Lab activity in preparation for

practical exam
AS level biology

Reducing and
non-reducing
sugars test
 A Food Test  is a chemical test
which determines the presence
of nutrients/biological molecules
commonly found in food.

These biological molecules

include sugars - reducing and
Food Tests non-reducing,  starch, protein
and lipids. 

Food tests use reagents which

react predictably with the
biological molecules  to give
observable color changes.
Food Tests

For each of the food tests you should be able to:

• Name at least 3 foods for which you would predict a positive

result
• Name the reagents used
• Outline the steps in sequential order
• Explain the principle or basis of the test
• Interpret the results obtained
What is Benedict's Test for Reducing
Sugars?

Benedict's Test for non-

The principal reagent in
reducing Sugars  is a test
Benedict's Test for Reducing
which determines the
Sugars is Benedict's Solution
presence of non-reducing
which contains
sugars in a test solution.
• copper(II) sulphate
• sodium carbonate
• sodium citrate
Sugars are classified as reducing or non-
reducing based on their ability to act as a
reducing agent during the Benedict's Test.

A reducing agent donates electrons

during a redox reaction and is itself
oxidized.
What are
The aldehyde functional group is the
reducing
reducing agent in reducing sugars. sugars?

Reducing sugars have either an aldehyde

functional group or have a ketone group -
in an open chain form - which can be
converted into an aldehyde.
 Reducing sugars are simple sugars and
include all monosaccharides and most
disaccarides.

Some examples of monosaccharides are

glucose, fructose and galactose.

Reducing
sugars Examples of reducing disaccharides are
lactose and maltose.

Note that the disaccharide sucrose is

not a reducing sugar. In fact, sucrose is
the most common non-reducing sugar.
What is the procedure for the Benedict's Test for
reducing sugars?
What is the procedure for the Benedict's
Test for reducing sugars?
• Add 2 cm3 of the sample solution to a test tube. 

• Add an equal volume of Benedict's solution to the test tube and swirl
or vortex the mixture.
• Leave the test tube in a boiling water bath for about 5 minutes, or
until the colour of the mixture does not change.
• Observe the colour changes during that time as well as the final
colour.
• To prepare a control, repeat the steps above using 2 cm3 of distilled
water instead of sample solution
What are the expected observations for the
Benedict's Test for reducing sugars?

Observations Interpretations

No Colour Change No reducing sugars present

(Blue)

Green Trace amounts of reducing sugars present

Yellow Low amounts of reducing sugars present

Orange Moderate amounts of reducing sugars present

Brick-red Large amounts of non-reducing sugars present

 Reducing Sugars  have an
aldehyde functional group
which can reduce soluble
What is the copper (II) ions  - in copper (II)
principle of sulphate -  to insoluble copper
(I) ions - in copper (i)oxide.
the
Benedict's
Test for The copper (I) oxide is seen as
a precipitate.
reducing
sugars?
 Reduced Species. The blue
copper(II) ions from copper(II)
sulphate are reduced to red
copper(I) ions by the aldehyde
groups in the reducing sugars.
State the
role of
copper
sulphate in This accounts for the colour changes
observed.
Benedict's
Solution. The red copper(I) oxide formed is
insoluble in water and is
precipitated out of solution. This
accounts for the precipitate formed.
Benedict Test • https://youtu.be/Lt7RCIfudYQ
Reducing
Sugars
 Benedict's Test for non-reducing
Sugars  is a test which
determines the presence of non-
reducing sugars in a test
solution.
What is
Benedict's
Test for non-
Non-reducing sugars are sugars
reducing which do not have an aldehyde
sugars? functional group  - the reducing
species. As non-reducing sugars
do not have the aldehyde group,
they cannot reduce copper (I)
(blue) to the copper(II) (red).
 Sucrose is the most common
disaccharide non-reducing sugar.

Sucrose is a non-reducing sugar

 • Many students confuse the Benedict's
Test for Non-Reducing Sugars with the 
Careful Benedict's Test for Reducing Sugars,
 perhaps because some of the steps in
the procedure are similar.
 Add 2 cm3 of the sample solution to a test tube.

What is the Add 1 cm3 of dilute hydrochloric acid and boil for

procedure one minute.

for the
Benedict's Allow the tube to cool and then neutralize the acid
Test for with sodium hydrogen carbonate. Exercise caution
due to 'fizzing' or effervescence.
Reducing
Sugars? Check with blue litmus paper or pH paper. Do not
worry if the resulting solution is slightly alkaline;
Benedict's solution is also alkaline since an alkaline
medium in needed for oxidation of the copper (II)
ions by the sugars
 Leave the test tube in a boiling
water bath for about 5
minutes, or until the colour of
What is the the mixture does not change.
procedure
Observe the colour changes
for the during that time as well as the
Benedict's final colour.
Test for
Reducing
To prepare a control, repeat
Sugars? the steps using 2 cm3 of
distilled water instead of
sample solution.
Observations Interpretations

No Colour Change No reducing sugars present

(Blue)

Green Trace amounts of reducing sugars present

Yellow Low amounts of reducing sugars present

Orange Moderate amounts of reducing sugars present

Brick-red Large amounts of non-reducing sugars present

 The test for non-reducing
sugars is often conducted on
a food sample which
What results tested negative for reducing
are expected sugar.
if a food
sample has
already been If reducing sugars have been
tested for shown to be present, a
reducing heavier precipitate is often
sugars? observed when the test for
non-reducing sugar is
conducted.
 What is the • Disaccharides are hydrolyzed to their
principle of constituent monosaccharides when
the boiled in dilute hydrochloric acid. 

Benedict's The monosaccharide products of

hydrolysis are reducing sugars i.e. have
Test for non- the aldehyde functional group and can
reducing reduce copper in the presence of alkali 
producing the colour changes.
Sugars?
 Why was
sodium • Neutralization. Sodium hydrogen
carbonate is necessary for
hydrogen neutrallization as the reduction of the
carbonate copper(II) ions will not take place in
acidic conditions - of excess acid is
added to the present.
mixture?
Benedict Test
Non- • https://youtu.be/QU0VBcHnQOk
Reducing
Sugars
 The Iodine Test for Starch is used to
determine the presence of starch in
biological materials. The test can be
qualitative or quantitative.

As a Biology Student, you will be testing for

the presence of this complex carbohydrate
in foods or in leaves as part of a 
What is the photosynthesis experiment.
Iodine Test The sole reagent required for the test
for Starch? is bench iodine solution  (0.1 M potassium
triiodide solution).
 What is the
procedure • The procedure  for the iodine test for
starch depends on whether the test
for the iodine sample is a solid or liquid.
test for
starch?
What is the procedure for the iodine test
for starch?

SOLID Peel off the skin of any vegetables e.g. potato and fruits
as these are often impermeable. Use a clean spatula to
SAMPLE remove samples of powdered food. Avoid cross
contamination with other foods.

Add a few ( 2-3) drops of bench iodine solution

potassium to a piece of solid food on a white tile.

Make observations.
What is the procedure for the iodine test
for starch?

LIQUID Add 10 cm3 of the liquid food sample to

SAMPLE a clean, dry test tube.
Add about 5 drops of iodine solution to
the test tube.
Note any colour changes.

To prepare a control, perform steps 1 -3

for de-ionized water.
What are the expected observations and
interpretation of the iodine test for starch?

Observation Interpretation

No change (Iodine remains Starch is not present

brown)
Starch
A blue-black colour
develops
Describe the structure of starch and state
which structural feature is key to the
colour change in the iodine test for starch.

• 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 molecule
 Linear - both straight and helical - molecules of starch are referred to as Amylose.

Where branched molecules of starch are called Amylopectin.

 Natural starches - from plants -
consist of a mixture of amylose
(10 - 25%) and amylopectin (75-
90%).The structure of the helical
amylose is key to the Iodine-
starch reaction. A helix is a coil or
a spring.
Potassium
Iodide The tri-iodide and penta-iodide
ions formed are linear and slip
inside the helix of the amylose
(form of starch).
 • Iodine on its own (small non-polar molecule)
is insoluble in water. Therefore Potassium
triiodide solution - Iodine dissolved in
Describe the potassium iodide solution - is used as a
reagent in the test.
composition of
the To be more specific, potassium iodide
iodine/potassium dissociates, and then the Iodide ion reacts
reversibly with the Iodine to yield the
tri-iodide triiodide ion. A further reaction between a
reagent in the triiodide ion and an iodine molecule yields
the pentaiodide ion.
iodine test for
starch. Since molecular iodine is always present in
solution, the bench iodine solution appears
brown; the iodide and triiodide pentaiodide
ions are colourless.
 • The starch-iodide complex is formed as
Explain the charge - recall electrons are charged particles
- is transferred between the starch and
principle or iodide ions  - tri-iodide or pentaiodide.
the basis of The transfer of charge between the starch
the colour and the iodide ion changes the spacing
between the energy levels/ orbitals.
change in
This change results in the starch-iodide
the  Iodine complex absorbing light at a different
Test for wavelength - than any other species
aforementioned - resulting in an intense
Starch. purple colour; Biologists call this colour blue-
black.
 Explain the
principle or the
basis of the
colour change in
the  Iodine Test
for Starch.
Potassium Iodide test
https://youtu.be/J5eaJc854uQ
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