12.3.

2018

5.1. THE EFFECT OF TEMPERATURE ON MEMBRANE PERMEABILITY

PLANNING

- HYPOTHESIS
As we increase the temperature, the kinetic energy increases meaning phospholipids
move further apart from each other, losing the membrane structure and increasing
its fluidity and permeability.
Now, using this scientific knowledge, I can suggest that as we increase the
temperature from 0 to 60 the permeability of the membrane increases so the
structure of the membrane is lost and the betalain (pigment in vacuole) comes out of
the cell, meaning more concentrated colour each time, which will mean a higher
absorbance value in the colorimeter.
As temperature increases absorbance increases, and therefore the transmission
value instead decreases.

- EQUIPMENT
 Water baths at 15ºC, 30ºC, 40ºC, 50ºC, and 60ºC containing test tube racks
 Colorimeter
 Thermometer
 10 beetroot cylinders (30 mm long)
 Knife, scalpel
 Ruler
 White tile
 Paper towel
 5 test tubes
 5 cuvettes
 Distilled water
 10 cm3 syringe
 Marker pen and stickers
 Forceps
 Timer

- METHOD
1. Take the 6 test tubes and label each one with its temperature (0ºC, 15ºC,
30ºC, 40ºC, 50ºC, 60ºC).
2. Add 10 cm3 of distilled water to each test tube using a syringe.
 3. Place each test tube to its corresponding water bath for 5 minutes, record
the actual temperature using the thermometer and leave the tubes in the
water baths.
4. Collect 10 beetroot cylinders and trim them all to 30mm using a knife and a
ruler on the white tile.
5. Rinse the cylinders under a running tap and dry them using lab paper tissue.
6. Add 2 cylinders to a tube in each temperature and leave for 15 minutes.
7. Label 6 cuvettes with temperatures from 0ºC to 60ºC.
8. Remove the test tubes from the water bath, swirl gently and use forceps to
remove the cylinders.
9. Pour the remaining liquid in each test tube into the cuvette with the
corresponding temperature.
10. Use the colorimeter to measure the absorption for each temperature and
record this information in a table (use green filter in the colorimeter and set
to zero using the 0ºC cuvette each time before measuring a new cuvette).
11. Plot a graph of temperature against absorption and draw best fit line.

- VARIABLES
 Independent: temperature (to see how it affects the membrane fluidity
and absorption).
 Dependent: light absorption of each cuvette.
 Control: cylinders of same length and same number of cylinders in each
test tube from the same beetroot (this is because different beetroots
could have different concentrations of betalain in them and also very
similar concentration of betalain will be present in the same length of
different cylinders), same volume of distilled water in each test tube,
leave beetroot in test tubes for the same amount of time (so the betalain
has same time in each of them to get out the cell and go to the liquid), dry
all the cylinders (to remove excess of water and have same mass for all of
them).

- RISK ASSESSMENT
 There are not any toxic or hazardous substances in this experiment but
use lab coat, gloves and goggles as we are working in the lab, also be
careful while handling with the water bath ºC, as we are working with high
temperatures and could cause burns, use tongs to take test tubes out of
the rack.
IMPLEMENTING

- TABLE

- How to calculate the mean, example at 15ºC

(0.54+0.61+0.62)/3 = 0.59
- How to calculate the standard deviation, example at 30ºC

( 0.67−1.13 )2 + ( 1.40−1.13 )2+(1.32−1.13)2 = 0.4004

√ 3−1

ANALYSIS

- GRAPH
  The gradient is 0.0357.
 The intercept of this graph is 0.1265.
 This graph shows a linear trend and a positive correlation, so as the
temperature (independent) increases, the absorbance (dependent) also
increases.
 Percentage change from 15ºC to 30ºC =
(1.353-0.590)/0.590 = 1.293
1.293 x 100 = 129.3%

EVALUATION

- CONCLUSION

As temperature increases the colour intensity of the liquid in the test tubes increases
and the absorbance also increases, we can deduce from this that as temperature
increases the kinetic energy of the molecules in the plasma membrane increases,
moving faster and apart from each other, this increases the fluidity and permeability
of the cell membrane and therefore the betalain comes out of the cell to the
solution, increasing the colour intensity.
This confirms my original hypothesis.

- ANOMALOUS RESULTS

We didn’t get any anomalous results in this experiment.

Results are very similar between them

- UNCERTAINTY

The syringe we used has a margin of error of 0.2 mL so the uncertainty is 0.1 mL.

- PERCENTAGE ERRORS

Percentage error of syringe used is 0.1/10 x 100 = 1%

- ACCURACY
 My results have a high level of accuracy as it is very close compared with the true
results, which the teacher and technician had.
I used a quantitative method rather than a qualitative (colorimeter) so this increases
the accuracy a lot.
I repeated the absorbance measurement using the colorimeter 3 times for each
temperature and then I calculated the mean, this gives also a good accuracy.

- PRECISION

 The equipment used was in good condition and some of them like the syringe
had a great precision.
 I calculated the standard deviation for each temperature and the error bars
in the graph show smaller variation, this means the repeated measurements
are very close to each other and to the mean value, so more precise.
 I used small units, for example in the syringe the volume of distilled water
was 10 mL, also the ruler used to cut the beetroot pieces was in mm.
 I used a good technique for the water bath, they had a good static control
and I left each test tube for at least 5 min in the water bath to adapt to the
new temperature conditions.
 In general, I had a good level of precision.

- REPEATABILITY/ RELIABILITY

 I took enough repeat measurements of the absorbance values (3).

 I got similar data each time I repeated the measurements.
 Measurement results were produced over a short period of time (1 h) and by
the same group and using the same equipment in the same place
(classroom).

- REPRODUCIBILITY

I compared my results with other group’s results, who followed the same procedure
and using the same apparatus and materials, and we got very similar results
between us, so my experiment has a high level of reproducibility.

- VALIDITY
  My results of this experiment are suitable to answer the questions set out in
the investigation.
 All the variables were controlled along the experiment procedure (like size of
the beetroot cylinders, using the same beetroot, same number of cylinders,
same volume of distilled water, etc).
 I repeated the measurements 3 times and also calculated the mean.
 I carried out a statistical test using standard deviation to identify possible
anomalous results.
 I used a large range of temperature values, using 6 in total from 0ºC to 60ºC.
 All these techniques give my experiment a good validity.

- LIMITATIONS

 Room temperature of the laboratory may fluctuate the real temperature in

the water baths.
 The stop watch was used by different persons in the group and by eye reflex,
so this could lead to misreading in the time.
 Not sufficiently large range of temperature values
 Inconsistent stirring of the solution in the test tube containing distilled water
and betalain.
 Impossible to put all the test tubes in the water bath at the same time and
start stop watch at the same time.
 Not all the beetroot cylinders were cut in the exact same length as different
people realised the cutting each time.

- IMPROVEMENTS

 We could have increased the range of values even more for example instead
of 50ºC to 60ºC having 52ºC, 54ºC, 56ºC, 58ºC and 60ºC.
 Stop watch used each time by the same person.
 Stir the solutions each time properly and for the same amount of time.
 Same person could cut the beetroot cylinder each time and looking more
carefully that is exactly 30 mm.