Harvey Rodgers

the effect of ethanol on

membrane permeabIlIty In
beetroot
Introduction
The aim of this experiment is to find out how the concentration
of ethanol effects the permeability of beetroot cell membranes.
This practical in particular uses different concentrations of
ethanol to affect the cell membranes, in order to see if different
concentrations allow more or less Anthocyanin to flow out of
the cell down the concentration gradient. Once diffusion has
occurred, the absorbance of the solution can be measured by a
colorimeter, to see how much Anthocyanin left the cell.

Equipment List
Beetroot
5 Boiling Tubes
250cm3 Beaker
Bungs
Colorimeter
Corer
5 Cuvettes
Distilled Water
Ethanol
Glass Marker
Graduated Pipette
Measuring Cylinder
Pipette Filler
Ruler
Scalpel/Vegetable Knife

Harvey Rodgers

Spatula
Test Tube Rack

Method
Serial Dilution Preparation 1. Collect the 5 boiling tubes and label them 100%, 50%, 25%,
12.5% and 6.25% using the glass marker. place them in the
test tube rack.
a) Using the measuring cylinder, put 10cm3 of distilled water in
boiling tubes 50%, 25%, 12.5% and 6.25%.
b) Then, put 20cm3 of ethanol in the boiling tube labelled 100%.
c) Using the graduated pipette and the pipette filler, remove
10cm3 of ethanol from the 100% boiling tube and add it to
the 50% boiling tube. Invert 3 times after sealing with bung.
d) After that, take out 10cm3 of the solution, and add it to the
25% boiling tube.
e) Repeat the steps until all of the tubes contain a solution of
ethanol and water.
2. Cut the beetroot in half using the scalpel/vegetable knife.
3. Using the corer, retrieve 5 cylinders of beetroot from the
centre. Use the spatula to gently remove the cylinder from
the corer.
4. Measure each cylinder using the ruler and cut them into
more similar lengths using the scalpel/vegetable knife.
5. Fill the 250cm3 beaker with distilled water.
6. Place the beetroot cylinders into the beaker to remove any
anthocyanin released due to the usage of the corer.
7. Remove the beetroot cylinders after about 10 - 20 minutes.
8. Place each beetroot cylinder into a boiling tube containing an
ethanol solution. Leave them in there for approximately an
hour.
9. While waiting for diffusion to occur, collect the 5 cuvettes
and calibrate the colorimeter.
10. After an hour has passed, remove 3cm3 of each solution
and place it into a cuvette for absorbance testing.
11. Place each cuvette into the colorimeter and test the
absorbance. Record the result. If possible, complete more
than 1 trial to ensure errors are minimised.

Harvey Rodgers

Risk Assessment
Item

Hazard

Ethanol

Irritant

Ethanol

Highly Flammable
Flash Point 12C

Glass

Very Sharp When

Broken

Caution
Wear goggles at all
times. Exercise
extreme caution
when handling
without gloves/ arm
protection.
Avoid any open
flames or sparks
that may be present
during any
experiment.
Hold all glassware
with a firm grip, to
avoid dropping and
shattering.

Results Table
Diluted
Ethanol
Concentratio
n (%)
100
50
25
12.5

Absorbance
1

Mean

Harvey Rodgers

6.25

Analysis
In conclusion, as the concentration of the ethanol solution
increases, so does the rate of diffusion, proven by all of the
beetroot samples. From a short change in concentration (6.25%
25%) not much change occurred. Higher changes in
concentration though, (25% - 100%) proved very effective for
increasing the rate of diffusion, and that when a larger quantity
of ethanol is used, the impact on the plasma membrane is
much more severe.

Limitations

Improvements

Beetroot cylinders were not

The beetroot could be
100% accurate, since they
observed with extreme
were removed and adjusted
precision, ensuring that the
by the naked eye, rather than
pigments were as close as
a machine. Also, their original
possible, by laying the
pigments will have been
fragments side by side and
different, even if only a little.
comparing closely.
Distilled water in the boiling
The evaporation can be
tubes may have evaporated
controlled better by putting
while waiting for diffusion to bungs on the boiling tubes, to
occur. This can affect the
allow no distilled water to
concentration of the ethanol
escape.
and cause the amount of
absorbance to change.