UNIVERSITI TEKNOLOGI MARA INTERNATIONAL EDUCATION CENTRE (INTEC)

BIOLOGY LABORATORY REPORT A LEVEL PROGRAMME

Name: Farhani Binti Othman IC Number: 950316-06-5134 Group: 11 SC 3 SID Number: 1311171403 Title: The Effect of Temperature on Membranes Lecturer: Puan Zakiah Binti Zakaria

Title : Determining the effect of temperature on membranes

Objectives : To determine the effect of different temperature on the membranes of the beetroot To compare the intensity colour of the dye solution after the emmersion of the beetroots To investigate what happen to the reading of absorbency and permeability of the membrane of beetroot with the different temperature.

Introduction : The cell membrane is made up mainly of phospholipids bilayes and proteins which contribute to its selectively permeable nature. The function and permeability of the cell membrane depends on its intact structure. When destroyed, the permeability of the cell membrane is distracted causing cellular contents to leak out. The cell membrane can be destroyed by physical damage, chemicals and high temperature. The cell membrane will mechanically ruptured when the cell are cut. High temperature interferes the structure of proteins and certain chemicals such as fat solvents dissolve the phospholipids, leading to damage of the membrane. Thus, it increased permeability. In this experiment, we are using beetroots as the subject matter. It is a favourite salads in 1970's British (served cooked and pickled in vinegar), beetroot is a root vegetable with dark, purple skin and pink/purple flesh. It has also enjoyed something of a deserved comeback in recent years, its earthy, rich and sweet flavour and vibrant colour lends itself to a variety of both sweet and savoury dishes. Beetroot is a close relative of spinach and chard and has an earthy flavour and a good nutritional content - it's also reckoned to be a good detoxifier.

Beetroot contains a red pigment called betacyanin, which is located in the large central vacuole of the beetroot cells. The vacuole is enclosed by a single membrane called the tonoplast and the whole cell is enclosed by the cell membrane made up of phospholipids and proteins. Betacyanin will remain inside the vacuoles of intact cells. However, if the membranes are damaged, betacyanin will leak out and produce a red/dark pink colour in the surrounding water. Cut cylinders of beetroot are used in this experiment. The beetroot cylinders were repeatedly washed following cutting until not more colour appeared in the wash water.

Problem Statement : How does the temperature effect the cell membrane structure and its permeability. This problem is occur as referring to recipe for cooked beetroots, it will usually recommended that you do not remove the outer skin of the beetroot and do not cut off all stalk ang root if you want to avoid getting lots of red dye in the cooking water.

Hypothesis : The higher the temperature, the higher the reading of absorbency

Variables : Manipulated variable : The temperature of water bath For 30oC water bath, just remain the boiling tube for room temperature. For 40,50 and 60oC, the boiling tube filled with 5ml water bath are heat up in the controlled machine.

Responding variable : The reading of absorbency The 3cm3 of dye solution are place into the cuvette before measuring its absorbency by using the colorimeter. The light must be shine through the smooth side. The colorimeter are adjust read zero absorbance for clear water every time before measuring the dye solution.

Constant variable : The volume of distilled water for beetroot section to be dispersed in. By using a 10ml measuring cylinder, 5ml of distilled water is measure and pour into each 12 test tubes.

Apparatus : Waterproof marking pen, small measuring cylinder, pipette for 2cm3, stopwatch, cuvettes, colorimeter, thermometer, boiling tubes, boiling tube racks, forceps, beaker, ruler, knife, white tile, size 4 cork borer, water bath at 30, 40, 50 and 60oC,

Material : Raw beetroot, distilled water

Technique : Measuring the absorbency of the dye solution by using the colorimeter and record the absorbency reading.

Procedure : 1) A raw beetroot is cut into section by using size 4 cork borer, with length of 1cm to get 12 section of it. Be careful not to spill beetroot juice on your skin or clothing as it will stain very badly.

2) 12 boiling tube are filled in with 5cm3 distilled water each. Each 3 of them then are prepared to be at the temperature of 30, 40, 50, and 60oC. When the water bath reach the required temperature, place one of the beetroot section into each of the boiling tube. Leave for 30 minutes in the water bath.

3) After that, the beetroot section are removed and the boiling tube is shaked to disperse the dye in the solution. 4) The colorimeter then is turned on is set to read % of absorbance. 5) The filter dial is set to the blue or green filter. 6) By using the measuring cylinder, 3cm3 of distilled of water is measured and poured in a cuvette. Place the cuvette into the colorimeter , and make sure that the light is shining through the smooth side. 7) The colorimeter then adjusted to zero absorbance for clear water.

8) The only place the cuvette filled with 3cm3 of the dye solution and take the reading of its absorbency. Repeat the reading for all dye solution for all temperature. 9) Record the data gained in table and appropriate way of presenting data.

Result :

Temperature (oC) Trial 1 30 40 50 60 0.269 0.17 0.42 0.432

Reading of absorbency Trial 2 0.231 0.241 0.44 0.498 Trial 3 0.231 0.192 0.318 0.268 Average 0.244 0.201 0.393 0.405

Table 1

0.45

0.4

0.35

0.3

Reading of absorbency

0.25

0.2

0.15

0.1

0.05

0 0 10 20 30 40 50 60 70 Temperature

Graph 1

Discussion : The experiment is carried out in order to investigate the relationship between the temperature of distilled water, which are prepared for 30oC, 40oC, 50oC and 60oC and the permeability of the cell membrane of the beetroots, which is measured by its reading of absorbency. The reading of absorbency for dye solution, gained from immersion of beetroots section. It was immerse in constant volume of distilled water in each test tube in order to get the accurate result. Table 1 shows the result for reading of absorbency for all dye solution. For each temperature, 3 trial are conducted to get the average. This way may increase the accuracy of the gained result. For distilled water with temperature of 30oC ,the reading of absorbency is 0.244. Meanwhile the reading of absorbency for dye solution in distilled water with temperature of 40oC, 50oC and 60oC are 0.201, 0.393 and 0.405 respectively. The best fit line in the line graph is obtained from the average reading of absorbency of thrice repeated trial of each temperature of the distilled water. From the recorded result in the Table 1, we found that the dye solution from the highest temperate of distilled water has the highest reading of absorbency compared the the lower temperature. The matter that exactly happen is when heat energy given is increase, the kinetic energy of the water molecule will increase too. At the same time, the water will expand too. This will have a disruptive effect on any membrane of its way. Furthermore, the membrane became more fragile due to structure of the membrane itself. The lipid in the phospholipid bilayer will turn to more fluid as the temperature rises up. One more thing is, the protein which is a part of membrane will untangle and break apart if they are heated too much. This situation will form holes within the membrane structure at once. Now, any pigment from the innermost compartment will spill out. Beetroots cells contain betacyanin which are red pigments. Thus, when the temperature of the distilled water increase, more holes on the cells membrane will be form and allow the releasing of betacyanin, increasing the intensity colour of dye solution. The kinetic energy generated by heat will also facilitate this process as the betacyanin will have more energy, making them move faster.

The result for experiment conducted at temperature of 30oC is higher result for 40oC experiment. It should be less than result for 40oC actually. This result may be obtained with some error or mistakes. Somehow we expect that the resut obtained is due to the incomplete rinse of beetroot before conducting the experiment. When the beetroot is cut into section from the raw beetroot, the outermost cell membrane is damaged and ruptured so that the betacynin had been release. That is why the section of beetroot must be rinse first to ensure the accurate result of effect of the temperature on the membrane. For experiment using 30oC of distilled water. The beetroot section have some dye left at their surface, that is why the absorbency reading obtained is higher than the experiment conducted for 40oC.

Further study : Beside using the temperature in order to investigate the effect on the permeability of the cell membrane, other method is by using ethanol. Ethanol is permeable to cell membrane. Because of the nature characteristic of ethanol as a chemical is it able to get into the cell very easily. This interferes the balance of the cells' osmotic environment. Osmosis is the movement of water. Ethanol interrupt this flow. It has also been proven that ethanol can disturb the ion channels in the plasma membrane. These channels in the cell membrane help pump ions like calcium and potassium in and out of the cell which directly control how much water is pumped in and out of the cell. If you are extracting pigments, ethanol is also the perfect solute for pigments and therefore helps in the extraction process.

Safety precaution : Individual: 1) Use a scalpel blade to cut the beetroots accurately and to avoid from cutting yourself, keeps your free hand as far as possible away from the blade. 2) Wears a rubber gloves would help more in protecting our hands from cut dy the blade. 3) To prevent the damages to the table, use a cutting mat when using the blade.

Experimental: 1) In an experiment, it will always an error or else says, it is hard to get the accurate result. To improve the result obtained, experiment is repeated two or three times and the average is taken as the final result. 2) The distilled water in all boiling tube must be constant at their required temperature. So along the experiment time, it will kept in the water bath for their required temperature. 3) Each time before measuring the absorbency of dye solution, the colorimeter must be adjust to zero again by using cuvette filled with distilled water. So that all the reading have standardize to be based on absorbency of the distilled water.

Coclusion : The higher the temperature, the more the beetroots cell membrane will be damaged, releasing more dye. Thus, the higher the temperature, the higher the reading of its absorbency. Meaning that, the higher its membranes permeability. Hypothesis is accepted.

References :
1. http://www.bbcgoodfood.com/glossary/beetroot 2. http://www.writework.com/essay/beetroot-cell-membrane-permeability-experiment 3. http://www.madsci.org/posts/archives/2006-03/1143666101.Cb.r.html 4. http://nutritiondata.self.com/facts/vegetables-and-vegetable-products/2348/2 5. http://www.coursework.biz/Essays/GCSE/Biology/550/