SEMESTER 1 SESSION JUNE 2023

FOUNDATION IN SCIENCE

CENTRE OF FOUNDATION STUDIES

XBO 013

BIOLOGY 1

LAB REPORT 1 :

IDENTIFICATION OF ORGANIC MOLECULES

NAME : SYAFIQA ZULAIKHA BINTI SHAHRULNIZOM

MATRIC NO. : M231012331

GROUP MEMBER’S NAME :

-> ALYAA IRDINA BT MOHD AFANDI (M231012031)

-> FATINI MUSLIHAH BT NOR HAYAZI (M231013831)
-> MINHALINA BINTI MOHAMAD AZMI (M231013631)

EXPERIMENT DATE : 24 JULY 2023

SUBMISSION DATE : 3 AUGUST 2023

INSTRUCTOR’S NAME : MADAM HAZIRAH BT ABDUL RADZAK

INTRODUCTION

Based on our everyday appetite, there are four major types of large biological molecules
that we consume: carbohydrates (such as sugars), lipids (such as fats), proteins, and nucleic
acids (such as DNA and RNA). Large biological molecules perform a wide range of jobs in
an organism such as metabolites, energy carries and messengers. For example, some
carbohydrates store fuel for future energy needs, and some lipids are key structural
components of cell membranes. Nucleic acids store and transfer hereditary information, much
of which provides instructions for making proteins. Proteins themselves have perhaps the
broadest range of functions: some provide structural support, but many are like little
machines that carry out specific jobs in a cell.

These large biological molecules will undergo chemical reactions which can build up and
break down. Most large biological molecules are polymers, long chains made up of repeating
many molecular subunits, called monomers. Carbohydrates, nucleic acids, and proteins are
often found as long polymers in nature. While lipids don’t generally form polymers. Large
biological molecules often assemble via the dehydration process or condensation process, in
which one monomer forms a covalent bond to another monomer, releasing a water molecule
in the process. However polymers are broken down into monomers via hydrolysis reactions,
in which a bond is broken, by addition of a water molecule. This is the reverse of a
dehydration process, and it releases a monomer that can be used in building a new polymer.
Dehydration reactions build molecules up and generally require energy, while hydrolysis
reactions break molecules down and generally release energy. Carbohydrates, proteins, and
nucleic acids are built up and broken down via these types of reactions. While certain types
of lipids are assembled via dehydration process even though the lipids are not polymers.
However, we can determine their presence by using a chemical test.

OBJECTIVES

1. To determine the presence of starch by chemical test.

2. To analyse solutions for the presence of simple reducing sugars.
3. To analyse a sample of vegetable oil for the presence of lipids.
4. To analyse solutions for the presence of protein.
5. To analyse and identify organic molecules found in other substances.
MATERIALS AND APPARATUS

Dropper, test-tube, test-tube rack, test-tube holder, brown paper and pencil, vegetable oil,
glucose solution, soluble starch solution, biuret reagent, benedict’s solution, Bunsen burner,
95% ethanol, 2% gelatin solution, iodine solution, distilled water, sticker label, test samples.

METHODS

3.1 Test for Carbohydrates

3.1.1 Test for Starch

The experiment began by labelled three test tubes as “1”, “2”, and “3” respectively. Then,
the three test tubes were placed in the test tube rack. 10 drops of soluble starch solution was
added to test tube 1, 10 drops of glucose solution was added to test tube 2, and 10 drops of
water to test tube 3, by using separate droppers for each solution. After that, 3 drops of iodine
solution was added to each test tube. The colour of each tube’s contents after addition of the
iodine was recorded. Be careful when handling the iodine solution, if iodine is spilled, rinse
with water and inform the instructor immediately.

3.1.2 Test for Simple Reducing Sugars

For this experiment, the water bath was heated to boiling on the hot plate. Three test tubes
were labelled as “1”, “2”, and “3” respectively. 10 drops of soluble starch solution was added
to test tube 1, 10 drops of glucose was added to test tube 2, and 10 drops of water was added
to test tube 3, by using separate droppers for each solution. The initial colour of each tube’s
contents was recorded. Then, 20 drops of Benedict’s solution was added to each of the three
test tubes and was placed in a boiling water bath for three minutes. The three test tubes were
removed from the water bath by using a test-tube holder and were placed in a test tube rack to
cool. The colour of each tube’s contents was recorded. The contents of the test tubes were
discarded and cleaned. Be careful when handling Benedict's solution, if Benedict’s solution is
spilled, rinse with water and call the instructor immediately. Be careful not to burn yourself
when removing the test tubes from the water bath. Benedict’s solution tests for the presence
of simple reducing sugars (monosaccharides and some disaccharides, but not
polysaccharides). Thus, a colour change might or might not occur when Benedict’s solution is
added to a carbohydrate and heated. A change from blue to green, yellow, orange, or red
occurs if monosaccharides or certain disaccharides are present. The original blue colour will
remain after heating if polysaccharides or certain other disaccharides are present.
3.2 Test for Lipids

3.2.1 Brown Paper Test for Lipids

A drop of water was placed on a small piece of brown paper. Followed by a drop of oil
was placed on the same piece of paper. A circle around the drop was drowned with a pencil.
Then, the paper was allowed to dry for a few minutes. The piece of paper was held up to the
light. The appearance of each spot was recorded. If a semi-transparent (translucent) spot is
evident, the sample contains lipids.

3.2.2 Solubility Test for Lipids

Two test tubes were labelled as “1” and “2” , respectively. 20 drops of 95% ethanol was
added to test tube 1 and 20 drops of water was added to test tube 2 by using separate
droppers. 5 drops of oil was added to test tube 1 and 2 and each test tube was shaken well, let
them settled, and the observation whether the oil is soluble in either solvent was recorded.
The contents of the test tubes were discarded and cleaned. LIpids are soluble only in nonpolar
solvents because lipids, themselves, are nonpolar. Water is polar; ethanol is not.

3.3 Tests for Proteins

Three test tubes were labelled as “1”, “2”, and “3”, respectively. 30 drops of 2% gelatin was
added to test tube 1, 30 drops of glucose solution was added to test tube 2, and 30 drops of
water was added to test tube 3, by using separate droppers. The initial colour of each tube's
contents was recorded. Then, 10 drops of biuret reagent was added to each test tube. The
colour of each tube’s contents was recorded after adding biuret reagent. The contents of the
test tubes were discarded and cleaned. Be careful when handling biuret reagent because it is
extremely caustic to the skin and clothing. If a biuret reagent is spilled, rinse with water and
call the instructor immediately. When biuret reagent is mixed with a protein, it will produce
lavender to violet colour.

3.4 Identification of Organic Molecules in Test Samples

All of the tests above on the test samples were run and the organic molecules present on
the test samples was determined.
OBSERVATIONS
Figure 1.
Showing the colour changes obtain
after iodine solution was added to substances
to
(from left to right as test tube 1,2 and 3
respectively). Test tube 4 was a test sample.
Figure 3.
Showing the translucent spot after oil was
dropped on the brown paper.
dropped.
Figure 2.
Showing the colour changes obtain
after benedict’s solution was added
substances (from left to right as
test tube 1,2 and 3 respectively).
Test tube 5 was a test sample.
Figure 4.
Showing the brown paper without
translucent spot after water was
Figure 5. Figure 6.

Showing the brown paper without the Showing the solubility of lipids after
translucent spot after test sample was oil was added ( test tube right to left
added. As test tube 1 and 2).

Figure 7. Figure 8.

Showing the violet colour after biuret Showing the yellow colour after
biuret
reagent was added to the gelatin. reagent was added to the glucose.
Figure 9. Figure 10.

Showing the colourless solution after Showing the yellow colour after the
a biuret reagent was added to the water. biuret reagent was added to the test
sample.

RESULTS

3.1

3.1.1 : Test for Starch

Test tube Substance Initial colour Colour after Starch present

adding iodine (Yes/No)

1 Starch Cloudy Blue black Yes

2 Glucose Colourless Brown No

3 Water Colourless Brown No

4 Test sample Colourless Yellow No

3.1.2 : Test for Reducing Sugar

Test tube Substance Initial colour Colour after Reducing sugar

adding present
Benedict’s (Yes/No)
solution

1 Starch Cloudy Blue No

2 Glucose Colourless Orange Yes

3 Water Colourless Blue No

4 Test sample Colourless Red Yes

3.2

3.2.1 : Brown Paper Test for Lipids

Substance Translucent on brown Lipids present

paper? (Yes/No)

Water Not translucent No

Oil Translucent Yes

Test sample Not translucent No

3.2.2 : Solubility Test for Lipids

Test tube Substance Dissolve? Lipids present

(Yes/No) (Yes/No)

1 Oil in ethanol Yes No

2 Oil in water No Yes

3.3 : Test for Protein

Test tube Substance Initial colour Colour after Protein present

adding biuret (Yes/No)
reagent

1 Gelatin Colourless Violet Yes

2 Glucose Colourless Yellow No

3 Water Colourless Colourless No

4 Test sample Colourless Yellow No

DISCUSSION

As we know, organic molecules can be found in a wide range of food. We identified and
determined the present and type of organic molecules by these experiments. The experiment
went well. In this test, to determine the presence of starch, we used iodine solution. After
adding the iodine solution to the substance that contains starch, we observed the colour of the
solution turns from brown solution to blue black solution. To be more specific, starch is a
type of carbohydrate . Starch can be classified as polysaccharides. It consists of two different
types of polysaccharides that are made up of glucose units which are the linear amylose and
the other is the amylopectin. Amylose is the compound that is responsible for the blue colour.
Its chain forms a helix shape, and iodine can be bound inside this helix. This explains why
the colour turns to blue black when there is a present of starch.

In the meantime, we used Benedict’s solution to test the presence of simple reducing
sugars. After adding Benedict's solution to the substance that contains small monomers of
carbohydrates such as monosaccharides and certain disaccharides, which have free ketone or
aldehyde functional groups. We observed that the colour of the solution turns from blue
solution to orange and red solution but some remain blue. The colour varies from green to
brick red , depending on the amount of and type of sugar. Benedict’s solution also can be
used to test for the presence of glucose in urine. Some sugars such as glucose are called
reducing sugars because they are capable of transferring hydrogens (electrons) to other
compounds, a process called reduction. When reducing sugars are mixed with Benedict's
reagent and heated, a reduction reaction causes the Benedict's reagent to change colour.
 Next, we also determined the presence of protein by adding the solution with biuret
reagent. We observed the colour turns to violet when protein is present. This test also is
widely used in determining the protein amount in urine. The negative Biuret test can be
determined by the colour of the solution, remaining blue. This means there is no protein
present in the solution. Proteins may be either a single polypeptide or more. Polypeptides are
a particular arrangement of amino acids synthesised into a polymer by condensation
reactions. The bond formed is a covalent bond, called a peptide bond. The violet colour is
present because of the peptides bond. The peptide bonds are formed at the ribosome.

Moreover, we also used brown paper which tested for lipids presence. The brown paper
becomes translucent after oil is added. This is because when fat is absorbed into the paper's
pores, the grease-stained part of the paper which is normally white due to the scattering of
light that shines through it allows less scattering, and the light passes through it, appearing
darker in colour and translucent. We also test the solubility of lipids and based on
observation, lipids do not present in ethanol but lipids present in lipids. This shows that lipids
can be dissolved in organic solvent but not in water. This is because lipids are nonpolar
molecules. The same goes to organic solvent such as ethanol, organic solvent is a nonpolar
molecule, while water is a polar molecule. Non-polar molecules will dissolve in nonpolar
molecules. The same concept can be applied when greasy food is spilled on clothing. It will
be difficult to clean because grease does not have charge and is not attracted to the water
molecule that has charge. We can replace the water with other organic solvent such as
alcohol. For example, we can use things that easily get at home such as hand sanitizer which
also contains alcohol.

Furthermore, the role of water being tested in this experiment : to act as a control. It is
because distilled water basically contains nothing in it, since it is inert, it won't affect the
outcome of tests. In this experiment, we must be concerned with the precaution steps, such
as when handling the biuret reagent. Biuret reagent will turn the skin to brownish-purple.
This is because there is a type of protein found on epithelial cells, which line the inside and
outside surfaces of the body called keratin. Keratins help form the tissues of the hair, nails,
and the outer layer of the skin. Thus, the biuret reagent will react with the protein at our skin.

CONCLUSION

In toto, the experiment conducted went successfully, even though there were some flaws in
the outcome of the process as there might be some mistakes in calculating the drops of
substances and measuring the time along the process. Thus, this miscalculation can be
improved in the future. Students can conclude that every biological molecule can be tested to
determine their presence. Each biological molecule plays an important role in our body. At
the end of this experiment, students can conclude that the test sample was actually part of
carbohydrates. Students determined that the test sample was sucrose. This is because the test
sample turns yellow after adding biuret reagent which is the same colour as glucose after
adding biuret reagent. Meanwhile, when students added Benedict’s solution to the test
sample, it turns orange, which shows that the test sample contains a high amount of reducing
sugar.

REFERENCE

❖ Website

> Chemistry of Life - What is Chemistry of Life? Retrieved from BYJUS website :

https://byjus.com/chemistry/chemistry-of-life/#:~:text=What%20is%20the%20Chemi
stry%20of,%2C%20 messengers%2C%20and%20energy%20 carriers.

> Skin : Layers, Structure and Function. Retrieved from Cleveland Clinic website :

https://my.clevelandclinic.org/health/articles/10978-skin#:~:text=Your%20
epidermis%20is%20the%20 top,together%20to%20 form%20this%20 layer.

> What are macromolecules? Retrieved by Khan Academy website :

https://www.khanacademy.org/science/ap-biology/chemistry-of-life/introduction-to-bi
ological-macromolecules/a/introduction-to-macromolecules.