AP Lab 1: Diffusion and Osmosis

Objectives:
Use dialysis tubing to model diffusion across the cell membrane
Investigate the influence of solute concentration on osmosis

Background
The cell membrane is a cells interface with its surroundings. In one sense, this
membrane must function as a barrier: it must keep together in one bundle the enzymes,
DNA and metabolic pathways that make life possible. The cell membrane must also
function as a gateway: waste products must be discharged through it and essential
materials (oxygen, water, etc.) must enter through it. A membrane that allows some
molecules to pass through while blocking the passage of others is said to be
semipermeable (or selectively permeable). Molecules pass through the cell
membrane either through processes that require the cell to expend energy (active
transport), or through processes driven by kinetic (thermal) energy of molecules
(passive transport).
In these lab activities, you will investigate the passage of materials through a
semipermeable membrane by passive transport. The membrane you will use, dialysis
tubing, is semi permeable because it has submicroscopic holes through it. Molecules
are in constant random motion. By chance, a molecules motion may move it toward
the membrane. If it collides with the membrane wall, it rebounds. If its motion takes it
toward a pore, it may either pass through the pore, or it may rebound, depending upon
the size of the molecule relative to the diameter of the pore. Molecules that are small
enough to pass through the poes can pass through in either direction. Notice that on
one side of the membrane solute molecules have displaced some of the water
molecules. Thus, there is a higher concentration of water molecules on the opposite
side of the membrane. More water molecules are available to collide with the
membrane on the side having the higher concentration of water. Thus, although water
molecules will move in both directions across the membrane, more will move from the
side having the higher concentration to the side having the lower concentration. The
movement of molecules from areas of higher concentration to areas of lower
concentration is called diffusion.
The diffusion of water molecules across a semipermeable membrane is termed
osmosis. A process that depends upon random motion might seem insufficient, but so
many water molecules are involved and they move so fast, that it's estimated that a red
blood cell floating in blood plasma gains an amount of water equal to 125 times its own
volume every second. It also loses the same amount of water each second, all by
osmosis. This occurs because the concentration of solutes in the blood plasma is the
same as the concentration of solutes in red blood cells. Solutions that have the same
solute concentration are isotonic. If we took a sample of whole blood and added salt to
the plasma, increasing its solute concentration, the plasma becomes hypertonic to the
solution in the red blood cells, and the cells lose water and shrink. If we add water to
the blood plasma, decreasing its solute concentration, the plasma becomes hypotonic
to the solution in the red blood cells. The cells gain water, swell, and may even burst.

Demonstration A: Diffusion
In Demonstration A, we will explore the diffusion of different molecules through dialysis
tubing, a semipermeable membrane. We will use glucose test strips to check for the
presence of glucose as well as IKI solution to test for the presence of starch. IKI reacts
with starch to give a dark blue, almost black color.

1. Indicate on Figure 2 the initial locations (inside or

outside of the bag) of all the kinds of molecules
that are available for diffusion through the
dialysis membrane.
a. Glucose (inside the bag)
b. Starch (inside the bag)
c. Iodine-Potassium (outside the bag)

2. For each of the molecules you listed on Figure 2, predict in the space below their
direction of net(overall) diffusion: into the bag, out of the bag, both into and out of
the bag equally, or none (will not diffuse across the dialysis membrane). Give a
reason for each prediction.
a. Glucose (none the bag because glucose is necessary for cellular
energy; it is also very large so it may not be able to pass through
easily)
b. Starch (none because starch is also necessary for cellular energy; it
is also larger so it may not be able to pass through easily)
c. Iodine-Potassium (into the bag because the molecules are pretty
small (only 3 per set))
Table 1. Demonstration A
Solution Color Glucose Test Results

Location Solution Initial Final Initial Final

Dialysis Glucose/starch Clear Purple/Black Positive Positive

Bag

Cup IKI Amber Amber Negative Positive

Analysis of Results - Demonstration A: Diffusion

1. Compare your results with your predictions. Do you find any conflicts that would
cause you to revise your predictions? If so, explain. The results of the
experiment showed that there is the presence of iodine within the bag,
which is what I predicted. However, the results also proved that some
glucose had entered the iodine-potassium solution, albeit not by much.

2. Does this activity account for the diffusion of all the molecules that you listed on
Figure 2 and in your predictions? If not, what data could have been collected to
show the net direction of diffusion of this molecule or molecules? This lab did
not show the net direction of diffusion of the IKI solution, it only showed
the diffusion of the glucose/starch solution. Perhaps by including a step in
the experiment that uses a chemical indicator to show the presence of the
solution would be great at showing IKIs diffusion.

3. What does your data tell you about the sizes of the molecules relative to the pore
size of the dialysis tubing? This lab tells us the sizes of the molecules is
slightly smaller than the pore size of the dialysis tubing. This is due to the
fact that while there was diffusion, the amount was not a lot, suggesting
that the size of the molecule may only be enough to pass through but not
enough to go at a rapid pace like a small molecule would.

Activity B: Osmosis
Materials
Dialysis tubing, plastic cups, distilled water, funnel, sucrose solutions, paper towels,
balance, tape, marker, calculator (optional)

Introduction
In Activity B, you will investigate the influence (if any) of solute concentration on the net
movement of water molecules through a semipermeable membrane. The solute the
class will use is sucrose (cane or table sugar) in the following molar concentrations: 0.0
M (distilled water), 0.2 M, 0.4 M, 0.6 M, 0.8 M, and 1.0 M. Your group will be
responsible for testing two of these sucrose solutions.

Procedure
Complete the following steps for each sucrose solution that you are assigned to test:

1. Label each cup with the concentration of sucrose that you will test as well as your
group name. Pour 160-170 ml of distilled water into each plastic cup.

2. Obtain a piece of dialysis tubing that has been soaked in water. The tubing
should be soft and pliable. Roll the tubing between your thumb and index finger
to open it. Close one end of the tube by knotting it or tying it off with string. This
will form a bag.

3. Using a small funnel, pour 15 ml of sucrose solution into the dialysis bag.
Smooth out the top of the bag, running it between your thumb and index finger to
expel the air. Tie off the open end of the bag. Leave enough room in the bag to
allow for expansion.

4. Dry the bag on paper towels and then determine its mass. Record this as the
initial mass in Table 2.

5. Immerse the dialysis bag in the water in the cup. Make sure that the portion of
the bag that contains the sucrose solution is completely covered by the water in
the cup at all times.

6. Repeat steps 1 - 5 with the other sucrose solution.

7. Wait 30 minutes before continuing to the next step.

8. After 30 minutes, remove the bag from the cup and dry it on paper towels. Mass
the bag and record the final mass in Table 2. Finally, determine the change in
mass of the bag and record this data in Table 2.

Table 2. Activity B
 *Only complete the first 4 columns for the sucrose solutions you are testing. You will fill
in the Class Average columns completely.
Contents in Initial Final Change Percent Class Class average -
Dialysis Bag Mass Mass in Mass Change in average - Percent Change
Mass Change in in Mass
Mass

0.0 M sucrose 14 12.3 -1.70 g 12.14% -0.85 6.07%

(distilled water) 0.00 g 0.00%

0.2 M sucrose -4.45 g 30.27% -1.225 22.54%

2.00 g 14.81%

0.4 M sucrose 2.15 g 14.33% 2.215 14.665%

2.28 g 15.00%

0.6 M sucrose 11.3 g 18.9 g 7.60 g 67.25% 6.925 66.625%

6.25 g 66.00%

0.8 M sucrose 12.5 g 21.2 g 8.70 g 69.60% 10.95 102.145%

5.8 g 13.20 g 134.69%

1.0 M sucrose 15.2 g 19.7 g 7.80 g 56.52% 6.15 43.065%

4.50 g 29.61%

Analysis of Results - Activity B: Osmosis

1. Calculate the percent change in mass. Use the following formula. Record the
results in Table 2.

% Change in Mass = (Change in M ass I nitial M ass) 100

2. Obtain and record the class averages for change in mass and percent change in
mass.

3. What does the change in mass indicate? The change in mass indicated that
water had entered the dialysis bag.

4. Write a hypothesis that this experiment is designed to test. If a sucrose solution

filled dialysis bag is immersed in water, then the mass of the dialysis bag
will increase with time.

5. What variable is being tested in this experiment? Mass of the dialysis bag
6. List at least 3 variables (other than the one listed in your answer to #5) that could
influence the outcome of this experiment. Briefly describe the method of control
used for each of these variables (a.k.a. How did you keep them from influencing
the outcome of the experiment?)

a. How secure the dialysis bag was: We made sure that the dialysis bag
was tight as possible to avoid leaks. This way the osmotic reactions
occurring between the dialysis bag and the water solution were controlled
to just between the molecules and the pores, not with other crevices on
the bag.

b. Amount of space allowed in each bag: We made sure equal amount of

space was allowed in each bag so they have equal opportunity for
expansion. This gives a baseline at which to determine changes in mass
with each bag.

c. How submerged the dialysis bag was in the water: We made sure that
the bag was fully submerged in the water for the whole 30 minutes in
order to ensure that osmotic reactions could happen with the whole bag,
not simply in parts. This gives a better chance for more osmotic reactions
to occur rather than just one portion of the bag.

7. Graph the percent change in mass for the class averages. Title the graph and
supply the following information:

a. The independent variable is: Molar concentrations of sucrose solution

b. The dependent variable is: Percent change in mass

Plot the independent variable on the x-axis, and the dependent variable on the
y-axis.
8. Do your results support your hypothesis, refute it, or require that you modify it?
The results required that I modify my original hypothesis to reflect the
relationship between molar concentrations and change in mass rather than
time and change in mass. This is due to the fact that the results of the
experiment show a stronger correlation with the former.

9. On the basis of your results, write a statement that expresses the relationship of
solute concentration and direction of net movement of water molecules in
osmosis. The higher the solute concentration, the more water molecules will
move inside the dialysis bag.

10. In which, if any, of the experimental setups were the solutions in the bag and
outside of the bag isotonic to each other? The experimental setup with the 0.0
M solution had one instance where the solutions in the bag and outside the
bag were isotonic in relation to one another.

11. When you drink a glass of water, most of it is absorbed by osmosis through the
cells lining your small intestine. Drinking seawater can actually dehydrate the
body. How? When you drink seawater, it has a higher solute concentration
than that of the cells. To reach equilibrium between it and the surrounding
seawater, the cell will expel out water until the concentrations are equal.
This is why you become dehydrated, because the cell is shriveling up as it
outputs more water.