Austin Stults September 29, 2011 Ms.

Kellogg/AP Biology Osmosis/Diffusion and Water Potential Lab PART 1: Diffusion Introduction: The movement of particles into and out of a cell through the plasma membrane is a highly regulated process involving many different proteins, movements, and cellular energy. One of these forces is known as diffusion. Diffusion is defined as the random movement of molecules, atoms, or ions from an area of relative high concentration to an area of relative low concentration. Diffusion is an inclusive term that is applicable to all movement of particles through a plasma membrane; this process is extremely important for the function of a cell. Only certain particles can diffuse across a membrane without any help from proteins. In this lab, we will focus on the factors that inhibit or promote diffusion by using dialysis bags, mixed glucose and starch solution, and Iodine solution to determine which molecules will move through the permeable membrane, which in this case is the dialysis bag. The hypothesis for this lab is that the glucose will move through the dialysis tubing, but the starch will not because of its large, complex 3D shape. Materials: Solution of soluble starch and 15% glucose Solution of Iodine Glucose test tape stripes

 Procedure:

Dialysis tubing, string Small plastic beakers, graduated cylinders, pipettes Distilled water

The first step in this lab was to take a strip of dialysis tubing and cut a piece of about six inches off. The tubing was dipped in water to make it easier to open. Once the tubing was opened, one end was tied off with string and 30 ml of soluble starch and glucose solution was poured into the tubing, and then the other end of the tubing was tied with some more string. Before pouring the glucose solution into the dialysis tube, a glucose test tape strip was used to confirm that there was glucose in the solution by putting the strip into the solution and waiting a few minutes. The strip turned a dark shade of brown to indicate the presence of glucose. A cup was filled with water, and then 20 ml of Iodine solution was added to the water, which turned the water a gold color. The purpose of adding Iodine to the water is that Iodine detects starch in a solution, and causes a chemical reaction that turns the solution blue. By using the Iodine, observers can detect if the starch moves through the dialysis membrane. After adding the Iodine to the water, the dialysis tubing was put into the water and left for five minutes. Results: After letting the dialysis tubing sit in the water for a few minutes, the first observable change was that the solution inside the tubing had turned a color of blue. This indicated that the Iodine had moved through the membrane into the extracellular fluid, which meant that the tubing was permeable to Iodine. The water remained a gold color, which indicates that the starch present inside the solution did not move out of the tubing. This means that the pores of the membrane were not big enough to allow the

diffusion of starch. We then used another glucose test tape strip to determine if any glucose had moved into the water. The glucose strip turned brown for a second time, indicating that the membrane pores were big enough to allow transport of glucose molecules, and that the glucose did diffuse into the water. Based on our results, I would rank the following items in order from smallest to largest as follows: water, Iodine, glucose, membrane pores, starch. If the experiment was repeated but sucrose was used instead of glucose, the results would most likely stay the same. Sucrose is a disaccharide containing the monosaccharides glucose and fructose. Compared to starch, which is a long chain of glucose molecules bound together, sucrose is much smaller than starch and relatively similar to glucose, so it should be able to move through the membrane. Possible sources of error include the string tied to the tubing being too loose and allowing molecules to flow through the open tube, instead of through the membrane; also, the tubing might not have been submerged in the water long enough for the diffusion to reach equilibrium, or the point where no net movement of molecules has been met. PART 2: Osmosis Introduction: Osmosis is defined as the diffusion of water molecules across a selectively permeable membrane. Water is an essential molecule for cellular function, and can exist in different concentrations in and out of a cell. Human cells function best in a state where the water to solute concentrations are equal inside and outside of the cell, also known as isotonic. Plant cells are best suited for an environment that is known as hypertonic, where the solute concentration is higher inside of the cell than outside of the cell, causing water

to move inside the cell. In other words, when plant cells have more solutes inside of them than outside, water will rush into the cell. If a cell has less solute than outside of the cell, water from inside the cell will rush out of it, and the cell is referred to as hypotonic. Tonicity refers to the concentration of solutes, not of water. In the purpose of this lab, dialysis tubing is filled with different molarities of sucrose solution and put inside a cup full of distilled water. The movement of water into and out of the cell, or osmosis, is determined by the concentration of solutions inside the dialysis bag. The masses of each solution will be recorded before and after being submersed in water, and the percent changes will be calculated to represent a correlation in the solute concentration and the movement of water into and out of the tubing. Materials: Solution of sucrose, 0.2-1.0 molarity Unknown molarity solution Dialysis tubing, string Small plastic beakers, graduated cylinders, pipettes Distilled water Scale

Procedure: The first step in this lab was to take a strip of dialysis tubing, open it, and pour 10 ml of the designated solution into the tube. For the sake of time, each group did a different molarity solution and the results were shared amongst each other. After pouring the solution into the tube and tying the ends while leaving some space for the expansion of the contents in the bag, the tubes were dried and weighed, and the masses were recorded. Then the tubes were put into a flask of distilled water and left to sit for approximately thirty minutes. After thirty minutes, the tubes were removed, dried, and weighed again. The new masses were recorded on the computer.

Results: The following shows the initial masses, final masses, and percent changes of all the solutions of sucrose found throughout the class: Treatment 0.0 M sucrose 0.2 M sucrose 0.4 M sucrose 0.6 M sucrose 0.8 M sucrose 1.0 M sucrose ? M sucrose Initial Mass 10.1 g 10.12 g 10.59 g 10.12 g 9.99 g 11.39 g 10.66 g Final Mass 10.08 g 10.7 g 11.81 g 11.73 g 11.9 g 14.33 g 11.7 g % Change -.198% 5.731% 11.52% 15.91% 19.119% 25.812% 9.756%

Graphing this data creates a visual correlation in the percent change, or dependent variable, relative to the molarity, or independent variable, of the sucrose solution. A trend line, the trend line equation, and the R2 value have been included.

Percent Change in Mass of Osmosisy = 0.251x Induced Solutions + 0.0037 R = 0.9917

Percent Change

Solution Molarity

As the graph shows, as the molarity of the sucrose solution increased, the percent change in the mass also increased. This is because the water had a tendency to diffuse across the dialysis membrane into the glucose solution, because the contents of the bags became steadily more hypertonic, causing more water to move into the bags. When the dialysis bags were tied, a space was left so that water can move into and out of the tube without it bursting. Using the trend line and the corresponding equation, as well as the percent change of the unknown solution, we can calculate the molarity of the unknown solution. y = 0.251x + 0.003 (y = percent change, x = solution molarity) .09756 = .251x + .003 x = .3767 M The following table describes the changes that would happen if all of the dialysis bags had been placed in a .4 M solution instead of distilled water. Treatment in Tube 0.0 M sucrose 0.2 M sucrose 0.4 M sucrose 0.6 M sucrose 0.8 M sucrose 1.0 M sucrose Reaction Tube is hypotonic, water moves out Tube is hypotonic, water moves out Tube is isotonic, no net movement Tube is hypertonic, water moves in Tube is hypertonic, water moves in Tube is hypertonic, water moves in

PART 3: Water Potential Introduction:

Water potential is known as the prediction of which direction water will move in a living cell. Water potential is measured by finding the sum of the solute potential and the pressure potential of an object. Water will move into and out of a cell depending on the concentration of solutes inside and outside of the cells. If there is more solute outside of a cell, water will move out of the cell into the solvent. If there is more solute inside the cell, water will move into the cell. Water will always move from an area of higher water potential to an area of lower water potential. The purpose of this lab is test water potential in potato cells by submerging them in various solutions, including distilled water and five other sucrose solutions with different molarities. To save time, the different solutions were split among the groups of the class and the results were shared. The percent change in the mass of the potatoes after being submerged in the solution can be used to describe a trend in the molarity of a solution and water potential. Materials: Solution of sucrose, 0.2-1.0 molarity Unknown solution One potato Cork borer Small plastic beakers, graduated cylinders, pipettes Distilled water Scale

Procedure:

The first step in this lab was to pour 50 ml of a sucrose solution with a 0.6 molarity into a beaker. Next, a cork borer was used to cut two potato cylinders out of the available potato. The cylinders were approximately 3 cm long. The skin on the

potato cylinders was cut off to give accurate results. The mass of the two cylinders was recorded, and then they were placed in the beaker with the sucrose solution and left overnight. After approximately 24 hours, the cylinders were removed from the beaker, dried and weighed, and their new masses were recorded. Results: The following table lists the various sucrose solutions, the initial masses, final masses, and the percent change:

Treatment 0.0 M sucrose 0.2 M sucrose 0.4 M sucrose 0.6 M sucrose 0.8 M sucrose 1.0 M sucrose ? M sucrose

Initial Mass 4.27 g 4.25 g 4.02 g 4.53 g 4.21 g 4.72 g 4.3 g

Final Mass 5.23 g 4.53 g 3.65 g 3.9 g 3.15 g 3.1 g 4.1 g

% Change 22.48% 6.59% -9.20% -13.91% -25.18% -34.32% -4.65%

The following graph shows the trend line of the percent changes (dependent variable) in relation to the solution molarity (independent variable). The trend line equation, as well as the R2 value have been included.

Percent Change in Mass of Potato Cells

Percent Change

y = -0.5486x + 0.1851 y R = 0.9749 = -0.5486x + 0.1851 Solution Molarity = 0.9749 R

As the graph shows, as the molarity of the sucrose solution increased, the amount of water in the potato cells steadily increased. This is due to the fact that as the solutions molarity steadily increased, the potato cells became more and more hypotonic relative to the solute concentration outside of the cells, and so the water inside the potato cells moved out. 1. To determine the molarity of the sucrose solution, which is isotonic, the trend line equation should be set equal to zero. y = -.548x + 0.185 (y = percent change, x = solution molarity) 0 = -.548x + 0.185 x = .338 M 2. To calculate the osmotic potential of the isotonic sucrose solution from the last equation the following formula needs to be used:

s = - iCRT
i = ionization constant of the solute (1 for sucrose) c = osmotic concentration of the isotonic solution r = pressure constant (.0831 liter bars/mole oK) t = temperature oK (273 + oC of solution)

s = -(1)(.338 M/liter)(0.0831 liter bar/M oK)(295 oK)

= -8.29 bars 3. If a potato cylinder were allowed to dehydrate by sitting in the open air, the water potential of the potato cells would become more negative because the water would be evaporating from the potato. When water leaves a cell, the osmotic potential becomes more negative.

4. Based on the results of this experiment, spraying water on fruits and vegetables would help maintain their freshness. Water will dehydrate from the plant as time progresses, and so the plant cells with start to become flaccid. If water is sprayed on the plants, the cells should absorb the water and become turgid again. If you think of this in terms of the graph and the trend line, when the potato was put in distilled water, the greatest percentage change in mass occurred, and it was a positive change.

5. Adding too much fertilizer with mineral salts in it can cause plants to wilt. The salts in the fertilizer create a hypertonic environment outside of the cell, so the water in the cells will diffuse out. The loss of water in the plant cells will decrease the turgor pressure that the plant cell walls give off, which will consequently cause the plant to wilt.

6. The value of the solute potential of the unknown solution in the potato experiment was .338 molarity. The value of the solute potential of the unknown solution in the potato experiment was .378 molarity. The values are relatively close to each other, which shows that the experiments were completed correctly.