CLASS GROUP : PI080S38

CENTRE OF FOUNDATION STUDIES

FOUNDATION BIOLOGY
LABORATORY REPORT
Biology 091
Experiment 1 : Identification Of Biological Molecule In Food
Lab Instuctor:
NURUL HUDA BINTI MUSA
Lab Partners: Group 3
MUHAMMAD AFIQ DANIAL BIN ZARUNI
(2020835758)
NURDEENA AQILAH BINTI AHMAD ZAIDI
(2020868426)
MAISARAH BINTI ALAUDIN
(2020481516)
AININ SOFEA BINTI MUHAMMAD NADHIRIN
(2020616474)
NUR FATINI BINTI MOHAMAD LATIF
(2020891618)
EXPERIMENT 1: IDENTIFICATION OF BIOLOGICAL MOLECULES IN FOOD

Introduction
Carbon, hydrogen and oxygen are the main organic molecules that are some of the common
organic compounds in the living organism. Each of these macromolecules is made up of smaller
subunits. Macromolecules and smaller subunits can be known as polymers and monomers,
respectively.Bonds between these subunits are formed by dehydration synthesis. Energy is needed to
make this process happen. A water molecule is removed (dehydration) and a covalent bond is
formed between the subunit, thus breaking the bond is called hydrolysis, which involves the release
of energy and the addition of water molecules.
Macromolecule has various types of properties and is structured. For example, lipids that are
made up of fatty acids. Lipids have a lot of C-H bonds and very little oxygen. Whereas proteins are
made up of amino acids. Proteins have classes of amino(-NH3 +) and carboxyl(-COOH). These
subunits and chemical groups of portraits give different properties to the macromolecules. For
example, monosaccharides such as glucose are polar and water-soluble, while lipids are non-polar
and insoluble in water.
Objectives
1. To carry out tests for reducing sugar, starch, proteins and lipids.
2. To determine types of biological molecules present in a food sample given.
Materials Apparatus
Distilled water Calibrated dropper
Glucose Test tube
Albumin Test tube rack
Starch Water bath
Vegetable oil Mortar and pestle
Benedict’s solution Spatula
Biuret reagent Vortex mixer
Lugol’s solution
Sudan IV solution
Peanuts

Procedure
1. Heat water bath to 96°C.
2. Get 8 test tubes, place them in a test tube rack.
3. Take 10 peanuts and grind them using mortar and pestle.
4. Label two test tubes: (i) glucose (ii) peanut
 In test tube labelled glucose, put in 2.0 ml glucose solution.
 In test tube labelled peanut, put half a spatula of ground peanut.
 Take note of the color of Benedict’s reagent.
  Use a calibrated dropper to take 2.0 ml Benedict’s reagent and place it into both test
tubes.
 Place both test tubes in water bath.
 Leave for 5 minutes.
 Observe color change.
5. Label two test tubes: (i) albumin (ii) peanut
 In test tube (i), put 2.0 ml albumin solution.
 In test tube (ii), put half a spatula of peanut.
 Take note of the color of Biuret reagent.
 Use a calibrated dropper to take 2.0 ml of Biuret reagent and place it into both test
tubes.
 Observe color change.
6. Label two test tubes: (i) starch (ii) peanut
 In test tube (i), put in 2.0 ml starch solution.
 In test tube (ii), put in half a spatula of ground peanut
 Take note the color of Lugol’s solution.
 Add 10 drops of Lugol’s solution in each of the test tubes,
 Observe color change.

7. Label two test tubes: (i) vegetable oil (ii) peanut

 Put in 2.0 ml of distilled water into each test tubes.
 In test tube (i), put in 2.0 ml vegetable oil.
 In test tube (ii), put in half a spatula of ground peanut.
 Take note the color of Sudan IV solution.
 Add 10 drops of Sudan IV solution into each test tubes.
 Use vortex to mix.
 Add another 2.0 ml of distilled water to each test tubes.
 Observe change in solution in test tubes.
Results

Reagent Food sample Observation Inference

Benedict’s Glucose Blue color changed to brick It contains high amount of

red color reducing sugar. Copper (II) ions
are reduced to Copper (I) ions.
Peanuts Blue color changed to It contains small amount of
greenish color reducing sugar. Copper (II) ions
are reduced to Copper (I) ions
Biuret Albumin Blue color changed to It contains high amount of
purplish blue color protein. Albumin reacts with the
Biuret reagent and formed the
complex of the dipeptide bonds
with the cupric ions.
Peanuts Blue color changed light It contains small amount of
blue color protein. Copper (II) ions are
reduced to Copper (I) ions in the
present of peptide bond.
Lugol’s solution Starch Brown color changed to Starch is present. Lugol’s will stain
blue black color starches due to iodine’s
interaction with the coil structure
of the polysaccharides.
Peanuts Brown color changed to Starch is present. Lugol’s will stain
black color starches due to iodine’s
interaction with the coil structure
of the polysaccharides.
Sudan IV Vegetable oil Reddish brown color to It contains lipid. Sudan IV soluble
reddish-orange color, two in lipids and lipids are not soluble
layers formed in water.
Peanuts Reddish brown color It not contains lipid. Sudan IV
remained the same insoluble in water.
(i) Benedict’s reagent + glucose
(ii) Benedict’s reagent + peanuts

(i) Biuret reagent + albumin

(ii) Biuret reagent + peanuts
(i) Lugol’s solution + starch
(ii) Lugol’s solution + peanuts

(i) Sudan IV solution + vegetable oil

(ii) Sudan IV solution + peanuts
Discussion

Peanut is used as a food sample for this experiment. The results show types biological that
present in the peanut are reducing sugar (glucose), protein (albumin), carbohydrates (starch ) and
lipids (vegetable oil ).

Benedict’s solution are used to determine reducing sugar in peanut and glucose. Blue color of
Benedict’s solution changed to greenish color in the test tube that contains peanut. It means that
peanut contains small amount of reducing sugar. Copper (II) ions are reduced to Copper (I) ions. In
the other hand, colour of Benedict’s solution in the test tube that contain glucose changed from blue
to brick-red colour which mean that glucose contains high amount of reducing sugar. Copper (II) ions
are reduced to Copper (I) ions still. We can see here that the changes in colour are different because
the concentration of presence of reducing sugar are different in each test tubes. Benedict's test is a
test for the presence of monosaccharides or certain disaccharides in a solution or to determine
reducing simple sugar or smaller carbohydrates like glucose. Reducing sugars are simple sugars and
include all monosaccharides and most disaccarides. Some examples of monosaccharides are glucose,
fructose and galactose.Examples of reducing disaccharides are lactose and maltose.Benedict's
solution has copper ions (Cu2+) that have a light blue color. When a solution containing these sugars
is mixed with Benedicts reagent and heated, a reduction reaction causes the copper ions change, the
Benedicts reagent to change color. The color varies from yellow to green to dark red, depending on
the amount of and type of sugar.  A change in color indicates the presence of reducing sugars.

Biuret reagent are used to determine protein in peanut and albumin. The colour of Biuret
reagent changed from blue to light blue in the test tube that contains peanut. This means that it
contains small amount of protein. Copper (II) ions are reduced to Copper (I) ions in the present of
peptide bond. In the other hand, blue color of Biuret reagent changed to purplish blue or violet color
in the test tube that contains albumin. It contains high amount of protein. Albumin reacts with the
Biuret reagent and formed the complex of the dipeptide bonds with the cupric ions. The different
concentration of protein present in peanut and albumin influenced the changes in colour of Biuret
reagent. Biuret test is a chemical test used for detecting the presence of peptide bonds. It is based
on the biuret reaction in which a peptide structure containing at least two peptide links produces a
violet color when treated with alkaline copper sulfate. The copper atoms of Biuret solution
(CuSO4 and KOH) will react with peptide bonds, producing a color change. Biuret solution is initially
blue; mixing it with proteins causes a violet color changed. The intensity of the color change is
related to amount of proteins insolution. A more intense color (darker violet) indicates higher protein
concentration. If proteins are present a chemical reaction caused between the copper ions and the
protein molecules.The chemical reaction causes the Biuret solution to turn from a light blue to purple
if proteins are present. For reaction to occur, the protein must contain 4-6 linked amino acids. A
deep violet color indicates the presence of proteins and a light pink color indicates the presence of
peptides. Also, reaction only works in an alkaline (basic) environment. Therefore biuret contains
sodium hydroxide, which produces and alkaline solution.

Lugol’s solution are used to determined starch in peanut. The colour of Lugol’s solution is brown
and this colour changed to to black color when peanut was being tested by Lugol’s solution. Can we
conclude that starch is present. Lugol’s solution will stain starches due to iodine’s interaction with
the coil structure of the polysaccharides while in the test tube that contains starch the changes of the
Lugol’s solution colour also same as result of Lugol’s solution test on peanut. Starch is a carbohydrate
found in plants. It consists of two different types of polysaccharides that are made up of glucose units
which are connected in two different ways. One is the linear amylose and the other is the branched
amylopectin;
Starch is a polysaccharide consisting of glucose units joined together by glycosidic bonds.The chains
formed during the condensation reaction are either linear or highly branched molecules.

Whereas branched molecules of starch are called Amylopectin.

To determine lipids in the peanut and vegetable oil, Sudan IV solution are being used. The dark
red or reddish brown colour of Sudan IV solution in the test tube that contains vegetable oil change
to reddish orange colour shows that there are presence of lipids in vegetable oil and it also formed 2
layers. This can be seen that Sudan IV soluble in lipids and lipids are not soluble in water. In the other
hand, the results of the Sudan IV on peanut were just remain the same which the colour of Sudan IV
(reddish brown) did not change when dropped into the test tube that contains peanut. Lipids are
nonpolar and therefore do not dissolve in water. Emulsifiers are molecules have both polar and
nonpolar parts and thus are capable of dissolving in or interacting with both lipids and water. When
emulsifiers are mixed with lipids and water, they may act to suspend small droplets of the lipid in
water. The Sudan IV test will test positive for lipids. The test procedure involves adding a few drops of
Sudan IV to the test solution. Sudan IV is a dye that will stain lipids. If no lipids are present then the
dye will sink to the bottom of the test tube. The lipid is not dissolved in water, but is broken into
smaller fragments that may remain suspended for long periods of time. Sudan IV solution contains a
fat-soluble dye. When Sudan IV is combined withlipids, the lipids absorb the dye particles, causing a
red color at the site of contactbetween lipids and the dye. Lipids do not readily dissolve in water; bc
of this when placed in water, they formdistinctive fat layers in the solution. Any noticeable red layer
is most likely causedby a reaction of Sudan IV solution and lipids.

https://www.chemistryviews.org/details/education/10128441/Why_Does_Iodine_Turn_Starch_Blue.
html

https://en.wikipedia.org/wiki/Iodine-starch_test

https://microbenotes.com/biuret-test-for-protein/

http://brilliantbiologystudent.weebly.com/biuret-test-for-protein.html

https://sites.google.com/a/wrps.net/cns-ontl/cns-2nd-semester-weblinks/unit-7-resources---
lab/chemical-tests-to-identify-biomolecules?tmpl=%2Fsystem%2Fapp%2Ftemplates%2Fprint
%2F&showPrintDialog=1

https://www.nthurston.k12.wa.us/cms/lib/WA01001371/Centricity/Domain/2585/BioMols%20Lab
%2018-19.pdf

http://faculty.uncfsu.edu/jraynor/BIOL%20200%20Online%20Lab/Biological%20Molecules
%20Lab.htm
CONCLUSION

This experiment indicated positive results as the colour of food sample which was peanut changed
in Benedict reagent test, Biuret solution test, Iodin test and Sudan IV solution test. Glucose , albumin,
starch and vegetable oil acts as a set of samples that tested during the experiment to determine the
biological molecules in food. The results showed specific colour changes in every macromolrcule
presence based on its concentration.

Glucose reacted with Benedict reagent, as did the peanut to a lesser degree showing the presence
of reducing sugar.

Albumin and peanut are confirmed contain protein through Biuret solution test.

Starch and peanut reacted with Lugol’s solution along with the peanut proving the presence of
carbohydrates.

Vegetable oil and peanut turned out not contain lipid through Sudan IV solution test. The resut
proved that peanut contains reducing sugar, carbohydrates and protein.