Bicol University

COLLEGE OF SCIENCE
Legazpi City

SEPARATION TECHNIQUES
Activity No. 2

Bs Chemistry 1A
Group 5 – Lab 2

Elouisa Olaybal
Hyna Lyza Ocampo
Mizhelle Kiana Moises
Precious Jiannah Faye Moreno
Mary Joy Nate

October 13, 2020

INTRODUCTION
Mixtures are combination of different substances. It could be homogenous and
heterogenous mixtures. Separating their components are important in the field of science in order
for it to be studied and understood. The purpose of this activity is to demonstrate the various
techniques of separating mixtures which are important techniques in the laboratory.
MATERIALS USED
In Activity A, the apparatus and equipment used in the experiment were watch glasses,
evaporating dish, glass stirrer, a cotton ball, glass funnel, a Bunsen burner, spatula, beakers, filter
paper, wired gauze, a tripod, iron clamp along with the iron stand, and a wash bottle. Chemicals
and components used in the experiment were distilled water, Ammonium chloride, sand, and salt.
In Activity B, the apparatus and equipment used in the experiment: separatory funnel,
funnel, iron ring stand, iron ring, nozzle, stopper, Erlenmeyer flasks. Chemicals used in the
experiment: iodine, water, cyclohexane.
In Activity C, Apparatus and equipment used in the experiment: 250 mL and 100mL round
bottom flasks, heating mantle, vacuum adapter, distilling head, beaker, thermometer adaptor,
thermometer, water condenser, iron stand, and iron stand. Chemicals and other materials used
in the experiment: mixture of acetone and water, water and porcelain chips.

In Activity D, Apparatus and equipment used in the experiment: Chromatography paper,

beaker and a thin metal stick. Chemicals used in the experiment: Wet erase marker and Water
(H2 O).

In Activity E, there are two experiments; the chemical reaction involved between lead
nitrate and potassium chromate and the centrifugation separation method. In the first experiment,
the apparatus and equipment used are: test tube, test tube rack and dispensing bottles. The
chemicals used are lead nitrate and potassium chromate. In the second experiment, the
apparatus and equipment used are: beaker, centrifuge, and centrifuge tube. The chemicals used
in the experiment: muddy water mixture and calcium carbonate in water mixture.

EXPERIMENTAL PROCEDURE
In the experiment of Activity A: Separating components of mixture with different
solubilities, the Ammonium chloride, sand, and salt were set on separate watch glasses and was
mixed together in an evaporating dish with the use of the glass stirrer. With the cotton ball, the
base of the glass funnel was clogged, then was used to cover the evaporating dish. As the Bunsen
burner, tripod, and wire gauze was set up, the evaporating dish was placed on top to be heated
until white fumes were observed. The evaporating dish was then removed from the heat and was
set to cool down. Once cooled, the ammonium chloride that was sticking on the walls of the funnel
were removed with a spatula, onto a watch glass. The remaining mixture of sand and salt on the
evaporating dish was then transferred to a beaker and water was added. The contents were then
stirred with the use of the glass stirrer. Filter paper was placed in the mouth of the funnel and was
attached to the iron clamp on the iron stand, with a beaker placed underneath the funnel. The
contents of the beaker were slowly poured into the funnel, filtering the solution. After filtration, the
sand on the filter paper was transferred on a separate watch glass using a spatula. The solution
of water and salt from the beaker was then transferred into an evaporating dish and was placed
on a Bunsen burner until the water completely evaporated and salt was left as residue in the
evaporating dish. The salt was then transferred into a watch glass using a spatula. The three
components were then carefully labeled in each of their corresponding watch glasses. The
documentation for each procedure is contained in Appendix A.
In performing Activity B: Liquid-liquid extraction, the first thing to do is to add some water
to the separatory funnel to make sure that the stop cock does not leak as well as the stopper.
Second, labeled each Erlenmeyer flask with aqueous phase and organic phase. Third, add iodine
in water using funnel to the separatory funnel for about 2/3 of the separatory funnel. Fourth, add
the cyclohexane (extracting solvent) to the separatory funnel. Fifth, seal the separatory funnel
then carefully lift from the iron ring stand. Sixth, invert the separatory funnel pointing away then
gently tip and open the stop cock. Seventh, close the stop cock and shake the separatory funnel
then again, open the stop cock and shake it until the layers mixed. Eight, invert the separatory
funnel, swirl it and gently place it back to the iron ring stand. Ninth, slowly open the stop cock to
let the aqueous phase out through the bottom into the Erlenmeyer flask (labeled aqueous). Tenth,
slow down as the meniscus of the top layer reaches the neck of the separatory funnel. Eleventh,
pour the extraction into the labeled Erlenmeyer flask (organic). Twelfth, return the aqueous phase
into the separatory funnel. Thirteenth, use a second volume of cyclohexane for another extraction
and add it to the separatory funnel. Fourteenth, tip the separatory funnel again, open the stop
cock, vent, and shake it again until the layers mixed. Fifteenth, let it sit for about 2-3 minutes until
both phases have gone to their appropriate positions. Sixteenth, drain again the aqueous phase
and slow down when the meniscus gets to the top of the stop cock. Seventeenth, transfer and
combine the extracted solute with the other organic phase. Eighteenth, return the aqueous phase
again in the separatory funnel and do one more extraction with the cyclohexane.
In Activity C: Separation of Components of Mixture with Different Volatilities, take a mixture
of acetone and water, pour it into a 250 ml round bottom flask. Then, clamp the round bottom
flask to a stand and placed it in a heating mantle. Take some porcelain chips, and put them into
the round bottom flask. Connect the distilling head to the round bottom flask. Then, connect the
thermometer adaptor into the distilling head and place the thermometer in it. Next, connect the
water condenser to the outlet of the distilling head and connect the vacuum adapter to the water
condenser and then connect the 100 ml round bottom flask to the vacuum adapter and place it in
a beaker containing water. Connect the rubber tube between the tap and the inlet of the condenser
to allow cold water to enter, similarly, connect another rubber tube to the outlet of the condenser
to pour the hot water from the condenser to the sink. Open the tap and allow water to flow through
the tube and fill the condenser and then comes out through the outlet. Set the temperature of the
heating mantle and heat the mixture to boil. Observe the thermometer as it reaches 56oC, acetone
vaporizes, then it condenses in the condenser and the acetone is collected to the receiver flask.
The water was left behind the 250 ml round bottom flask.

Plate 1: Simple Distillation Set-up

 Plate 1 shows how apparatus and equipment are connected to each other. Ensuring that
apparatus are properly connected to another will make distillation procedure safe and will gain
accurate result.
The experiment of Activity D: Separation of Minute Components of a Mixture by Paper
Chromatography, started at preparing the materials needed which are a wet erase marker whose
pigments are soluble to water, chromatography paper and water. Then, a line was drawn at the
lowermost part of the chromatography paper which determined the starting spot of the mixture.
The chromatography paper was attached in a thin metal stick and was placed into a beaker with
a small volume of water in it. After a couple of minutes, the water begun rising up to the
chromatography paper and later separated the pigments of the marker. The water continued to
rise up until all of the pigments were completely separated. Finally, with the used of ruler the
maximum distance travelled by the water was determined. The documentation for paper
chromatography is contained in Appendix A.

In the first experiment of Activity E: Effective Separation of Precipitate from its Mother
Liquor, prepare first the apparatus and chemicals needed. Dispense the lead nitrate into the test
tube. After that, dispense small amount of potassium chromate into the same test tube. Observe
the reaction between the two chemicals. In the second experiment, the Centrifugation Separation
Method, prepare the chemicals and apparatus needed. Take the sample of muddy water mixture
from the beaker into the centrifuge tube and place it on the centrifuge. Same process with the
sample of calcium carbonate in water mixture, take the sample of the mixture from the beaker into
the centrifuge tube and place it on the centrifuge. Thereafter, rotate the centrifuge for a few
minutes. Stop the rotation. Remove the centrifuge tube and observe.

RESULTS AND OBSERVATION

Figure 1: Separation of components of a solution with different solubility based on the experiment.
 As shown in figure 1, the schematic diagram shows the different processes of separating
techniques performed in the experiment, and their end products.
In consideration of the properties of the components present, the separating techniques
most suitable to be done in the experiment were through the process of sublimation, evaporation
and filtration.

Cyclohexane Iodine
Organic Phase
(extracting solvent)

Iodine in water

Separatory Mixing of Extraction

Funnel Phases Process Separatory
Funnel

Aqueous Phase

Figure 2: Schematic Diagram of Liquid-Liquid Extraction

Figure 2: Schematic Diagram of Liquid-Liquid Extraction illustrates the process of
extraction performed in the experiment. The cyclohexane was added in the separatory funnel with
iodine in water. The layers were mixed and then the solute was extracted from the aqueous phase.
The result showed that the iodine was extracted from the aqueous phase into the organic phase
using cyclohexane as organic/extracting solvent.

Mixture of acetone
and water

Heating of
mixture

at 56oC, (Boiling point water

of Acetone)
vapors

condenser

Condensation
Figure 3: Schematic diagram of the separation of
components of a mixture with different volatilities

acetone
 Figure 3 shows how the process of simple distillation take place in the experiment. The
mixture is composed of two components, namely acetone and water. The mixture of acetone and
water was put in a heating mantle. Upon heating the mixture and reached 56oC, vapors are
observed and it passes through the condenser. The vapor then condenses and acetone was
collected from the receiving flask while water was left behind the round bottom flask that was in
the heating mantle.

The result of the Activity D performed was shown below. The results were obtained
through the procedures indicated on the experimental procedure.

Plate 2: Separated of pigments from the marker

The Plate 2: Separated of pigments from the marker, showed the result of the experiment.
After performing the separation of the pigments of the marker using paper chromatography, it is
clearly shown in the figure above that the originally black ink of the marker had been separated
into different pigments which have different colors.

Observation
Solution of lead (II) nitrate Colorless solution
Solution of potassium chromate Yellow solution
Reaction between the two solution A yellow insoluble solid is formed that
precipitates to the bottom of the test tube.
Table 1: Qualitative observation of the first experiment
Table 1 is the qualitative observation of the reaction between lead (II) nitrate and
potassium chromate wherein it described the physical appearance of the lead (II) nitrate solution
and potassium chromate solution as well as the reaction between the two.
 Mixtures Centrifuge

Muddy water mixture

centrifugation
Calcium carbonate in Centrifugal force is
applied
water mixture

Precipitates Centrifugate

Mud particles Water from

both mixtures
Calcium carbonate
particles

Figure 5: Schematic Diagram of Centrifugation

Figure 5 is the schematic diagram for the second experiment to demonstrate centrifugation
to increase the rate of sedimentation of light and small particles from muddy water mixture and
calcium carbonate mixture. The experiment results into the separation of precipitates which are
the mud particles and calcium carbonate particles as well as water from both mixtures called the
centrifugates.
DISCUSSION
Activity A: In the experiment, the solution was composed of components of different
solubilities, hence separating them would require to take into consideration their physical and
chemical characteristics in order to perform the appropriate separating technique. It was important
to know the characteristics and behavior of the components before undergoing the process of
separation in order for it to work. When heat is added to Ammonium chloride, it decomposes and
forms ammonia gas (NH3) and hydrogen chloride gas (H2). Once it cools down, they recombine
to form back to ammonium chloride. This gives the impression that ammonium chloride undergoes
sublimation which is when solid substances are converted into gas and vice versa. Through this
process, the ammonium chloride evaporates from the mixture as white fumes rising from the
evaporating dish. The cotton-stoppered inverted glass funnel slows down this process to let the
gas cool down, allowing the crystals of ammonium chloride deposit along the walls of the glass
funnel. Since sand can be separated from a mixture through filtration while salt can be separated
through evaporation, by adding water to the solution, these separating techniques can be
executed. In the process of separating the sand, the solution was poured into a filter paper placed
inside the glass funnel. This process filters the sand hence separating it from the remaining
solution of salt + water. As mentioned before, salt can be separated from water through
evaporation. The salt + water solution was therefore put to a boil until the water completely
evaporates leaving only the salt as residue inside the evaporating dish.
Activity B: In the experiment, the iodine in water was added into the separatory funnel as
well as the cyclohexane, which served as the extracting agent. When cyclohexane, a non-polar
solvent, was added into the separatory funnel a dynamic equilibrium was established, wherein
the solute distributes itself between the two phases. The iodine was distributed in the aqueous
phase as well as in the organic phase. Iodine is a non-polar molecule which is likely to be
dissolved in a non-polar solvent, the reason why iodine molecules moved into the organic phase.
As a result, the iodine (solute) was extracted from the aqueous phase into the organic phase.
The goal of the liquid-liquid extraction is to isolate compound from a mixture and move
things between two phases. These two liquid phases are usually an organic phase and aqueous
phase. The experiment used the chemicals iodine, water and cyclohexane to illustrate the process
of the liquid-liquid extraction. Iodine is not very soluble in water since it is a non-polar molecule
and water is a polar molecule. The hydrogen bond of water is stronger than the intermolecular
bond of iodine and polar bond of water. Thus, instead of allowing the iodine molecules to come
between the water molecules, the latter would remain hydrogen bonded to each other. The
cyclohexane and water have different densities also considered as two immiscible liquids due to
their different polarities. Cyclohexane has a density of 0.774 g/mL and it is a non-polar molecule.
The carbon and hydrogen in cyclohexane is in a neutral relationship and remained neutral after
performing covalent bond. On the other hand, water has a density of 1g/mL and it is a polar
molecule. The water molecule has a partially negative oxygen and partially positive hydrogen
making it a polar molecule. The density allowed the liquids to sit on their appropriate positions.
The cyclohexane is less dense than water and it would be the top phase of the extraction. If the
organic phase is denser than water, it would be at the bottom phase. Iodine crystals completely
dissolved in cyclohexane because non-polar molecule like iodine dissolves in a non-polar solvent
like cyclohexane. The solubility of iodine in cyclohexane produced a purple solution while in water
it produced a yellow-brown solution since iodine is only slightly soluble in water.
In the experiment, extraction is defined as the transfer of solute from one phase to another.
The safe amount of aqueous phase and organic phase when added to a separatory funnel is 2/3
and 1/3 respectively. The Erlenmeyer flasks in the experiment were labeled with aqueous phase
and organic phase. It is important to labeled the containers to make sure that nothing valuable
gets thrown away and to avoid confusions inside the laboratory. The tipping and opening of stop
cock of the separatory funnel is important in liquid-liquid extraction to release any pressure that
might develop with the dissolution of the two phases. The multiple extraction of the liquid in the
experiment is to make sure that all the iodine solute had been removed from the aqueous phase
into the organic phase.
Activity C: Distillation is defined in the activity as a method of separating components of a
mixture containing two miscible liquids that boil without decomposition and have sufficient
difference in their boiling point. On the other hand, distillation can also be defined as a process
used in separating components of a mixture with different volatilities by heating it to boil and
vaporize the most volatile component in the mixture. Volatile is a term that refers to a substance
that vaporizes readily whereas volatility is a measure of how quickly or readily a substance
vaporizes or transfers from a liquid phase to a gas phase.
The two substances that made up the mixture are water and acetone. The main difference
between them is their volatilities. As acetone is more volatile than water, their molecules begin to
move more quickly when heat energy was applied. Water and acetone have a large difference
between their volatility and boiling point. Acetone has a boiling point and is vaporized at 56oC
while water at 100oC. The water was left in the round-bottom flask as water has a high boiling
point and is less volatile than acetone. Since acetone has a low boiling point and is most volatile
between water, it will evaporate easily and more quickly than water.
The distillation process involves physical state changes to the mixture. The type of mixture
used in the activity can be classified as a solution, specifically a liquid solution. The mixture was
heated until it boils, making the liquid particles turn into a vapor (gas phase) because heat energy
is transferred to the molecules. Upon reaching the condenser, the vapor is cooled down and the
condensation occurs making the vapor into liquid again.
liquid boiling gas condensation liquid

The simple distillation can separate acetone and water, however, acetone and water forms
azeotrope or a mixture of two or more liquids in which the proportions of it cannot be altered or
modified by simple distillation, thus, to obtain the components into their pure state, the process
must be repeated. Moreover, in the experiment, porcelain chips were utilized and are put inside
the round bottom flask for it will prevent the distillation of the liquid from overheating and trigger a
more regulated boiling process, thus eliminating the risk that the liquid in the round-bottom flask
will bump into the condenser and allowing smooth boiling process.
Activity D: Using paper chromatography the pigments of the wet erase marker were
separated by water. The water rises up to the chromatography paper because of capillarity action.
Capillarity action refers to the ability of a liquid to flow in narrow spaces due to adhesive and
cohesive forces interacting between liquid and surface without the assistance of or even in
opposition to, external forces like gravity. The cohesive and adhesive properties draw the water
into the chromatography paper which was used to separate the pigments of the marker.

Different solvents dissolve different substances. A polar solvent like water dissolved polar
substances like the water-soluble ink. The different properties of the pigments contributed to the
separation of it. Some of it are heavier and some are lighter and some of it are more soluble in
water and some are less. Because of these properties, the pigments with its distinctive colors
travelled at different speeds which result to its separation from one another.

The figure below shows the state of the ink of the marker (M**) before and after the
process of separation using paper chromatography. As the water begun to rise up the
chromatography paper, the pigments of the ink begun separating until it become fully separated
as shown at the end of the experiment. The figure also shows that as the water continued to travel
its distance, the distance of the pigment individually was decreasing. The first pigment to be
separated had the farthest distance meaning that the pigment was more soluble in water and was
lighter therefore, it can be easily carried by the water compared to the other pigments.

Figure 6: Schematic Diagram of

Paper Chromatography
 After the experiment, the originally black ink from the starting spot of the mixture was
completely separated into different pigments as shown in Figure 1. With the use of a ruler, the
distance travelled by the different pigments will be identified and also the maximum distance
travelled by the water which was approximately 7.2 cm.

Activity E: The reaction between lead (II) nitrate and potassium chromate is called
precipitation. Precipitation or double replacement reaction occurs when combining two solutions
and switching of two reactants occur, creating an insoluble solid called precipitates. For the first
experiment, Pb(NO3)2 (aq) + K2CrO4 (aq) → PbCrO4 (s) + K2 NO3 (aq), lead (II) chromate
precipitate was formed in the reaction. Precipitation is used to obtain the lead (II) chromate from
the reaction between the solution as it precipitates to the bottom of it. Therefore, separating the
lead (II) chromate from the solution. Precipitation creates a new compound from the reactants
and slowly settle at the bottom compared to other separation techniques.

The centrifugation separation technique is a technique used for separating small and
lightweight particles from a solution with different densities of the component. It speeds up the
rate of sedimentation using the centrifugal force. The muddy water mixture and calcium carbonate
mixture were samples used to demonstrate how centrifugation separates precipitate from its
mother liquor. Mud particles and calcium carbonate particles were deposited at the bottom of the
centrifuge tube, these insoluble particles are called precipitates while the water that was
separated from both mixtures is called the centrifugate.
Decantation and Filtration are effective separation techniques for separating the liquid
from the precipitate. Decantation is the process of separating solid-liquid components of mixtures
by pouring out the liquid out of the container and let the solid particles settle at the bottom.
Filtration is a separation technique that used a porous filter that only the liquid component mixtures
can pass leaving the solid particles on the porous filter therefore separating solid and liquid
component of mixture.
CONCLUSION
A solution containing components of different solubilities can be separated through the
combination of the different separation techniques, depending on their physical and chemical
characteristics.
The Liquid-Liquid Extraction can extract the iodine solute from the aqueous phase into the
organic phase. Also, the solubility and the density of liquid have an important role in liquid-liquid
extraction.
The mixture of acetone and water can be separated by simple distillation because the
mixture is made up of substances that have different volatilities. To obtain the components in the
pure state, the process must be repeated. Using porcelain chips in simple distillation helps to
have smoother and controlled boiling process.
Paper chromatography mainly serves as a tool for examination and separation of minute
components of a mixture. Using capillary action and the right solvent to use in dissolving a certain
substance, the separation of the mixture will be dealt. In the case of this experiment, with the used
of water as the solvent to dissolved a water-soluble ink, the different pigments were separated
and recorded a maximum distance travelled by the water of approximately 7.2 cm.
 Precipitation leads to the formation of an insoluble solid particles that slowly deposits at
the bottom of the container. Lead (II) nitrate and potassium chromate demonstrated precipitation
as it had formed the insoluble lead (II) chromate solid particle during reaction which slowly settles
at the bottom of the test tube. Centrifugation is a useful separation technique to speed up the
sedimentation process of these types of mixtures. It uses centrifugate that applies centrifugal
force during the rotation of the apparatus to the sample and results to the solid particles being
settled at the bottom of the container and separated from the centrifugate, the liquid portion
separated in the process.

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Separation Techniques. Activity No. 2. Chem 100.1 – Principles of Chemistry Laboratory.

Semester 1 S/Y 2020-2021.

Separation Techniques. Activity No. 1. Chem 100.1-Principle of Chemistry Laboratory.

Semester 1 S/Y 2020-2021.

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APPENDICES
Appendix A

Plate 3: The compounds on separate watch glasses

Plate 4: Mixing the three compounds in an evaporating dish

Plate 5: Separating the ammonium chloride from the mixture through sublimation

Plate 6: Transferring the crystalized ammonium chloride into a separate watch glass

Plate 7: Placing the remaining mixture in a beaker and adding water to it

Plate 8: Mixing the solution and filtering it through the filter paper

Plate 9: Transferring the sand into a watch glass

Plate 10: Heating the remaining solution

until the water completely evaporates
 Plate 11: Transferring the salt residue from the evaporating dish on to a watch glass

Plate 12: The three compounds placed and labeled separately on different watch glasses

Plate 13: Paper Chromatography