Edexcel IAL Biology A Level

Core Practical 1
Use a semi-quantitative method with Benedict’s reagent to estimate
the concentrations of reducing sugars and with iodine solution to
estimate the concentrations of starch, using colour standards.

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Benedict’s test for reducing sugars
Independent variable: Concentration of reducing sugar
Dependent variable: Colour change of Benedict’s reagent

All sugars can be classified as being reducing or non-reducing. A test with​ ​Benedict’s reagent​ ​can
not only reveal if it is a reducing sugar or not, but also ​how strong​ of a reducing agent it is.

Equipment list
● Benedict’s reagent
● Test tubes
● Water bath set to 90 °C
● Pipette(s)
● Test tube rack
● Stop watch

Method
1. Pipette 5 cm³ of the ​solution being tested​ into the test tube followed by 2 cm³ of
Benedict’s reagent.
2. Place the test tube into a test tube rack and then into the ​water bath​ ​and leave it for exactly
2 minutes.
3. Remove the test tube and observe its ​colour.

This test is semi-quantitative because by observing the colour change on a scale from blue to red
it’s possible to estimate the ​concentration of the reducing sugar. ​A ​blue colour is a negative
result​ ​since this is the colour of the Benedict’s reagent, while ​colours closer to brick-red​ ​give a
positive result ​with an ​increasing​ ​concentration of reducing sugar.

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Iodine test for starch
Method
1. Pipette 2 cm³ of the test solution into a test tube and add 2 drops of​ ​potassium iodide
solution.
2. A colour change from​ ​brown to blue-black​ ​indicates the presence of ​starch.

Use of colour standards

The concentrations of both starch and reducing sugars can be estimated using ​colour standards
which is when the benedict’s test and iodine test for starch are carried out on ​known
concentrations of starch​ ​and ​reducing sugars. ​The colours produced are known as colour
standards to which the results of testing unknown solutions can be compared to estimate their
concentrations.

Risk assessment
Hazard Risk Precaution

Liquids Spillage that could cause Wipe up any liquid spillages as

surfaces to be slippery soon as they occur
leading to an accident Put lids on bottles
Put bottles away once used
Keep away from edge of desk

Hot water bath Scalding Take care in removing and

replacing the water bath lid and
have a first aid kit nearby
Remove test tubes from the water
bath with tongs
Keep away from edge of desk

Glassware Cuts from sharp objects Take care when handling glass
objects
Keep away from edge of desk

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Edexcel IAL Biology A Level
Core Practical​ 2

Investigate the vitamin C content of food and drink.

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Equipment list
● 1% DCPIP solution
● 5 cm³ syringes
● Test tubes
● Test tube rack
● Pipettes
● 4 different fruit juices
● 1% vitamin C solution

Method
1. Use a pipette to transfer 1 cm³ of DCPIP solution to a test tube.
2. Then fill a 5 cm³ syringe with 5 cm³ of vitamin C solution.
3. Add the vitamin C solution from the syringe ​one drop at a time ​to the test tube containing
the DCPIP, being sure to ​gently shake​ ​the test tube after each
addition.
4. Stop adding the vitamin C once the ​blue DCPIP​ has turned
colourless.​ ​Record the​ ​volume ​of the solution that has been
added to cause the colour change.
5. Repeat steps 1-4 to obtain 2 further results for the ​vitamin C
solution​ and then calculate a ​mean volume​ added, first
discarding any anomalous results.
6. Then repeat steps 1-5 using each of the ​4 fruit juices​ in place
of the vitamin C solution.

Risk assessment

Hazard Risk Precaution

DCPIP Could irritate skin Avoid skin contact and always

transfer it using pipettes/ a
syringe
Wear eye protection

Fruit juices Could have been kept in the Do not drink it

school lab for a long time.

Glassware Cuts from sharp objects Take care when handling

glass objects
Keep away from edge of desk

Liquids Spillage that could cause Wipe up any liquid spillages

surfaces to be slippery leading as soon as they occur
to an accident Put lids on bottles
Put them away once used
Keep away from edge of desk

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Results table

Drink used Volume 1 Volume 2 Volume 3 Mean volume needed to

(cm³) (cm³) (cm³) decolourise DCPIP (cm³ )

Vitamin C
solution

Fruit juice 1

Fruit juice 2

Fruit juice 3

Fruit juice 4

Conclusion
This experiment can be used to estimate the ​concentration of Vitamin C​ ​in fruit juices since
vitamin C is an​ ​antioxidant ​and DCPIP is an ​oxidising agent.​ ​When vitamin C is added to DCPIP
the DCPIP is reduced which eventually causes a colour change from ​blue to colourless​.​ Since
the volume and concentration of both the vitamin C solution and DCPIP are known, the
concentration of vitamin C in each fruit juice can be calculated by comparing it to the standard
solution.

The calculation
1 cm³ of 1% vitamin C solution ​contains ​10 mg of vitamin C. ​Using this information you can
calculate the mass of vitamin C needed to ​decolourise a given volume of DCPIP​ (for instance if
1.6 cm³ were needed that would be 16 mg). From there you can calculate the concentrations of the
fruit juices.

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 Edexcel IAL Biology A Level
Core Practical​ 3

Investigate membrane properties including the effect of alcohol and

temperature on membrane permeability.

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Beetroot can be used to investigate the ​permeability​ of cell membranes, since when its cell
membranes are damaged a ​coloured pigment,​ that gives beetroot its purple colour, ​leaks out.
The higher the permeability of the membrane, the more pigment is released outside of cells. The
permeability of cell membranes is affected by a number of factors including ​temperature, alcohol
and solvent concentration.

Independent variable: Temperature or alcohol concentration

Dependent variable: Absorbance reading of liquid

Equipment list
● Beetroot
● Scalpel
● 4 test tubes
● Colorimeter
● Cuvettes
● Stopwatch
● Water baths
● Pipette

Method

The following method describes how to investigate how cell membrane permeability changes with
temperature; the effect of alcohol can also be investigated by placing the beetroot cubes in
solutions containing ​differing concentrations of ethanol​ instead, and then following the same
colorimetry technique.

1. Use the scalpel to cut 4 equal pieces of beetroot from the same beetroot and ​rinse each
piece with water ​to remove any pigment already released from the cutting.
2. Use a pipette to fill 4 test tubes with 5 cm³ of water and place a cube of beetroot in each
test tube.
3. Place each of the test tubes in a​ water bath​ at the following temperatures - 0°C, 20°C,
40°C, and 60°C. Remove the test tubes from the water baths after ​exactly 5 minutes​ and
remove the beetroot pieces ​leaving only
the coloured liquid.
4. Set up a colorimeter by giving it 5 minutes
to ​stabilise ​and by first measuring its
absorbance with​ pure water in a cuvette.
Then use a pipette to fill a cuvette with
liquid from each of the 4 test tubes to
measure and ​record their absorbance
with the colorimeter.
5. Plot a graph of temperature against
absorbance.

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Risk assessment

Hazard Risk Precaution

Broken glass Cuts from sharp object Take care when handling
glass objects
Keep away from edge of desk

Ethanol Highly flammable and volatile Keep away from naked flames
and extreme heat
Put the lid on once used
Keep away from edge of desk

Results table

Temperature (​°C) Colorimeter absorbance

20

40

60

Alcohol concentration (mol/dm³) Colorimeter absorbance

Graph

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Conclusion

Temperature
● Below 0 °C - ​Ice crystals ​form in the membrane, ​piercing it​ and allowing molecules
including beetroot pigment to leak out.
● 0°C - 40°C - As temperature increases the ​phospholipids​ in the membrane ​gain kinetic
energy​ and ​move more​, increasing the permeability.
● Over 40°C -​ ​Proteins​ ​in the membrane ​deform at high temperatures​ meaning they
cannot control​ ​what goes in and out of the cell - increase membrane permeability.
Furthermore, the ​phospholipids ​start to ​melt​ and ​expanding water inside the cell​ puts
increasing pressure​ on the membrane.

Alcohol
Both ​ethanol ​and the ​phospholipid bilayer​ that makes up the cell membrane are ​non-polar.​ This
means the ​lipids dissolve in solutions containing ethanol​, thus disrupting the membrane. The
higher the concentration of lipids the more the membrane will be disrupted allowing more pigment
to leak out.

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 Edexcel IAL Biology A Level
Core Practical​ 4

Investigate the effect of temperature, pH, enzyme concentration and

substrate concentration on the initial rate of enzyme-catalysed
reactions.

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This experiment investigates the effects of different factors on the rate of the digestion of milk by
the enzyme ​trypsin​. As the milk protein called casein is digested the ​white, opaque milk colour
becomes ​more pale and translucent​, eventually turning ​colourless. More light​ passes through
the ​transparent and lighter solutions​ so a colorimeter can be used to measure the absorbance
of the solution which in turn indicates the rate of reaction of the experiment. The faster the rate of
the reaction, the ​lower​ the initial absorbance reading will be - ​as paler solutions absorb less
light.

Equipment list
● Colorimeter
● Cuvettes
● Water baths at the following temperatures: 20°C, 30°C, 40°C, 50°C and 60​°​C
● pH buffers at the following pHs: 5, 6, 7, 8 and 9
● Distilled water
● Stopwatch
● Test tubes
● Test tube rack
● Protease enzyme solution - 1% trypsin enzyme solution
● 2% semi-skimmed milk
● Pipettes
● Tongs

Method

Factor 1 - the effect of temperature

1. Prepare the 5 water baths and ​monitor their temperatures​ using thermometers.
2. Use a pipette to add 2 cm³ of trypsin solution to 5 test tubes and place one in each water
bath.
3. Then use a pipette to add 2 cm³ of milk to 5 test tubes and again place one in each water
bath.
4. Start the stopwatch and time for 5 minutes, ​allowing the contents of each test tube to
reach the temperature of the water bath.

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 5. Pipette 2 cm³ of trypsin and 2 cm³ of distilled water into a cuvette and take a colorimetry
reading to set the reference absorbance of the colorimeter to ​zero.
6. Now remove the milk test tube and the trypsin test tube from the 20°C water bath and add
the contents of each to a single cuvette, shaking it and then immediately placing it in the
colorimeter.
7. Take an initial reading and then record the absorbance reading at 15 second intervals for 5
minutes or until there is only a ​slight change ​in absorbance between readings.
8. Repeat steps 6-7 for the remaining 4 temperatures, being sure to​ rinse the cuvettes
between each temperature and setting the ​absorbance to zero with the water and
trypsin cuvette each time.

Factor 2 - pH

1. Prepare the reference cuvette by adding 1 cm³ of buffer solution, 1 cm³ of trypsin and 2 cm³
of water to it and setting the colorimeter absorbance of this to ​zero.
2. Prepare 5 cuvettes each containing 1 cm³ of a ​different pH buffer solution​ and 1 cm³ of
trypsin solution. Prepare a further 5 cuvettes all containing 2 cm³ of milk solution.
3. Add the contents of 1 of the milk cuvettes to the cuvette containing the pH 5 buffer solution
then ​immediately​ place it in the colorimeter.
4. Take an initial reading and then record the absorbance reading at ​15 second intervals​ for
5 minutes or until there is only a ​slight change​ in absorbance between readings.
5. Repeat steps 3 and 4 with the remaining 4 pH buffer solutions.

Factor 3 - Enzyme concentration

1. Prepare these 10 cm³ solutions of different enzyme concentrations using the following
quantities of distilled water and 1% trypsin solution:

Solution Volume of trypsin Volume of distilled

concentration (%) (cm³) water (cm³)

1 10 0

0.8 8 2

0.6 6 4

0.4 4 6

0.2 2 8

2. Pipette 2 cm³ of trypsin and 2 cm³ of distilled water into a cuvette and take a colorimetry
reading to set the reference absorbance of the colorimeter to ​zero.
3. Add 2 cm³ of milk to a cuvette, then add 2 cm³ of the 1% enzyme solution to the cuvette.
Mix quickly then place in the colorimeter.

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 4. Take an initial reading and then record the absorbance reading at ​15 second intervals​ for
5 minutes or until there is only a ​slight change​ in absorbance between readings.
5. Repeat steps 3 and 4 with the remaining 4 enzyme solutions.

Factor 4 - substrate concentration

1. Prepare these 10 cm³ solutions of different substrate concentrations using the following
quantities of distilled water and 2% milk solution:
○
Solution Volume of milk (cm³) Volume of distilled
concentration (%) water (cm³)

1 5 5

0.8 4 6

0.6 3 7

0.4 2 8

0.2 1 9

2. Pipette 2 cm³ of trypsin and 2 cm³ of distilled water into a cuvette and take a colorimetry
reading to set the reference absorbance of the colorimeter to zero.
3. Add 2 cm³ of milk to a cuvette, then add 2 cm³ of the 1% milk solution to the cuvette. ​Mix
quickly ​then place in the colorimeter.
4. Take an initial reading and then record the absorbance reading at ​15 second intervals​ for
5 minutes or until there is only a slight change in absorbance between readings.
5. Repeat steps 3 and 4 with the remaining 4 milk substrate solutions.

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Risk assessment

Risk Hazard Precaution

Liquids Spillage that could cause Wipe up any liquid spillages

surfaces to be slippery leading as soon as they occur
to an accident Put lids on bottles and put
them away once used
Keep away from edge of desk

Hot water bath Scalding Take care in removing and

replacing the water bath lid
Have a first aid kit nearby
Remove test tubes from the
water bath with tongs
Keep away from edge of desk

Glassware Cuts from sharp objects Take care when handling

glass objects
Keep away from edge of desk

Trypsin May cause an allergic reaction Wear gloves when handling to

or respiratory problems if avoid skin contact, if it gets on
inhaled skin rinse thoroughly with cold
water

Results table

Time (seconds) Absorbance reading

15

30

45

60

...

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 Edexcel IAL Biology A Level
Core Practical​ 5

Use a light microscope to make observations and labelled drawings

of suitable animal cells.
Use a graticule with a microscope to make measurements and
understand the concept of scale.

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Equipment list
● Light microscope
● Microscopic slide, specimen and cover slip

Method
1. Begin by securing the prepared
slide to the stage of the
microscope, using the
accompanying clip.
2. Rotate the revolving nosepiece to
line up the ​lowest power​ objective
lens.
3. Look through the eyepiece and use
the coarse adjustment knob to
lower and raise the stage until an
image comes into focus.
4. Then use the fine adjustment knob
to focus the image further.
5. In order to see and study cells
steps 3 and 4 may need to be
repeated using a ​higher power objective lens​ - 3 are usually found on the nosepiece.
6. Produce a biological drawing of cell(s) observed.

Biological drawings

When producing biological drawings, in this case of cells being observed under the microscope,
certain rules should be followed:
● Draw in pencil only
● Use blank white paper
● Do not sketch - draw clear, unfeathered
lines only
● Include labels
● Label lines should be drawn with a ruler
and not cross over each other
● Do not shade the drawing - for instance
when indicating the thickness of
membranes / cell walls instead draw 2
lines
● Include the magnification of the
microscope in the title
● Make diagrams as simple as possible and draw them an appropriate size so everything can
clearly be seen
● Include a scale bar where possible

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Using a graticule

By using an eyepiece graticule and a stage micrometer the ​size of cells​ and ​potentially their
organelles​ can be accurately calculated. An eyepiece graticule is a ruled line with regular intervals
that appears when you look down the microscope and a stage micrometer is a microscopic slide
with an accurate scale and is used to calibrate the eyepiece as follows:

1. Place the stage micrometer on the stage and ​line up its scale with the eyepiece ​graticule.
On the stage micrometer 1 division is equal to 0.1mm.
2. Count how many eyepiece divisions are equal to 1 stage division. This can be used to
calculate the​ length​ of each eyepiece division at this magnification - for instance, if 10
eyepiece divisions are equal to 1 stage division (0.1 mm) then each eyepiece division
measures 0.1 ÷ 10 = 0.01 mm.
3. The stage micrometer can now be replaced with the microscopic slide containing the
specimen. It is important​ not to change any adjustments or objective lenses at this
point.
4. When observing cells now, their length or size of organelles can be calculated accurately by
counting the number of eyepiece divisions - for instance a cell that is 3 eyepiece divisions

wide would be 0.03 mm wide.

The following formula can also be used to calculate ​magnification​ and​ object and image sizes
when observing specimens:

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Risk assessment

This experiment is low risk however it is important to keep in mind that microscopes can be heavy
so should be handled carefully and not put on the edges of workbenches where they could be
knocked off. Furthermore, when preparing various samples to observe there is a risk of someone
being allergic, for instance where some species of plant are being handled in the lab before being
cut to use as specimens.

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 Edexcel IAL Biology A Level
Core Practical 6

Prepare and stain a root tip squash to observe the stages of

mitosis.

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Equipment list
● Root
● Scalpel
● Ruler
● Boiling tube
● Hydrochloric acid
● 25 cm³ measuring cylinder
● Pipette
● Distilled water
● Paper towels
● Microscopic slide
● Cover slip
● Mounted needle
● Stain
● Optical microscope

Method

1. The ​ends of a root or shoot​ are where growth and therefore ​mitosis​ are occurring, so to
observe mitosis the plant tissue to be observed must be cut from these areas. Use a
scalpel to measure and cut the end 1 cm of the root.
2. Measure out 10 cm³ of 1 mol/dm³ hydrochloric acid into a boiling tube and place it in a
water bath at 60°C.
3. Transfer the root tip section into the boiling tube and leave it for 5 minutes.
4. Remove the tip and use a pipette filled with distilled water to rinse it. Leave it to air dry on a
paper towel.
5. Now cut the end 2mm of the root tip and place this on a microscopic slide. Use a mounted
needle to spread the specimen out forming a thin layer that will allow light to pass through
when observing it under a light microscope.
6. Use a pipette to apply a couple drops of stain to the specimen, the stain binds to
chromosomes making them easier to see. There are several stains that can be used
including Toluidine Blue O and Feulgen Stain.
7. Finally, place a cover slip on top of the specimen on the microscopic slide. ​Press down
firmly​, ​further spreading and thinning ​the specimen for observing. The slide can now be
viewed under an optical microscope.

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Risk assessment

Hazard Risk Precaution

Scalpel Cuts from sharp edges Take care when using it When
carrying it around the lab
place on a white tile
Cut the root tip on a chopping
board

Glassware Cuts from sharp edges if Have a dustpan and brush in

broken the lab to sweep up and
dispose of any broken glass
immediately
Avoid placing glassware on
the edges of the lab benches
Keep away from edge of desk

Hydrochloric acid and stain
Toxic if ingested or enters
body; HCl is corrosive on skin.
Wear goggles and gloves
when handling these
Replace stoppers immediately
after use
Keep away from edge of desk

Conclusion

You need to be able to

recognise these stages of
mitosis when viewing cells under
a microscope.

The mitotic index can be

calculated by ​counting the
number of cells with visible
chromosomes ​- these are the
number of cells undergoing
mitosis - and also counting the
total number of cells visible.

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 Edexcel IAL Biology A Level
Core Practical​ 7

Use a light microscope to make observations, draw and label plan

diagrams of transverse sections of roots, stems and leaves
Use a light microscope to make observations, draw and label cells
of plant tissues
Use a light microscope to identify sclerenchyma fibres, phloem,
sieve tubes and xylem vessels and their location

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Equipment list
● Large leaf
● Root
● Plant stem
● Light microscope
● Microscopic slides
● Cover slips
● Distilled water
● Pipette
● Toluidine blue O stain
● Razor
● White tile
● Tweezers
● Paper towel

Method
1. Take the plant stem and place it on a white tile and fill a petri dish with a ​shallow level​ of
distilled water.

2. Lightly run the razor edge down the length of the stem several times to give ​several thin
specimens​, using tweezers to place them in the petri dish. Select the​ ​thinnest​ ​specimen
and place it on a microscopic slide.

3. Use a paper towel to dab the edges of the specimen to ​absorb any excess water.

4. Put on the eye protection and gloves and use a pipette to add 2-3 drops of the stain to the
specimen. Leave it to soak in for 3 minutes and then again use a paper towel to ​absorb
any excess liquid.

5. Lay the coverslip onto the slide, pressing firmly to ​thin the specimen​ and ​remove any
air bubbles​ between the coverslip and the slide.

6. Observe the specimen under the light microscope:

○ Rotate the revolving nosepiece to line up the
lowest power​ ​objective lens.
○ Look through the eyepiece and use the ​coarse
adjustment knob​ to lower and raise the stage
until an image comes into focus.
○ Then use the ​fine adjustment knob​ to focus
the image further.
○ Produce a biological drawing using either the ​low
or medium objective lens.​Repeat steps 1-6
using the root section leaf.

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 7. Use the figure below to identify the different plant tissues in the stem.

Risk assessment

Risk Hazard Precaution

Toluidine blue O stain
Could cause an allergic Use eye protection and
reaction, may irritate the skin if disposable gloves when
contact occurs handling
If it comes into contact with
skin then rinse with cold,
running water

Stem, root and leaf Could cause an allergic Minimise contact, wear gloves
reaction if an allergy is present

Razor Cuts from sharp objects Take care when using; keep
away from the edge of the
desk
Carry it on a white tile when
moving around the lab
Keep away from edge of desk

Microscope Heavy object Do not use on the edge of the

desk as it could be knocked
onto body parts such as the
feet

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Biological drawings

When producing biological drawings, in this case of cells being observed under the microscope,
certain rules should be followed:

● Draw in pencil only

● Use blank white paper
● Do not sketch - draw clear, unfeathered lines only
● Include labels
● Label lines should be drawn with a ruler and not cross over each other
● Do not shade the drawing
● Include the magnification of the microscope in the title
● Make diagrams as simple as possible and draw them an appropriate size so everything can
clearly be seen
● Include a scale bar where possible

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Edexcel IAL Biology A Level
Core Practical​ 8

Determine the tensile strength of plant fibres.

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Independent variable: The length of plant string used
Dependent variable: The mass added before the string breaks

Equipment list
● Plant to test - stinging nettle stems or celery are good ones to use
● Cotton wool
● 2 retort stands with clamps
● Small weights - 10g is a good mass
● Chopping board or white tile
● Scalpel
● Gloves
● Ruler

Method
1. Use the scalpel to remove 9 fibrous strings from your plant sample, examining each one to
check there are ​no breakages​ along its length and that its ​diameter is even.
2. On the chopping board or white tile use a ruler and scalpel to cut the 9 strings into three 10
cm, three 15 cm and three 20 cm lengths.
3. Set up the clamps and retorts as shown in the diagram with the first 10 cm string.
4. Ensure the string is properly secured with the cotton wool cushioning directly beneath it,
then begin to add weights to the string, 10g at a time ​until the string breaks.​ Record the
mass added in your results table.

5. Repeat step 4 with each of the other two 10 cm strings and calculate a mean mass added
before the string breaks. Then repeat for the 15 cm and 20 cm lengths.
6. Plot a graph of your results.

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Risk assessment

Risk Hazard Precaution

Plant samples Potential allergic reaction Wear gloves when handling

Wash hands after the practical

Scalpel Cuts from sharp edges Take care when using, always
use on a chopping board and
carry on a white tile when
transporting it around the lab
Keep away from edge of desk

Weights The accumulation of weights Only stack weights above the

that fall when the string breaks cotton wool padding and work
could cause injury if it falls on on a clear workbench
any body parts

Results table

String length Piece 1 (g) Piece 2 (g) Piece 3 (g) Mean mass added before
(cm) the string broke (g)

10

15

20

Conclusion
Different factors affecting the tensile strength of plants can also be investigated, such as comparing
the strengths of plant fibres which have the same length but are from different species of plant. The
strength of plant fibres is down to their ​chemical composition​, such as the strong cellulose chains
and links that form plant cell walls and the addition of chemicals like lignin which add strength and
support to vessels in the vascular bundle.

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 Edexcel IAL Biology A Level
Core Practical​ 9

Investigate the antimicrobial properties of plants, including aseptic

techniques for the safe handling of bacteria.

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Independent variable: The species plant used
Dependent variable: Area of inhibition zone

Equipment list
● Disinfectant spray
● Paper towels
● Syringe
● Agar plate already covered with bacteria
● Filter paper cut into 4 equally sized discs
● Incubator at 25 °C
● Sterile forceps
● Pestle and mortar
● Marker pen
● Tape
● Plant samples - mint and garlic
● Measuring beaker - 50 cm³
● Ruler

Method

1. Begin by using the disinfectant spray to clean

the workbench and wash your hands
thoroughly ​minimise the risk of
contamination​ ​in the experiment.
2. Use the marker pen to split the seeded agar
plate into 4 equal quarters and mark the
letters A-D on the edge of the plate, one in
each quarter.
3. Take the first plant sample - mint - and place it
into the mortar. Use the pestle to grind the
plant into a ​fine paste​ and then use a syringe to add 10 cm³ of ethanol.
4. Use the sterile forceps to soak a filter paper disc in the solution created for 20 seconds and
then place the disc on to the sterile agar plate to dry.
5. Once dry, use the forceps to transfer the disc to the second agar plate which has been
seeded with bacterial culture and ​quickly replace the lid​ to the agar dish.
6. Wash the pestle and mortar and repeat steps 3-5 with the second plant sample - garlic.
7. Then fill the 50 cm³ beaker with a small volume (around 5-10 cm³) of alcohol. Use clean
forceps to soak the last 2 filter paper discs in the alcohol, allow them to dry as before and
finally transfer them to their relevant quarters on the seeded agar plate.
8. Tape the lid on at 4 points making it secure but also allowing oxygen to enter so the
bacteria may respire aerobically​.​ Then incubate the dish at 25 °C for 48 hours.
9. Disinfect the work surface again and wash your hands.
10. After the 48 hours have passed use a ruler to measure the ​diameter of the inhibition
zone (clear zone)​ ​created around each disc.

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Risk assessment

Risk Hazard Precaution

Microorganisms If not handled safely and Store bacterially safely,

appropriately the users could
become infected
minimise the time spent with
the lid off and incubate at
temperatures below 30°C.
Don’t eat or drink during the
course of the experiment.
Wear eye protection and wash
hands before and after
experiment.
Do not open any inoculated
plates.

Disinfectant, agar jelly Could cause skin irritation Avoid skin contact, wear
gloves if necessary

Plant samples Potential allergic reaction Wear gloves when handling

Ethanol It is flammable and volatile Do not use it near a naked

flame
Replace stopper immediately
after use

Results table

Substance Diameter of clear zone Area of clear zone

(mm) (Radius² x π) (mm²)

A - Mint

B - Garlic

C - Control 1

D - Control 2

Conclusion

The effectiveness of the antimicrobial properties of each plant can be compared by looking at the
area of the inhibition zones created.​ The one with the​ ​largest inhibition zone​ has ​killed the
most bacteria​ ​and is therefore the most effective. The discs dipped in alcohol only acts as a
control​ to show that the plants alone are causing the death of bacteria and not any other factor.

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