Subtopic 4.

1
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Biological molecules
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4.1.1 () Contents
4.1.2 () 4.1.0 The big picture ()
4.1.1 Carbohydrates, fats and oils, and protein ()
4.1.3 ()
4.1.2 Practical: Testing for biological molecules ()
4.1.4 () 4.1.3 DNA (E) ()
4.1.4 Summary and key terms ()
4.1.5 ()
4.1.5 Checklist ()

Section 4.1.0

The big picture

Have you ever heard the phrase, ‘you are what you eat’? In many ways that phrase is true. The contents of the food we
eat are used by our bodies as energy, as well as the building blocks for the new cells we constantly produce by the
million.

Macronutrients that include carbohydrates, fats and oils, and proteins make up the majority of our food, while
micronutrients such as vitamins and minerals make up only a tiny minority. However, all are required for our bodies to
function properly and be as healthy as possible.
 You can see the amounts of these different nutrients in many of the foods you eat if you read the nutrition facts labels
found on the packaging (Figure 1).
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Figure 1. A food nutrition facts label.

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Source: U.S. Food and Drug Administration (https://commons.wikimedia.org/wiki/File:FDA_Nutrition_Facts_Label_2016.png), Public domain, via

4.1.4 () Wikimedia Commons

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How do we know that these nutrients are in the food? Food scientists can carry out a variety of tests to find out which
types of biological molecules are in different types of food. In this subtopic, you will learn about some of these
biological molecules, and will also have an opportunity to carry out some of the tests for them.

Before you move on to the next section, read through the learning outcomes below and answer the section questions to
check that you have the prior knowledge required to understand this subtopic.

 Learning intentions
By the end of this subtopic you will be able to explain the chemical composition of carbohydrates, fats and oils, and
proteins, and how to carry out tests for starch, reducing sugars, fats and oils, proteins and vitamin C.

If you take the extended course, you will also be able to describe the structure of DNA.

To see the full list of learning objectives, go to the Checklist (/schoolstaff/app/cambridge-igcse-biology-

fe2023/book/biological-molecules/biological-molecules/checklist/).
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Section questions
- Hide 3 questions
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Question 1
4.1.3 () Fill in the blanks

All living things require 1 nutrition to provide a supply of materials for energy, growth and development.
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Accepted answers and explanation

#1 nutrition
nutrients

Nutrition is a characteristic of life. All living organisms require a source of nutrition to provide them with the materials necessary for energy, growth and
development.

Question 2
Multiple choice

Hydrogen, carbon and nitrogen are examples of …

Choices

Correct choice #1

elements

Answer explanation
 Hydrogen, carbon and nitrogen are elements, which are pure substances consisting only of atoms that all have the

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same numbers of protons in their atomic nuclei.

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#2
compounds
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#3
minerals
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#4
gases
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Question 3
Short text

True or false?

DNA is found in the nucleus of all cells.

Correct answers

False
F
Answer explanation

Bacteria do not have a nucleus therefore their DNA can not be found in a nucleus. DNA is found in the nuclei of plant
eukaryotic cells.
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4.1.0 () Section 4.1.1

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Carbohydrates, fats and oils, and proteins
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4.1.4 () Carbohydrates
4.1.5 () Carbohydrates are compounds of:

carbon, C
hydrogen, H
oxygen, O.

One of the simplest carbohydrates is glucose, a simple sugar. Figure 1 shows the structure of glucose. Although the
chemical structure is beyond the requirements of the syllabus, it can help to build your understanding.
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4.1.3 () Figure 1. The structure of glucose.

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4.1.5 () Some carbohydrate molecules (like the sugar we normally eat) consist of two simple sugar molecules chemically joined
together. 

Larger carbohydrate molecules like cellulose, starch and glycogen are made from many glucose molecules joined
together. Table 1 describes three of these complex carbohydrates.

Table 1. Three complex carbohydrates made from glucose.

Name Function Source

starch energy and nutrient storage chloroplasts and cytoplasm in plant cells

cellulose structural support plant cell walls

glycogen energy and nutrient storage liver and muscles

 Exercise questions
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+ Show 1 question

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Fats and oils
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Like carbohydrates, fats and oils are compounds of:
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4.1.4 () carbon, C

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hydrogen, H
oxygen, O.

Fats and oils are very similar chemically, but fats are in the solid state at room temperature whereas oils are in the liquid
state. Fats tend to be found in animals, while oils tend to be found in plants. These substances are made from two
smaller molecules, glycerol and fatty acids. 

A molecule of a fat or oil is formed by the reaction of a glycerol molecule with three fatty acid molecules. Depending
on the fat or oil, the three fatty acid molecules can be the same or different. Figure 2 is a ‘block diagram’ to show the
general structure of fats and oils.
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Figure 2. The general structure of fats and oils.
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Proteins
Like carbohydrates, fats and oils, proteins contain:

carbon, C
hydrogen, H
oxygen, O.

However, all proteins also contain nitrogen atoms, N. Some proteins may also contain very small amounts of sulfur and
selenium.

Proteins are very large molecules made from smaller molecules called amino acids.

Exercise questions
 + Show 1 question

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4.1.0 () Amino acids

4.1.1 () There are over 20 naturally occurring types of amino acid. Each molecule contains two reactive groups of atoms that

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allow amino acids to join end to end. The very long molecules produced by this process are proteins. Figure 3 shows
the general structure of an amino acid. Although the chemical structure is beyond the requirements of the syllabus, it is
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can help to build your understanding.
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Figure 3. The general structure of an amino acid.

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Section questions
- Hide 3 questions

Question 1
Short text

State which smaller molecule starch, glycogen and cellulose are made from.

Correct answers

glucose
 Answer explanation

Starch, glycogen and cellulose are made from many glucose molecules, chemically joined together in different ways.
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4.1.0 () Question 2
Multiple choice
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Which two smaller substances are fats and oils made from?
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Choices
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Correct choice #1
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Glycerol and fatty acids
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Answer explanation

Fats and oils are made from two substances, fatty acids and glycerol. Starch, glycogen and cellulose are made from
glucose. Proteins are made from amino acids.

#2
Glycerol and amino acids

#3
Glucose and fatty acids

#4
Glucose and amino acids

Question 3
Multiple choice

Which elements are found in fats and oils?

 Choices

Correct choice #1
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Hydrogen, oxygen and carbon
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Answer explanation
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Like carbohydrates, the elements that are found in fats and oils are hydrogen, oxygen and carbon.
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Hydrogen, nitrogen and oxygen
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Hydrogen, carbon and nitrogen

#4
Nitrogen, carbon and oxygen

Section 4.1.2

Practical: Testing for biological molecules

Food samples can be analysed to determine whether certain biological molecules are present.

Aim
 To test for the presence of different biological molecules including reducing sugars, starch, fats and oils, protein, and 
vitamin C.
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Safety
4.1.2 () A full risk assessment must be carried out before starting this practical activity (see section 0.0.1
(/schoolstaff/app/cambridge-igcse-biology-fe2023/book/introduction/introduction/introduction-and-key-features/) for
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more information).
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4.1.5 () Benedict’s solution and biuret solution are irritants. Wear eye protection. Do not get biuret solution on your skin.
Take care with the hot water in the water bath.
Ethanol is highly flammable, harmful and a health hazard. Make sure there are no flames nearby.
Always ensure the opening of test tubes is pointed away from your face and anyone else’s.

Setting up your experiment

You will need the following equipment:

a variety of food samples

10 test tubes
two test tube racks
10 rubber bungs to fit test tubes
water bath set to 90 °C
Benedict’s solution
biuret solution
 ethanol
iodine solution
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DCPIP
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water
4.1.1 () dry erase pen
4.1.2 () glass stirring rod
dropper
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mortar and pestle.
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Running your experiment
1. Place five test tubes into each of your test tube racks and number each set from 1 to 5 with the pen.
2. Place 2 cm3 of your first food sample into each of the test tubes in the first rack. If food is solid, grind with a
mortar and pestle and mix with water to form a liquid.
3. Place 2 cm3 of water into each of the test tubes in the second rack.
4. To each test tube labelled 1, add four to five drops of Benedict’s solution. Swirl to mix and or stir with a glass
stirring rod then place into the hot water bath for up to five minutes.
5. Observe the colour changes and record your results (Figure 1).

 Study skills
In chemistry:

oxidation is gain of oxygen or loss of electrons

reduction is loss of oxygen or gain of electrons.

 In a chemical reaction involving these processes, one substance is oxidised while another is reduced. Some
carbohydrates are described as reducing sugars. They can cause the reduction of other substances (and become
4.1 () oxidised during this process). Glucose is an example of a reducing sugar.

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Figure 1. The results of testing for reducing sugar using Benedict’s solution.

Exercise questions
- Hide 1 question

Question 1

Multiple choice
 Which colour would you expect to see as result of a Benedict’s test for reducing sugar, if there were only trace amounts
of sugar in the solution?
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Choices

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Correct choice
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yellow or green

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Answer explanation

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Trace amounts of reducing sugar will produce a green or yellow colour in a Benedict’s test.

red or orange

red or blue

purple or black

    6. To each test tube labelled 2, add four to five drops of iodine solution.

    7. Place a bung in the test tubes and shake both gently for 10 seconds.

    8. Observe the colour changes and record your results (Figure 2).
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Figure 2. Negative (orange-brown) and positive (blue-black) iodine tests for starch.

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      9. To each test tube labelled 3, add 2 cm3 of ethanol.

    10. Place bungs on the test tubes and shake gently.

    11. Filter both solutions using filter paper into new test tubes. Discard the residue.

    12. Carefully add 2 cm3 of water to both mixtures, watching closely when the water hits the solutions.

    13. Record your observations (Figure 3).

Figure 3. A positive (white/milky colour) ethanol emulsion test for a fat or oil. The negative result will show no colour change.
     14. To each test tube labelled 4, add four to five drops of biuret solution.

    15. Place bungs on the test tubes and shake gently for 15 seconds.

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    16. Observe the colour changes and record your results (Figure 4).
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Figure 4. Negative (blue) and positive (violet/lilac) biuret tests for proteins.

    17. To each test tube labelled 5, add five drops of DCPIP, which is blue.

    18. Gently swirl the mixtures for 30 seconds.

    19. Observe the colours and record your results (Figure 5).
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4.1.3 () Figure 5. The DCPIP test for vitamin C. A positive result for vitamin C is a change from blue to colourless. If the blue colour
remains, there is no vitamin C.
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Analysing your results

Test for reducing sugars

If a reducing sugar is present, the final colour can range through green, yellow, orange and finally red (Figure 1). The
range of colours can make this test semi-quantitative, in that the more the colour changes, the more of the Benedict’s
solution has reacted and the more reducing sugar was present in the original sample.

If no reducing sugar is present, the mixture remains blue.

Test for starch

As shown in Figure 2, if starch is present, the mixture will become blue-black.

If no starch is present, the mixture will remain orange-brown.

 Test for fats and oils
4.1 () If fat or oil is present, when the water is added it will create a white or milky cloud as it hits the solution (Figure 3). If
4.1.0 () the food sample does not contain fat or oil, there will be no change. Fats and oils are less dense than water, so if it is left
to settle you may see a layer of oil or fat on the surface.
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Test for protein
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If a protein is present, the mixture will become light purple (Figure 4). If no protein is present, the mixture will remain
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light blue.
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Test for vitamin C

When vitamin C is present in a food sample the blue colour from the DCPIP will disappear (Figure 5). If a large
amount of vitamin C is present, the blue colour may disappear permanently from the liquid DCPIP. If only a small
amount of vitamin C is present, the blue colour may disappear slightly from the liquid that is in contact with the food
sample.

 Making connections
The DCPIP test can be used to determine amounts of vitamin C in different foods. To do this, the scientist first
needs to create a calibration curve for reference. This is done by repeating the DCPIP test on several solutions that
contain different, but known concentrations of vitamin C.

Once you have a calibration curve, you can test food samples by adding DCPIP drop by drop, using a graduated
pipette or a burette, until the blue colour no longer disappears. Record this volume of DCPIP. Plot this volume on
the calibration curve and where it intersects is the concentration of vitamin C in your sample.
 This method is used to test foods such as fruit juices or multivitamin tablets.

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Conclusion
4.1.2 () The video shows the tests being carried out using a similar method to that described here and the results you should
expect to see.
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Food Tests - Iodine, Biuret, Benedict's, Ethanol, DCPIP
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Discuss any difficulties you had in making your measurements. Can you suggest ways to change or expand the
practical?

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 Section questions
- Hide 4 questions
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Question 1
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Multiple choice

4.1.1 () Which of the following turns blue-black with iodine?

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Choices
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Correct choice #1

4.1.4 () Starch
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Answer explanation

Only starch turns blue-black in the presence of iodine.

#2
Glucose

#3
Fats

#4
Proteins

Question 2
Multiple choice

Which of the following tests requires a boiling water bath?

Choices
 Correct choice #1

Benedict’s test
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Answer explanation
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Only the test for reducing sugars, using Benedict’s solution, requires a boiling water bath.
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#2
Iodine test
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#3
Biuret test
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#4
Testing with DCPIP

Question 3
Short text

In a positive ethanol emulsion test for fats and oils, state which layer (top or bottom) is cloudy white?

Correct answers

top
the top
Answer explanation

Fats and oils are less dense than water, so the cloudy white emulsion forms in the top layer.

Question 4
Multiple choice
 What colour is seen if protein is present in a biuret test?

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Choices

4.1.0 () Correct choice #1

4.1.1 () A violet/lilac colour

4.1.2 () Answer explanation

4.1.3 () Biuret turns from pale blue to violet/lilac in the presence of proteins.

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#2
4.1.5 () A green colour

#3
A blue colour

#4
A brick-red colour

Section 4.1.3

DNA (E)
Extended 
 DNA  (deoxyribonucleic acid) is the genetic material in living cells. It contains the instructions for each living
organism. Nearly every cell in your body has a complete copy of all of your DNA. Your DNA came from your
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parents; half from your mother and half from your father.
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4.1.1 () DNA molecules are very large molecules containing many atoms. DNA consists of two strands that coil
together to form a double helix (two spirals that wrap around each other). Each strand contains three main
4.1.2 ()
chemicals: phosphates, sugars called deoxyribose and nitrogen-containing bases (Figure 1). The phosphates
4.1.3 ()
and sugars form the backbone of the strands, while the bases are located in the centre of the molecule.
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4.1.5 ()

Figure 1. Part of a molecule of DNA.

Pairs of bases form hydrogen bonds between the two strands. This keeps the double helix structure together.

There are four bases: A, T, C and G. They always pair up this way:
 A with T

4.1 () C with G.

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These are called complementary base pairs. This complementary pairing is key to how DNA functions and
4.1.1 () how it is used in cells, as you will learn in subtopic 17.1 (/schoolstaff/app/cambridge-igcse-biology-
4.1.2 () fe2023/book/inheritance/chromosomes-genes-and-proteins/the-big-picture/).

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4.1.4 ()  Study skills 

4.1.5 () You do not need to know the full names of the bases. If you are curious to know, they are called adenine,
thymine, cytosine and guanine. Also, the small units that form DNA, containing a phosphate, a sugar and a
base, are known as nucleotides (again, this term is beyond the requirements of the syllabus).

 Activity 

Use the following drag and drop activity to check your understanding of complementary base pairing in
DNA.
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 Check
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 Activity 

Design and build your own DNA model to show your understanding of DNA structure. You can use lots of
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different materials for this, candies and sweets, modelling clay, paper, pipe cleaners, etc.
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Watch the following video for an overview of DNA structure. It goes into a bit more detail than you need to
know, but is very clear and informative.

DNA Structure
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4.1 () Section questions

- Hide 3 questions
4.1.0 ()

Question 1
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Multiple choice

4.1.2 ()
Extended
4.1.3 ()
What shape does a DNA molecule have?
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4.1.5 () Choices

Correct choice #1

Double helix

Answer explanation

The shape of a DNA molecule is a double helix.

#2
Single helix

#3
Triple helix

#4
Double spiral

Question 2
Multiple choice
 Extended
4.1 ()
Which are the correct complementary base pairs in a molecule of DNA?
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Choices
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Correct choice #1
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A with T, G with C
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Answer explanation
4.1.4 ()
Base A always pairs with base T, while base G always pairs with base C.
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#2
A with A, G with G

#3
G with T, A with C

#4
T with T, C with C

Question 3
Fill in the blanks

Extended

DNA is the  1 genetic material of a living cell.

Accepted answers and explanation

 #1 genetic
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DNA is the genetic material of a living cell, containing the instructions for each living organism.
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Section 4.1.4
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Summary and key terms
Carbohydrates are composed of the elements carbon, hydrogen and oxygen. Small carbohydrate molecules called
sugars, like glucose, form large carbohydrate molecules such as starch, glycogen and cellulose.
Fats and oils are also composed of the elements carbon, hydrogen and oxygen. The small molecules glycerol and
fatty acids are combined to form the larger fat and oil molecules. Most commonly, one glycerol is combined with
three fatty acids.
Proteins are composed of the elements carbon, hydrogen, oxygen and nitrogen. They also sometimes have sulfur
and selenium. Proteins are made of smaller molecules known as amino acids.
A variety of tests can be used to identify biological molecules. Benedict’s solution will test for reducing sugars,
iodine tests for starch, biuret solution is used to test for protein, ethanol is used for the emulsion test for fats and
oils and DCPIP is used to test for the presence of vitamin C.

Extended 
 DNA structure is composed of a double strand coiled together in the shape of a double helix. Each strand
contains bases which form bonds that hold the strands together. These bases always pair in the
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same complementary way. A with T and C with G.
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4.1.2 ()  Key terms

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Review these key terms. Do you know them all? Fill in as many gaps as you can using the terms in
4.1.4 () this list.

4.1.5 ()
1. Glucose is an example of a . Plants store energy in the form of ,
while animals use to store energy.

2. Plant cell walls are made of the carbohydrate .

3. are made of glycerol and .

4. are made of chains of many .

5. [Extended] The molecule which is the genetic material of a cell is .

fatty acids DNA Fats carbohydrate Proteins cellulose starch amino acids

glycogen

 Check

 Reuse
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Section 4.1.5

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Checklist
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4.1.5 ()
 What you should know
After studying this subtopic, you should be able to:

Core

List the chemical elements that make up: carbohydrates, fats and proteins.
State that large molecules are made from smaller molecules, limited to:
starch, glycogen and cellulose from glucose
proteins from amino acids

fats and oils from fatty acids and glycerol.

Describe the use of:
iodine solution test for starch
Benedict’s solution test for reducing sugars

biuret test for proteins

ethanol emulsion test for fats and oils

DCPIP test for vitamin C.

 Extended

4.1 () Describe the structure of a DNA molecule:

two strands coiled together to form a double helix
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each strand contains chemicals called bases
4.1.1 ()
bonds between pairs of bases hold the strands together
4.1.2 () the bases always pair up in the same way: A with T, and C with G.

4.1.3 ()

4.1.4 ()

4.1.5 ()

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