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To investigate the effects of the organic solvents (10% ethanol, 30% ethanol, 50% ethanol, chloroform and paraffin on the permeability of cell membrane of beetroot.

Organic solvents will destroy the cell membrane and make it permeable to the red pigment. The higher the concentration of the alcohol, the greater will be this effect.

Principle of design:
Cell membrane consists of protein and phospholipids. Anthocyanin is water soluble red pigment that presents in the vacuole of beetroot cell and do not diffuses out. Beetroot discs were placed in different organic solvents for a certain time span. The cell membranes may destroy so that the anthocyanin diffused out and the solution turned red. The colour intensity of the solution is measured by a colorimeter. Then, the effect of the organic solvents (10% ethanol, 30% ethanol, 50% ethanol, chloroform and paraffin on the permeability of cell membrane of beetroot is investigated.

Independent variable:
The independent variable is the different organic solvents. It is changed by immersing the beetroot tissue into the different solvents for a fixed period of time.

Dependent variable:
The independent variable is the permeability of beetroot cell membrane. It is measured by relative colour intensity of red solution, calculated as % light absorption of solution

Controlled variable:
Same number of beetroot discs used in tubes of different solvents Same time span of immersion in the different solvents Same volume of organic solvent and water used Same thickness of beetroot discs Same strength of shaking the tube with beetroot discs Same colorimeter used for measuring different red solution

Procedure & precautions:

1. 2. 4 cylinders of tissue from a beetroot, each with a length of 2cm long, were cut using a cork borer. The cylinders of beetroot were cut into about 30 discs of 2mm thick using a razor blade. This would shorten the path and increase the surface area for diffusion. Care must be taken while using the razor blade. The discs were rinsed in distilled water several times to remove anthocyanin that leaked out


4. 5. 6. 7. 8. 9.

from the damaged cells as the resulting of cutting The discs were left inn a beaker if of water to avoid drying out, 5cm3 of 10% ethanol, 30% ethanol, 50% ethanol, chloroform and paraffin and distilled water were added separately into labeled test tubes. The control was set up using the distilled water 5 pieces of beetroot discs were added into each of the test tubes. Care must be taken to prevent damage of the beetroot discs in transferring. The tubes were left for ___ minutes and were swirled occasionally with same strength to enable even distribution of anthocyanin in thee water. The discs from each test tube were removed carefully after ___ minutes. Care must be taken to prevent damaging the beetroot discs and leakage of anthocyanin into solution. The relative colour intensity of each tube was measured using a colorimeter in term of % light transmission in each solution. Light absorption% = 100 - %light transmission

A table showing the effect of different solvents on the permeability of beetroot cell membrane Solution 10% ethanol 20% ethanol 30% ethanol chloroform paraffin oil water Permeability of beetroot cell membrane = relative colour intensity of solution %light transmission (T) light absorption% (100-T)

1. Interpretation of result Organic solvent chloroform and ethanol will dissolve phospholipids bilayer and destroy the membrane, so it becomes permeability to anthocyanin. The cell membrane of beetroot tissue will be destroyed by the organic solvents, thus releasing the red pigments in their cell saps. Beetroot tissue treated with organic solvents will give a more intense red colour than that treated with water. Ethanol dissolves the lipid component of the membrane and destroys the cell membrane. Membrane loses its selectively permeability. Red pigment leaks out. There will be increasing colour intensity in the solutions in which the beetroot tissue is treated with increasing concentration of alcohol.

2. Source of error
a) b) c) d) e)

The beetroot tissues might be damaged during handing, some pigment leaked out. Beetroot discs were not blotted dry when transferred from paraffin oil / chloroform to the distilled water to allow time for outward diffustion. Beetroot discs were obtained by freehand sectioning and they might be of different size The amount of anthocyanin present in each test tubes might not be the same initially The cuvettes holding the red so1ution might be scratched and this would affect the measurement of % light absorption by the colorimeter.

a) b)

3. Improvement of result Handle the beetroot discs carefully so that anthocyanin would not leaked out accidentally Blot dry the discs before transferring the discs

4. Topic for further investigation