Biology Practical

1-The effect of temperature on membrane permeability

1. The independent variable: Temperature of water
2. The dependent variable: % transmission of light through resulting
solution
3. Control variables: a) volume of distilled water
b) Time left in water
c) Size of beetroot piece
4. Validity: a) The same diameter core is used to ensure the surface
area of all the beetroot pieces are of the same size.

b) When the beetroot has been cut some of the cell membranes are
broken, which means some anthocyanin will leak out. This must be
completely washed off in order to maintain the reliability of the
results.
c) Distilled water should be used so that there will be a reliable
substance to test with.
d) Beetroot must be raw, not cooked. Use a size 4 cork borer and cut
with care using a cutting board. Leave the cores overnight in a
beaker of distilled water. The pigment from any cells that have been
cut by the cork borer will leak into the water. Rinse away any
pigmented water in the morning and replace with fresh water.
5. Reliability: Repeat the experiment at least thrice and compare
them.

Risk assessment
Risk
1. Cutting tool like knives and
scalpels can be dangerous and
cut the skin.

Method to control the risk

1. Always carry cutting tools like
knives and scalpels in a small
tray. Replace the cutting tool in
the tray when not in use. Take
care while carrying scalpels or
knives around the laboratory.

2. Beetroot juice will stain

clothing (and, temporarily, skin)
but is not hazardous.

2. Students can wear lab coats

to protect their clothing from
stains.

Method: Using a cork borer and knife, cut pieces of beetroot into 1
cm length cylinders. Place in distilled water overnight to remove any
dye released on preparation. Wash and blot dry. Place 8 boiling
tubes of distilled water into 8 water baths of different temperature.
Then add a piece of beetroot to each and leave for 30 minutes.
Remove the beetroot and shake tubes to disperse dye. Set
colorimeter to % absorbance on blue/green filter. Calibrate using
distilled water in a cuvette first then add 2cm3 of beetroot solution
from the first temp to a new cuvette. Place into colorimeter to read
% absorbance. Repeat for all other pieces.
Calculations & outcome: to calculate % transmission = 100%absorbance.
As temperature increased, % transmission slightly increased to a
point at which it greatly increased due to membrane molecules
gaining more heat energy, vibrating more to a point where the
vibrations caused large gaps in the membrane enabling the release
of dye also proteins in membrane denatured leaving large pores.

2- The effect of alcohol concentration on membrane

permeability
1. The independent variable: Alcohol concentration
2. The dependant variable: % transmission of light through resulting
solution
3. Control variable: a) Size of beetroot piece
4. Validity: a) The same diameter core is used to ensure the surface
area of all the beetroot pieces are of the same size.

b) When the beetroot has been cut some of the cell membranes are
broken, which means some anthocyanin will leak out. This must be
completely washed off in order to maintain the reliability of the
results.
c) The volume of the alcohols should be the same.
d) Distilled water should be used so that there will be a reliable
substance to test with.
e) Beetroot must be raw, not cooked. Use a size 4 cork borer and cut
with care using a cutting board. Leave the cores overnight in a
beaker of distilled water. The pigment from any cells that have been
cut by the cork borer will leak into the water. Rinse away any
pigmented water in the morning and replace with fresh water.
5. Reliability: Repeat the experiment at least thrice and compare
them.

Risk Assessment

Risk

Method to control the risk

1. The compounds used in this

experiment are flammable and
poisonous. Methanol, sometimes
referred to as wood alcohol, can
cause blindness and death.
Propanol is fatal if consumed.
One possible reason why they
are so dangerous to living
organisms is that they might
damage cellular membranes.
2. Some alcohols are flammable.

1. The compounds used in this

experiment are flammable and
poisonous. Avoid inhaling
vapours. Avoid contacting them
with your skin or clothing.

2. Be sure there are no open

flames in the lab during this
experiment.

Method: In this experiment, you will test the effect of three

different alcohols (methanol, ethanol, and 1-propanol) on
membranes. Ethanol is found in alcoholic beverages. Methanol,
sometimes referred to as wood alcohol, can cause blindness and
death. Propanol is fatal if consumed. One possible reason why they
are so dangerous to living organisms is that they might damage
cellular membranes. Methanol, ethanol, and 1-propanol are very
similar alcohols, differing by the number of carbon and hydrogen
atoms within the molecule. Methanol, CH3OH, is the smallest,
ethanol, CH3CH2OH, is intermediate in size, and 1-propanol,
CH3CH2CH2OH, and is the largest of the three molecules.
If beet membranes are damaged, the red pigment will leak out into
the surrounding environment. The intensity of colour in the
environment should be proportional to the amount of cellular
damage sustained by the beet.
To measure the colour intensity, you will be using a Colorimeter or
Spectrometer. In this device, blue light from the LED light source will
pass through the solution and strike a photocell. The alcohol
solutions used in this experiment are clear. If the beet pigment leaks
into the solution, it will colour the solution red. A higher
concentration of coloured solution absorbs more light and transmits
less light than a solution of lower concentration. The device monitors

the light received by the photocell as either an absorbance or a

percent transmittance value.
You are to prepare five solutions of differing alcohol concentrations
(0%, 10%, 20%, 30%, and 40%) for each of the three alcohols. A
small piece of beet is placed in each solution. After ten minutes,
each alcohol solution is transferred to a cuvette that is placed into
the Colorimeter or Spectrometer. The amount of light that
penetrates the solution and strikes the photocell is used to compute
the absorbance of each solution. The absorbance is directly related
to the amount of red pigment in the solution. By plotting the percent
alcohol vs. the amount of pigment (that is, the absorbance), you can
assess the amount of damage various alcohols cause to cell
membranes.