Important laboratory techniques made for the extraction of caffeine

Villanueva H. Carlos (20180513F), Candia T. Paul(20184154K)

Organic Chemistry Laboratory I, PQ-212B, FIP
karlozvh04@[Link], poweryc@[Link]

Completed: April 29, 2019

Presented: May 6, 2019

SUMMARY

The work done in the laboratory seeks the extraction of a chemical substance from its natural source.
in our case, isolation of natural products such as alkaloids (caffeine) through pure tea. The process was carried out
extraction, purification, and identification of caffeine through tea. First, 3 extractions were made with
20mL of dichloromethane was shaken to extract the organic phase, after extracting the 3 phases, a ...
extraction more but this time with 30mL of NaOH, then anhydrous sodium sulfate was added to remove them and
We filter to remove the excess sodium sulfate. Then we eliminate the solvent by evaporating it.
bell and finally it sublimated in a kitasato with a test tube filled with ice in order to obtain the
caffeine in solid state stuck to the test tube.

Palabras clave:Alcaloides, Cafeína,Emulsiones, Embudo de extracción, Sublimación.

THEORETICAL ASPECTS

1.1. The problem to be solved

The work done addresses a problem:

1.- Currently, there are two common ways in industries to obtain caffeine from alkaloids.
the main difference lies in the liquid separation, which can also be done through a method of
Distillation. We must prove that the simplest way to separate solid-liquid is through a funnel.
separator in a simple extraction process.

2.-Due to the fact that alkaloids have similar chemical properties, we will analyze whether it is possible to use them
same solid-liquid extraction methods, liquid-liquid extraction, purification by sublimation and use
from the void for all alkaloids.

To carry out this experiment, it is necessary to understand some concepts and laboratory techniques such as
alkaloid, caffeine, extraction, purification by sublimation, emulsions, so that, when making them you understand, for
what each technique is for and why it is necessary to understand each concept. To start the discussion, the following are proposed:
next questions:

What is an alkaloid?
What is caffeine?
What is an emulsion?
How is the solid-liquid and liquid-liquid extraction technique performed?
How is the purification process by sublimation carried out?

1.2. Theoretical Framework

1.2.1. Alkaloid

Alkaloids are nitrogenous compounds, with nitrogen usually being part of them.
of a heterocycle and, in some cases, forming part of an open chain. They are also composed of
by carbon and hydrogen, many contain oxygen, which gives them a series of physical properties (solid,
crystallisable), and they rarely contain sulfur.

Physical-chemical properties of alkaloids

Most alkaloids are found in dicotyledonous plants in the form of salts (malates,
tanatos, citrates, etc.). Except for some like nicotine, which are liquids, the others are solids,
they have a bitter taste and alkaline reaction, generally insoluble in water, more so in ether and
easily soluble in alcohol. With acids, they form salts and with certain substances, they produce colored reactions.
Oh no, but characteristics. They commonly have the nature of tertiary amines, and are mostly poisons.
very violent; strong tea infusions can be used as an antidote; the tannin of the infusion
it precipitates the alkaloid and inhibits its absorption in the digestive tract.

Most important alkaloids (Illustrated in Figure 1)

CocaineAddictive stimulant of the central nervous system, specifically of the dopaminergic system. It
extract from the sheet of thecocaIt can be used in surgery (as an anesthetic).

Heroine(diacetylmorphine): Synthesized from morphine. Like it, it is analgesic, but also

It has certain stimulating effects. It is very addictive and acts quickly. More potent than morphine but less
durable.

NicotnoIt is extracted from tobacco. It is a powerful poison used as an insecticide in greenhouse spraying.
low doses, it's stimulating. Causes tobacco addiction.

TheobromineIt is found in the cacao tree, and therefore, in chocolate (especially, chocolate
black). It is a stimulant of the central nervous system (less than caffeine) and a bronchodilator. It causes
possibly chocolate addiction. In dogs, cats, and other animals it is toxic.

Quinine: It was originally extracted from the bark of the cinchona tree, a tree native to South America. Its name comes from...
from the Quechua language. Famous for being used against malaria.

MorphineIt is extracted fromopioit has strong narcotic and anesthetic properties. That's why it is the most used in
medicine for pain, especially severe pain. Highly addictive.

Figure 1. Most consumed alkaloids tobacco and coca leaf

1.2.2. Caffeine

Caffeine, also known as 1,3,7-trimethylxanthine, is illustrated with its formula in Figure 2; it is an alkaloid.
belonging to the family of xanthines. It has stimulant properties on the central nervous system.
It can be obtained from various sources, such as coffee beans or tea leaves, or synthetically.

It is considered a psychoactive drug, although with mild side effects if consumed properly. But
As such, it can become addictive. It does not provide nutritional value, and can be used to promote certain
areas, such as sports and the performance of cognitive tasks.

It can affect mood and cause physical side effects, mostly positive ones, especially.
in the sports field.

Generally, a daily caffeine intake of about 400-600 mg is estimated for individuals.

such as doses that are not associated with health risks or toxicity.

Figure 2.- Developed chemical formula of caffeine

1.2.3. Emulsions

Emulsion is a process that involves mixing two different liquids that cannot be mixed.
to say, that they are immiscible with each other. One of the liquids will be forming the phase, known as the
name of the dispersant, which will be constituted by the liquid that will be dispersed within the other liquid
what makes up the mixture, which will be the one forming the phase known as continuous, or also, dispersant.
A large part of the emulsions are made between water and oil, or other commonly used fats, as is the case.
of mayonnaise, milk and its cream, butter and margarine, etc. For example, in the case of the
butter/margarine, the fat is surrounding the water droplets, so it is partly an emulsion
of water in oil.

We can distinguish three types of emulsions:

Flocculation, unstable emulsion where particles join together forming a mass.

Cremation, unstable emulsion where the particles tend to concentrate more on the surface.
from the mixture that is formed, although remaining separate (they can also accumulate at the bottom of the
mix).
Coalescence (union) is the type of unstable emulsion, where the particles that constitute it merge.
turning into a liquid layer.

When we talk about water-in-oil emulsions (very common) or vice versa, it greatly influences the
volume of each phase, as well as the type of emulsifier used. Thus, a rule known as the
name of 'Bancroft's rule', where the emulsifiers, as well as the emulsified particles, tend to usually
to increase the dispersion of the phase in which an optimal solution is not usually made. A classic example.
for the purpose of emulsions, it may be the case of proteins, which dissolve easily in water,
but not in oil, which causes them to form oil emulsions in water.

The color that emulsions usually have is white. When an emulsion is diluted, light is dispersed.
following the Tyndall effect. In the case that we are talking about a concentrate, the most frequent color
it will be yellow, as the color will have distorted towards this tone.
We talk about emulsifiers shown in Figure 3, also known as emulsifying agents, when we
we refer to a substance that manages to stabilize an emulsion, generally it involves substances
surfactants.

Figure 3.- Most common emulsion, water with oil

1.2.4. Solid-liquid extraction

Definition: Operation by which a soluble component is extracted from a solid using a solvent.
Other denominations: Leaching, "Leaching", Washing (particular case).
Generally, it is divided into 3 parts:
1) Change of phase of the solute (If it is liquid, there is no change of phase) when dissolving in the solvent.
2) Diffusion of the solute through the solvent, through the pores of the solid and towards the periphery of the solid particle.
3) Transfer of the solute from the solution in contact with the particles to the body of the solution.

Although any of the 3 parts affect the extraction speed, the first takes place quickly.
Therefore, its effects are negligible. If the solid has a cellular structure (for example: seeds
oilseeds, coffee beans, beet, etc.), the cell walls provide additional resistance to the
mass transfer as shown in Figure 4. Therefore, the extraction rate is relatively low.
 Figure 4. Solid-liquid extraction by distillation

1.2.5. Liquid-liquid extraction

Liquid-Liquid Extraction is, along with distillation, the most important basic operation in separation.
homogeneous liquid mixtures. It consists of separating one or more substances dissolved in a solvent by means of
its transfer to another solvent that is insoluble, or partially insoluble, in the first one. The transfer of
matter is obtained through direct contact between the two liquid phases. One of the phases is dispersed in
the other one to increase the interfacial area and increase the flow of transferred matter.

In a liquid-liquid extraction operation, the feed is called the solution whose components are
intends to separate, extraction solvent from the liquid that will be used to separate the desired component
We noted in Figure 5, refining the already treated feed and extracting the solution with the solute.
recovered

Applications

Separation of inorganic compounds such as phosphoric acid, boric acid, and sodium hydroxide.
Recovery of aromatic compounds.
Refining of lubricating oils and solvents
In the extraction of products that containsulfur.
Obtaining ofcerasparaffinics
Desulfurization of petroleum products
Pharmaceutical products Example in the acquisition of penicillin
Food industry
Obtaining expensive metals, such asuranium-vanadium.

Factors that affect extraction

Composition of the diet,temperature, pressureyspeedof flow.

The desired degree of separation.
Choice of solvent.
 Operating temperature and pressure.
The formation of emulsions and foams.

Figure 5. Liquid-liquid extraction using a separatory funnel

1.2.6. Purification by sublimation

Purification by sublimation is a very simple laboratory technique that allows obtaining a product very
pure. The necessary condition is that at a given temperature the vapor pressure of the solid equals
the external pressure (atmospheric). Under these conditions, the solid turns into gas directly without going through the phase
liquid, this process is known as sublimation.

This phenomenon is very useful in devices such as fire extinguishers loaded with carbon dioxide.
solid carbon (dry ice). The vapor pressure is so great that it far exceeds 760 mmHg before
reaching the melting point, causing a very rapid sublimation.

Although many solids have a vapor pressure lower than atmospheric pressure at their melting point, it can be
sufficient for slow evaporation to occur from an open container, such as naphthalene
or camphor, generating its characteristic odors. The process is accelerated by gently heating (without reaching the
melting point) or reducing the pressure with or without heat (sublimation at reduced pressure)[3].

Sublimation under reduced pressure, cold finger

The equipment necessary for this technique is somewhat more complex and allows for different possibilities, with some
For some substances, the pressure reduction will be sufficient, while for others it will be necessary to heat and use a refrigerant.
of specific design, the cold finger. It is very convenient to consult the phase diagram of the substance that
We want to purify to know the best temperature and pressure conditions that we can apply.

The solid to be purified is introduced into a collector tube with a lateral outlet for the vacuum as shown to us.
Figure 6 and ground mouth to adjust the cold finger. Water circulates through the interior of this piece to
provide a cold surface that condenses the vapor generated at the base of the collector.
 Figure 6. Test tube connected to vacuum
1.2.7. Melting point

The normal melting point of a crystalline solid is the temperature at which the solid turns into a liquid.
under atmospheric pressure. For pure substances, the phase change from solid to liquid occurs within a range
small variation in temperature and from there its relative value to identify a substance by its point of
fusion

2. HYPOTHESIS AND OBJECTIVES OF THE EXPERIENCE

2.1. Working Hypothesis

If sublimating with equipment connected to a vacuum, then caffeine will be obtained.

purified with a considerable amount of purity.

2.2. Objectives of the research

Isolate the alkaloid caffeine using liquid-liquid extraction as a technique from a mixture.
of tea, and subsequent purification of the compound by the sublimation technique.

Getting acquainted with or learning to implement the different laboratory techniques in this case the
liquid-liquid extraction is a widely used methodology in extraction processes of
compounds with dissolution from a mixture.

3. EXPERIMENTAL PART

The first step of the experiment done in the laboratory was to introduce 5 bags of pure filtering tea into a glass.
precipitate adding 100mL of boiling distilled water afterwards. Then let it rest for 5 minutes to
afterwards extract the bags by squeezing well to obtain the largest amount of water without breaking them
small bags, once cooled as we see in Figure 7, we can proceed to the next step.
 Figure 7.- The 100mL of distilled water mixed with pure tea

We poured the solution with 20mL of dichloromethane into the extraction funnel shown in Figure 8.
to obtain the substance that is found at the bottom after moving the mixed funnel
funnel with what remains at the top we add 20mL of dichloromethane again and so once more
in order to obtain 3 proportions of the resulting organic phase to then pour back into the funnel but
this time with 30mL of a NaOH solution, when collecting the organic phase in a flask we dry it with
sodium sulfate, when everything dries we filter the material by gravity to remove the excess sodium sulfate.

Figure 8.- Pure tea solution with 20mL of dichloromethane

Then we place the obtained material in the evaporation hood to eliminate the solvent after passing through a
We will slow down time to be able to sublime it in a Kitasato with a test tube filled with ice.
obtaining the purified caffeine in solid state stuck on the walls of the test tube as we see in the
Figure 10. then use a spatula to remove it and place it on a filter paper to present it.
 Figure 10.-Obtaining caffeine stuck on the walls of the test tube
Flowchart of the experiment
The following Figure 11 shows the main operations performed during the experiment.
Figure 11. Main operations carried out in laboratory No. 2
4. RESULTS AND CALCULATIONS

Obtained Data:
Weight of the tea bags: 9.587g
9.047g
Density of caffeine: 1.23
Mass of impure caffeine = 0.0615g
Mass of pure caffeine: 0.002g

I. Caffeine extraction

Table 1. Results of 'Caffeine Extraction from Tea Bags'

Tea mass (g) Caffeine mass (g) Performance (%)

9,047 0.0615 0.679

% Rendimiento= 0.679

II. Sublimation of caffeine

Table 2. Masses of caffeine

Caffeine masses Mass of pure caffeine (g) Yield (%)

impure
0.615 0.002 3.25

Sublimation yield = 3.25

5. DISCUSSION OF RESULTS

Extraction is based on the solubility of the substance to be extracted and the solvent used, in the case of caffeine.
an apolar solvent such as dichloromethane was used.

To extract caffeine, a solid-liquid extraction was performed, where all substances were dissolved in water.
present in the tea bags that were soluble in this solvent, to then perform a liquid extraction
liquid in which there is an aqueous phase and an organic phase, water and dichloromethane, respectively.

By having both phases interacting, the solute begins to migrate to the organic phase until reaching a
equilibrium between the two phases, consequently the concentration of solute will increase in the organic phase and
it will decrease in the aqueous phase, distributing the total solute concentration between both phases. The distribution of
solution depends on its solubility in both solvents and can be interpreted with the coefficient of
distribution(KD) and is constant for each solute in certain solvents. In this case:
 KD[S Dichloromethane] / [S Water] KD10g/100ml / 2.1g/100ml = 4.8

The partition coefficient of caffeine in water and dichloromethane shows that the solute will be distributed in
mostly in the organic phase of dichloromethane.

Liquid-liquid extraction was carried out in a separating funnel where the organic phase is the
denser remains at the bottom and the aqueous phase at the top, when making constant extractions with
dichloromethane (3 extractions) allowed the caffeine that was dissolved in the water to be extracted, as according to the
distribution coefficient, caffeine would distribute into the organic phase, so in each extraction, the phase
aqueous had less dissolved caffeine.

By adding sodium hydroxide, an alkaline extraction was made, where the acidic substances present in the
organic phase forms water-soluble salts.

After the organic solvent has been in contact with the aqueous phase, it will be saturated with water, although
the amount of water is very small so a drying agent was added to prevent dragging of
impurities with water. The drying agent used was anhydrous sodium sulfate, which binds the water to it, as
It can be seen in the following equation:

Na2SO4 Na2SO4x 10 hours2O

When filtering the solution, Na was separated.2SO4x10H2O, leaving the organic phase without water residues, so that to the
Using the rotary evaporator, the dichloromethane was evaporated, leaving only a white solid.
impure.

The amount of impure caffeine obtained was 0.0615g as indicated in Table 1, so when taking out the
percentage of extraction yield:

%R = (mass obtained experimentally (g) / initial mass (g)) x 100

%R= 0.679 %

A 0.679% caffeine was obtained with respect to the mass of tea, so the amount of caffeine present in
five tea bags is very small, or there may have been errors in the experimental procedure
operational top in the extraction with the decantation funnel when not properly separating the organic phase from the
aqueous, it is recommended to shake properly due to possible emulsions.

Sublimation is a process in which substances in a solid state pass into a gaseous state without going through
liquid state. This technique consists of heating the solid mixture causing it to change to a state
gaseous, the vapors solidify again upon hitting the cold finger and thus, they are deposited at the base
from the cold surface in the form of crystals. During the practice, the sublimation of caffeine was carried out.
in such a way that starting from 0.0615 g of impure caffeine as stated in Table 1 (extracted in the previous practical),
0.002 g of pure caffeine was obtained. As indicated in Table 2, sublimation was performed because in the first
Some of the formed crystals fell from the cold finger due to their own weight, because of mistakes.
operational when removing the cold finger or were left on the walls of the tube due to a large part of this
The yield of this experience was 3.25%, this value may be due to sample losses beforehand.
mentioned or that the sample had a high percentage of impurities.
6. CONCLUSIONS

We consider that, based on the results obtained on the procedures done with the
caffeine, the following conclusions correspond:

If we obtain purified caffeine, but due to different factors as we have already seen, we do not
by properly separating the organic phase from the aqueous phase, we obtain a very small mass stuck to the
test tube.

We must take into account the compounds we deal with when breaking emulsions due to
that those possible compounds when trying to extract the desiccant and leave the caffeine with it
dichloromethane as pure as possible, and then set it up on the rotary evaporator as much as possible is that
we will extract everything we added when breaking the emulsion, thus generating greater impurity
to caffeine and it is what we want the least.

The procedure that has been carried out for the extraction of caffeine can be generalized with
the other alkaloids since they also have a basic character and are soluble in
organic substances.

6. QUESTIONNAIRE

6.1. Investigate properly and answer: What other alkaloid is found in tea?
Are idealistic the people who drink tea "because coffee makes them nervous"? Why?

Another alkaloid found in tea is theophylline, which is a substance that is present in smaller proportions in the
caffeine, its effect is essentially vasodilatory, that is, it dilates veins and coronary arteries, improving by
both the circulation.

If those people are idealists, why do they create a reality based on that idea that coffee puts them in?
nervous if in the end both tea and coffee contain caffeine, then if supposedly coffee makes them nervous.
then he should do it too.

6.2. Alkaloids are often obtained from plants by extraction with acids. Why in the
the procedures described do not use an acid extraction?

Because caffeine is a base and if we perform the procedure with an acidic extraction then
we would obtain an acidic solution instead of getting the result in the laboratory.

6.3. What is the function of NaOH in this practice?

The function of NaOH is to neutralize acids so that the obtained aqueous solution is a base, thus reaching the
caffeine in solid state.
6.4. The vapor pressure of a certain compound is 265mmHg at its melting temperature (109-110°C)
Will the compound sublimate?

The compound does not sublimate because the pressure is much lower than atmospheric pressure, making it impossible for it to do so.
sublime.

6.5. Caffeine is extracted from tea leaves, obtaining 200mL of an aqueous solution containing
2g of caffeine. This alkaloid is extracted from the aqueous solution using volumes of 100mL of
chloroform. If the distribution constant of caffeine between chloroform and water at 25°C:

A) What is the name of the team required for this operation?

To carry out the procedure, a separating funnel is needed to separate the solutions after shaking.

B) What size would you request from the warehouse?

I would request a measure of 500mL because two extractions of 200mL are made and 100mL is added.
chloroform

C) What is the maximum number of extractions that you would perform in practice?

2 extractions because when obtaining them we know it has better miscibility

D) What is the amount of caffeine that would remain in the aqueous solution after extracting it 2 times with
chloroform, as indicated?

Upon making the first extraction with the relationship:

K = C1 / C2

K: ALLOCATION COEFFICIENT

C1Y C2:CONCENTRATIONS

10
100(2 x)

x=1.7 in aqueous

So it results in 0.3 from the first extraction

The second extraction:

x
100 10
0.3 x
200
X=0.25 in aqueous

Therefore, the caffeine will have 0.5 in quantity.

7. BIBLIOGRAPHIC REFERENCES

[Link]
Lidia Galagovsky, Organic Chemistry Practical Fundamentals for the Laboratory, pages 117 and 118
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44a3042a281d/2/organic_chemistry_lab_SCORM.zip/page_16.htm
Lidia Galagovsky, Organic Chemistry Practical Fundamentals for the Laboratory, page 19