David Solinger AP Biology November 3, 2011 Lab 1: Diffusion and Osmosis Objectives: Measure the water potential of a substance

ance in a controlled experiment. Determine the osmotic concentration of living tissue or an unknown solution from experimental data Describe the effects of water gain or loss in animal and plant cells Release osmotic potential to solute concentration and water potential

Introduction: Diffusion is the process that allows particles or molecules to move from an area of high concentration to an area of low concentration. This process accounts for the movement of many small molecules across cell membranes. The rate of diffusion of particles will be affected by the steepness of the concentration gradient or the difference in concentration between two areas. Diffusion can only occur across a membrane if there is a concentration gradient and the membrane is passable. If neither one is present, diffusion wont occur. The size of the tiny pores in a membrane, among other things, decides which particles can diffuse across the membrane. The process of a solute moving through a semipermeable membrane is known as dialysis. Osmosis is a specialized case of diffusion that involves the passive transport of water. In osmosis, water moves through a semipermeable membrane from a region of higher concentration of water to a region of lower concentration. When two solutions have the same concentrations of a solute they are said to be isotonic. When one solution has more solute than another solution it is said to be hypertonic to the other. When one solution has less solute than another solution it is said to be hypotonic. In osmosis, the concentration of water is more important than the concentration of solute. If two solutions of unequal concentrations of a solute are separated by a semipermeable membrane the water can pass through but the solute cannot, the net flow of water will be from the hypotonic solution to the hypertonic solution. This is osmosis and will continue until equilibrium is reached, meaning the rate of flow going both ways across the membrane will be equal and the net flow will be zero. Water potential is used to describe the tendency of water to leave one place in favor of another. Water always moves from an area of higher water potential to an area of lower water potential. Water potential is affected by pressure and the amount of solute. If a red blood cell dropped into distilled water, water will move into the red blood cell and cause the cell to expand, because it has a lower water potential. Water potential is the sum of pressure potential and solute potential. At standard pressure the pressure potential is always 1. Water potential can be positive, negative, or zero. Tonicity is a measure of the osmotic pressure gradient of two solutions separated by a semipermeable membrane and can be described using water potential. It is only affected by solutes

which cannot pass through a membrane because those solutes create an osmotic pressure. Tonicity refers to the relative solution concentrations and can be described using the terms isotonic, hypertonic, and hypotonic defined earlier. Water will move from a hypotonic solution to a hypertonic solution through a water permeable membrane, given that the membrane doesnt allow the solute to diffuse. Normally cells in our body and the solutions theyre in are isotonic and changes in their relative tonicity can cause them to fill with water and burst or shrink; both cases cause harm to the cells. Plasmolysis is osmotically induced shrinkage of a cells cytoplasm. It occurs when a cell is placed in a hypertonic solution causing the water in the cell to be pulled out due to osmosis and as a result the cytoplasm shrinks. For example if a solution with a high concentration surrounds a plant cell the outside solution is hypertonic to the cytoplasm of the cell resulting in water escaping the cell and cytoplasm shrinking. Part A: Table 1.1 Initial Contents Bag Beaker 15% glucose & 1% sucrose HO + IKI Solution Color Initial clear golden brown Final blue/black golden brown Presence of Glucose Initial Final positive negative positive positive

1) Lugols iodine entered the bag, because the solution within the dialysis bag changed from a clear color to a blue/black color. We know because the solution within the bag contains starch that the blue/black color is caused by the diffusion of Lugols iodine into the back, which then reacted with the starch present. Glucose diffused out of the bag, an area of high glucose concentration, into the beaker solution, an area of low glucose concentration. We know this because after the experiment was conducted we tested the beaker solution for glucose using Benedicts solution. This indicates that glucose molecules are small enough to diffuse through the membrane. 2) The movement of Lugols iodine resulted from the relative high concentration of it outside the bag compared the solution within the bag. It also occurred because it is small enough to diffuse through the pores in the membrane. The glucose was in higher concentration in the bag compared to outside the bag resulting in it to diffuse through the bag. Glucose was able to diffuse through the bag because it is small enough, but we know the starch didnt diffuse, even though there was a concentration gradient, because the outside solution after the experiment was conducted didnt react to Lugols iodine. This means the starch was too large for the membrane

3) This experiment can be conducted quantitatively by measuring the percent change in mass of the dialysis bag at the end of the experiment compared to the beginning. The change in mass can show the net movement of water through the bag during the experiment. 4) Water molecules, IKI molecules, glucose molecules, membrane pores, and starch molecules. 5) I would expect the glucose and IKI molecules the diffuse out of the bag as a result of the higher concentration of the two of them inside the bag in relation to the outside of the bag. When the IKI diffuses out of the bag I would expect it to react with the starch present outside of the bag and turn the solution blue/black. I would also expect the solution outside the bag to react positively to Benedicts solution at the end of the experiment due to the glucose that diffused into it. The starch is too large to diffuse, so I would expect it not diffuse at all. Part B Table 1.2 Contents in Dialysis Bag 0.4 M 0.6 M 0.8 M 1M Table 1.3 Final Mass (g) 30.63 31.54 33.79 33.93 Mass Difference (g) 3.32 3.75 5.03 4.99 Percent Change in Mass 12.16 13.49 17.49 17.24

Initial Mass (g) 27.31 27.8 28.76 28.94

Group 1 Distilled Water 0.2 M 0.4 M 0.6 M 0.8 M 1M 1.24 4.19 6.7 11.3

Group 2

Group 3 -9.204 5.766

Group 4 10.97 -8.587 5.86 7.971

Group 5

Group 6 0.5347 1.481

Total 22 11.4 31 42.26 54.4 50

Class Average 5.49 3.78 7.75 10.56 13.61 12.51

12.16 13.49 17.49 17.24

14.43 9.351

6.257 9.496 11.906 11.381

10.61 12.06

Graph 1.1

Effect of Molarity on Percent Change in Mass

16 14

Percent Change in Mass

12 10 8 6 4 2 0 0.2 M 0.4 M 0.6 M 0.8 M 1M Class Average Percent Change in Mass Linear (Class Average Percent Change in Mass)

Molarity
1) The percent change in mass of the sausage increases as the molarity of the sucrose solution is increased; this is a direct relationship. This relationship is caused by an increase in the steepness of the concentration gradient and as a result the net movement of water increases. 2) If the bags were placed in a 0.4 M sucrose solution rather than distilled water the percent change in mass for the distilled water sausage and the 0.2 M sausage should actually be negative, because they would both be hypotonic to the outside solution. Change in mass for the 0.4 M sausage should be zero, because it would be isotonic to the surrounding solution. The percent change in mass for the 0.6 M, 0.8 M, and 1 M solutions would still be positive, because they would all be hypertonic to the beaker solution. The order of the sausages from smallest change in mass to largest change in mass should not change just the percent change in mass for each should decrease. 3) We calculate the percent change in mass rather than just the change in mass because the initial masses for all of the sausages were not necessarily the same, which would make it difficult to compare the change in mass. The percent change in mass allows us to compare the changes in mass easily. 4) Percent change in mass = final mass-initial mass x 100 Initial mass Percent change in mass = 18g 20g x 100 20g Percent change in mass = -10%

Part C Table 1.4 Contents in Beaker Distilled water 0.2 M Sucrose 0.4 M Sucrose 0.6 M Sucrose 0.8M Sucrose 1 M Sucrose Initial Mass (g) 11.88 12.01 8.43 8.77 9.94 8.63 Final Mass (g) 13.4 12.34 7.07 6.2 6.87 5.3 Mass Difference (g) 1.52 0.33 -1.36 -2.57 -3.07 -3.33 Percent Change in Mass 12.8 2.7 -16.1 -29.3 -30.89 -38.6

Graph 1.4

Effect of Molarity on Percent Change in Mass

30

Percent Change in Mass of Potato

20 10 0 -10 -20 -30 -40 0M 0.2 M 0.4 M 0.6 M 0.9 M 1M Percent Change in Mass of Zucchini Linear (Percent Change in Mass of Zucchini)

Molarity of Sucrose Solution in Beaker

Part D Calculation for potato cores: s=-iCRT s = -(1)(0.25 mole/liter)(0.831 liter bars/mole K)(296 K) s = -5.9 bars

1) If a potato core was allowed to sit in the open and dry out, its water potential would increase and the concentration of water inside the cells would decrease. The cores would become dehydrated causing them to more readily absorb water than before. 2) A plant cell that has a lower water potential than its surroundings means that is hypertonic to its surrounds and the net flow of water will be into the cell. The cell will gain water. 3) The pressure potential is zero. The pressure potential is always zero if the container is open to the atmosphere at standard atmospheric pressure. 4) The dialysis bag has the greater water potential. 5) Water will diffuse from the inside of the bag out to the solution in the beaker. This is because the water potential within the bag is higher than the water potential in the beakers solution and water always flows from a higher water potential to a lower water potential until equilibrium is reached.

6/7)

Effect of Molarity on Percent Change in Mass

30

Percent Change in Mass of Zucchini

20 10 0 -10 -20 -30 -40 0M 0.2 M 0.4 M 0.6 M 0.9 M 1M Percent Change in Mass of Zucchini Linear (Percent Change in Mass of Zucchini)

Molarity of Sucrose Solution in Beaker

b) The molar concentration of the zucchini cells based on the graph is 0.358 M. 8) a) s=-iCRT s = -(1)(0.358 mole/liter)(0.831 liter bars/mole K)(300 K) s = -8.92 bars b) = p + s = (0 bars) + (-8.92 bars) = -8.92 bars 9) Adding more solute to a solution decreases its water potential meaning that water will more readily diffuse into it. Adding solute increases its molarity thereby increasing s and making the water potential a smaller number (larger negative number). 10) a) Distilled water has a higher concentration of water molecules compared to a red blood cell.

b) Distilled water also has a higher water potential, because its molarity is zero meaning its water potential is also zero, while the molarity of a red blood cell is a positive number, causing its water potential to be negative. c) The red blood cell would flood with water and possibly burst. Because the cells have such a low water potential in relation to distilled water and water flows from a higher potential to a lower potential water will flow into the cells.

Part E 1) The onion cells initially look like rectangles clearly defined by cell walls and full with a purple/pink color.

2) After plasmolysis has occurred the cell walls look like they havent changed, but the purple/pink colored parts of the cell (cytoplasm) has shrunk significantly in all the cells.

3) After water was added back to the cells, they regained their former full purple/pink pigment.

1) Plasmolysis is osmotically induced shrinkage of a cells cytoplasm. It occurs when a cell is placed in a hypertonic solution causing the water in the cell to be pulled out due to osmosis and as a result the cytoplasm shrinks while the cell wall maintains the cells shape. 2) In this case the NaCl solution had a relatively high concentration compared to the onion cell. The solution is hypertonic to the cytoplasm of the cell, resulting in water escaping the cell (high water potential) and going into the outside solution (low water potential). 3) Plasmolysis is the cause of grass dying near roads that were salted. The melting ice turns into a highly concentrated solution of NaCl with a low water potential relative the cells in the grass. The concentration gradient between the grass and the melted ice causes water to flow out of the grass cells and into the solution create by melted ice. If the grass is left in this relatively hypertonic solution and doesnt regain its normal amount of water within its cells, they will die.