Part I.

Diffusion

Objectives
Demonstrate the process of simple diffusion
Demonstrate the influence of molecular size on diffusion through a selectively permeable membrane

Experiment 1. Membrane selective permeability; the effect of

molecular size on diffusion through a membrane

Procedure:
1. Obtain a piece of dialysis tubing. Soak in tap water until softened.
2. Using label 4 test tubes: 0 G (Glucose), 15 G, 30 G, 60 G and 4 more: 0 S (Starch), 15 S, 30 S, 60 S.
Place these test tubes in the test tube rack.
3. Roll up and clamp one end of the dialysis tubing. Gently tease open the opposite end, forming a bag.
4. Combine 10 mL each of the starch and glucose solutions in a plastic mixing bottle. Mix thoroughly by
shaking the bottle for at least 5 seconds..
5. Carefully pour the combined solution from the mixing bottle into the bag.
6. Carefully squeeze the bag to remove most of the air and seal with a clamp, leaving space for the bag to
expand.
7. Thoroughly rinse the outside of the filled bag to remove any spillage of the organic solution. Pat the filled
bag dry with a clean paper towel, and gently squeeze the bag to confirm it is completely sealed. If leakage
occurs re-seal, re-test for leaks and rinse the outside again.
8. Place the bag into the 400 mL beaker.
9. Add 75 mL of warm tap water (approximately 40oC). Make certain the as much of the bag is submerged
as possible. The clamps do NOT need to be submerged.
10. Immediately (time 0) take a sample of the water surrounding the bag and distribute 20 drops of this
sample into each of 2 test tubes labeled 0 minutes.
11. Set your timer for 15 minutes. Repeat the sampling and analysis procedures on the beaker water after 15
minutes, 30 minutes, and 60 minutes of incubation in the warm water.
12. While you wait for your second sample, set up Positive and Negative Controls, as indicated below, to be
used in analyzing for the presence of the starch and glucose in the products of your diffusion tests.

13. Use the reagent solutions to analyze the sample for the presence of starch and glucose in the water
samples at time 0,15,30,60 minutes. Follow the negative control procedure utilizing sample water
from the beaker rather than distilled water. For example: At time 0, place 10 drops of sample water
in each 0 minute test tube. Then add 5 drops of iodine to one test tube to test for starch; 20 drops
of Benedicts to the other test tube to test for glucose.
14. Record the results of your tests in Table 1. Record a - for negative test results, a +for positive test results,
and ++for stronger positive test results.

Table 1.
Elapsed Time (min) Starch Glucose
0 Negative Negative
15 Negative Positive
30 Negative Positive
60 Negative Positive

15. After 60 minutes, remove the bag from the beaker and place in a large weighing boat. Blot dry with a paper
towel.
16. Drop 10 drops of iodine reagent on the surface of the dialysis bag. Observe. Note the approximate time
required for a change to occur.

Questions:
1
 1. Use the timeline below to indicate the time required for the iodine, glucose and starch to diffuse into or out of the
dialysis bag. Use an I to represent the time required for iodine to diffuse into or out of the bag. Use a G for
glucose and an S for starch. Hint: The iodine reagent consists of crystalline Iodine (I) and potassium iodide, KI.
The formula for the reagent is IKI and which consists of 3 atoms. The formula for glucose is C 6H12O6; it consists of 24
atoms. Starch is a polymer consisting of multiple glucose monomers.

_________________________________________
0 15 30 60 minutes

2. Which of the following DID diffuse: iodine, glucose, starch? Did they diffuse at the same time? Why or why not?

3. Which molecule did NOT diffuse? Why did it not diffuse?

Part II. Osmosis

Osmosis is the diffusion of water (or other substance) across a differentially permeable membrane. Water, like
other molecules, diffuses from a region of relatively high concentration (higher free energy) to a region of lower
concentration (lower free energy). Although water molecules move in both directions across a selectively permeable
membrane, when the concentration of water molecules is higher on one side of a membrane their net movement is toward
the side where they are less concentrated. The molecules will continue to diffuse down the concentration gradient until
they reach equilibrium on both sides of the membrane, i.e. the concentration of water molecules is the same on both
sides. In terms of energetics the free energy level within the system is minimal.

Objectives
Demonstrate the process of osmosis using dialysis tubing
Demonstrate the process of osmosis through the biological membrane of a shelless chicken egg

Materials: Equipment
solutions, miscellaneous items 600 mL beaker
400 mL beaker with corn syrup dialysis tubing
shelless chicken egg tubing clamps
Recycled corn syrup digital balance, small weighing boat

Experiment 2: Osmosis demonstration using an artificial membrane

Procedure:
1. Prepare a 20 cm piece of dialysis tubing by soaking it in tap water for a few minutes.
2. After soaking the dialysis tubing in water to soften it, open the tubing and close one end using a tubing clamp to
form a bag.
3. Fill the bag about full with recycled corn syrup solution, gently squeeze it to displace the air, and close the
end with a dialysis tubing clamp. Carefully rinse the outside of the filled bag with water, and gently pat it dry with a
paper towel. Immediately (time 0) weigh the bag (to minimize the effects of evaporation), record the weight in
Table 2, and immediately place the bag into a 600 mL beaker filled with 500 mL tap water.
4. After 15 minutes remove the bag from the water, dry and weigh it as before, and quickly returning it to the beaker
of water. Record the weight as before.
5. Repeat the weighing and recording process after 30, 45 and 60 minutes.
6. Using the weights you have recorded calculate the net change (gain or loss) in weight for each time period. Net
increase = (New weight in g Initial weight in g). Record the net net weight changes in their respective spaces in
Table 2 (p. 4).
7. Using the weights you have recorded calculate the percentage of weight change at each interval as follows:

% change = (new wt in grams- initial wt in grams/initial wt in grams) X 100

8. Plot the percentage results on graph 2 (Figure 2, p.5). Note that the net change may be either a positive number
(gain) or a negative number (loss).

2
 Table 2 Weights for Dialysis Tubing
Time Intervals (min.)
0 15 30 45 60
Initial Weight (g) 32 32 32 32 32
(Does not change)
New Weight (g) ----- 36 39 42 44
Net Gain or Loss (g) ----- +4 +7 +10 +12
% Change in -----
mass* 12.5% 21.87% 31.25% 37.5%
*Percent change in mass = (new wt initial wt) / initial wt X 100

Graphing: You will create two graphs for Experiment 2. Remember, if the bag gained/lost weight, your graph
should reflect that visually. Adjust zero as necessary to create a graph that depicts what occurred in lab.

Create a line graph by plotting the dialysis tubing net gain or loss in weight (use numbers from the third row in
Table 2) over time in the graph below. Label both axes and add a title to the graph.

Figure 1:

Plot the percent change in mass over time in Figure 2. Label each axis and create a title for the graph.

Figure 2:

3
Questions:
1. Was there a change in the weight of the dialysis bag? Briefly explain what happened and why using the
terms hypertonic, hypotonic and osmosis.

2. The diagram below represents the dialysis bag and the beaker of water in which it was placed. Label the
solution inside the dialysis bag and the solution inside the beaker as hypertonic or hypotonic. Using
an arrow, indicate the direction in which water flowed: into, or out of the model cell (dialysis bag).

Beaker

Dialysis Bag

Experiment 3. Osmosis demonstration using a biological membrane

Each group will use a shelless chicken egg. The shell has been dissolved away leaving the shell membrane and the
rest of the egg intact. The membrane selectively permits some kinds of molecules to pass through while preventing
others from doing so.

Procedure:
1. Carefully (fragile!) remove the shelless egg from the container and gently pat it dry to remove any excess surface
water. Immediately weigh the egg, record the weight and place it into the beaker containing corn syrup.
2. Repeat the drying and weighing process after 15, 30, 45 and 60 minutes in the corn syrup. Record the results of
each weighing in Table 3 (below).
Note: When the egg is first placed in the corn syrup, half of the egg will not be submerged. GENTLY roll the egg with the plastic spoon to make
certain that all of the membrane is in contact with the syrup.

Table 3 Weights for Shelless Chicken Egg

Time Intervals (min.)
0 15 30 45
Initial Weight (g) 98 98 98 98
(Does not change)
New Weight (g) ----- 90 84 79

Net Gain / Loss(g) ----- -8 -14 -19

% Change in mass* ----- -8.16% -14.28% -19.38%

*Percent change in mass = (new wt initial wt) / initial wt X 100

4
Create a line graph by plotting the shelless egg net gain or loss in weight over time in the graph below. Label
both axes and add a title to the graph. Remember, if the bag gained/lost weight, your graph should reflect that visually.
Adjust zero as necessary to create a graph that depicts what occurred in lab.

Figure 3:

Plot the percent change in mass over time using the graph below. Label both axes and add a title to the graph.

Figure 4:

5
Questions:
1. Was there a change in the weight of the egg? Briefly explain what happened and why using the terms
hypertonic, hypotonic and osmosis.

2. The diagram below represents the egg and the beaker in which it was placed. Label the solution inside the
egg and the solution inside the beaker as hypertonic or hypotonic. Using an arrow, indicate the
direction in which water flowed: into, or out of the cell (egg).

Beaker

Egg