INTERNATIONAL UNIVERSITY – HCMC

School of Biotechnology

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BIOLOGY
LABORATORY

REPORT FOR PRACTICAL 2

ORGANIC COMPOSITIONS OF THE CELL

Course name: Biology

Course ID: BT155IU
Instructor: Mr. Quang
Group number: 2
Group members: Nguyễn Ngọc Phương Anh BTBCIU19034
Nguyễn Thị Ánh Hồng BTBTIU19015
Nguyễn Anh Thư BTBTIU19041
Phan Hải Nam BTBCIU19050
Date of submission: 04/12/2020

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I/ CARBOHYDRATES
1. Introduction
Carbohydrates (CnH2nOn or (CH2O)n) are a group of compound that presents in
plant and animal cells. They are divided into monosaccharide, disaccharides, and
polysaccharides. In this experiment, we will learn how to detect the starch in the
given samples by Lugol solution and understand the effect of temperature to the
structure of the starch.
2. Procedure
a) Task 1 – Microscopic Observation of Starch Granules.
‒ Prepare the clean glass slide and coverslip.
‒ Cut the potato and scratch potato at the edge of this cut.
‒ Collect the scratching and place on the slide, add a Lugol drop, and cover
with a coverslip. Then observe the phenomenon under the microscope.
b) Task 2 – Chemical Detection of Starch.
‒ Add 5ml of starch suspension into a test tube.
‒ Take out one drop of rice starch suspension and put it onto the glass slide.
Add 1 drop of Lugol solution and mix well. Observe the color change of this
suspension (compare with the original color of Lugol solution).
‒ After that, add 5 drops of concentrated HCl solution into that test tube
containing 5ml of starch suspension. Mix well.
‒ Take out 1 drop of starch-HCl mixture onto the slide and test the color with 1
drop of Lugol solution.
‒ Place the test tube containing starch and HCl into the rack which has been
already submerged in the hot water (the water bath is set to 100 oC) and boils
this suspension. Every 5 minutes, take out one drop of hydrolyzed starch-HCl
mixture using Pasteur pipette and put onto a glass slide, let it cool down for a

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 while and add 1 drop of Lugol solution. Observe the change of color
intensity.
‒ Continue to boil and test with Lugol solution until no color change is
detected. Mark the time that the color does not change.
3. Results
a) Microscopic observation

Starch granules

Potato cells

b) Effect of temperature to the structure of starch

Color of spot

Lugol Light yellow

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Lugol + starch Dark blue

Starch + HCl
mixture

0 min Dark blue

5 min Dark purple brown

10 min Brown

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15 min Dark yellow

20 min Yellow

25 min Light yellow

30 min No color change

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4. Discussion
a) The phenomenon when adding Lugol solution to potato starch granules?
The dark blue start to appear when the Lugol solution is added to potato starch
because the structure of Lugol is Iodine and Kali Iodine. When starch react with
Iodine in normal condition, it makes starch has a blue dark color.
b) Explain the different color in starch – HCl mixture after time boiling and
why can we say that the structure of Starch is affected by temperature?
‒ Over time, the dark blue color fades. Only the golden brown color of Lugol.
‒ Because when we add HCl to starch and mix, over time and boil the starch gets
broken down into glucose. Glucose does not work with lugol so it will not
appear dark blue. The longer it is boiled, the more the starch is broken down,
so there is no more starch that reacts with Lugol, the dark blue color fades.
When the starch is fully decomposed, the dark blue color will not appear.
II/ PROTEINS
1. Introduction
‒ Proteins are large biological molecules, or macromolecules, consisting of one
or more long chains of amino acid residues. Proteins perform a vast array of
functions within living organisms, including catalyzing metabolic reactions,
replicating DNA, responding to stimuli, and transporting molecules from one
location to another. Proteins differ from one another primally in their
sequence of amino acids, which is dictated by the nucleotide sequence of
their genes, and which usually results in folding of the protein into a specific
three–dimensional structure that determines its activity.
‒ In this lab, we will know about Biuret reaction on Protein and the function of
Biuret reaction.

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2. Procedure
‒ Prepare 2 clean and dry test tubes.
‒ Pipette 2 ml of egg albumin and 2 ml of fresh cow milk into 2 test tubes.
‒ Adding 10 drops of 10% NaOH solution into each test tube. Mix well.
‒ Adding 3 drops of 0.5% CuSO4 solution into each test tube.
‒ Observe the color.
3. Results

Protein solutions Original Color After 10% NaOH After 0.5% CuSO4
Egg albumin Light yellow Percipitation Dark Purple
Fresh cow milk White Light pink Light Purple

Original 10% NaOH 0.5% CuSO4

Egg albumin

Original 10% NaOH 0.5% CuSO4

Fresh cow milk

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4. Discussion
a) Explain the function of 10% NaOH and 0.5% CuSO4 in Biuret reaction?
Biuret test uses a reagent consists of potassium hydroxide and copper sulfate.
The function of 10% NaOH is to create a strong basic environment. After that, two
nitrogen atoms from two adjacent peptide bonds can coordinate with the other two
nitrogen atoms on other peptide chains in the conformation with copper ions form
a violet-colored complex.
b) After adding 10% NaOH, the phenomenon in egg white is different from
in cow milk, why?
Because egg whites can undergo gelation at extreme pH. When we drip 10%
NaOH solution on the egg whites, it begins to gel and gradually appears a
coagulant. In milk, it has a specific protein called calcium caseinate, when adding
10% NaOH into milk, salt and milk will divide into two parts is emulsion and
watery liquid.
c) Why is the color intensity in egg white different from in cow milk?
Because of specific protein, the salt of milk convert to ions and stick around
charged region protein so it has a purple cloudy color but egg don't have that
specific protein so it has a purple color.
III/ LIPIDS
1. Introduction
‒ Lipids, any of a diverse group of organic compounds including fats, oils,
hormones, and certain components of membranes that are grouped together
because they do not interact appreciably with water. The membranes of cells
and organelles (structures within cells) are microscopically thin structures
formed from two layers of phospholipid molecules. Membranes function to
separate individual cells from their environments and to compartmentalize

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 the cell interior into structures that carry out special functions. The three
main types of lipids are triacylglycerols (also called triglycerides),
phospholipids, and sterols.
‒ Lipids perform three primary biological functions within the body: they serve
as structural components of cell membranes, function as energy storehouses,
and function as important signaling molecules.
‒ In this experiment, we will detect and observe the lipid in plant cells using
red Soudan III solution.
2. Procedure
‒ Slice the peanut thinly.
‒ Place it on the slide, add a drop of Soudan III solution and wait 10 minutes.
‒ Wash the slide with 20% Ethanol.
‒ Add a drop of water or immersion oil to the sample and put the coverslip on.
‒ Put the sample on the stage, and use the microscope to observe it with
different magnifications.
3. Results

Lipids

Peanut slice

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4. Discussion
a) Why is Soudan III used to detect lipid?
Soudan III is used to stain nonpolar molecules because it is fat-soluble (can be
dissolved in fats and oils). This means that is can stain fats and oils even when
water is used to wash the fats and oils away (the fats and oils are insoluble in
water).
b) Why do we have to wash the stained sample with 20% Ethanol before
observation under microscope?
We wash the stained sample with 20% Ethanol before observation in order to get
the lipids to be a copper color. Soudan III is soluble in ethanol, therefore, we can
use ethanol to get rid of the excess Soudan III while keeping the lipids present with
their copper staining from the Soudan III.

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