BIO 091

LAB REPORT 1 (S33)

DATE : 10th AUGUST 2020
EXPERIMENT 1
IDENTIFICATION OF BIOLOGICAL MOLECULES IN FOOD

GROUP 2:

1. MUHAMMAD AZFAR BIN JAFRI 2020887592

2. NOOR NAIM ZUHAIR BIN NOOR ROSLY 2020824714
3. NUR SAIDATUL HUSNA BINTI ANUAR 2020451868
4. NURIN NISA’ BINTI AHMAD ZAMRI 2020867922
5. SHAZWINA AYNA BINTI MOHD FAHMI 2020877016

LECTURER’S NAME :
NHAWAL AMINIE BT SAIDON
1.0 Abstract

In this lab, we have done tests for macromolecules like reducing sugar, protein,
starch and lipids by using different reagents for each test. A different colour change of the
reagents would verify the presence of macromolecules.

For reducing sugar, Benedict reagent is used to analyse the presence of reducing
sugar in glucose solution and peanut . A calibrated dropper was used to take 2.0 ml of
Benedict reagent and placed it into two test tubes that contained glucose solution and
peanut each. As a result, both food samples have reducing sugar but differ in amount. It was
observed by changes of colour in both glucose solution and peanut.

For protein, the reagent that we used is Biuret’s reagent. Biuret test is used to detect
the presence of protein. The theory that we have was that, if the food sample contained
protein such as albumin, it would somehow react with Biuret’s reagent when it is added in it.

For starch, Lugol’s solution was used as the reagent. The result is that, if the food
sample contains starch, the colour of the reagent would change from brown to blue-black.

For lipid, Sudan IV solution was used as the reagent. The result that we obtained is
that the food sample that contains lipid such as vegetable oil would change the Sudan IV
solution colour while the food sample that does not have lipid will undergo no changes.
2.0 Introduction

Food contains different amounts of macromolecules, such as carbohydrates,

proteins,and lipids . All of these macromolecules are important to all living things.
Macromolecules polymers consist of a long chain of monomers. Different reagents can
detect the presence of some monomers.

Carbohydrates contain carbon, hydrogen and oxygen. There are four groups of
carbohydrates which are monosaccharide, disaccharide, oligosaccharide and
polysaccharide. Monosaccharide is simple sugar. Two monosaccharide joined by a covalent
bond form a disaccharide. Three or more disaccharide joined together forming the
polysaccharide. The presence of simple sugar can be tested by Benedict’s test while starch
can be tested by Lugol’s solution.

Proteins consist of one or more polypeptides. Amino acid is the monomer for proteins
that join together by a covalent bond that is called peptide bond. There are four levels of
organization for protein. Primary structure, secondary structure, tertiary structure and
quaternary structure. Proteins can be classified by their structure, composition and function.
Biuret’s test can identify the presence of protein.

Lipids contain carbon, hydrogen and oxygen. There are 3 important groups of lipid
which are triglyceride, steroid and phospholipid. Lipids are hydrophobic because they mostly
contain hydrocarbons that form nonpolar covalent bonds. So, they only dissolve in organic
solvents. Lipids can be tested by Sudan IV reagent.
3.0 Objective

In this lab we demonstrated the experiments to identify biomolecules in food samples.

First, we did the experiment to detect the presence of reducing sugar. Second, the
experiment to detect the presence of protein. Then, the experiment to detect the presence of
starch. Lastly, the experiment to detect the presence of lipids. The result was collected by
observing the colour changes of reagents after the reagent’s were put into the test tubes with
a food sample.
4.0 MATERIALS & METHODS

Firstly, a water bath is heated to 95°C or using a hot plate . Half of a 250ml beaker is filled
with water and boiled. 8 test tubes are prepared and placed in a test tube rack . Next, 10
peanuts are prepared and grinded using mortar and pestle.

4.1 BENEDICT TEST

Firstly , in a test tube labelled glucose, put in 2.0 ml glucose solution and in the test tube
labelled peanut, put in half a spatula of ground peanut. After that, the colour of benedict’s
reagent is observed . A calibrated dropper to take 2.0 ml Benedict reagent is used and
placed into both test tubes. Next , both test tubes are placed in a water bath or boiling bath.
Leaves for 5 minutes and colour change is observed.

4.2 BIURET TEST

Firstly , in test tube labelled albumin, put in 2.0 ml albumin solution and in test tubes labelled
peanut, put in half a spatula of ground peanut. The colour of Biuret reagent is observed . A
calibrated dropper to take 2.0 ml Biuret reagent is used and placed into both test tubes.
Lastly ,colour change is observed.

4.3 LUGOL TEST

Firstly, two test tubes are labelled with starch and peanut. Secondly, a 2.0 ml starch solution
is put in the test tube labelled starch followed by half a spatula of ground peanut is put in
another test tube labelled peanut. Next, the colour of Lugol’s solution is observed.Then 10
drops of Lugol’s solution is added in each test tube and the change of colour is observed.

4.4 SUDAN IV TEST

Firstly, two test tubes are labelled with vegetable oil and peanut. Secondly, 2.0 ml of distilled
water is put into each test tube. Secondly, 2.0 ml vegetable oil is put in the test tube labelled
vegetable oil and half a spatula of ground peanut is put in the test tube labelled peanut.
Next, the colour of Sudan IV solution is observed followed by 10 drops of Sudan IV solution
are added to each test tube. Then, the vortex is used to mix the substance in each test tube.
After that, another 2.0 ml of distilled water is added to each test tube. Lastly, the change of
colour in each test tube is observed.
5.0 Result of experiment

FOOD TEST TEST TUBE COLOUR CONCLUSION

INITIAL FINAL

BENEDICT’S Glucose Blue Brick red with High amount of

precipitate reducing sugar is
present

Peanut Blue Green Traceable amount of

reducing sugar is
present

BIURET Albumin Blue Purple Protein is present

Peanut Blue Light purple Low amount of protein

is present

LUGOL’S Starch Brown Blue-black Carbohydrate is

present

Peanut Brown Blue-black Carbohydrate is

present

SUDAN IV Vegetable oil Dark red Redish Lipid is present

orange layer
forms

Peanut Dark red Light red with Low amount of lipid is

precipitate present

FIGURE 5.0 : Observation of the experiments.

6.0 Discussion

The results of the first experiment supported the hypothesis that glucose contained
reducing sugar as shown in the result, the colour of Benedict’s solution changed from blue to
brick red precipitate. ​Some sugars such as glucose are called reducing sugars because they
are capable of transferring hydrogens (electrons) to other compounds, this process is called
reduction. When reducing sugars are mixed with Benedict's reagent and heated, a reduction
reaction causes the Benedict's reagent to change color. The color varies from green to dark
red (brick) or rusty-brown, depending on the amount of and type of sugar. This colour also
explains the result in the second test tube for this experiment. In the second test tube with
ground peanuts as a food example, green colour formed when it mixed with Benedict’s
solution to show that a very low amount or only traceable amount of reducing sugar is
present in ground peanuts.

Based on the result of our second experiment, the hypothesis is proven that there is
presence of protein in albumin and peanut as the Biuret reagent changed its colour from
blue to purple.The normal colour of Biuret reagent is blue. The reagent turns purple in the
presence of peptide bonds. The proteins detected must have at least three amino acids,
which means that the protein must have at least two peptide bonds. The reagent’s copper
ions, with a charge of +2, are reduced to a charge of +1 in the presence of peptide bonds,
causing colour change. However, there is a slight difference between the purple colour
intensity from both test tubes containing albumin and peanut depending on the amount of
protein found in the food samples. Amount of protein in albumin solution is high due to its
darker colour of purple formed while the light purple formed from the mixture of peanut and
Biuret reagent indicates low amount of protein found in the peanut.

Next, for the third experiment, we can conclude that the hypothesis is also proven in
this test. This is shown in the result, when a test tube labelled A, which contains 2.0 mL of
starch solution reacts when iodine is added. ​Using iodine to test for the presence of starch is
a common experiment. A solution of iodine (I​2​) in water has a light orange-brown color. If it is
added to a sample that contains starch, the color changes to a deep blue. ​Amylose in starch
is responsible for the formation of a deep blue color in the presence of iodine. The iodine
molecule slips inside of the amylose coil. Iodine - KI Reagent: Iodine is not very soluble in
water, therefore the iodine reagent is made by dissolving iodine in water in the presence of
potassium iodide. This makes a linear triiodide ion complex which is soluble that slips into
the coil of the starch causing an intense blue-black color. The same reason is also applied to
the result in test tube B, which contains ground peanuts. This shows that both starch solution
and peanuts contain carbohydrates.
 For the last experiment that we conducted in this session, the result that we obtained
supported the hypothesis that stated lipids are present in both test tubes. When Sudan IV
solution is dropped into a test tube containing vegetable oil mixed with distilled water, the
colour changes from dark red to reddish orange layers formed on the top of the mixed
solution. This is because, Sudan IV solution is only soluble in vegetable oil and not soluble in
distilled water. in the second .Meanwhile, in test tube B contained ground peanuts, dark red
colour changes to light red with precipitate. This shows that only a low amount of Sudan IV
solution is dissolved in peanuts due to its low amount of lipids.
7.0 CONCLUSION

In a nutshell, we have achieved our objective for this experiment as we were able to figure
out the nutrient in glucose, albumin, starch, vegetable oil and peanuts by identifying which
test tube has changed colours after adding the reagents and resulted positively . Peanut is a
great source of nutrients . This lab proved our hypothesis is correct and did change colours
to their reactive colours when specific nutrients were presented . To illustrate, glucose has a
high amount of reducing sugar compared to peanuts since in benedict's test,it showed a
positive result. On the other hand, proteins are present in albumin and peanut since both of
them turn the blue colour of biuret solution into purple ( albumin) and light purple (peanuts).
Furthermore, starch and peanut contain carbohydrates since in Lugol’s test, both of them
showed a positive result . Lastly, lipids are present in vegetable oil and peanut but differ in
amount .
REFERENCE

1.​https://education.seattlepi.com/biuret-test-mean-biology-4659.html