%

Calculation of RF (Eugenol).

fig.1 plate with eugenol samples

A
RF=
B

A = Distance walked since the

origin for the composed

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 B = distance traveled from the origin by the reference compound.

3.2
R F1= =0.64
5
3.0
RF2= =0.6
5
2.8
R F3 = =0.56
5

Sample value A Value B value of RF

1 3.2 5 0.64

2 3.0 5 0.6

3 2.8 5 0.56

Identification of phenol

Ferric chloride + acetone + eugenol

Green color identifies phenol

Analysis of results

7.43 mL of eugenol was obtained.

To identify phenol, green or blue colors were expected.
observed an olive green color.
RF Tests

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 On the chromatography plate, the samples can be observed.
identified at the same point, for which it can be considered the
eugenol with high purity.

Conclusions
Using steam distillation, the separation of the oil was achieved.
essential oil of clove, eugenol, which was later identified as phenol
content in the molecule of eugenol and chromatography tests were carried out
in a thin layer to observe the impurities that the oil may contain
extracted.

References
1. Furniss, BS. Hannaford, AJ. Smith, PW. Tatchell, AR. Textbook of
practical organic chemistry. 5th ed. Essex (UK): Longman Scientific &
Technical;1989.
Hardegger. E. Introduction to Organic Chemistry Practices. Spain:
Reverté Editions; 1965.
3. Solomons. T.W.G. Organic Chemistry

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