Investigating the effect of different ethanol concentrations on the cell membrane

permeability in beetroots

How does the concentration of ethanol (2%, 4%, 6%, 8%, 10% 99%) effect beetroot to
determine the permeability of its cell membrane measured using a colorimeter to determine the
transmission of light passing through the ethanol whilst keeping the time( (10 ± 0.005 minuets),
the volume of the ethanol solution (40mL), the mass of the beetroot slices( (10 ± 0.01g), size of
cuvette from the colorimeter( +/- 0.05cm) and color of light when using the colorimeter (blue
450nm) the same?

Introduction
1.3 membrane structure - fav topic in biology
And in interest on how alcohol can damage the cell membrane
Sanitizers work through disrupting the cell membrane and therefore damages the
components of the virus making it futile

Hypothesis

Ethanol being a non-polar solvent meaning that it is a solvent that does not have a dipole
movement and therefore has the ability to dissolve non-polar substances such as lipids. Therefore
placing beetroot within ethanol allows its membrane to become much more permeable and it
becomes susceptible to the leakage of substances in and out of the cell - in this instance the
pigment of the beetroot will leak. As concentration of ethanol increases so does the membrane
permeability therefore making the two directly proportional.

Null hypothesis
Forming a null hypothesis, I predict that there will be no connection between the concentration of
the various ethanol solutions and the permeability of the cell membrane of the beetroot.

Graph of the hypothesis

From this graph we can

understand that the transmission
of light would consistently
decrease as the concentration of
ethanol increases due to the
increase of red pigment of the
beetroot leaking out as a result of
the ethanol penetrating the cell
membrane of the beetroot.

Variables
Independent variable -
The independent variable for this experiment will be the concentration of ethanol. This will vary
from 2%, 4%, 6%, 8%, 10% 99% which will be modified through dilution of the desired volume of
distilled water (as shown in the sample results table). This will be changed in order to see the
various effects each concentration has on the cell membrane permeability within the beetroots.

Dependent variable-
The dependent variable fo this experience is the permeability of the beetroot cell membrane which
is inferred from light absorbance (transmission of light) - Au. This variable will be what I am
measuring within my experiment. With the colorimeter being able to find the absorbance of a
solution it means that the higher the absorbance of the solution and the more pigment the end
product solution is the more permeable the membrane is depending on the ethanol solution.

Control variables Why should it be controlled ? How will it be controlled?

Time (10 ± 0.005)
This will be controlled due to the This will be controlled by keeping
fact that if the duration that the the time constant with a timer
beetroot is kept in the ethanol and placing the beetroot slices
solution is too short then the into the ethanol solutions for ten
solution will not fully penetrate minuets and then will be taken
and interact with the cell out at the exact same time.
membrane, however if the
duration that the beetroot reacts
with the ethanol solution is too
long then the solution would have
a longer time to penetrate the cell
membrane of the beetroot
resulting in varied and
inconsistent results as a result of
inconsistent timings.

Mass of beetroot slices (10 ±
0.01g)
Not controlling the mass of the
beetroot slices could affect the
amount of time the ethanol
solution is able to penetrate the
cell membrane.
This will be controlled by cutting
the beetroots the same size in
order for them to be the same
mass then using a calibrated
scale (±0.1g) to measure the
mass of the beetroot slices to
ensure they are the same.

Type of beetroot The type of beetroot should be
kept the same due to the fact that
different types of beetroots have
different types of structures and
different types of pigments, for
example the red beet contains
betacynanins pigments, where as
the golden beets contain
betaxanthins.
This will be controlled through
using the same type of beetroot
throughout the experiment. I will
be purchasing a big piece of
beetroot then using that piece to
cut up the 5 slices of 10g of
beetroot being used within my
experiment to ensure the same
type of beetroot is being used.

Volume of ethanol solution (+/- This is vital to be controlled in
0.05cm3) 
 order to have reliable results as
well as to ensure that we are
testing the correct concentration
of ethanol with reaches the cell
membrane of the beetroot.
This will be kept constant by
measuring each volume of the
ethanol solution through a syringe
for accuracy.

Colorimeter wavelength Controlling the colorimeters This will be kept constant by

(450nm) wavelength would mean that
either less or more light could
possibly be absorbed by the
solution therefore effecting the
values given to us- giving us an
inaccurate result.
programming the colorimeters
wavelength to 450nm in order to
match the wavelength of the color
blue.
Control variables Why should it be controlled ? How will it be controlled?

Size of cuvette from the
colorimeter (uncertainty of +/-
0.05cm)
This needs to be controlled due
to the fact that many cuvettes are
made from various materials and
are made in different sizes and
could affect the results of the
experiment.
This will be controlled by using
the same size cuvette for all trials
as well as the same type of
cuvette.

Time (10 ± 0.005) Not keeping the time as a Keeping the time of how long
constant causes the data of the each beetroot piece is kept in the
experiment to be invalid. If the ethanol solution will be controlled
time isnt kept the same and each by using a timer. The timer will be
piece of beetroot stays in the started as soon as the beetroot is
solution for different amounts of placed in the solution and as
times it would affect how much soon as the ten minuets has
the various ethanol solutions come to an end the beetroot will
react with the cell membrane. Too be taken out of the solution.
much time would mean that the
cell membrane would interact too
much with the ethanol and too
little time would mean that the
cell membrane hasn’t reacted
enough with the ethanol.
Therefore keeping the time
constant is vital for the reliability
of the experiment.

Uncontrolled variables
Uncontrolled Why are they not controlled?
variables

Temperature The temperature will not be controlled due to the fact that my school has a
central AC, meaning that it is automated and cannot be controlled and changed
for the individual classrooms- therefore controlling the temperature would not be
possible. This has a possible effect on my experiment due to the fact that
permeability of the membrane is known to be affected by temperature. Increasing
the temperature of the room has a possibility of making the membrane unstable
as well as more fluid where as decreasing the temperature causes the membrane
to slow down. However with the temperature staying the same throughout my
experiment will allow for similar results throughout using the various ethanol
concentrations.

Environmental considerations
Throughout the duration of this experiment there will be no use of chemicals nor equipment that is
harmful to the environment. The use of ethanol isn't harmful to the environment as long as it is
disposed of properly as well as the use of beetroot which isn't anywhere near harmful to the
environment as it is an organic product which is produced from our environment. The majority of
the equipment is reusable and therefore is not harmful to the environment as they will continue to
be put to use, however there will be paper used throughout the experiment to write down my
findings and the data I collect. I will transfer the data onto my laptop in order to have a digital set
of the data and when the paper is of no use I will ensure to recycle the paper as it aids in the
reduction of greenhouse gas emissions which contribute to climate change therefore affecting our
environment.

Equipment
- 6 test tubes (10cm3)

- 3 beakers (100mL)

- Pre-prepared diluted ethanol solutions (2%, 4%, 6%, 8%, 10% 99%)

- 250mL of distilled water

- Colorimeter (± mg/L) x1

- Cutting board

- Test tube rack

- 6 Pipettes

- Stopwatch (± 0.001s)

- Forceps

- Vegetable knife or scalpel

- Petri dish

- 6 syringes

- Scale (± 0.1g)

- Whiteboard marker

- Ruler

- 6 Cuvettes

- 2 untouched store-bought beetroots

Concentration table
Test tube A Concentration % Volume of ethanol Volume of distilled Ratio of ethanol
(+/- 1) (cm3) water to water
(+/-1) (cm3)
(+/-1)

A 2 1.0 4.0 n/a

B 4 2.0 3.0 2:3

C 6 3.0 2.0 3:2

D 8 4.0 1.0 4:1

E 10 5.0 0.0 n/a

F 99 5.0 0.0 n/a

Safety
This experiment is medium-risk, meaning that it is essentially safe as long as precautions are
followed-

• always wear your goggles as well as a lab apron to protect your eyes and clothing as if the acid
goes into your eyes it will cause damage to the cornea and result in blindness and if the
solutions come on your clothes it will stain

• If you get any sort of chemical solution in your eyes you must react immediately and gently
flush the eye out with water with tap water for roughly twenty minuets, go to a medic to get the
injury checked out and if necessary go to the hospital, to be safe continue washing the eye
during the ride to the ambulance

• Make sure everybody involved in the experiment is aware of the dangers and understands ways
or methods to be safe

• If you swallow the solutions wash out the mouth with drinking water but do nothing else, do not
provoke yourself to vomit and immediately consult a doctor.

• Do not touch any chemicals as it can react with your skin and if you do touch any go to your
teacher and make sure to wash it off carefully

• Don't play around with the glass materials or any of the materials as if the glass breaks it can
injure someone and cause severe damage

Alongside these standard procedures, there are also safety precautions that must be followed in
specific to this experiment:

Hazards Why are they hazardous
Glass objects As glass is fragile and can easily
be broken if mishandled it can
lead to glass shards and sharp
edges that could easily cause
skin injury. With glass also being
transparent it isn't the easiest to
see when broken, therefore being
a hazard in disguise.
Ethanol Ethanol being an alcohol solution
makes us weary that it can cause
skin irritation and rashes if the
solution comes into contact with
your skin and if the solution
comes into contact with your
eyes then serious eye damage
can occur as well as eye irritation.
scalpel/knife A scalpel is known to be sharp
and can cut through skin
therefore being able to cause
injuries and it also has the
capability to harm others.
Method
Results and data collection
Processed data
Calculations
Qualitative data
Results graph
Conclusion
How to stay safe
To stay safe whilst handling glass
objects it is important to tread
carefully around them. However if
a glass object is broken then do
not pick up the broken pieces
with bare hands, instead use cut-
resistant gloves to pick up and
dispose of the broken glass. It is
also vital to not run around with
the glass and to be careful with it
as it is not only a hazard to you
and others but it is the schools
material and therefore should not
be broken.
Ethanol being a flammable
solution, it is vital to keep it away
from heat. It is also crucial to
wear safety goggles to protect
your eyes and avoid showing skin
to reduce contact with your skin
and the ethanol. This can be done
through wearing a lab coat and
wearing gloves when handling
ethanol.
Keep a safe distance whilst
holding the scalpel and ensure to
cut away from your body to avoid
harm to yourself. Do not put more
pressure and cut fast with the
scalpel if you want to speed up
but in fact take your time to avoid
the scalpel harming you. Ensure
to hold the scalpel firmly
whenever you are using it as well
as holding the beetroot nice and
tight to avoid the slipping of the
beetroot which could result in
cutting your own finger. Make
sure to dispose or keep the
scalpel away safely after use.

Evaluation