Loneker 1 Abby Loneker Mrs. Kuntz AP Biology 19 October 2011 AP Lab 1: Diffusion and Osmosis Purpose: 1.

Explain the effects of solute and concentration gradient on diffusion across a selectively permeable membrane 2. Determine the osmotic concentration of a living tissue or unknown solution from experimental data 3. Measure the water potential of a solution in a controlled experiment 4. Describe the effects of water gain or loss in animal and plant cells 5. Relate osmotic potential to solute concentration and water potential Exercise 1A: Diffusion: Table 1.1
Initial Contents 15% Glucose & 1% Starch H2O & IKI Initial Solution Color Clear Brownish Red Final Solution Color Black Brownish Red Initial Presence of Glucose Yes No Final Presence of Glucose Yes No

Bag Beaker

1. Which substances are entering the bag and which are leaving the bag? What experimental evidence supports your answer? The Lugols solution entered the bag. This was evident because Lugols will turn black when it reacts with starch and the solution inside the bag, which contained starch, turned black at the end of the experiment. There is no evidence that anything left the bag because the solution

Loneker 2 outside of the bag did not react with any starch and the glucose test showed no evidence of glucose in the beaker after the experiment. 2. Explain the results you obtained. Include the concentration differences and pore size in your discussion There was a higher concentration of Lugols solution outside the bag. This caused the solution to want to enter the bag to reach equal concentrations on both sides of the membrane. There was a higher concentration of glucose and starch inside of the big. This caused the solution to enter the bag as well to dilute the solutes as the glucose and starch molecules were too big to leave the pores of the membrane. 3. Quantitative data uses numbers to measure observed changes. How could this experiment be modified so that data could be collected to show that water diffused into the dialysis bag? Instead of using Lugols solution, submerging the bag in water would allow observations of how water diffused into the bag. A change would be calculated by weighing the bag before submerging it as well as after submerging it. If it weighed more after, it would prove that water had diffused into the bag. 4. Based on your observations, rank the following by relative size, beginning with the smallest: glucose molecules, water molecules, IKI molecules, membrane pores, starch molecules. IKI Molecules, water molecules, membrane pores, glucose molecules, starch molecules 5. What results would you expect if the experiment started with glucose and IKI solution inside the bag and only starch and water outside? Why? The water would diffuse into the bag to try and dilute the IKI solution while the IKI would diffuse out of the bag to try to reach equilibrium. The glucose would remain trapped inside the

Loneker 3 bag while starch would remain outside the bag as both are bigger than the pores. The Lugols would exit the bag and react with the starch molecules, turning the solution black. Exercise 1B: Osmosis Table 1.2
Contents in Dialysis Bag Distilled Water 0.2 M Sucrose 0.4 M Sucrose 0.6 M Sucrose 0.8 M Sucrose 1.0 M Sucrose Initial Mass 15.97g 16.25g 15.75g 15.64g 18.05g 17.23g Final Mass 16.95g 18.64g 23.30g 26.68g 31.19g 35.41g Mass Difference 0.98g 2.37g 7.55g 11.04g 13.14g 18.18g Percent Change in Mass 6.14% 14.6% 47.9% 70.6% 72.8% 105.5%

Table 1.3
Contents in Dialysis Bag Distilled Water 0.2 M Sucrose 0.4 M Sucrose 0.6 M Sucrose 0.8 M Sucrose 1.0 M Sucrose Group 1 -1.49 10.6 35.93 47.63 44.79 92.69 Group 2 2.97 13.71 14.02 28.67 50.75 73.08 Percent Change in Mass of Dialysis Bag Group Group Group Group Total 3 4 5 6 4.54 6.14 1.41 -8.89 4.68 2.46 14.71 -9.43 5.87 37.92 39.5 47.95 -10.99 .203 126.62 8.1 70.59 1.21 60.19 216.39 3.97 72.8 34.01 -34.25 170.07 6.6 105.51 -5.67 14.69 287.9 Class Average 0.78 6.32 21.1 36.1 28.35 47.98

Graph 1.1

Concentration of Solution v. Percent Change in Mass

Percent Change in Weight of Bag 120 100 80 60 40 20 0 0 0.2 0.4 0.6 Concentration of Soultion 0.8 1 Class Data Group Data

Analysis Questions:

Loneker 4 1. Explain the relationship between the change in mass and the molarity of sucrose within the dialysis bag. As the molarity of the sucrose inside the bag increased do did the change in mass. This is because more water had to enter the bag to reach almost equal concentrations on both sides of the bag. 2. Predict what would happen to the mass of each bag in this experiment if all the bags were placed in 0.6 M sucrose instead of distilled water. Explain. The bags with distilled water, 0.2 molar, and 0.4 molar solutions would lose mass as water would leave the bag in an attempt to dilute the hypertonic solution. The bag filled with 0.6 molar solution would be isotonic to the solution and would therefore experience no net change in mass. The bags with 0.8 and 1.0 molar solutions would gain mass as water would enter the bag in an attempt to reach equal concentrations on both sides of the membrane. 3. Why did you calculate the percent change in mass rather than the change in mass? The starting mass would have an effect on how much the mass changed. This adds a separate variable that needs to be accounted for. The way to account for variability in the starting weights is to calculate the percent change. This stops the relative starting masses from changing the data. 4. A dialysis bag is filled with distilled water and placed in a sucrose solution. The bags initial mass is 20 g. and its final mass is 18g. Calculate the percent change in mass. = = -10%

5. The sucrose solution in the beaker would have been hypertonic to the distilled water in the bag.

Loneker 5 Exercise 1C: Water Potential What is the water potential of a potato cell that has a concentration of 0.32 M (assume room temp. is 25o C)?

s = -iCRT s = -1(0.32)(0.0831)(25+273) s = -7.92 p = 0 = s+ p = -7.92+0=-7.92 bar

1. If a dehydrated potato cell was placed in a cup of distilled water, would the water potential of the potato cell increase or decrease? Why? The water potential of the potato would have to increase to reach equilibrium. This is because a dehydrated potato cell would have a lower water potential than distilled water because of the presence of solutes in the potato. The water potential would have to increase to level with that of the distilled water. 2. If a plant cell has a greater water potential than its surrounding environment and if pressure is equal to zero, is the cell hypertonic (in terms of solute concentration) or hypotonic to its environment? Will the cell gain or lose water? Explain. A cell with a greater water potential will have less solute than its surroundings. A solution with less solute is a hypotonic solution. This means that the cell will lose water because the water will flow from + to -. 3. Figure 1.5 is open to the atmosphere. What is the pressure potential of the system. The pressure potential is 0 because it is an open system. 4. In figure 1.5 where is the lowest water potential? Beaker

5. Water will diffuse out of the bag. Why? Water will always flow from areas of higher water potential to areas of lower water potential.

Loneker 6 6. Zucchini cores placed in a sucrose solution at 27oC resulted in the following percent changes after 24 hours:
% Change in Mass Sucrose Molarity 30 20 10 -3 -17 -25 Distilled Water 0.2 M 0.4 M 0.6 M 0.8 M 1.0 M

7. Graph the results

Percent Change in Mass

40 Percent Change in Mass 30 20 10 0 -10 -20 -30 Molarity of Solution 0 0.2 0.4 0.6 0.8 1 1.2

8. Where the graph crosses the x axis is where the solution would be isotonic to the zucchini core. This is where there would be no net movement of water. What is the molar concentration of the solutes within the zucchini cells? Approximately 0.56 molar 9. Refer to the procedure for calculating water potential. a. Calculate the solute potential of the sucrose solution in which the mass of the zucchini cores does not change.

s = -iCRT

s = -1(0.56)(0.0831)(300) s = -13.96 bar

Loneker 7 b. Calculate the water potential for the zucchini cores.

= s+ p

p=0

= -13.96+0=-13.96 bar

10. What effect doe adding more solute have on the solute potential component? Why? Adding more solute will decrease the solute potential component because a larger number will be multiplied by a negative 1 resulting in a lower solute potential. 11. Consider what would happen to a red blood cell placed in a hypertonic solution. a. Which would have the higher concentration of water molecules? RBC b. Which would have a higher water potential? RBC c. What would happen to the red blood cell? Why? The red blood cell would shrink because the water would have to flow from the red blood cell (higher ) to the solution (lower ). Exercise 1D: Water Potential- An Inquiry Based Approach Procedure: Record the mass of 6 approximately evenly sized pieces of potato; they should be no more than a cm2 in size. Place 1 piece of potato in each of 6 beakers. The beakers should be filled with 100 ml of solution. Fill one beaker with distilled water, another with 0.2M sucrose, another with 0.4M sucrose, another with 0.6M sucrose, another with 0.8M sucrose, and the last with 1.0M sucrose. Leave the potato pieces in for 30 minutes. When the time is up, remove the potato pieces and pat them dry. Then mass the pieces and record the values in the data chart. Use the collected values to calculate the percent change in each solution and graph. Then calculate the x-intercept to find the concentration where the net mass change is zero. This is the correct concentration for an isotonic solution.

Loneker 8 Data: Data Table

Molarity of Sucrose Distilled Water 0.2 M Sucrose 0.4 M Sucrose 0.6 M Sucrose 0.8 M Sucrose 1.0 M Sucrose Initial Mass 0.96g 1.62g 1.22g 1.43g 1.56g 1.26g Final Mass 0.99g 1.58g 1.22g 1.23g 1.25g 1.10g Percent Change in Mass 3.125 -2.469 0 -7.692 -19.87 -12.70

Graph 1.3

Percent Change in Mass v. Sucrose Molarity

5 Percent Change in Mass 0 -5 -10 -15 -20 -25 Sucrose Molarity 0 0.2 0.4 0.6 0.8 1 1.2

Conclusions & Sources of Error: According to the graph, the molarity of the potato cells is approximately 0.18M. This is the x-intercept: the place where the net mass change is equal to 0 and the solution is therefore isotonic to the cells. One source of error in this experiment is the variation in the size of potato pieces. Some were larger than others and this could have inhibited water from diffusion into the cell because of a poor surface area to volume ratio. Another source of error is the amount of time the potato

Loneker 9 pieces were soaked in the water. A longer time soaking would have made sure the cell was able to reach equilibrium and would have clarified the results. Exercise 1E: Onion Cell Plasmolysis Procedure: (For pictures of all 3 slides see last page)

1. Prep a wet mount slide of a small piece of the epidermis of an onion. Observe the cells under 100 x magnification. Describe the appearance of the onion cells. The cells are rectangular in shape with a thick outer cell wall and a thinner cell membrane within it. 2. Add NaCl to the cover slip. Explain what happened to the cells. The membrane shrunk while the cell wall remained in place. This is because the solution was hypertonic to the cell. The water within the onion cell rushed out of the membrane, shrinking it. 3. Remove the cover slip and flood the cell with fresh water. Describe and explained what has happened. The membrane swelled against the cell wall. This is because the solution was hypotonic to the cell. The water outside of the cell rushed in to try to reach equilibrium, expanding the cell. Analysis of Results: 1. What is plasmolysis? Plasmolysis is the process in plant cells where the plasma membrane pulls away from the cell wall due to the loss of water through osmosis. 2. Why did the onion cells plasmolyze? The cells plasmolyzed because they were placed in a hypertonic solution that caused water to rush out of the cell. This made the cell membrane retreat from the cell wall.

Loneker 10 3. In the winter, grass often dies near roads that have been salted to remove ice. What causes this? The salt on the road attracts the water in the grass due to the concentration gradient. This causes water in the grass to leave and without water the grass dies very quickly.