The effect of temperature on membrane permeability:

Introduction:

Beetroot contains high concentrations of betalin. This is a purple pigment found inside the vacuoles
of the cells. The pigment cannot move across the undamaged plasma membranes. The permeability
of a plasma membrane can vary in different conditions. In this experiment I will investigate the
effect of temperature on the amount of pigment leaking through beetroot plasma membranes

Hypothesis:

There will be no significant correlation between the temperature of the beetroot and the amount of
pigment that leaks from the beetroot.

There will be a significant correlation between the temperature of the beetroot and the amount of
pigment that leaks from the beetroot.

As the temperature increases the cell membrane will become more permeable and therefore more
pigment will leak out the increase in temperature will cause proteins in the phospholipids bilayer to
denature these proteins such as carrier proteins and channel proteins are no longer able to carry out
their function and therefore there will be gaps left in the membrane allowing the pigment to diffuse
out.

Variables:

variable Why? How?

Dependant- the % absorbency
of light
Independent- the temperature
of the water the beetroot is
placed in
Controlled- volume of distilled
water
Controlled- the size of each
beetroot piece
Controlled- the length of time
the beetroot is left in each
temperature
This method will give us
quantitative data which is more
accurate than describing the
colour of the solution which is
qualitative
To see the effect of
temperature on cell membrane
permeability
If the volume was to vary the
pigment that leaked out of the
beetroot would be more dilated
and results would be less
accurate
The larger the pieces of
beetroot the more pigment that
could leak out of the larger
surface area or volume making
the results unfair and inaccurate
If temperature has an increasing
effect on membrane
permeability then the longer the
beetroot is left the more
pigment will leak out causing
inaccurate results ( unfair test)
The % absorbency will be
measured using a colorimeter
A thermometer is used to
maintain the temperature of a
water bath with boiling water
Measure 5cm^3 in a cylinder
and pour it into a boiling tube
and place the beetroot in the
same volume of water for each
temperature
Use a size 3 cork borer so that
all the pieces have the same
width(volume) and cut them all
so that they’re 1cm in length
Let the beetroot remain in the
water bath for 30 minutes for
each temperature
Apparatus:

Key apparatus Justification

Cork borer ,knife ,white To cut(knife)the beetroot to an equal length (ruler) and volume (cork
tile, forceps and ruler borer) on the tile and hold it with the forceps
thermometer To measure the temperature of the water baths
Colorimeter and To measure the % absorbency of the pigment that seeped into the
cuvettes distilled water
Stop watch To ensure the beetroot is in the water bath for 30 minutes exactly
Boiling water To maintain the temperature of the water baths
Distilled water To add to the beetroot in the boiling tube
Pipette To measure the small equal volume of distilled water to put i the boiling
tubes

Method:

Wear eye protection at all times

1. label each of the six test tubes with a different temperature

2. use a syringe to add 5cm^3of distilled water to each of the six test tubes
3. place each of the test tubes in the corresponding water bath for five minutes
4. check the temperature is correct using a thermometer
5. cut six beetroot cylinders using a cork borer and using a knife ruler and white tile trim them
down to 1cm long each
6. wash the cylinders with distilled water and pat them dry with a paper towel
7. add one beetroot cylinder to each of the six test tubes and leave them for 30 minutes in the
water bath
8. shake the test tubes and using forceps remove the beetroot cylinder and discard it
9. using a pipette remove enough of the liquid to fill one cuvette
10. set the colorimeter to the blue filter of wavelength 470nm
11. set the colorimeter to measure absorption rather than transmission
12. zero the colorimeter using a cuvette filled with distilled water
13. place each cuvette into the colorimeter and read the absorption recording your results in a
table
14. plot a graph of absorption against temperature
Risk assessment:

Potential hazards Hazard Actions to minimise risk Risk score (/5)

Scalpel, knife and cork Cut skin from sharp metal Cut away from the fingers and 3
borer use forceps to hold the sample
whilst cutting and keep all
equipment away from the edge
of the desk
Broken glass Cuts skin or shoes Take car whilst handling glass 2
ware and keep away from edges
of surface
Boiling water Burns the skin with scalding Use tongs to remove boiling 2
water tubes from the water and wear
eye protection handle the water
with care and keep away from
edges
Purple betalin pigment Stains skin and clothes Wear a lab coat and gloves if if 0
touches the skin just wash it off

Results:

Temperature (c) Colorimeter Reading (% absorbance of light)

Sample A Sample B Sample C Mean
20 98.5 96.0 96.0 99.2
30 93.9 96.0 96.0 95.0
40 80.1 76.9 76.9 78.0
50 26.3 31.0 31.0 29.1
60 0.7 0.7 1.0 0.8
70 0.0 0.1 0.0 0.0
 Colorimeter Reading (% absorbance of
light) Mean
120

100

80
Axis Title

60 Colorimeter Reading (%
absorbance of light) Mean
40

20

0
1 2 3 4 5 6

Analysis and conclusion:

Evaluation what went wrong? :

Bibliography: