Core Investigation :- The effects of

temperature or alcohol on the

membrane structure on Beetroot
pigments.
Planning
Introduction.
We are investigation the effects of temperature or alcohol on the membrane of
the Beetroot cells. The aim of this practical is to use beetroot to examine the
effect of temperature or alcohol on cell membranes and relate the effects
observed to membrane structure. To function correctly a cell needs to be able
to control transport across the partially permeable cell membrane. Another way
of looking at this practical is the investigation of the destruction of the cell
membranes caused by different temperatures. We will try to find a correlation
between the temperature and the destruction of the cell membranes. The way
we will prove that the cell membrane has been destroyed will be by using a
colorimeter to measure the light intensity that is able to pass through the dye
solution of the beetroot after being in different temperatures for half an hour.
Even thought we are investigating the effects of temperature on the cell
membrane, the investigation can also be made using alcohol, so this will also
be explained further on.

Prediction.
I predict that in the temperature investigation, as temperature increases, the
more cell membranes will be destroyed, with means that the dye solution left
over will become more concentrated. This is due to the cell membrane lipid
bilayer structure with embedded proteins, as it has been shown in the fluid
mosaic model. Proteins are denatured at high temperatures, as enzymes. The
heat affects the lipid bilayer considerably, preventing it from recovery after
broken. There will be pores formed in the membrane that will effect in the
structural breakdown of the cell membranes.
I also predict that in the alcohol investigation, the more alcohol solution used
will increase the cell membranes destroyed. This should be due to the fact that
alcohols are known to be modulators of lipid bilayer properties. They will affect
the lipid bilayer and the proteins round and inside the lipid bilayer. The lipid
bilayer will cutoff and won't be able to rejoin to form again the cell membrane,
this means it will be broken and the dye will be formed in the distilled water
surrounding the beetroot, increasing the dye solution.

Fair Testing.
Independent Variables: These are the factors that are being changed, that we
are investigating. The independent variable would be the temperature in which
the cut beetroot would be placed for 30 minutes. The different temperatures
that will be used will be : -8oC, 22oC (RTP), 40 oC , 50 oC, 60 oC.

We will vary the size cut of the beetroot; we will use three different beetroot
sizes and will investigate if the size of the cut beetroot will affect is any way the
destruction of the cell membranes when introduced in different temperatures.
Using different sizes will ensure that we can ensure that different surface areas
are used in the experiment, and this way will help us to have more reliable
results.
When doing the investigation that testes the alcohol effect on the beetroots
cell membrane, the independent variable will be the concentration of alcohol in
the solution in which the cut piece of beetroot will be placed. Different sizes of
beetroot will also be used in this investigation because of the same reason as
the one in the investigation of the temperature effect on the cell membrane.
Control variables: The control variables are the factors that we will try to
maintain the same, and that may affect the final results. One of the control
variables will be the size of beetroot that each group will be using, this means
that one group or person will only work with one size of beetroot so that no
errors are made and to prevent having anomalies results that should have
being prevented of that should be found in another table of results.
We will also be controlling the amounts of diluted water that we put in the test
tubes that contain the cut beetroot; this is because the concentration of dye
will be different with different amounts of diluted water. And this is why we will
put the same amounts of diluted water in each test tube before putting it in the
water baths, which will have different temperatures.
When doing the alcohol investigation, we will try to maintain the same
temperature trough out the whole investigation, as the temperature changes
may also have a negative effect in the investigation and may vary the final
results. This will be controlled by trying to do the investigation as followed as
possible and at the same time, so that the room temperature will vary the less
possible.
In both investigations we will use the same type of test tubes, as different test
tubes may have effects on the results, as they may increase the pressure put
on the cut beetroot. And this is also why we will use the same beetroot
throughout all the investigations, to try and have the most accurate results as
possible, as different beetroots will have different characteristics and some can
always be rotted, which means that the final results will be completely
different, as the cell membrane wont be in a considerable good state or
conditions.
Another control variable would be the colorimeter used at the end of the
investigations, as different colorimeters will work in different ways and will
have different units or range of results.
Dependent variables: In both investigations we will be trying to find how the
independent variables have an effect on the cell membranes and if they would
be destroyed. This will be controlled by putting 2cm3 of the dye solution that

had been left after the heating, and placing it in a colorimeter to calculate the
change it has had after the heating.

Reliability, Accuracy and Precise:

The measurements we will be using will be taken from the colorimeter. We will
also have to record the temperatures in which the beetroot was put in, so that
we can have a organised rage of results and to prevent errors from occurring.
The colorimeter will tell us the light intensity that was able to pass through the
beetroot dye solution.
Two groups will take one set of measurements of each temperature, and then
share their results with the group that used the same size cut beetroot. This
means that everyone will end up with two sets of results of the light intensity
that passed through each solution that was before in a different temperature.
We will have results on five different temperatures which means we will have a
good range of different results in different conditions.
We will use colorimeters, to try to be as much precise as possible, as
colorimeters are machines and should be calibrated to work properly to have
accurate, precise and reliable answers. The only error that can be made is if
before using the colorimeter there has been problems with the investigation,
but this has a low probability of happening.
As mentioned before there is a low probability of having anomalies results, but
if this happens we will be able to figure it out as we will have two sets of results
on each temperature, and if there isn't a line of best fit in a line graph with the
results, then there is no proportionality between the cell membrane destruction
and the temperature. If this was to happen then the most logical response is
that the results are as they are and there is no proportionally in the results, or
all the investigation has been done incorrectly and that all the investigation
was just to prove that there is no relationship between the temperature and the
cell membrane destruction.

Equipment:

Raw beetroot
Size 4 cork borer
White tile
Knife
Ruler
Water baths at -8oC, 22 oC (RTP), 40 oC, 50 oC, 60 oC or alcohol.
Plastic beaker, about 250 cm3
boiling tube racks
Crushed ice
8 boiling tubes
Thermometer (one per water bath)
Colorimeter
Cuvettes
Stop clock
Distilled water
Pipettes for measuring 2 cm3 and 5 cm3
Small measuring cylinders

If alcohol concentration is investigated several water baths and ice will not be
required. Pipettes and alcohol will be needed.

Safety Precautions
In this investigation we are not using harmful substances, neither corrosive or
flammable substances. This means that no real precautions have to be made.
We will follow the raw safety precautions, that include wearing goggles and lab
coats. The beetroot stains the clothes and will not go off, this means that if
some is spilled on the clothes, they will stay with a red stain forever, and that's
why we wear the lab coats, to prevent our clothes from end up with red stains.

Method:
The method for the temperature investigation:
1. Cut sections from a single beetroot using a size 4 ( we used a size 3) cork
borer. Cut five, 1cm length slices from these section. Be careful not to
spill beetroot juice on your skin or clothing as it will stain very badly.
2. Place the slices of beetroot in a beaker of distilled water to rinse extra
dye that has been left after the cut.
3. Place five boiling tubes (test tubes) each containing 5cm3 distilled water
into water baths at 8oC, 22 oC (RTP), 40 oC, 50 oC, 60 oC. Leave for five
minutes until the water reaches the required temperature. Place one of
the beetroot sections into each of the boiling tubes. Leave it for thirty
minutes in the water baths.
4. Decant the liquid into a second boiling tube or remove the beetroot
section using a technique that does not squeeze the slice e.g. spear with
a pointed seeker. Shake the water/solution to disperse the dye.
5. Switch on the colorimeter and set it to read % absorbance.
6. Set the filter dial to the blue/green filter.
7. Using a pipette accurately, measure 2cm3 distilled water into a cuvette.
Place the cuvette into the colorimeter, making sure that the light is
shining through the smooth sides.
8. Adjust the colorimeter to read 0 absorbance for clear water. do not alter
the setting again during the experiment.
9. Place 2cm3 of the dye solution into a colorimeter cuvette and take a
reading for absorbency. Repeat the readings for all the temperatures.
10.
Present your results in an appropriate way.
11.
Identify any trends or patterns in your results.
12.
Explain any trends or patterns, supporting your statements with
evidence from your data and using biological knowledge. You can find out
more about the biochemistry of the main components of the cell
membrane in the textbook and in the interactive tutorials on lipids and
protein structures.
13.
Describe how you could have improved this experiment to give
more reliable results.

The procedure to investigate the effect of alcohol in the cell membrane:

1. Cut sections from a single beetroot using a size 4 cork borer. Cut eight,
1cm length slices from these section. Be careful not to spill beetroot juice
on your skin or clothing as it will stain very badly.
2. Place the slices of beetroot in a beaker of distilled water to rinse extra
dye that has been left after the cut.
3. Place one of the beetroot sections into a boiling tube containing 5cm3
distilled water. This is 0% alcohol concentration. Repeat with seven test
tubes containing 10%, 20%, 30%, 40%, 50%, 60% and 70% alcohol.
Leave the boiling tubes for 30 minutes.
4. Decant the liquid into a second boiling tube or remove the beetroot
section using a technique that does not squeeze the slice e.g. spear with
a pointed seeker. Shake the water/solution to disperse the dye.
5. Switch on the colorimeter and set it to read % absorbance.
6. Set the filter dial to the blue/green filter.
7. Using a pipette accurately, measure 2cm3 distilled water into a cuvette.
Place the cuvette into the colorimeter, making sure that the light is
shining through the smooth sides.
8. Adjust the colorimeter to read 0 absorbance for clear water. Do not alter
the setting again during the experiment.
9. Place 2cm3 of the dye solution into a colorimeter cuvette and take a
reading for absorbency. Repeat the readings for all the alcohol
concentrations.
10.
Present your results in an appropriate way.
11.
Identify any trends or patterns in your results.
12.
Explain any trends or patterns, supporting your statements with
evidence from your data and using biological knowledge. You can find out
more about the biochemistry of the main components of the cell
membrane in the textbook and in the interactive tutorials on lipids and
protein structures.
13.
Describe how you could have improved this experiment to give
more reliable results.

Reference list:

Yahoo answers (2006) "Science and Mathematics - Biology".

https://answers.yahoo.com/question/index?
qid=20070221021128AA9ZRfB
[Accessed on 23 November
2014]
Biophysical journal (2011) "Alcohol's Effects on lipid bilayer properties".
http://www.cell.com/biophysj/abstract/S0006-3495%2811%2900843-5
[Accessed on 23 November 2014]