Lab 04: Diffusion and Osmosis

Introduction:
The Diffusion and Osmosis lab introduced many different concepts that are
explored in a series of three experiments. Firstly, on a larger level than just this
experiment, diffusion and osmosis has to do with cells functions. Atoms and molecules
are constantly bumping into each other, also known as kinetic energy. This concept of
kinetic energy (motion) results in diffusion. A cells membrane is selectively permeable
which allows this movement to be regulated.
A cell has a membrane and a cytoplasm, which it moves materials in and out of to
uphold homeostasis. Also the cells membranes, which are phospholipid bilayers, have
proteins embedded into them, and because the fatty acids in the bilayer are hydrophobic,
they limit the movement of water. A cells environment is aqueous, meaning the solvent
is water. This also means that water can pass freely in and out of cell through osmosis or
by specialized proteins called aquaporins.
Diffusion means solutes move from a high concentrated area to an area of a lower
concentration. Also diffusion does not require any energy; however if a solute was
moving from a low to high concentration, ATP and pumps (protein carriers) would be
needed. Water moving down its concentration gradient is called osmosis. The solute
concentration and the resistance to water movement, called turgor pressure, in a cell
affect osmosis.
Hypertonic, hypotonic and isotonic describe solutions, which are separated by
selectively permeable membranes. A hypertonic solution is when there is a high solute
concentration and lower water potential compared to the other solution and this causes
the cell to appear as shriveled up. A hypotonic solution is when there is a lower solute
contraction and higher water potential than the solution outside the membrane. An
isotonic solution is when both solutions have equal water potential.
Water potential shows which way water will diffuse. In non-walled cells the
movement of water in and out of the cell is affected by the solute concentration on both
sides of the membrane. However, in walled-cells, the cell wall prevents the cell from
bursting as water enters. This causes pressure to build up inside the cell, which affects the
rate of osmosis. Water potential is free energy per mole and has two major components:
the osmotic/solute potential (dependent of solute concentration) and the pressure potential
(dependent on exertion of pressure).
Water Potential = Pressure Potential + Osmotic Potential

Water moves from higher water potential area/free energy to one of lower water
potential/free energy. Water potential measures the tendency of water the diffused from
one space to another. Looking at water potential of pure water in an open beaker, it would
be zero. This is because both the osmotic and pressure potentials are 0. In general, an
increase in (+) pressure will result in a higher pressure and water potential. Then, the

addition of solute to water lowers the osmotic potential and because of that, decreases the
water potential. The osmotic potential is:

i=ionization constant, C = molar concentration, R = pressure constant and T =

temperature in K
If a selectively permeable membrane separates the cytoplasm, the water moves
from the surrounding area (high water potential) to the inside of the cell (lower water
potential) since the solutes in the cytoplasm. The movement of water into the cell
(osmosis) causes the cell to swell and therefore also causes the pressure to increase inside
the cell because the cell membrane is pushing against the cell wall. This is called turgor
pressure counteracts the diffusion. After some time, the turgor pressure increases to
oppose to negative solute potential of the cell. This causes the water potential of the cell
to equal the water potential outside the cell. When this happens, a dynamic equilibrium is
reached and the movement stops.
When solute is added to the water outside the cell, the water potential outside the
cell decreases. When enough solute is added the water potential outside the cell equals
the water potential inside the cell, which causes there to be no movement of water. Then,
if more solute is added to water outside the cell, water will leave the cell and move to the
area of lower water potential (outside of cell). This causes the cell to lose turgor pressure
and cause the membrane to shrink.
Lab 1: Surface Area to Volume Ratio (Diffusion)
Experimental Question: If you put agar cubes each with a different surface area to
volume ratio into a solution which cube would that solution completely diffuse through
the fastest?
Hypothesis: If the agar cubes are put into beaker filled with vinegar, then the cube with
the largest surface area to volume ratio (1:1) will diffuse through the fastest.
Materials List
1. Agar Cubes (1 cm3, 2 cm3, 3 cm3)
2. 1 200 mL beaker
3. Vinegar
4. Stop Watch

Procedure:
1. Get the three Agar cubes: 1 cm3, 2 cm3, 3 cm3
2. Record each ones surface area and volume
3. Take the 200 mL beaker and put the 3 agar cubes into it
4. Pour vinegar into the same beaker and make sure all the cubes are fully
submerged in the vinegar

5. With the stopwatch, record the time it takes for each cube to completely lose its
color

Results:
Length of Cube
Side

Surface Area
(cm2)

Volume (cm3)

Surface Area:
Volume Ratio

1 cm
2 cm
3 cm

1
4
9

1
8
27

1:1
1:2
1:3

Time Until
Complete
Diffusion
22.08 minutes
74.54 minutes
109.5 minutes

Conclusion: If the agar cubes are put into beaker filled with vinegar, then the cube
with the largest surface area to volume ratio (1:1) will diffuse the fastest. In this

experiment, the hypothesis turned out to be true. The cube with the length of cube side 1
cm, surface area of 1 cm2 and volume of 1 cm3 had a surface area: volume ratio of 1:1
and diffused completely in 22.08 minutes. The cube with the length of cube side 2 cm,
surface area of 4 cm2 and volume of 8 cm3 had a surface area: volume ratio of 1:2 and
diffused completely in 74.54 minutes. The cube with the length of cube side 3 cm,
surface area of 9 cm2 and volume of 27 cm3 had a surface area: volume ratio of 1:3 and
diffused completely in 109.5 minutes. Looking at this data, it shows that the cube with
the smallest surface area of volume ratio (1:1), diffused the fastest.
Since cells rely a lot on diffuse to transport its molecules; large cells would not be
able to transport substances and water in and out fast enough. This is why there are not
huge cells in organisms. This experiment proved that diffusion happens faster and more
efficiently with smaller cells because of their larger surface area to volume ratio.

Lab 2: Effects of Osmotic Potential Differences Across a Membrane

Experimental Question: If different molarities of the 6 different sucrose solutions are put
into pre-soaked dialysis tubing and submerged into water, which of the bags will have the
greatest % change in mass?
Hypothesis: If each of the 6 different sucrose solutions are put into pre-soaked dialysis
bags and submerged in water for 20 minutes, then the sucrose solution with the greatest
molarity (1.0 M) will have the greatest % change in mass due to the diffusion of water
into the tubing.
Material List:
1. 6, 20 cm, Pre-soaked dialysis tubing
2. Fishing line
3. 0.0 M, 0.2 M, 0.4 M, 0.6 M, 0.8 M, 1.0 M sucrose solutions - 20 mL each
4. 6 plastic cups
Procedure:
1. Get the 6, 20 cm pre-soaked dialysis tubing
2. Tie one end of each of the 6 tubes with fishing line
3. Pour 20 mL of each of the 6 sucrose solutions (0.0 M, 0.2 M, 0.4 M, 0.6 M, 0.8 M,
1.0 M) into separate tubes
4. Remove some of the air from each bag, and then tie that end also with the fishing
line
5. Rinse the bags, and blot each of them carefully with a paper towel
6. Weight each bag and record their initial mass
7. Put each bag into each of the 6 plastic cups and fill the cups with distilled water
until the bags are completely submerged into the water
8. Wait 20 minutes and then take each of the bags out and blot them carefully
9. Weigh each bag and record the final masses

Results:

Contents of
beaker
0.0 M
0.2 M
0.4 M
0.6 M
0.8 M
1.0 M

Initial Mass

Final Mass

12.71
12.94
13.57
14.65
17.22
13.91

12.83
14.32
15.53
17.35
20.31
17.95

Mass
Difference
.12
1.38
1.96
2.7
3.09
4.03

% Change in
Mass
.94%
10.66%
14.44%
18.43%
17.94%
29.04%

Class Data:

Conclusion: If each of the 6 different sucrose solutions are put into pre-soaked
dialysis bags and submerged in water for 20 minutes, then the sucrose solution with the
greatest molarity (1.0 M) will have the greatest % change in mass. In this experiment the
hypothesis was proven true. The 0.0 M solution had a .94% change in mass. The 0.2 M
solution had a 10.66% change in mass. The 0.4 M solution had a 14.44% change in mass.
The 0.6 M solution had an 18.43% change in mass. The 0.8 M solution had a 17.94%
change in mass. The 1.0 M solution had a 29.04% change in mass. The water diffused
into the membrane in order to reach equilibrium. The water was moving from an area
outside the cell of low concentration of solution and a high water potential, to an area
inside the cell membrane of high concentration of solution and a low water potential.
Since the 1.0 M of sucrose solution had the greatest solute concentration, more water
moved into the membrane in order to increase its water potential in the cell.
Looking at the classs data as a whole, all the groups had varying data except for
groups 1,2 and 3 which all have the trend of the % mass change moving upward as the

molarity increased within each dialysis tubing. These sets of data all seem to be correct
and the others (groups 4,5 and 6 have experimental errors). This is because that the
osmotic potential is suppose to move from a hypotonic to hypertonic solution. Therefore,
as the molarity inside the bag increases, the % change inside the more saturated bag
should increase.
The classes group data however is very unreliable. Looking at the mean graph in
the graph above can see this. If the data was reliable, a line of best fit would be formed
and be linear. However, a line of best fit if very off in this graph and the data is very
varied. Making the data unclear and not accurate. Luckily we know what is supposed to
happen regarding the data.
The osmotic potential, which is the potential of water molecules to move from a
hypotonic solution to a hypertonic solution across a semi-permeable membrane, is greater
in the solution with the 1.0 M because the water is moving from a hypotonic solution to
the most hypertonic solution. Since the osmotic potential is dependent on the solute
concentration also is why the % change in mass was the greatest in the 1.0 M of sucrose
solution. The 0.0 M of sucrose solution had the smallest % change in mass because there
was a lower osmotic potential. The solute did not have a high molarity, and therefore
there was not a high solute concentration. This meant that equilibrium was for the most
part already reached.

Lab 3: Determining the Osmolarity of Different Apples

Experimental Question: How will putting the Gingergold apples in different molarities of
sucrose solution affect the mass of the apples?
Hypothesis: If the Gingergold apple cylinders are placed in a solution with a greater
molarity of sucrose (1.0M), then those apples will have the least change in mass.
Materials List:
1.
2.
3.
4.
5.

0.0 M, 0.2 M, 0.4 M, 0.6 M, 0.8 M, 1.0 M sucrose solutions

Core border
Ruler
6 plastic cups
2 apples (same kind)

Procedure 3:
1. Use the cork borer to cut the 2 apples into 24 apple cylinders (each cylinder has to
be the same size)
2. Separate the 24 cylinders into 6 groups of 4 and determine the mass of each of the
6 groups
3. After recording each of the masses, take the 6 cups and pour each of the sucrose
solutions into separate cups
4. Place the 6 groups of 4 apple cylinders into each of the 6 cups
5. Cover the beakers with plastic wrap

6. Keep overnight
7. The next day record the temperatures of the sucrose solutions
8. Remove the cores from each of the cups (blot gently) and weigh their new final
masses
Group 1 Results:
Contents of
Temperature
beaker
(o C)
0.0 M
24
0.2 M
24
0.4 M
24
0.6 M
24
0.8 M
24
1.0 M
24

Initial Mass

Final Mass

4.97
4.77
5.17
5.14
4.8
5.13

5.96
6.07
6.49
6.3
4.96
4.8

Mass
Difference
.99
1.3
1.32
1.16
.16
.33

% Change in
Mass
19.9
27.25
25.5
22.6
3.33
-6.43

Conclusion: If the Gingergold apple cylinders are hypertonic in a hypotonic

solution, then four apple cylinders in the sucrose solution with the greatest molarity (1.0
M) will have the least change in mass. In this experiment the hypothesis was proven true.
The apple cylinders that were left overnight in the 1.0 M sucrose solution had the least
percent change in mass, -6.43%, and a .33 weight difference. Then in the 0.0 M sucrose
solution had a mass change of 19.9% and a .99 mass difference. This shows the
significant difference between the two different sucrose solution molarities and their
effects on the apples mass change. All of the other apple cylinders gained mass from the
sucrose solution surrounding it; however, the apple cylinders in the 1.0 M of sucrose
solution lost mass since the apples were more hypotonic than the saturated sucrose
solution.

Osmolarity is when the apples water potential is equal to the sucrose solution.
This happens at around .9 M because the line of best fit reaches 0 at around this point. To
calculate the solute potential of the apple, the ionization constant = 1, molar
concentration is .9, the pressure constant is .0831 and the temperature is 24 o + 273K.
Using the formula solute potential = - iCRT, the solute potential is -22.21. This means
that if a Gingergold apple was put into a concentrated solution of .9 M, there would be no
diffusion between the apple and the environment. This means that there would not be any
mass change.
Since the environment of 1.0 M of sucrose solution surrounding the apples was
hypertonic in comparison to the apples, which were hypotonic, the movement of water
was different than all of the other sucrose solution and apple trials. Instead of the solution
surrounding the apple moving into the apple because the solution was hypotonic and had
greater water potential to the hypertonic apple cylinders with less water potential, the
apples juice moved out of the apple and into the hypertonic solution that had a lower
water potential in order to reach equilibrium.