Name:

Nicholas Mack

Block: 3

Group Members names: Jake Sasfai, Salem Belay, Maya G.

Testing the Properties of Diffusion and Osmosis

Introduction:
The purpose of this lab was to observe and test the properties of osmosis and diffusion in three separate
experiments. All the experiments in this lab involved the determination of the osmotic potential and the
diffusive properties of water. In modern understanding of biology, all living cells consist of a cytoplasm
and a semi-permeable membrane. All cells need to allow certain products into their cytoplasm in order
to perform basic functions within the organism. The membrane of every cell is selectively permeable,
meaning that the cell only allows certain molecules to enter and function within the cell. The
membranes selectivity is largely dependent on whether the molecule attempting to enter is hydrophilic
or hydrophobic. Hydrophobic molecules are non-polar molecules that are resistant to water. These types
of molecules have little trouble passing through the membrane because they can tolerate its hydrophobic
interior. On the other hand, hydrophilic molecules are polar molecules that are attracted to water. They
cannot pass through the membrane easily because they are resistant to its hydrophobic interior. Though
they cannot pass through the membrane directly, Hydrophilic molecules have other methods of going
across the membranes phospholipid interior. Examples include the use of transport proteins and iongated channels. Another important example is the use of aquaporin. An aquaporin is a protein that has
the capability to transport water across the membrane to the organelles of the cell. Since water is a
necessity for most cells, there are a number of ways besides the use of aquaporin that allow water
molecules to enter the cell. One of the most important properties of water in regards to cell biology is its
ability to diffuse across the membrane through osmosis. Diffusion is the process of in which solutes
move from an area of high concentration to low concentration. This process requires no energy because
it is the natural state for molecules, such as water, to move to an area of low concentration (low free
energy) until equilibrium is reached on both sides. In an extracellular, aqueous environment, water is the
solvent where its rate of diffusion is largely determined by the amount of solute concentration on both
sides of the cellular membrane. This causes the cell to be hypertonic, hypotonic, or isotonic towards its
environment. In hypertonic environments, more water exits the cell then enters because there is a lower
concentration of solute on the exterior of the cell. In isotonic environments, the rate at which water
enters the cell is equivalent to the amount of water leaving the cell, otherwise known as equilibrium.
Most cells cannot operate in these types of environments because when equilibrium is reached, no work
can be accomplished. On the other hand, in hypotonic environments, the rate at which water entering the
cell is greater than the amount of water leaving the cell. Cells without cellular walls will, in this type of
environment, explode if too much water enters the cell. Plant cells, however, notably prefer hypotonic
environments because its cell walls prevent the cell from exploding when a large quantity of water
enters. Large quantities of water entering a plant cell cause a certain amount of pressure to be exerted,
leading this discussion to water potential. Water potential is the measure of the diffusion of water
between two compartments (outside the cell and the membrane). This rate is determined by the sum of
pressure potential and solute / osmotic potential. Pressure potential is the amount of pressure exerted on
the cell. In plant cells, this pressure is called turgor pressure because when water enters the cell
(assuming that the environment is hypotonic) the cell membrane will push against the cell wall, causing
this type of pressure to be exerted. Solute potential, on the other hand, is the measure of the amount of
solute in the solution relative to the cell. At atmospheric pressure, if more solute is added to the water,
the solute potential decreases and therefore the water potential decreases. Solute potential is equal to
iCRT. The variable i is the ionization of the constant, C is the molar concentration (osmo. potential), R is
the pressure constant, and T is the temperature in K. With this information, in this lab, students

conducted several experiments to analyze and test the properties of diffusion and osmosis. These three
experiments are explained in more detail below.
The purpose of experiment (or part) 1 was to analyze the diffusion of a solution across agar cubes with
different volume to surface area ratios. Each cube was placed in a solution that was mostly comprised of
vinegar (acidic). Using a ruler, students measured the volume and surface area of each cube and
recorded those measurements. The agar cubes were then placed in the solution and the students recorded
the amount of time the solution was able to diffuse through each cube. The goal of this particular
experiment was to see whether the size of a solute affect the solutions rate of diffusion across that
solute. The purpose of Part 2 was to test the diffusive properties of water (distilled) with different
concentrations of sucrose. Students were asked to make a model of a cell membrane by using pre-soaked
dialysis tubing. In each tube, students poured a different amount of sucrose solution. These tubes were
then placed in beakers of distilled water. The goal of this experiment was to see which tubes contained
the most water after diffusion and which exhibited the greatest mass percent change. This was
determined by measuring the mass of each tube at the conclusion of the experiment. Part 3 tested the
osmolarity of an apple. Four pieces of an apple were measured and cut and each set of pieces was placed
one of the six sucrose solutions. At the conclusion of the experiment, students then measured the final
mass of apple piece and the temperature of each solution. The experiment determined which sucrose
solution diffused through the apple the most. These three experiments collectively analyzed the
properties of diffusion and osmolarity and gave the students a better idea of how the cell membrane
works in aqueous environments.
Problem:
Problem 1: Will the volume and surface area of sugar cubes placed in a solution
have any affect on the rate of the solutions diffusion across the agar cubes?
Problem 2: Does the rate of diffusion differ on the amount of sucrose concentration
in the model membrane?
Problem 3: Does the amount of sucrose solution in water affect its diffusion across
pieces of an apple? What factors are an apples osmolarity determined by?
Hypothesis:
Procedure 1: If there is a larger surface area to volume ratio in an agar cube, then
it will take longer for the solution to diffuse across the cube.
Procedure 2: If there is more sucrose concentration in a dialysis tube, then less
water will be able to enter the model cell, causing no change in the final mass.
Procedure 3: If there is more sucrose concentration in a solution, then the rate at
which that solution diffuses across a 3 x 1 apple piece will decrease.
Materials:
Procedure 1: The materials used include: sugar cubes of different sizes (3), beaker, 200 mL of vinegar,
and a timer.
Procedure 2: The materials used include: six different sucrose solutions (0.0, 0.2, 0.4, 0.6, 0.8, 1.0 M)
[20 mL of each], six 20 cm strips of pre-soaked dialysis tubing, distilled water, three 250-mL beakers,
string, marker, scale, and a timer.

Procedure 3: The materials used include: 100mL of each of the six sucrose solutions, six 250 mL
beakers, 5 mm cork borer, ruler, scale, thermometer, paper towel, and plastic wrap.
Procedure:
Procedure 1 / Part 1 (derived from information on lab handout):
1. Obtain three different sized agar cubes (1 cm3, 2 cm3, 3cm3).
2. Use ruler to measure length, width, and height of each cube in cm.
3. Using those measurements, determine the surface area (cm2) and volume (cm3)
of each cube and, subsequently, determine the volume surface area ratio of
each cube using those results.
4. Fill beaker with 200 mL of vinegar and place each cube inside.
5. Using a timer, record the amount of time it takes for the solution to diffuse
across each cube. The student will know the solution has completely diffused
across when the cell loses its color.
Procedure 2 / Part 2:
1. Obtain six, 20 cm strips of pre-soaked dialysis tubing.
2. Using strings, tie one end of each tube to form 6 bags.
3. Pour 20 mL of each of the following sucrose solutions into separate bags 0.0,
0.2, 0.4, 0.6, 0.8, 1.0 M.
4. Remove excess air for each bag.
5. Tie the other end of each bag.
6. Rinse each bag to remove sucrose from the string and outside surface.
7. Blot outside of each bag and record the initial mass of each bag.
8. Place each bag in one of three 250mL beakers and fill with 200 mL of distilled
water.
9. Let stand for approximately 20 minutes.
10.

Remove bags and carefully blot each.

11.

Determine the mass of each of the bags and the percent change. Record all.

Procedure 3 / Part 3:
1. Obtain 100 mL of each of the sucrose solutions and pour each solution into a

separate, labeled 250 mL beaker.

2. Use cork borer to cut 24 apple cylinders. Each cylinder should be 3 cm. in
length. Remove any skin found on the cylinders.
3. Determine mass of four cylinders together and record. Put these 4 cylinders
into one of the sucrose solutions.
4. Do the same for four other cylinders together and record. Put these four
cylinders into one of the sucrose solutions.
5. Do the same for all of the other cylinders (in sets of four) and place in
remaining sucrose solutions.
6. Cover beakers with plastic wrap and let beakers stand overnight.
7. Next day, record temperature of each of the sucrose solutions
8. Remove plastic wrap from beakers, dry all cylinders using a paper towel, and
record the final mass of each of the sets of apple cylinders tested. (Note that
there are 4 cylinders in a set).
9. Record percent change.

Results:
PART I:
Length of Cube
Side
1 cm
2 cm.
3 cm.

Diffusion of Agar Cubes According to their Volume and Surface Area

Surface Area
Volume (cm3)
SA : V Ratio
Time Until
2
(cm )
Complete
Diffusion
2
3
6 cm
1 cm
6:1
23 min.
24 cm2
8 cm3
3:1
53 min.
2
3
54 cm
27 cm
2:1
83 min.

Figure 1.1: Diffusion of Cubes in Relation to Side Length

90
80
70
60
50

Time (min)

Time Until Complete Diffusion (min) 40

30
20
10
0
1

Length of Cube Side (cm)

Images 1.1 & 1.2: Agar Cubes Before (Left) and After (Right) Diffusion

PART II:
A. Masses of Dialysis Tubes With Different Amounts of Sucrose
(M) Group 4 Data
Contents of
Initial Mass
Final Mass
Mass Difference
Beaker
0.0M
15.82 g
15.83 g
.01
0.2M
16.58 g
16.87 g
.29
0.4M
18.43 g
19.41 g
.98
0.6M
13.73 g
14.55 g
.82
0.8M
20.0 g
20.62 g
.62
1.0M
19.92 g
21.50 g
1.58

Concentration
% Change In
Mass *
0.06%
1.75%
5.32%
5.97%
3.10%
7.93%

*Percent Change In Mass = [(Final Mass)-(Initial Mass) / (Initial Mass)] x 100

Figure 2.1: Amount of Sucrose in Relation to Percent Change in Mass (Group 4)

9.00%
8.00%
7.00%
6.00%
5.00%
% Change In Mass 4.00%
3.00%
2.00%
1.00%
0.00%
0.0M

0.2M

0.4M

0.6M

0.8M

1.0M

Amount of Sucrose In Beaker (M)

Image 2.1: Group 4 Dialysis Tubes (Each Color Represents a Different Sucrose
Solution)
Image 2.2: All Six Sucrose Solutions (Right)

Concentrati
on

Group
1

0.0 M
sucrose

0.94%

0.2 M
sucrose

10.66
%

0.4 M
sucrose
0.6 M
sucrose
0.8 M
sucrose

14.44
%
18.43
%
17.94
%

1.0 M
sucrose

29.04
%

B. Class Data for Mean Percent Change in Dialysis Tubes

Group Grou Grou Group Grou Mean
Standar
2
p3
p4
5
p6
d
Deviati
on
0.9% 0.06
2.91% -4.56% 0.212
0.51%
%
31.7
%
4.46% 3.2% 1.6% 14.23 2.525% 0.106
%
19.0
%
5.14% 9.5% 5.43
11.12 2.64
8.045% 0.040
%
%
%
7.00% 10.1
5.97
5.4% 7.403% 0.062
%
%
2.48%
14.15 12.0
3.1% 22.12 11.342 0.081
%
%
%
1.26
%
%
16.0% 20.1
7.93
12.002 0.11
%
%
0.83% 0.23
%
%

Standa
rd Error
-1.86%
1.03%
3.28%
3.02%
4.63%
4.90%

Figure 2.2: Amount of Sucrose in Relation to Mean Percent Change In Mass

0.0M

0.2 M

0.4 M

0.6 M

0.8 M

1.0 M

Amount of Sucrose (M)

PART III:
Percent Change & Temperature of Apple Cylinders in Different Amounts
Concentration **
Contents of
Temperature Initial Mass
Final Mass
Mass
Beaker
Difference
0.0M
20 C
3.58g
3.16g
-.420g
0.2M
19 C
3.58g
4.66g
1.08g
0.4M
19 C
3.75g
4.77g
1.02g
0.6M
20 C
3.10g
3.96g
.860g
0.8M
20 C
2.47g
2.73g
.260g
1.0M
10 C
2.50g
2.55g
.050g

of Sucrose
% Change in
Mass
-11.7%
30.1%
27.2%
27.7%
10.5%
2.0%

Apple Variety: Granny Smith Green Apple

**Note: The initial and final masses were taken 24 hours apart. The temperature of each
solution was also taken 24 hours after experiment commencement.

Figure 3.1: Amount of Sucrose in Relation to Percent Change of Mass of Apple Cylinders
35.00%
30.00%
25.00%
20.00%
15.00%
% Increase in Mass
10.00%
% Decrease In Mass
5.00%
0.00%
0.0M
-5.00%

0.2M

0.4M

0.6M

0.8M

1.0M

-10.00%
-15.00%
Sucrose Molarity (M)

Image 3.1: Apple Used in Experiment and Apple Cores (Cylinders)

Molar Concentration of Apple Cores: -9.31M (no net gain or loss of water from the
tissue)
(Note: Linear Regression equation is y=1.457x+13.57 substitute 0 for y to find molar
concentration of cores).
Solute Potential:
Formula: -iCRT
(-1) (-9.31) (1.496) (18C +273) = 4052.44

(?)

(*Note: 18 is the average temperature for the sucrose solutions)

Error Analysis:
Although all of the experiments in this lab went relatively well, there were a few minor errors that
affected the consistency and accuracy of each part of this experiment. In Part I, the main error was a
measurement mistake. While the instructions specifically said to fill the beaker containing the agar
cubes with 200 mL of vinegar, the students filled the beaker with almost double that amount. This could
have affected the accuracy of the results, including the rate of diffusion. In Part II, the main error was
that much of the sucrose solution in each dialysis tube leaked because the students had a difficult time in
tying and securing the tubes in the distilled water solution. This leakage of sucrose solution from the
model cells could have affected the accuracy of the data and may have drastically shifted the students
conclusions on the permeability of cell membranes. As with the class data, the main error was that at
least two groups miscalculated their mass percent changes, which is especially evident when Group 6
stated that for 0.0 sucrose concentration, there was a -31.7% mass percent change. Miscalculations such
as this may have effected the standard deviation and standard error calculations leading to inaccurate
results and conclusions. This also led to an inaccurate regression line for the data that is especially
evident in the Results. Lastly, for Part III of this lab, the main error inflicted was that it took two
unplanned trials for the students to get their results. When the group first placed their apple cylinders in
the sucrose solutions, they did not wait 24 hours before taking their mass, causing them to start the
experiment over. This led to inadequate results, which could have effected their conclusions. Also,
during the second attempt, some of the apple cylinders that were already massed initially were placed in
the wrong sucrose solutions, causing a bit of confusion toward which one corresponded to which. This
mistake is evident in the students results. Despite these errors, the three parts of this experiment
collectively demonstrated the properties of water potential, osmosis, and diffusion.
Conclusion(s):
This lab was divided into 3 parts: Effect of Surface Area-Volume Ratios on Diffusion, Effects of Osmotic
Potential Differences Across a Membrane, and Determining Osmolarity of an Apple. Therefore, each
paragraph of this section will be devoted to discussing, analyzing, and concluding a single part of this lab.
Part 1 The purpose of Part 1 of this experiment was to determine whether increasing the Surface Area
Volume ratios of agar cubes would effect the diffusion of solution across these cubes. As written in the results
section, the experiment called for 3 agar cubes whose sides were 1, 2, and 3 cm in length, respectively. The SAVolume Ratio, as calculated, decreased as the length of the side of an agar cube increased. In the experiment,
the students determined that as the surface area and volume of the cube increased, the amount of time of time it
took for the solution (in this case vinegar) took to diffuse across the cube also increased. For the smallest cube
with a SA:V ratio of 6:1 and a volume of 1 cm3 , the time it took for the solution to diffuse across and discolor
the cube was approximately 23 minutes. After those 23 minutes, the cube continued to discolor in the solution.
For the largest cube with a SA:V ratio of 2:1 and a volume of 27 cm3, the time it took for the solution to diffuse
across the cube was 83 min, almost four times as long as the smallest cube. The reason why rates of diffusion

were affected by the volume and surface area of a cube was because the more material the cube had, the harder
it was for the solution to penetrate through the surface. One way to visualize this is by viewing the cubes as
permeable membranes. If a membrane is dense and contains a lot of material, it is obvious that it will take
longer for solution to penetrate through the membrane and reach the cytoplasm. If, however, the membrane is
thin and contains less material, such as the case with the smaller cube, then it will less effort for the solution to
enter and penetrate the membrane. This experiment, therefore, demonstrated an important property of diffusion
in which the time it takes for a solution to penetrate across a membrane is reliant on the total surface area and
volume of that membrane.
Part 2: The second part of the experiment dealt with analyzing the masses of model cell membranes that
contained different sucrose concentrations during water osmosis. As written in the Results section, there were
six different sucrose solutions used in this experiment labeled as 0.0, 0.2, 0.4...1.0. Each amount corresponded
to the amount of moles of sucrose in each tube. In the experiment conducted by Group 4, the students saw an
increase in the percent change of initial and final masses as the sucrose concentration in the tubing increased.
For instance, in the tube containing 0.2M of sucrose, there was a 1.75% increase between the initial mass (taken
before diffusion commenced) and the final mass of the dialysis tube. However, in the solution containing a full
1.0M of sucrose, there was a 7.93% increase between masses, indicating that the higher the sucrose
concentration in the cell, the less permeable the membrane was toward the water attempting to diffuse into the
cell. The aqueous environment was hypotonic to the cell because, in this experiment, more solution diffused
into the cell then out, indicated by the gradual increase in mass. According to the understanding of this process,
the solvent (such as the water in this case) always has a natural tendency to diffuse to lower concentrations
either in or out of the cell, depending on the tonicity. In this case where the environment was hypotonic to the
cell, the water had a greater tendency to diffuse through the cell with the most sucrose concentration because
more sucrose concentration in the cell equaled less free energy. Because water moves to an area of high water
potential (or high free energy) to an area of low water potential (or low free energy), the water tended to
penetrate more into the cells with a greater molarity of sucrose. With this information, the students concluded
that the net osmosis rate would equal zero when the rate at which water enters the cell is equal to the rate at
which water leaves the cell. This can only happen if there is no solute in the solvent and the cell itself. Although
the group data provided a clear idea of waters diffusive properties, the class data failed to do so. Groups 1-5
successfully proved that water tends to move to cells with higher sucrose (non-solute) concentrations. However,
the other groups data was slightly incorrect because their data contained negative mass percent change for 0.0
M in which their initial mass was greater than the final mass. This occurrence, of course, is not logically correct
because water will always have a tendency to diffuse through a cell unless prevented by some outside force.
Therefore, that mistake caused an inaccuracy of data for the class, as seen in Figure 2.2. Yet, despite this
obstacle, the students were still able to successfully find the standard bars and calculate linear regression (see
Results page). This part of the experiment, in summary, was essential toward the understanding of the diffusive
properties of distilled water and its reaction with sucrose-filled cell membranes.
Part 3 Part three of this experiment, similar to that of part of two, was to analyze the diffusive properties of
water by observing the osmolarity of apples. As explained in the procedure, sets of 4 pieces of apple (Green)
were each put in one of the six sucrose solutions overnight and weighted twenty four hours after to see if the
solution had any effect on the osmolarity of the apples. Based on the results, it is safe to say that the
concentration of sucrose in the solution did have an affect of the apples in their masses. According to the
students results, an increase in the amount of sucrose in the surrounding solution caused a decrease in the
amount of solution the apple absorbed (calculated by taking the mass of the apple). The reason behind this was
because the surrounding solution was hypertonic to the cell, meaning that it had a lower solute concentration
than inside the cell, causing more water to be released then absorbed. Therefore, the apple cylinders in solutions
with the most sucrose (and therefore the most solute) tended to have little % change in mass than those in
solutions with little sucrose molarity (where the tendency was greater). The only major flaw in this part of the
experiment was in the 0.0M category where the data indicated that there was a -11.7% change in mass when, in
reality, this was unrealistic. As explained in the error analysis, this occurrence was probably due to some human
error. This mistake also effected the solute potential calculation because, with a negative value now inserted, the
resulting value for this equation came out to be negative, which definitely did not correspond to the other
groups data. This mistake also affected the accuracy of the molar concentration of the apple cores, whose value

also came out to be negative. Despite these setbacks, this experiment was important in establishing the
relationships between water potential, solute potential, and osmosis.
Sources
AP Lab Handout
Campbell Biology Textbook page 101