Investigating the effect of a named variable on the plasma

membrane

Research two methods for this practical. Evaluate the methods you have found. Reference the
methods.

Method 1

Website: Notes - RP 04 Investigating Cell Membrane Permeability - AQA Biology A-level

(physicsandmathstutor.com)
Date accessed: 6/11/22

Equipment list:

 Beetroot
 Scalpel
 Forceps
 Cutting board
 Ruler
 Tongs
 Distilled water
 Boiling tube rack
 Colorimeter
 Cuvettes
 Filter paper
 Timer
 Water bath
 Thermometer
 Ethanol

Method

1. Cut beetroot into 6-10 identical cubes using a scalpel.

2. Wipe/rinse to clean off any pigment released as a result.

3. If investigating temperature: place each of the cubes of beetroot in an equal volume of distilled
water (5-15ml).

4. Place each test tube in a water bath at a range of temperatures (30-80°C)

5. If investigating concentration of solvents: create a dilution series of ethanol using distilled water.
Ethanol concentrations should range from 0-100% ethanol.

6. Leave the samples for 20 minutes - the pigment will leak out of the beetroot.

7. Set the colorimeter to a blue filter and zero using a cuvette with distilled water.

8. Filter each sample into a cuvette using filter paper.

 9. Measure the absorbance for each solution. A higher absorbance indicates higher pigment
concentration, and hence a more permeable membrane.

Risk Assessment:

Graph
● Plot a
graph
of

absorbance against ethanol concentration/temperature.

Conclusion:

Temperature- As the temperature increases, the permeability of the cell-surface membrane also
increases. This is because the proteins in the membrane denature as the heat damages the bonds in
their tertiary structure. This creates gaps in the membrane, so it is easier for molecules to pass
through it. At low temperatures, phospholipids have little energy and are packed closely together to
make the membrane rigid. This causes a decrease in permeability and restricts molecules from
crossing the membrane. At very low temperatures, ice crystals can form which pierce the cell
membrane and increase the permeability.

Method 2
Website: Investigate the effect on membrane permeability - A Level Revision (weebly.com)
Date accessed: 6/11/22

Objectives

 Know how the effect of temperature on membranes of beetroot cells and how it can be
determined
 To be able to recognise quantitative variables that should be controlled in an investigation
Safety

 Water baths at temperatures above 50 degrees may scald. Take care when removing lids to
allow steam to escape away from the face or body
 Take care with sharp items such as the cork borer and scalpel. Try to cut with sharp items
away from your body
Variables

 Independent variable
 The temperature of the water bath
 Dependant variable
 The permeability of the plant membrane which can be inferred from light
absorbance
 Control
 Surface area of the beetroot, we cut the same size for each disc
 Control
 Volume of water in the test tube
Equipment

 Water baths pre-set at required temperatures

 Thermometers, one for each water bath
 Distilled water
 10cm3 syringe
 Beetroot
 Cork borer size number 4 or 5
 White tile
 Scalpel
 Ruler
 Pipette
 Test tubes
 Colorimeter
 Cuvettes, all with the same thickness / pattern
 Pen for labelling test tubes
 Forceps
 Pointing needle

Method

1. Prepare six water baths each at separate, predetermined temperatures. For our experiment
this was 35, 40, 50, 60, 75, 80
2. Use a syringe to add 10cm3 of distilled water top eight test tubes. Label each test tube with
a temperature from the pre-set range
3. Place each test tube in the water baths set to the corresponding temperature then label
each test tube
4. Check the temperatures of each bath making sure they are correct. It is unlikely to be exactly
the right temperature so in graphs and tables use the real value
 5. Cut eight beetroot cylinders using the cork borer. Then use the scalpel, ruler and white tile
to cut to the same length. Wash the cylinders thoroughly to get rid of the pigment that has
come out of the broken cells
6. Add one beetroot cylinder to each of the eight tubes and leave them in the water baths
7. Shake the tubes once. Working quickly, use forceps to remove the cylinders (making sure to
not pierce the disc as this will affect the result)
8. Set the colorimeter to blue / green filter and percentage transmission. Zero the colorimeter
then, using a blank cuvette filled with distilled water
9. Transfer liquid from each test tube into a cuvette, place into the colorimeter and read the
percentage transmission reading, recording your results in a suitable table
10. Plot a graph of transmission against temperature

Our class found that as temperature increased, transmission decreased. This means permeability of
the membrane increased as the temperature went up.

Analysis of results
The reason that the permeability increased is because the heat gives the membrane, which is fluid
and made up of phospholipids and proteins, energy. This makes the membrane ‘leakier’ by making
more holes in the membrane as it gets hotter. It also gives the molecules, or ion this case the
pigment, more energy allowing them to diffuse. Diffusion occurs as there is a high concentration of
pigment in the cells and a low concentration in the surrounding water, this means that the
molecules will passively diffuse through the membrane but the heat giving kinetic energy allows
more to diffuse in a quicker time.

Evaluation: One limitation of method 1, is that there are no variables stated such as what is the
independent, dependant and control variables within this experiment whereas method 2 clearly
states what is measured , changed and kept the same each for each temperature used therefore
allowing this method to be replicated more easily. A limitation of method 2, is that there is a very
brief description of a risk assessment by identifying only 2 hazards and with no further description of
instructions if there was an emergency with the hazards whereas method 1, the table clearly
identifies all hazards, precautions and instructions on what to do if something happens making this
more of a safe method to use. Another limitation of method 2, is that there is no specific timings
given of how long the test tubes should be in the water bath and how long to leave the sample
which method 1 gives these specific timings allowing there to be a greater validity. However both
methods 1 and 2, conclude on their findings at the end of the experiment and how these findings
were measured allowing there to be a clear result that needs to be shown if the methods were
replicated. A limitation of method 2, is that there is no clear instruction on how to draw a graph and
which labels are needed to be used whereas method 1, clearly explains to plot a graph of
absorbance against ethanol concentration/temperature further allowing the results to be clearly
demonstrated. A limitation of method 1, is that when mentioning the temperatures used for the
water baths, there is a brief range of temperatures given (between 30-80) whereas method 2 gives
precise temperatures of 35, 40, 50, 60, 75, 80- therefore making it more easier to follow the
method. Another limitation of method 1, is that there is no summary of what the experiment is
trying to find whereas method 2 provides a clear objective and what should be expected to perform
this experiment.