ABSTRACT

In experiment 4, a study was conducted in order to investigate the liquid- liquid extraction
technique between separation of benzoic and from two organic compounds mixture which were
naphthalene and benzoic acid. Besides, it was provide a better understanding in the usage and
application of separation funnel. In general, liquid- liquid extraction is a process where a solute
was extracted from a binary solution by having it to interact with another immiscible solvent
in which the solute is soluble. The experiment was started by weighing 0.5g of benzoic acid
and 0.5g of naphthalene and dissolve the mixture in 25 ml of dichloromethane. then, the
solution was put into a clean separatory funnel with suitable size and 25mL of 10% aqueous
sodium hydroxide (NaOH). Make sure the outlet was closed, so that the solution will not leak
from the funnel. In order to get two separated phases, which were aqueous phase and organic
phase, the funnel was swirl slowly and the outlet was opened for awhile to make sure the gas
will out and this step were repeated until the mixture fully separated and no gas available. In
order to know which one was the organic phase and aqueous phase, the one who was upper is
considered as aqueous phase. Next, when the mixture already separated , the outlet was opened
to get the upper or organic solution the wash it with 10mL of dichloromethane. After that, the
layer was poured into a small beaker and cool about 15°C. After reached 15 °C, ice- cold
concentrated hydrochloric acid (HCI) was added until a pH of 1 is obtained. Then, by using
Buchner funnel, the solid was vacuum to filtrate it. Lastly, the product was dried for a while,
and weighed. the mass of the product obtained was 2.16g.
In experiment 5, the experiment was conducted to see the isolation of caffeine from tea. The
objectives to achieve was to extract the caffeine from tea leaf. The experiment was started by
weighing 10g of tea leaf and 3g of calcium carbonate to a 200mL beaker then 100 mL of boiling
water was added. Be careful with not boiling water. Next, using ice bath , cool the aqueous
solution to room temperature about 24°C and by using the gravity filtration, the solution was
filtered by using muslin cloth and filter paper equipped together and vacuum it for a while.
Then, the filtrate was poured into a 200mL separatory funnel and the mixture was gently shake
and swirl for at least 5 minutes for two times, the first time 30mL and 25 mL with
dichloromethane and the extraction was obtained. Before poured into the separators funnel, the
outlet was closed to prevent any leaking while doing separation technique. After that, the
combined dichloromethane was extracted with 5g of anhydrous sodium sulphate until it
became hydrated with the water. The beaker filled with the combined mixture was placed on a
water bath at low -medium heat (80°C) until it dried. The mass of caffeine obtained was 0.12g
and the approximate percentage of caffeine in tea calculated using the mass of the theoretical
mass of 5% of crude caffeine in the tea leaf was 24%.
INTRODUCTION
The process of liquid - liquid extraction entails the transfer of specific compounds
between two solvents that are either immiscible or only slightly miscible. Liquid -liquid
extraction is also frequently used to wash an organic phase, such as to get rid of inorganic
compounds or to protonate or deprotonate bases or acids so they can dissolve in water.
Below is a flow diagram of a relatively common extraction process, which involves
quenching a reaction mixture with water, extracting it (many times), washing it with brine,
drying it, filtering it, and then evaporating it to produce a crude product or a pure product.

Liquid - liquid extraction is normally carried out with one aqueous phase (either pure
water, or an aqueous solution), and one organic phase. We will refer to it as extraction from
here on. It’s vital to remember that the desired component, which is often an organic molecule,
can exist in either phase theoretically. It depends on the characteristics of the substance as well
as the aqueous phase. We will first look at some typical extraction instances before talking
about some technical considerations.

There are three types of extraction. Firstly, the types of extraction is neutral chemical
extraction. The extraction procedure usually consists of repeatedly extracting with an organic
solvent of the target organic chemical is neutral (neither acidic nor basic ). secondly, if the
desired chemical is acidic, we can use an aqueous base to selectively deprotonate that
compound. the deprotonated chemical will be drawn into the aqueous phase as a result.
We can discard the organic phase, which now contains any byproducts and/or unreacted starti
ng ingredients, because relatively few organic molecules are soluble in water. Precipitation of
the desired product is possible through acidification of the aqueous phase. Thirdly, extraction
of fundamental substances. The procedure is quite similar to that for the acidic compound
above if the result is basic. Using an aqueous acid, we can protonate the basic chemical
Precipitation of the desired product is possible through acidification of the aqueous phase.
The basic component may precipitate after neutralising the acidic phase or it may need to be
extracted again using an organic solvent.
The separatory funnel is a common piece of extraction equipment. There are several
various sizes available, but 100ml is the most common quantity and is what you will find in
most PSU Labs. the funnel has a top and a nozzle and a stopcock can be used to close or open
the nozzle. we can precisely differentiate the two phases because of the structural property of
the funnel's tiny nozzle.

The formation of emulsions by some chemicals , a bothersome habit, is a typical issue

with extractions that is not caused by poor method. These typically form between the two layers
and are chemically made up of the two solvents as well as other substances present in the initial
solution. the emulsion may self-resolve in a short period of time or may be highly persistent.
In the latter situation, draining the emulsion out in the phase that doesn't contain the product
and again extracting it can occasionally be a good method.

In general, caffeine is an organic substance which is known by its bitterness. It is a

white crystalline and it is also a central nervous system stimulant that blocks the action of
adenosine which prevents the drowsiness induced by adenosine. Caffeine has a chemical
formula of C8H10N4O2 with its IUPAC name as 1,3,7-Trimethylpurine-2,6-dione. The
following figure shows the structure of the caffeine,

The pyrimidinedione and imidazole rings are fused together to generate caffeine, an
alkaloid. A pyrimidinedione ring has six members and two nitrogen atoms, whereas an
imidazole ring has five members and two nitrogen atoms. Caffeine commonly used stimulants
in athletes, caffeine creams are applied to the stain to reduce redness and itching in dermatitis
and caffeine is used as an ingredient in energy drinks, soft drinks and other beverages.

When tea or coffee is brewed, the caffeine is removed from the tea or coffee grounds
because it is soluble in hot water. Although caffeine is water soluble, dichloromethane has a
substantially higher solubility for caffeine. Since dichloromethane and water cannot mix, a two
layer mixture results from the mixing of the two substances. Because it is denser than water,
dichloromethane often makes up the bottom layer of the two part mixture.
Caffeine can be drawn out of the organic layer by brewing tea and adding methylene chloride
to it. Because the organic layer is insoluble in water, it may be removed when the water
separates from it and the solvent can then be expelled to yield practically pure caffeine.
It’s conceivable that throughout the extraction procedure, an emulsion will form. Spin
the tubes in the Centrifuge for two to three minutes to break the emulsion. Make sure the
centrifuge is balanced by placing two tubes in the rotor with almost equal masses opposite one
another. The mixture ought to be divided into two layers, with a dark upper layer and almost
colourless bottom layer. The tubes should be centrifuged once more if a third frothy green -
brown layer appears between the top and lower layers.
The choices of solvent used during liquid-liquid extraction involves several
perspectives in other to decide the suitability of a particular solvent. The solvent which was
chosen shall allow the existence of large density difference between two phases to ensure the
successful extraction. It should also be selective towards the solute being extracted and
contributes to the low interfacial tension between two phases during the extraction. Therefore,
the solvent used during the liquid-liquid extraction are water which is a polar universal solvent
and non-polar organic solvent. The difference in both of their properties allows the creation of
two immiscible separated layers as organic solvents does not dissolve very well in water. The
presence of two different solvents allows the solute to have a choice of two solvents which they
can dissolve in which is known as solvent partitioning. Higher solubility of solute in either
solvent allows it to be separated easily as the two solvents are immiscible that makes them easy
to be separated from one and another along with the solutes that dissolved in the solvent.
Objectives
Experiment 4
1) Separation of benzoic acid from two organic compounds mixture (naphthalene and
benzoic acid )
Experiment 5
1) To extract the caffeine from the tea leaf.
Reagents
Experiment 4
• Naphthalene , benzoic acid, dichloromethane, sodium hydroxide, distilled water,
hydrochloric acid.
Apparatus
• Beaker (4×50 ML), separation funnel, graduated cylinder, thermometer, ice bath, glass
rod , pH paper , vacuum filter, Buchner funnel, clamp , suction flask , filter paper ,
dropper.
Experiment 5
Reagent
• Calcium carbonate, tea leaf , ice, dichloromethane, anhydrous sodium sulphate.
Apparatus
• Beaker, hot plate, separation funnel, Erlenmeyer flask, filter paper, water bath, distilled
water, glass rod, muslin cloth.
 Methodology

A mixture of 0.5g of benzoic acid and 0.5g of naphthalene were dissolved in 25 ML of

dichloromethane.

the separatory funnel was make sure clean and the outlet was closed.

the solution was put into a separatory funnel of an appropriate size and 25ML of 10% aqueous Na
OH were added.

the organic phase and aqueous phase were determined. the two phases were extracted thoroughly
and separate completely.

the aqueous phase was washed with 10 mL of dichloromethane.

the aqueous layer was poured into a small beaker and cold until 15° C.

Ice-cold concentrated hydrochloric acid were added drop by drop until a pH of 1 obtained.

the solid were vacuum into a Buchner funnel and the precipitate were rinsed from the beaker into the
funnel
Experiment 5

The product were dried, weighed and the percent recovery was calculated based on the amount of the
initial mixture.

to a 200 ML beaker containing 10 g tea leaf and 3g of calcium carbonate , 100mL of boiling water was
added. The beaker was set on the hot plate and boiled gently for 15min.

The aqueous solution was cold to room temperature using an the bath and the solution was filtered
using gravity filtration ( using muslin cloth and filter paper equipped together ).

the filtrate was poured into a 200mL separatory funnel . The tea solution was extracted two times (
gently shake / swirl the mixture for at least 5 minutes ), first time 30 mL and the second time 25mL
with dichloromethane. Both extracts were combined in a 200ml Erlenmeyer flask.

The combined dichloromethane was dried with 5g of anhydrous sodium sulphate. The sodium
sulphate was swirled for several minutes to become hydrated with the water.

The dry dichloromethane solution was filtered into a pre-weight 100 mL beaker.

the beaker was placed on a water bath and the contents was evaporated to dryness at low-medium
heat (80°C). The flask was weighed to find the crude mass of caffeine.

the approximate percentage of caffeine in tea was calculated by using the mass of theoretical mass of
5% of crude caffeine in the tea leaf.
Results
Experiment 4
Phases Layer observed Chemicals obtained
Aqueous upper layer Naphthalene, benzoic acid
organic lower layer Dichloromethane

Reactant Mass of product

Naphthalene + benzoic acid 2.16g

Experiment 5
Reactant Mass
Caffeine 0.12g

Calculation
theoretical mass = (5% × 10) = 0.5g
% mass crude caffeine = (0.12g/0.5g) ×100% = 24%
Discussion
The experiment was conducted to investigate the topic of liquid-liquid extraction which
the application of separation funnel was involved and dichloromethane served as the organic
solvent that contributes to the organic phase of solute. The aim of the experiment carried out
was to observe separation of benzoic acid from two organic compounds mixture (naphthalene
and benzoic acid). Introduction towards the usage of separation funnel was carried out before
the experiment was conducted.
The experiment was started by weighing 0.5g of naphthalene and 0.5g of benzoic acid
and dissolved them in 25 mL of dichloromethane. then, the mixture were put into a separatory
funnel and 25mL of 10% aqueous NaOH were added. The funnel was then stoppered, shaken
and vented. The venting of the funnel was to release the gas produced in the funnel and this is
to prevent the built-up of pressure that might cause the separation funnel become broken. The
shaking of the funnel allows the interaction between two solvents and the separation can be
observed by the end of the shaking which two non-miscible layers were observed. The process
of adding dichloromethane and shaking of separation funnel can be repeated to maximize the
yield of product required.
After the sample solution was added with dichloromethane and shaken, two non-
miscible layers were observed as the funnel was clamped to the retort stand to settle down the
solution. From the results obtained by using aqueous NaOH, the upper layer in the non-miscible
layers consisted the naphthalene and benzoic acid solution while the lower layer is the
dichloromethane. The occurrence of non-miscible layers was due to the difference of densities
of the solvents. According to literature, water which acted as the solvent has a density of 1.00
g/cm3 while dichloromethane has a density of 1.13 g/cm3. Dichloromethane which has a higher
density will sink to the bottom of the separation funnel whereas the lighter part which is the
naphthalene and benzoic acid solution will stay at the top part of funnel. Besides that,
dichloromethane that occupied the bottom layer was in organic phase as it is an organic solvent
whereas the naphthalene and benzoin acid solution that was in the top of the funnel was in
aqueous phase as the solvent involved is water.
After the settlement of both layers, the bottom layer which was the organic phase of the
solute was drained out and transferred into a beaker. Then, repeated the same step, the solution
of mixture was drained out and transferred into a small beaker and cool it about 15°C. While
stirring, ice-cold concentrated HCI were added drop by drop until a pH of 1 is obtained. Next,
the solid obtained was vacuum into a Buchner funnel, and the precipitate was rinsed from the
beaker into the funnel. Last step in this experiment, the product was dried , weighed, and
calculated the percent recovery based on the amount of the initial mixture.
Dichloromethane was used as the solvent during this experiment as it has lower toxicity
and ability to selectively dissolve the mixture of naphthalene and benzoic acid. It also has a
higher density than the water which allows the extraction to become easier. Naphthalene and
benzoic acid also shows higher solubility in dichloromethane than in the water as it is a non-
polar organic compound and dichloromethane is a polar inorganic solvent. This property allows
the mixture to greatly dissolves in dichloromethane as it is able to bond itself through non-
polar Van Der Waals interactions.
 For experiment 5, the experiment was conducted to extract the caffeine from tea leaf.
This experiment we can observe the caffeine obtained from tea leaf using the same technique
as experiment 4, which is separation technique.
The lone pair of electrons that surround the nitrogen are what give caffeine its
fundamental properties. The presence of nitrogen regulates solubility. Because the caffeine in
tea is soluble in water, this is why it was boiled. Additionally, since organic chemicals are
immiscible in water, it is possible to distinguish inorganic from organic compound when water
is present.
The experiment was started by weighing 10g of tea leaf and 3g of calcium carbonate
and put into a 200mL beaker then 100mL of boiling water was added. The aqueous tea solution
was cooled to 25°6 after the heat bath because otherwise the dichloromethane would have
evaporated and the caffeine extraction would not have been
successful. Emulsions or solid, insoluble substances like cellulose were separated from the ca
ffeine and tannins during the extraction process.
After that, the filtrate was poured into a 200 mL separatory funnel and extracted gently
by shaking and swirling for at least 5 minutes. For first time extraction, 30mL of
dichloromethane were added and followed by 25mL for second time and both extracts were
combined in a 200 mL Erlenmeyer flask.
The aqueous layer needed a difference in solubility to separate the tannins into the
aqueous layer , which is why salt calcium carbonates was added to aid in breaking up any
emulsion. Tannins are acidic while sodium carbonates is basic. As a result, the tannin functions
as an electrophile and sodium carbonate as a nucleophile. The emulsions are eliminated when
the nucleophile attacks the electrophile in acid or base reaction.
Dichloromethane caused the formation of two layers: an organic layer and an aqueous
layer. Dichloromethane is found near the button of the separatory funnel because it is denser
than water. The density of its 1.325 g/ m. The organic layer is below the aqueous layer due to
the solute content in that layer. As a result, there is a high concentration of caffeine and little
water.
To get a pure sample of caffeine, the organic layer's water was removed using a drying
agent which is anhydrous sodium sulphate. When extra water was taken out, the sodium
sulphate ceased to clump. The solution was put in a steam bath to extract the dichloromethane,
which evaporated and left a green residue.
As the last step, the mass of caffeine obtained was 0.12g and the approximate
percentage of caffeine calculated was 24%.
Several errors might occur during the conduction of experiment. Over shaking of the
separation funnel will not leads to a proper interaction of both solvents but it might cause the
occurrence of emulsion in the mid-zone between two phases. Emulsion is known as an
intermediate solubility between two phases and its occurrence will cause the difficulty in
extracting one phase and leads to quantitative problems. Besides that, contamination of
extracted phase which is organic phase might also occur if the solution in separation funnel is
overly drained. This can be prevented by leaving a small portion of the phase required in the
funnel and more dichloromethane can be added for the next extraction. Precautions should also
be taken to ensure the safety and accuracy of the data collected. Both ends of the separation
funnel should be tightly closed in order to prevent spills.
CONCLUSION
In conclusion, liquid - liquid extraction can be used to acquire the caffeine found in tea leaves
and also in naphthalene and benzoic acid. The ability to extract caffeine from a binary solution
is due to caffeine's higher solubility in organic solvents like dichloromethane than it does in
water. Naphthalene and benzoic acid also shows higher solubility in dichloromethane than in
the water as it is a non-polar organic compound and dichloromethane is a polar inorganic
solvent. This property allows the mixture to greatly dissolves in dichloromethane as it is able
to bond itself through non-polar Van Der Waals interactions. The amount of caffeine in tea
leaves was significantly influenced by the way its made. In this extraction, the sample solution
displayed a separation layer, with the sample solution at the top and dichloromethane at the
bottom that contained caffeine dissolved in it. Immiscible solvents are used because they allow
the needed solute to dissolve in them while preventing water, the liquid-solid extraction solvent
from mixing them. Liquid-liquid extraction was used in a variety of processes and industries,
including fermentation, perfumery and others. It may be said that this technique is the most
popular first separation method for extracting closely related functional groups from specific
chemicals and organic molecules. However, the liquid- liquid extraction procedure will also
have some drawbacks. one of the drawbacks is the need for significant quantities of organic
solvents , whose recycling costs are rising. Additionally, the emergence of emulsion issues
makes it difficult for the extract to fully recover.
Questions
Experiment 4
1.Why dichloromethane were used as a solvent?
- Dichloromethane dissolves many organic solvents whose polarity is least or are nonpolar
forms. So, considering its volatility as well as hydrophobic natures dichloromethane is used as
a solvent.
2.Why NaOH and HCl were added and please write the equations?
- NaOH is base and HCI is an acid which when they mix together, it will be neutralisation
reaction.
NaOH + HCI → NaCl + H2O

3.How to test the organic and aqueous phase in the mixture?

- to determine which layer is which, one can simply add distilled water to the funnel. Whichever
layer increases in size must be the aqueous layer and the other is the organic layer.

4.Describe one industrial application of liquid extraction?

- Examples of extraction in food industry is in edible oil industry in which oil is extracted
from natural products such as peanut, soya beans and so on.
Experiment 5
1.Why is the ethyl acetate layer above the water layer?
- Ethyl acetate are less dense than water.
2. Do you think your final mass of caffeine represents all of the caffeine originally in your
tea leaves? List two places where you could have “lost” some of your final caffeine (not
Including human error).
- No, the results not 100% originally.
- 1) The material misfortune through move during the system.
-2) Theoretical measure of caffeine in tea sacks
3. Draw the structure of caffeine and state what is the other compound is present in tea
leaf.
 REFERENCES

▪ F. Melanie, (2019), Extraction of caffeine from tea, https://www.studocu.com/en-

us/document/new-mexico-state-university/organic-chemistry-ii/lab-report-extraction-
of-caffeine-from-tea/7345981

▪ ByJus, (2023), Caffeine chemical formula, https://byjus.com/caffeine-chemical-

formula/

▪ S. Alexander (Jun 21,2021), Liquid liquid extraction,

https://chem.libretexts.org/Bookshelves/Organic_Chemistry/Book%3A_How_to_be_
a_Successful_Organic_Chemist_(Sandtorv)/02%3A_COMMON_ORGANIC_CHEM
ISTRY_LABORATORY_TECHNIQUES/2.03%3A_LIQUID-
LIQUID_EXTRACTION

▪ W, Hayley, (Feb 27,2018) , Extraction,

https://www.academia.edu/37597453/Lab_report_5_Extraction

▪ Studylib, (2013), Extraction of benzoic acid from naphthalene,

https://studylib.net/doc/7467480/extraction-of-benzoic-acid-from-naphthalene

▪ J.D.Thornton. Extraxtion, Liquid-liquid. , https://www.thermopedia.com/content/752/

▪ Hannah. (30 March 2021). The process of liquid-liquid extraction.,

https://ecolink.com/info/the-process-of-liquid-liquid-extraction/
APPENDICES