Diffusion and Osmosis

Cori Michibata September 26, 2011 Period 5 AP Biology

I.

Introduction Purpose: The overall purpose of this lab was to study diffusion, osmosis, and water potential in a lab setting, based on experimental data. The purpose of the diffusion part of the lab was to observe the net movement of molecules across a selectively permeable membrane. The purpose of the osmosis part of the lab was to analyze and compare the movement of water molecules through a semipermeable membrane to solutions of different sucrose concentrations. The purpose of the potato core part of the lab was to determine the water potential of a potato cell. Background Information: Cellular transport is important to life in that it is crucial in maintaining the homeostasis of a cell; the cell membrane helps to preserve this internal balance of the cell by regulating the movement of molecules into and out of the cell through either passive or active transport. Passive transport is a spontaneous process (it does not rely on the cell's own energy) that involves the random movement of molecules such that the net transfer is from a high concentration to a low concentration; it includes diffusion, facilitated diffusion, and osmosis. Diffusion is the process of molecules distributing themselves through a semipermeable membrane to balance out the concentrations of the solutions on either side of the membrane. The molecules will still move around randomly once equilibrium is reached, but there will be no net movement; this is called a dynamic equilibrium. Osmosis specifically refers to the diffusion of water molecules across a semipermeable membrane from an area of higher concentration of water to an area of lower concentration. When comparing two solutions, the solution with the lower concentration of water is considered to be the hypertonic solution, while the solution with the higher concentration of water is the hypotonic solution; water molecules will move from the hypotonic solution to the hypertonic solution in order to achieve equilibrium. If the two solutions have equal concentrations of water, they are isotonic and are already at dynamic equilibrium. When comparing solutions, there is a direct correlation between the concentration of water and

water potential; that is, the hypotonic solution has higher free energy, and the hypertonic solution has lower free energy. Therefore, water molecules move from a region of high water potential (hypotonic solution) to a region of low water potential (hypertonic solution). Factors that affect water potential are changes in solute concentration and pressure. Water potential is an important concept in the study of cells; it determines the net movement of water between a cell and its environment. Water will move out of cell with a higher water potential (lower solute concentration) than its surroundings, causing the cell to shrivel. However, when the cell is in a hypotonic solution, water will move into the cell, causing it to swell; an animal cell will burst when the pressure of the additional water becomes too great, but the cell wall of a plant cell prevents the plant cell from bursting, causing the pressure within the cell to build. Water potential increases with physical pressure; the net movement of water is directly proportional to the amount of physical pressure that is applied on the system. Therefore, as water moves into the cell, the plant cell's water potential increases with the pressure within the cell, which affects the movement of water molecules between the cell and its surroundings. Expectations: In the diffusion part of the lab, the membrane pores of the dialysis tubing should contain the larger starch molecules within the bag while allowing molecules of glucose, water, and iodine potassium-iodide to diffuse between the solutions in the bag and in the beaker. It must be assumed that water has diffused into the dialysis bag because the procedure does not call for the documentation of quantitative data (such as the mass of the dialysis tube before and after standing in the H2O + IKI solution for 30 minutes); however, we should be able to see that the glucose molecules have diffused into the solution in the beaker and the IKI molecules have diffused into the bag. By the end of the experiment, the solution in the beaker should test positive for the presence of glucose, and the solution in the dialysis tubing will turn a dark blue-black color to indicate the presence of starch; because starch is one of the original contents of the

dialysis bag, the diffusion of IKI molecules into the bag must be responsible for the color change. In the osmosis part of the lab, the percent change in mass should increase with the original molarity of the solution within each dialysis bag. The varying concentrations of the original solutions in the bags will cause different amounts of water molecules to diffuse into each bag, depending on how much water needs to move into each bag to achieve dynamic equilibrium. No sucrose molecules should pass through the dialysis tubing into the solution in the beaker because the disaccharide molecules are too large to diffuse through the membrane. In the potato core part of the lab, the water potential of a potato cell should definitely be less than that of distilled water because the concentration of water in distilled water is obviously much higher than the concentration of water in the potato cores. The water potential of a potato cell may still be less than the water potentials of solutions with low concentrations of sucrose, but the water potentials of the solutions with higher concentrations of sucrose will definitely be higher than that of a potato cell. When the potato cores are soaked in a solution that has a higher water potential than the potato cells the water moves into the potato cells to achieve equilibrium; when the cores are soaked in a solution that has a lower water potential than the potato cells the water moves from the potato cells into the solution. Once equilibrium is reached, the solution and the cell are isotonic and there should be no net movement of water molecules. Variables: In the diffusion part of the lab, the independent variables were the different contents of the beaker and the dialysis bag; the beaker contained IKI, while the bag contained glucose and starch. The dependent variables were the color change in the bag due to the diffusion of IKI and the presence of glucose in the beaker, as indicated by Benedict's solution. The colors of the solutions acted as the controls because we knew that there would be a color change if either the glucose or the IKI molecules diffused across the dialysis tubing. In the osmosis part of the lab, the independent variables were the different concentrations of the solutions in the dialysis bags. The

dependent variables were the amounts of water that diffused into each bag, indicated by the changes in each bag's mass after standing in distilled water for 30 minutes. The dialysis bag containing distilled water should act as the control group because in that setup, the membrane separates solutions of equal concentrations of water; therefore, there should be no net movement of water. In the potato core part of the lab, the independent variable was the varying molarities of the solutions in the cups. The dependent variable was the movement of water molecules between the potato cores and each solution, indicated by amount of mass the potato cores gained or lost. The controls for this part of the experiment were the potato cores themselves; we used the same type of plant cell throughout the experiment, keeping the cell composition as consistent as possible from trial to trial. II. Methods In the diffusion part of the lab, we tied off one end of our dialysis tubing and filled it with 25 mL of 15% glucose/1%starch solution. We tied off the other end, making sure to squeeze out as much air as possible to allow room for water and IKI molecules to diffuse into the bag. We let it stand in a solution of water and IKI for 30 minutes to allow the glucose, water, and IKI molecules to diffuse through the membrane. When we saw that the solution in the bag had turned blue-black, we tested both solutions for glucose by heating them; they both turned orange, indicating that glucose had diffused into the beaker and was still present in the bag. In the osmosis part of the lab, we followed the same steps to fill six dialysis bags with six different solutions: distilled water, 0.2 M sucrose, 0.4 M sucrose, 0.6 M sucrose, 0.8 M sucrose, and 1.0 M sucrose. In order to remove any impurities from the outside of the bags, we rinsed them off after filling them. We then dried and massed each bag, then let them sit in cups of distilled water for 30 minutes to allow for the water molecules to diffuse across the membrane. We removed the bags from the cups after 30 minutes and dried and massed each. In the potato cores part of the lab, we used a cork borer to cut eight potato cores. We

cut off the skin from each core and cut them to make them as similar in size as possible, then massed them in two groups of four. We placed the two groups of potato cores in two sucrose solutions of different molarities (one 0.8 M and one 1.0 M) and covered them with plastic wrap to keep the water from evaporating. After letting the potato cores stand overnight, we patted them dry and massed them in the same groups of four. III. Results/Data Table 1: Qualitative Diffusion
Initial Contents Bag: 15% glucose & 1% starch Beaker: H2O + IKI Solution Color Initial Final Very slightly cloudy white Ruby (iodinecolored) Dark navy Ruby (iodinecolored) Presence of Glucose Initial Final Present Absent Present Present

Table 2: Percent Change in Mass of Dialysis Bags (Lab Group Data)

Contents in Dialysis Bag Distilled Water 0.2 M 0.4 M 0.6 M 0.8 M 1.0 M Initial Mass 24.9 g 25.5 g 25.5 g 20.0 g 23.6 g 29.0 g Final Mass 24.8 g 26.3 g 27.8 g 22.7 g 27.8 g 34.1 g Mass Difference 0.1 g 0.8 g 2.3 g 2.7 g 4.2 g 5.1 g Percent Change in Mass -0.4% 3.1% 9% 13.5% 17.8% 17.6%

Table 3: Percent Change in Mass of Dialysis Bags (Class Data)

Group1 -0.5% 4.16% 8.68% 14.14% 16.79% 16.93% Group 2 0.36% 8.93% 3.85% 13.32% 14.53% 16.44% Group -0.99% 4.61% 10.23% 17.03% 18.72% 18.9% Group 4 -0.387% 2.8% 8.17% 5.4% 14.98% 17.72% Group 5 -0.8 4.68 8.73 14.71 16.85 20.51 Group 6 -0.4 3.1 9 13.5 17.8 17.6 Average -0.45 4.71 8.11 13.02 16.61 18.02

Distilled Water 0.2M 0.4M 0.6M 0.8M 1.0M

Table 4: Percent Change in Mass of Potato Cores (Class Data)

Group 1 12.77% 1.57% -15.85% -21.85% -16.12% -13.96% Group 2 21.66% 1.57% -15.73% -22.73% -26.09% -21.74% Average 17.215% 1.57% -15.79% -22.29% -21.105% -17.85%

Distilled Water 0.2M 0.4M 0.6M 0.8M 1.0M

*Lab group data IV. Conclusions The diffusion part of the lab required us to explore the concept of diffusion qualitatively, using dialysis tubing as a model of a selectively permeable membrane. We were able to observe the random movement of glucose, IKI, and water molecules through the dialysis tubing as the molecules dispersed themselves between the solutions in the beaker and the dialysis bag to achieve dynamic equilibrium. The net movement of the glucose molecules was from the bag to the beaker because the concentration of glucose was originally greater in the bag than in the beaker. After 30 minutes, the solution in the beaker tested positive for the presence of glucose, proving that the glucose molecules had diffused from the bag to the beaker. The IKI solution must have also diffused across the membrane because although initially only the solution in the beaker contained IKI, the solution within the dialysis bag turned blue-black by the end of 30 minutes, indicating that the IKI had passed through the membrane pores; the color change was the result of the indicator coming into contact with the starch in the bag. This color change was only visible within the dialysis bag, proving that the starch molecules were too large to diffuse through the membrane pores; had this not been the case, the solution in the beaker would have also turned blue-black. The size of the pores in the dialysis tubing allowed the smaller molecules (water, IKI, and glucose) to pass through the membrane, while preventing the larger molecules (starch) from moving along the concentration gradient. The data from the osmosis part of the lab generally proved that the amount of water that moves into a system

corresponds to the original concentration of the solute; the higher the original molarity of the sucrose solution, the more water molecules that move into the dialysis bag to balance out the concentrations of the solutions on either side of the membrane. This was proven by the generally increasing percent changes in mass based on results from the bag of distilled water as compared to the results from the bag of 1.0 M sucrose solution. The only part of our data that was inconsistent with the rest of our findings was the fact that the percent change in mass of the 0.8 M sucrose solution was 0.2% greater than that of the 1.0 M sucrose solution; the percent change in mass should have been greater in the 1.0 M sucrose solution than in the 0.8 M sucrose solution. It is possible that we did not rinse the exterior of the bag containing 1.0 M solution thoroughly enough, or that we did not leave enough room in the bag for the complete expansion of its contents, or that the dialysis tubing was leaking. As expected, the potato cores absorbed the most water when placed in distilled water because their water potential was much lower than that of pure water. The cores still gained 1.57% in mass after soaking in 0.2 M sucrose solution, indicating that the water potential of the 0.2 M solution was still higher than that of a potato cell. The values for the percent change in mass became negative as the molarity of the solutions increased, starting with 0.4 M sucrose solution; that is, water molecules diffused from the potato cells to the solution because the water potential of the cells was higher than the water potentials of the 0.4 M, 0.6 M, 0.8 M, and 1.0 M sucrose solutions. The results for the percent change in mass for the 0.6 M, 0.8 M, and 1.0 M solutions look startlingly different from what was expected from the experiment. Soaking the potato cores in the 0.6 M solution yielded the largest percent change in mass, followed by the 0.8 M solution, followed by the 1.0 M solution; of these three solutions, the 1.0 M solution was expected to effect the greatest percent change in mass while the 0.6 M solution was expected to effect the least percent change in mass. These results are deceiving; because the masses of the potato cores are so small, any minor errors made in massing them before and after soaking them in the solutions create a large percent error.