CORE PRACTICAL 4

Purpose: To investigate the effect of alcohol concentration on membrane permeability.

Method:

1. Use a size 4 cork borer to cut cylinders of fresh beetroot tissue. Place on a tile and
cut into 1cm wide discs.
2. Place the slices in a beaker of distilled water. Leave overnight to wash away excess
dye.
3. Next day, 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 boiling tubes for
30 minutes.
4. Switch on the colorimeter and set it to read % absorbance.
5. Using a pipette accurately, measure 2 cm3 distilled water into a cuvette. Place the
cuvette into the colorimeter, making sure that the light is shining through the
smooth sides. Adjust the colorimeter to read 0 absorbance.
6. Place 2cm3 of the dye solution into a colorimeter cuvette and take a reading for
absorbency. Repeat the readings for all the alcohol concentrations.

SAFETY: Take care using cork borer and knife.

Factors to be controlled
- The storage conditions and age of beetroot must be taken into account
- The diameter and thickness of the discs must be standardised
- The volume of ethanol in the test tube must be controlled
- The number of discs in the test tube must be the same for all trials
- The duration of ethanol treatment must also be standardised
- The volume of red pigment in the cuvette must be standardised
- Temperature
Independent variable: Percentage of alcohol
Dependent variable: Size of beetroot
RESULTS
Ethanol% 0 10 20 30 40 50 60 70
Absorbance 0 0.08 0.09 0.02 0.48 0.79 0.84 0.83

Conclusion
As the ethanol % so does the absorbance, therefore more of the red liquid has diffuse out
when there has been a higher % of ethanol. This is because alcohol dissolves the lipids in the
cell membrane, therefore the membrane loses its structure and the red liquid can diffuse
out.
As the number of potato disks increases the amount of oxygen also increases’ therefore
suggesting that enzyme concentration is directly proportional to rate of reaction. The
more enzyme molecules there are in a solution, the more likely a substrate molecule is to
collide with one and form an enzyme-substrate complex.