TITLE

Biological molecules

OBJECTIVES
1) To test the presence of carbohydrates,lipids and proteins using various test for starch,
sugar, lipids and proteins.

INTRODUCTION
Biomolecules are molecules that occur naturally in living organisms. Biomolecules
include macromolecules like proteins, carbohydrates, lipids and nucleic acids but nucleic
acids not utilized by the body as nutrient.Biomolecules consists mainly of carbon and
hydrogen with nitrogen, oxygen, sulphur, and phosphorus. Biomolecules are very large
molecules of many atoms, that are covalently bound together.
All of this biomolecules are used to run our metabolic processes. Carbohydrates
include both sugar and polymers provide the body with source of fuel and energy, it aids in
proper functioning of our brain, heart and nervous, digestive and immune system. Deficiency
of carbohydrates in the diet causes fatigue, poor mental function.
Each protein in the body has specific functions, some proteins provide structural
support, help in body movement, and also defense against germs and infections. Proteins can
be antibodies, hormonal, enzymes and contractile proteins.
Lipids, the primary purpose of lipids in body is energy storage. Structural
membranes are composed of lipids which forms a barrier and controls flow of material in and
out of the cell. Lipid hormones help in mediating communication between cells.

METHODOLOGY

EXPERIMENT 2.1 : TEST FOR SUGAR USING BENEDICTS SOLUTION

MATERIALS

10 test tubes in test tube rack

Pippet
Test tube clamp
Hot plate
Sticker for labelling
Stock bottle of :
i.
Benedict s solution
ii.
Distilled water (dH2O)
iii.
Glucose solution
iv. Maltose solution
v. Lactose solution
vi.
Sucrose solution
vii.
Starch solution
viii.
Fructose solution
ix.
Lemon juice
x.
Unsweetened orange juice
xi.
Colorless non diet soda

PROCEDURES
1) The beaker was half filled with tap water and heat was applied with a hot plate or burner to
bring the water into a gentle boil.
2) 2ml of the correct stock solutions was pipeted into the test tubes as described in Table
1.Different pipet used for pippeting different solutions.
3) 2ml of Benedicts solution was added to each test tube;the mixture was agitated by shaking
the tubes from side to side.The initial colour was recorded.
4) The test tubes was heated in the boiling water bath for 3 minutes.By using the test tube
clamp, the tubes was removed and any colour changes in the column colour after heating
was recorded.The conclusions regarding the presence/absence of simple carbohydrate was
recorded.

EXPERIMENT 2.2 : TEST FOR STARCH USING IODINS SOLUTION

MATERIALS

Test tube in the test tube rack

Sticker for labelling
Pippet
Dropper bottles containing Iodin solution
Stock bottle of :
i.
Distilled water (dH2 O)
ii. Glucose solution
iii.
Maltose solution
iv. Lactose solution
v. Sucrose solution
vi.
Starch solution

PROCEDURE
1) 3ml of the correct stock solutions was pipeted into the test tube as described in table 2.The
test tubes were labelled using stickers.
2) The colour of each solution in the column initial color was recorded.
3) 9 drops of Iodins solution was added to each test tube,the mixture was agitated by shaking
the tubes from side to side.The colour after adding the Iodins solution was recorded.

EXPERIMENT 2.3 TEST FOR LIPIDS

MATERIALS

Test tubes in test tube racks

Sudan III dye
Marker pen
Small ruler
Egg white solution
Honey solution
Corn oil
Distilled water
Labelling sticker

PROCEDURE

1) 4 test tubes was marked at 2 cm measured from the bottom of the tube.The top end of the
test tube was labelled as 1,2,3 and 4.
2) Each test tube was filled up to the 2 cm mark with one of the following: distilled water
(tube 1),honey solution(tube 2),corn oil(tube 3) and egg white solution(tube 4).
3) 10 drops of Sudan III dye was added to each test tube.
4) The tube was swirled to mix.The tubes was examined for result.The result was recorded.
5) The contents of all tubes in the container designated for Sudan III disposal was
discarded.Do not pour Sudan III down the sink drain.The test tube must cleaned throughly.

EXPERIMENT 2.4 TEST FOR PROTEINS USING BIURET REAGENT

MATERIALS

Test tubes in test tube rack

Small ruler
Marker pen
Distilled water
Label sticker
Honey solution
Corn oil
Egg white solution
10% NaOH solution
1% CuSO4

PROCEDURE
1) 4 test tubes was marked at 2cm and 4cm measured from the bottom of the test tube.The top
end of the test tube was labelled as 1,2,3 and 4.
2) Each test tube was filled up to the 2 cm mark with one of the following: distilled water
(tube 1),honey solution(tube 2),corn oil(tube 3) and egg white solution(tube 4).
3) 10% NaOH solution was added to each test tube up to the 4cm mark.
4) 5 drops of CuSO4 solution was added to each test tube and swirled to mix.The tubes was
examined for the appearances of any violet colour and the result was recorded.
5) The contents of the tubes was discarded and washed thoroughly with detergent and rinsed
them well.The excess water was shaked out.

RESULT

EXPERIMENT 2.1: TEST FOR SUGAR USING BENEDICTS SOLUTION

Tube
1
2
3
4
5
6
7
8
9
10

Contents
dh2O
Glucose
Maltose
Lactose
Sucrose
Starch
Fructose
Lemon juice
Orange juice
Non-diet soda

Initial
colour
Blue
Blue
Blue
Blue
Blue
Blue
Blue
Blue
Green
Blue

Color after
heating
Blue
Brick red
Brick red
Brick red
Blue
Blue
Brick red
Green
Brick - red
Brick -red

Conclusion
Absence of carbohydrate
Presence of carbohydrate
Presence of carbohydrate
Presence of carbohydrate
Absence of carbohydrate
Absence of carbohydrate
Presence of carbohydrate
Absence of carbohydrate
Presence of carbohydrate
Presence of carbohydrate

EXPERIMENT 2.2: TEST FOR STARCH USING IODINS SOLUTION

Tube
1
2
3
4
5
6

Contents
dh2O
Glucose
Maltose
Lactose
Sucrose
Starch

Initial
colour
Colorless
Colorless
Colorless
Colorless
Colorless
Brown

Color after
heating
Yellow
Yellow
Yellow
Yellow
Yellow
Blue black

Conclusion
Absence of starch
Absence of starch
Absence of starch
Absence of starch
Absence of starch
Presence of starch

EXPERIMENT 2.3 :TEST FOR LIPIDS

Tube
1
2
3
4

Contents
dh2O
Honey solution
Corn oil
Egg white solution

Initial
colour
Colorless
Brown
Colourless
Bright
yellow

Color after adding

Sudan III
Colorless
Brown
Red cloudy
Bright yellow

Conclusion
Absence of lipid
Absence of lipid
Presence of lipid
Absence of lipid

EXPERIMENT 2.4 TEST FOR PROTEINS USING BIURET REAGENT

Tube
1
2
3
4

Contents
dh2O
Honey solution
Corn oil
Egg white solution

Initial
colour
Colorless
Brown
Colourless
Bright
yellow

Color after adding

CuSO4
Colorless
Brown
Cloudy
Violet

Conclusion
Absence of protein
Absence of protein
Absence of protein
Presence of protein

DISCUSSION.

At the end of the experiment, the presence of carbohydrates,lipids and proteins

could be tested by using various test for starch,sugar,lipids and proteins.
The sugar was tested by using Benedicts solution.The color of the obtained
precipitate gives an idea about the quantity of sugar present in the solution.A greenish
precipitate indicates about 0.5 g% concentration; yellow precipitate indicates 1 g%
concentration; orange indicates 1.5 g% and red indicates 2 g% or higher concentration.In the
experiment the result obtained for the presence of higher sugar is glucose, maltose, lactose,
fructose, orange juice and also non-diet soda which is the color of the solution turned brickred after heating.
Next ,the starch was tested by using Iodins solution. Iodine dissolved in an
aqueous solution reacts with the starch, producing a purple-black colour. However, the
intensity of the colour decreases with increasing temperature. Also, the test cannot be done at
very low pH values due to the hydrolysis of the starch under these conditions.In this
experiment,starch solution change from brown colour to blue black after adding Iodins
solution.
The lipids was tested by using Sudan III dye solution.The solution will change
into cloudy dark red solution with the presence of lipid.This was proved by the corn oil
solution that changed from colorless solution into cloudy dark red solution.

And lastly the proteins was tested by Biuret reagent. The Biuret test is done to show
the presence of peptide bonds, which are the basis for the formation of proteins. These bonds
will make the blue Biuret reagent turn violet.This color change is dependent on
the number of peptide bonds in the solution, so the more protein, the more intense the

change.Obviously in this experiment, egg white solution turned into violet colour after adding
CuSO4.

CONCLUSION.
At the end of this experiment, the presence of the carbohydrate, proteins, and
lipids could be determined by using various of test.In order to obtain a good result the steps
must be followed with handlecare and avoid the mistakes that can disrupt the result such as
not using the same pipet for pippeting different solutions.

REFERENCES
1) https://en.wikipedia.org/wiki/Benedict's_reagent
2) http://igbiologyy.blogspot.com/2012/12/33-biuret-test-for-proteins.html
3)http://brilliantbiologystudent.weebly.com/ibrodine-test-for-starch.html

POST LAB QUESTION.

The chemical bond causes the Biuret reagent to react and to give the characteristic violet
colour is peptide bond.This color change is dependent on the number of peptide bonds in the
solution, so the more protein, the more intense the change.When the peptides are very short,
the solution turns a pink color, rather than violet.