Palawan State University

College of Sciences

Bachelor of Science in Biology Major in Medical Biology

ORGANIC
MOLECULES
Laboratory Report
(Separation and Purification of Organic Compounds)

Buera, Estela L.
Gimongala, Janica B.
Maylanon, Marites B
Valones, Queen Crissel B.

Separation and Purification of Organic Compounds

 I -Abstract

The separation and purification of organic compounds from impurities are important laboratory
procedure before determining their purity and qualitative analysis for elements present in organic
compounds. Organic compound prepared in the laboratory are usually contaminated with other organic
substances or impurities due to side organic reactions that yield substance other than the desired
product. In addition, the organic compounds have the tendency to undergo partial decomposition or
rearrangement upon standing or when exposed to heat and light. The different methods of purification
and separation are solution and filtration, crystallization, decolorization, sublimation, extraction
distillation, chromatography, dialysis and electrophoresis.

To conduct this experiment, we started from “Solution and Filtration”. You have to mix
thoroughly 0.5 gram of naphthalene and 0.5 grams of brown sugar in an evaporating dish and divide
into two equal portions. Then, transfer one portion into a test tube and add 3-5 mL water. Shake well
and filter. Lastly, place the other portion in another test tube and add 2 ml ethyl alcohol. Shake well
and filter. Second method is “Crystallization”. You must mix in a beater 2 grams of impure urea and
10 mL water in a small beaker and boil the mixture gently for 5 minutes replacing occasionally the
water lost by evaporation. Next is filer the solution while hot into a test tube immersed in ice water.
Observe the Formation of crystals. Compare the crystals with the original sample. Lastly, filter off the
purified crystals and dry. Third one is “Decolorization”, where you have to dissolve 1 gram of brown
sugar into 10 mL water in a small beaker then, divide the solution to two equal parts (into two test
tubes). To one test tube, add ½ spatula of bone black or activated charcoal and boil for 5min (replace
the water lost by evaporation). Lastly, allow the hot mixture to cool and filter. Observe the color of the
filtrate. Compare it with that of the original sugar solution. Second to the last in the list is
“Sublimation”. Placing 0.5 gram of benzoic acid and 0.5 grams of sodium sulfate in a dry evaporating
dish is its first step. Next is covering the dish with a filter paper and tuck the edges tightly against the
dish. You must not also forget to place pinholes in the center of the filter paper and cover this holed
area with a funnel. Plug the top of the stem of the funnel with some cotton, heat the dish slowly with a
low flame. Collect the sublimate that deposits on the walls of the funnel. Last step is dissolving a small
amount of sublimate in barium chloride solution. Last method to complete this experiment is
“Distillation (Modified)”. First, you have to put 50 mL of 70% rubbing alcohol in a 250 mL Florence
flask. With a one-holed cork insert a bent glass tubing into the cork. Fit this tightly into a Florence
flask. Insert the end of a glass tubing into another cork. Fit the cork into a 250 ml Erlenmeyer flask as
receiver. For the last step, heat the mixture slowly until distillates are collected into the receiver. Test
the fluid inside the receiver by igniting the stirring rod which has been dipped into the distillate.

We concluded that after conducting these experiments, that organic compound is composed
of hundreds to thousands of individual molecules -this compound is covalently linked to atoms of
other elements most commonly the hydrogen, oxygen or nitrogen. In this experiment the separation
and purification of organic compound using the four method such as evaporation, purification,
crystallization and filtration. These four methods possess different ability to determine the properties
of compound as will recrystallization in which this impure compound was recrystallized and its
identity was determined by melting point, recrystallization determined to be water by observing
solubility in different condition. We also concluded that these separation techniques are useful to
separate constituent particles of a mixture of substances, to purify the compounds and check the purity
of organic compounds.
II – Introduction

The separation and purification of organic compounds from impurities are important laboratory
procedure before determining their purity and qualitative analysis for elements present in organic compounds.
Organic compound prepared in the laboratory are usually contaminated with other organic substances or
impurities due to side organic reactions that yield substance other than the desired product. In addition, the
organic compounds have the tendency to undergo partial decomposition or rearrangement upon standing or
when exposed to heat and light. The different methods of purification and separation are solution and filtration,
crystallization, decolorization, sublimation, extraction distillation, chromatography, dialysis and
electrophoresis.

III– Materials

Naphthalene

Brown sugar platform balance

Ethyl alcohol mortar & pestle
Urea 2- test tubes
Sodium sulfate, Na S02 3- macro test tubes
Barium Chloride, Baca test tube rack
Sodium chloride, NaCl evaporating dish
filter paper ml pipet
Activated charcoal 3- 50 mL beaker
short stem funnel hot plate
spatula stirring rod
test tube brush test tube holder
2- 250 ml beaker aspirator
Masking tape cotton
1-250 mL Erlenmeyer flask graduated cylinder
1-250 ml Florence flask glass tubing

2 corks
IV- Procedures

1. Solution and Filtration

 Mix thoroughly 0.5 gram of naphthalene and 0.5 grams of brown sugar in an evaporating dish. Divide
into two equal portions.
 Transfer one portion into a test tube and add 3-5 mL water. Shake well and filter.
 Place the other portion in another test tube and add 2 ml ethyl alcohol. Shake well and filter.

2. Crystallization

 Mix in a beater 2 grams of impure urea and 10 mL water in a small beaker.

 Boil the mixture gently for 5 minutes replacing occasionally the water lost by evaporation
 Filer the solution while hot into a test tube immersed in ice water. Observe the Formation of crystals.
Compare the crystals with the original sample.
 Filter off the purified crystals and dry. Hand in to your professor.

3. Decolorization

 Dissolve 1 gram of brown sugar into 10 mL water in a small beaker.

 Divide the solution to two equal parts (into two test tubes)
 To one test tube, add ½ spatula of bone black or activated charcoal and boil for 5min (replace the water
lost by evaporation)
 Allow the hot mixture to cool and filter. Observe the color of the filtrate. Compare it with that of the
original sugar solution.

4. Sublimation

 Place 0.5 gram of benzoic acid and 0.5 grams of sodium sulfate in a dry evaporating dish
 Cover the dish with a filter paper and tuck the edges tightly against the dish
 Place pinholes in the center of the filter paper and cover this holed area with a funnel
 Plug the top of the stem of the funnel with some cotton, Heat the dish slowly with a low flame.
 Collect the sublimate that deposits on the walls of the funnel.
 Dissolve a small amount of sublimate in barium chloride solution.

5. Distillation (Modified)

 Put 50 mL of 70% rubbing alcohol in a 250 mL Florence flask. With a one-holed cork insert a bent
glass tubing into the cork. Fit this tightly into a Florence flask.
 Insert the end of a glass tubing into another cork. Fit the cork into a 250 ml Erlenmeyer flask as
receiver.
 Heat the mixture slowly until distillates are collected into the receiver. Test the fluid inside the receiver
by igniting the stirring rod which has been dipped into the distillate.
V-Data/Results

1. Solution and Filtration

We mix 0.5 gram of naphthalene and 0.5

We transfer one portion into a test tube and
grams of brown sugar thoroughly in an
add 3-5 mL water. We then shake it well and
evaporating dish and divide it into two equal
filter.
portions.

Lastly, we place the other portion in another test tube and add 2 ml
ethyl alcohol, shake it well and filter.

*as a result of this experiment,

brown sugar has been dissolved and is left on the filter, naphthalene on the other hand dissolved in alcohol.
 2. Crystallization

First, we mix in a beater 2 grams of We boil the mixture gently for 5

impure urea and 10 mL water in a small minutes replacing occasionally the
beaker. water lost by evaporation.

Lastly, we filer the solution while hot

into a test tube immersed in ice water.
We also observe the formation of
crystals.

*the result of this experiment may vary depending on the temperature of the ice-cold water.
 3. Decolorization

First, we dissolve 1 gram of brown Then, we divide the solution to two

sugar into 10 mL water in a small equal parts (into two test tubes)
beaker.

We then added ½ spatula of bone black or We allow the hot mixture to cool and filter.
activated charcoal and boil for 5min in beaker We also observe the color of the filtrate and
(while replacing the water lost by compare it with that of the original sugar
evaporation). solution.

*using bone black or activated charcoal, functions to remove impurities from the solution. Making
the solution clearer than the other as a result.
 4. Sublimation

First, we place 0.5 gram of benzoic We cover the dish with a filter paper and tuck the edges
acid and 0.5 grams of sodium sulfate in tightly against the dish and place pinholes in the center of
a dry evaporating dish the filter paper and cover this holed area with a funnel.

*to be honest, we failed to get the result for this experiment, since we do not have cotton to use as
cover to the funnel. It also takes more time to make this method successful.

5. Distillation (Modified)

First, we put 50 mL of Then, we insert the end of

70% rubbing alcohol in a a glass tubing into another
cork. Fit the cork into a Lastly, we heat the mixture slowly
250 mL Florence flask.
250 ml Erlenmeyer flask as until distillates are collected into
With a one-holed cork
receiver. the receiver.
insert a bent glass tubing
into the cork. Fit this
tightly into a Florence
flask.
*as a result, we have collected the liquid gases vapor or the fluid that were received from boiling
alcohol
VI- Analysis

 1 Mixture naphthalene. brown sugar and water/ alcohol

o Which substance dissolved in water, sugar naphthalene? Brown Sugar
o Why? Because sugar and water are both polar that have enough forces to dissolved.
o Which substance dissolved in alcohol, sugar or naphthalene? Naphthalene
o What substance is left on filter paper? Sugar
o Why? Because alcohol is not pure polar that’s why alcohol have no enough force to
dissolve sugar.
o Define: solution is a homogeneous mixture of two or more substances.
Filtration is a technique used either to remove solid impurities from an organic
solution.
Filtrate is the product of filtration; a gas or liquid that has been passed through
a filter.

 2. Mixture: Benzoic acid and water

o 2.1 In procedure 2.b, what is the purpose of boiling the mixture? To let the water evaporate
o 2.2 In procedure 2.0, why should a solution be filtered while hot? Because when we let it cool,
the solute is no longer soluble in the solvent.
o 23 Compare the crystal formed in the filtrate with that of the original sample. To be honest, we
did not observe much crystals formed, the temperature of the ice-cold water, where we
immerse the test tube may be the reason.
o 24 Define crystallization. It is the (natural or artificial) process by which a solid form,
where the atoms or molecules are highly organized into a structure known as a
crystal. Some of the ways by which crystals form are precipitating from a solution,
freezing, or more rarely deposition directly from a gas.
 3. Mixture of brown sugar and water
o 31 What is the function bone black or activated charcoal? It is used in removing colored
impurities from liquids, especially Solutions of raw Sugar.
o 3.2 Compare the color of the filtrate with that of the original solution. The original solution
has a light brown while the filtrate is a colorless solution.
o 33 Define decolorization. The activated charcoal has the capacity of de-colorization; is the
change of color and visual appearance of a materials.
 4. Mixture of benzoic acid and sodium sulfate
o 4.1 What is the purpose of plugging the tic of the stem of a funnel with cotton? The purpose
was to trap the gas, and not let the vapor gas escape from top of the funnel.
o 42 Define sublimate. Is a process of forming a new product.
o 43 Describe the sublimate. The Solid that form by the vapor of Benzoic acid.
o 4.4 What is the composition of sublimate, benzoic acid or sodium sulfate? The composition is
like a colorless crystalline structure.
o 45 Define sublimation. It is a process in which the solid turns into a gas without turning
into liquid.
 5. Mixture. 70% rubbing alcohol
 o 5.1 What is the fluid called inside the receiver? Alcohol
o 52 How do you know? Because the smell of fluid is like alcohol, but have a strong odor.
o 53 Define distillate. The result of distillation process like the liquid gases vapor or the fluid
that receive from boiling alcohol.
o 5.4 Define distillation. Is a process of separating mixture by removing Soluble minerals
from the chemical.

VII- Conclusion

We concluded that after conducting these experiments, that organic compound is composed
of hundreds to thousands of individual molecules -this compound is covalently linked to atoms of
other elements most commonly the hydrogen, oxygen or nitrogen. In this experiment the separation
and purification of organic compound using the four method such as evaporation, purification,
crystallization and filtration. These four methods possess different ability to determine the properties
of compound as will recrystallization in which this impure compound was recrystallized and its
identity was determined by melting point, recrystallization determined to be water by observing
solubility in different condition. We also concluded that these separation techniques are useful to
separate constituent particles of a mixture of substances, to purify the compounds and check the purity
of organic compounds.