LAB REPORT 2

Course name (and course’s ID): Practice in Biology S1_2023-24_G06

Instructor: Ms. Phạm Hồng Điệp
Group number: 5 – Saturday afternoon
Group member (name and ID): Phạm Hữu Tuấn Anh BTBTIU23119
Bùi Ngọc Thảo My BTBTIU23134
Trần Bảo Ngọc BTBTIU23145
Trần Vũ Thanh Thảo BTBTIU23115
Date of submission: 04/11/2023

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I/ CARBOHYDRATES:
1/- Introduction:
- Carbohydrates are most abundant organic compounds found in living organism
and are composed of carbon, hydrogen, and oxygen. Carbohydrates play a vital
role as a primary source in providing energy for functioning of living organism
in the form of glucose. Most of carbohydrates are made of Cx(H20)y, which is
why it is called carbohydrate (hydrates of carbon).
- Carbohydrates (saccharides) are divided into three main types: monosaccharide
(single carbohydrate molecule), disaccharide (two monosaccharides are linked
together), and polysaccharide (many monosaccharides are linked together).
-They are synthesized in green plants through photosynthesis, a process that uses
energy from the sun and converts carbon dioxide and water into glucose and
oxygen.
2/- Procedure: Materials: potato, microscope, glass slides, coverslips, and Lugol
solution, test tubes, Pateur pipettes, concentrated HCl solution, test tube racks,
test tube clamps, toothpick and water bath.

Microscopic Observation of Starch Granules:

1. Prepare the clean glass slide and coverslip
2. Cut the potato and scratch potato at the edge of this cut
3. Collect all the scratching and place on the slide
4. Add a distilled water on the sample cover with coverslip
5. Observe under microscope (from low to high magnification)
6. Then, adding 1-2 drops of Lugol solution to the edge of the coverslip and
observe the changes

Chemical Detection of Starch:

1. Add 5ml of starch suspension into a test tube
2. Take out one drop rice starch suspension and put onto the glass slide. Then,
add 1 drop of Lugol, mix well with toothpick
3. Observe the color change (compare with the original Lugol color)
4. Then, add 5 drops of concentrated HCl solution into that test tube containing
5ml starch suspension. Mix well
5. Take out 1 drop of starch that mix with HCl solution, placing on glass slide
and test the color with 1 drop of Lugol solution
6. Place the test tube containing starch and HCl into the rack that place in 100
o
C and boil it.

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 7. Every 2 minutes take 1 drop of that starch-HCl mixture using Pasteur pipette
and put onto a glass slide. Let it cool down a bit and then add 1 drop of Lugol
solution. Observe the change of color intensity
8. 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: identify observed objective lense and starch granules

Figure I.1. Starch granules observed under microscope with 40x objective lens

b) Effect of temperature to the structure of starch: Observe the change of color

intensity:

Figure I.2, I.3. Lugol solution (right side of glass slide) and starch-HCl-
Lugol mixture (left side of glass slide) color change over time:
- Lugol solution’s color: brown
- Starch with Lugol solution color: deep blue, gradually turn into
light brown
- After 24 mins (12 times), there is no color change

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4/- Discussion:
a) Explain the phenomenon when adding Lugol solution to potato starch
granules?
When adding Lugol solution, potato starch granules have dark color (deep blue
color but not actually dark blue) due to amylose in starch. The change of color is
based on the reaction between amylose in potato starch granules and iodine ions
in Lugol solution.
b) Explain the different color in Starch – HCl mixture after a time of boiling.
Based on the color of spot, why can we say that the structure of starch is affected
by temperature?
-The color of spots becomes lighter and lighter until there is no change throughout
the time of boiling. The longer the boiling time the lighter the color will be. As a
result, the color changes from deep blue to light brown (same as Lugol).
-Starch includes chains of monomers as glucose. These monomers bonded to each
other by glycosidic linkage. When we add HCl, the polymer chain of starch
becomes shorter. After boiling, the glycosidic linkage are broken; then when
adding Lugol solution, starch and iodine cannot combine with each other, creating
a light brown color. Therefore, we say that the structure of starch is affected by
temperature.

II/ PROTEINS:
1/- Introduction:
- Proteins are large, complex molecules that are essential components of all
human tissues (e.g., bones, blood, hormones, enzymes, antibodies). Proteins
contain the elements carbon, hydrogen, and oxygen just as carbohydrates and
lipids do, but proteins are the only macronutrient that contains nitrogen.
- Proteins are macromolecules composed of amino acids. Amino acids are
commonly called the "building blocks" of protein. Amino acids contain five
elements. Each amino acid consists of a central carbon atom (C) connected to a
side chain (R), a hydrogen (H), a nitrogen-containing amino group (NH2), and a
carboxylic acid group (COOH).
- Amino acids are linked together by peptide bonds. Short chains of amino
acids are called peptides, and longer chains of amino acids are called
oligopeptides or polypeptides.

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2/- Procedure:
● Material: protein suspensions of egg white, test cubes, pasteur pipettes,
10% NaOH solution, 0.5% CuSO4 solution.
● Qualitative detection of Protein:
1. Pipette 2ml of protein suspension into a test cube.
2. Add 2ml (or 10 drops ) of 10% NaOH solution. Mix well.
3. Add one to 2-3 drops of 0.5% CuSO4 solution into the test cube.
4. Observe the color.
3/- Results:

Figure II.1. Test tubes containing milk (left) and egg white (right)
Putting 10% NaOH and 0.5% CuSO4 into the tube transform the color of both
tubes:
● Milk: from white to light purple
● Egg white: from transparent white to purple

4/- Discussion:
a) Explain the function of 10% NaOH and 0.5% CuSO4 in Biuret reaction?
-The NaOH is there to raise the pH of the solution to alkaline levels; the crucial
component is the copper II ion (Cu2+) from the CuSO4.
-When peptide bonds are present in this alkaline solution, the Cu2+ions will form
a coordination complex with 4 nitrogen atoms from adjacent peptide bonds
-The complex of Cu2+ ions and nitrogen atoms make the color of CuSO4 solution
changes from blue to violet.
-This color change is dependent on the number of peptide bonds in the solution,
so the more protein there are, the more intense the change is.

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b) After adding 10% NaOH, the phenomenon in egg white is different from in
cow milk, why?
After adding NaOH solution, the phenomenon of egg becoming transparent and
colorless because of proteins in eggs is albumin. However, the structure of this
protein has many hydrogen bonds which are easily affected in basic environment.
c) Why is the color intensity in egg white different from in cow milk?
There is a difference in protein between egg white and cow milk
+ Protein in egg (albumin) seems to look like glue.
+ Protein in milk (casein) has a hydrophilic side.
=> The color of this reaction in eggs is deep blue and looks like a precipitate and
the color of the solution of milk is purple.

III/ LIPIDS:
1/- Introduction:
Lipids are a group of naturally occurring molecules that include fats, waxes,
sterols, fat-soluble vitamins (such as vitamins A, D, E, and K), monoglycerides,
diglycerides, triglycerides, phospholipids, and others.
Biological lipids originate entirely or in part from two distinct types of
biochemical subunits or "building-blocks": ketoacyl and isoprene groups. Using
this approach, lipids may be divided into eight categories: fatty acids,
glycerolipids, glycerophospholipids, sphingolipids, saccharolipids, and
polyketides (derived from condensation of ketoacyl subunits); and sterol lipids
and prenol lipids (derived from condensation of isoprene subunits).
The main biological functions of lipids include storing energy, signaling, and
acting as structural components of cell membranes.
Lipids may be broadly defined as hydrophobic or amphiphilic small
molecules; the amphiphilic nature of some lipids allows them to form structures
such as vesicles, liposomes, or membranes in an aqueous environment.
2/- Procedure:
Qualitative Detection of Lipids:
1. Scratch the peanut which was soaked in water.
2. Place the scratching on a glass slide; add a drop of Soudan III solution. Keep
sample in this solution for staining in 5 minutes.
3. Wash the slide with 20% Ethanol.
4. Put the coverslip on.
5. Observe the lipid granules stained in peanut cells using microscope up to 40x.

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3/- Results:

Figure III.1: Lipid granules observed under microscope with 40x objective lens

4/- Discussion:
a) Why is Soudan 3 used to detect lipid?
Soudan III is a red soluble dye which is used to identify the presence of lipids.
It occurs by the reaction: Soudan III reacts with the lipids or triglycerides of
peanut. As a consequence, the color of the lipids turns out red or orange, or pink
depends on the concentration.
b) Why do we have to wash the stained sample with 20% Ethanol before
observation under microscope?
Because Ethanol is a solvent to dissolve away purple-iodine complex, so Ethanol
20% is used in order to remove it from the cell wall if its cells are gram-negative.