AP Biology

Daniela Guerrero
Block 7
Diffusion and Osmosis Lab

Investigating how different solutions with different molar concentrations affect
diffusion and osmosis in a solution.

Introduction
Research:
The plasma membrane plays a very important role within the cell. It functions
as a selectively permeable membrane, which enables or prohibits which
molecules move into the cell and which leave the cell. There are two main
processes by which molecules can be transported: passive transport and
active transport. The main difference between passive transport and active
transport is the energy used to enable the transport. Passive transport is
driven by kinetic energy of the molecules that are being transported. There
are two types of passive transport: diffusion and facilitated diffusion. On the
other hand, active transport depends on energy from ATP in order to move
molecules against the concentration gradient, this means from hypotonic
solutions to hypertonic solutions. (Bowen, 2000)

Diffusion is the net movement of particles from a region of high concentration
to a region of low concentration, this means molecules are moving with the
concentration gradient. Diffusion will continue until an isotonic solution, which
means both solutions are uniform, is attained. Within diffusion we have
osmosis, which involves the transport of water across the membrane. Water
moves with the water potential gradient, this means that water moves from a
hypotonic solution, which has a high water potential to a hypertonic solutions,
which has a lower water potential. Osmoregulation involves keeping the
concentration of the cells cytoplasm at a suitable concentration. Water
potential is the energy of water in order to do work by moving from a
hypotonic solution to a hypertonic solution. The formula for water potential is
the following:
Water Potential () = Pressure Potential (p) + Solute Potential (s)
Water potential enables you can calculate the diffusion of water molecules.
(IHW, 2005)

Dialysis bags are a type of semi-permeable membrane tubing, which
resemble the permeability of the plasma membrane in cells. This tubing bag is
made from regenerated cellulose and cellophane. It can be used for both
diffusion and osmosis. Potatoes, on the other hand, the cells in a potato
contain both a plasma membrane and a cell wall. Because the potato cells are
plant cells, in order for cell to be healthy it must be turgid. This means that the
cell is placed in a hypertonic solution compared to the cell causing water to
enter the cell and make the cell wall very firm. On the other hand, animal cells
are healthy when they are placed in a isotonic solution which will enable a
certain amount of water to enter the cell even though it doesnt make the cell
turgid because thanks to the lack of a cell wall, the cell would burst if it is
placed in a hypertonic solution. (IHW, 2005)

There are certain key characteristics, which enable molecules to diffuse
through the plasma membrane. Small nonpolar molecules diffuse easily
through the plasma membrane. These small non polar molecules include
glucose which is a monosaccharide small enough to pass through the
membrane, salt which is small compound that also diffuses through the
membrane, and water which even though it is a polar molecules it passes
through the plasma membrane. On the other hand, bigger molecules, such as
proteins and sucrose, which is a disaccharide, cannot diffuse through the
membrane due to their size and or polarity. (Gil, 2012)

Objective: How will different solutions present in different molar
concentrations affect diffusion across a semipermeable membrane?

Hypothesis: If a hypertonic cell is submerged into a hypotonic solution, then
water will diffuse into the cell thus increasing its mass because the hypotonic
solution has a high water potential compared to the cell, which has lower
water potential.



Experimental Design
Manipulated Variable Responding Variable Constants Control
Different solutions
types of solution
used inside the
dialysis bags:
Glucose
Sucrose
NaCl
Water

Concentration of
sucrose solution in
which the potatoes
where placed:
1 M
0.8 M
0.6 M
0.4 M
0.2 M
The amount of water
transported across
the plasma
membrane,
measured by the
change in mass (in
g) of the model cell
in the dialysis bags
and the potatoes.
Type of water:
distilled water
Solutions tested:
glucose, sucrose,
NaCl, and
proteins
Room
temperature at 25
o
C
Amount of time
left for the
solutions to
diffuse in the
dialysis bags 30
minutes and in
the potato
experiment 24
hours.
100 mL of protein
solution placed in
the beaker in
which the dialysis
bags would be
placed.
10 mL of different
solutions placed
inside the dialysis
bags.
100 mL of
sucrose solution
in which the
potatoes would
be placed
Initial mass of
potatoes 4 g
Size of potatoes
2.2 cm x 2.2 cm
Beaker containing
100 mL of 5%
ovalbumin protein
solution surrounding
the model cell
containing the
dialysis bag with 10
mL of protein inside.


# of trials: 1


Procedure

Materials:
100 mL of 1 M of sucrose
100 mL of 0.8 M of sucrose
100 mL of 0.6 M of sucrose
100 mL of 0.4 M of sucrose
100 mL of 0.2 M of sucrose
10 mL of glucose
10 mL of sucrose
110 mL of distilled water
10 rubber bands
10 pieces of 10 cm yarn
Tape
Aluminum paper
510 mL of 5% ovalbumin
protein
10 mL of NaCl
5 dialysis bags, 20 cm long
each
5 beakers of 250 mL
Balance
5 2.2 cm x 2.2 cm potato
cubes
Knife
Ruler
Balance

Procedure:
Dialysis Bags
1. Place 100 mL of 5% ovalbumin protein in each of the five beakers
2. In each of the 5 dialysis bags place 10 mL of each of the different
solutions: glucose, sucrose, water, NaCl, and 5% of ovalbumin protein.
3. Close each of the 5 bags from each side carefully with the rubber
bands and the yarn pieces.
4. Record the weight each of the five bags in grams and record you data
in a table under the column of initial mass.
5. Place each of the dialysis bags in one beaker and begin to record the
time.
6. The beaker containing the protein bag will be the control
7. Wait for 30 minutes in order to remove the bags from the beakers.
8. After those 30 minutes remove each of the bags from their beakers and
weigh each of the bags again
9. Record the data in the table under the column final mass.
10. In order to calculate the percentage difference, subtract the initial value
from the final value, divide by the initial value and multiply by 100.
11. Record the percentages in a data and graph the values.


Potatoes
1. In each of the 5 beakers place 100 mL of each of the five different
molar concentrations of sucrose: 1 M, 0.8 M, 0.6 M, 0.4 M, 0.2 M and 0
M (water)
2. Cut 5 cubes of peeled potato 2.2 cm by 2.2 cm
3. Weigh each potato and record the data
4. Place 1 potato cube per beaker
5. Cover the beakers with aluminum paper
6. Record the time and wait for 24 hours
7. After the 24 hours remove each potato from its beaker and record the
final weight. Record your data.

Results and Analysis
Results:

Table 1:
Relationship between the surrounding solutions and the solutions inside the
bag.

Water Sucrose NaCl Glucose Protein
Content inside the bag
Percent of
change in
mass
Percent of
change in
mass
Percent of
change in
mass
Percent of
change in
mass
Percent of
change in
mass
Water 3.8 -19.8 8.3 -10.6 3.7
Sucrose 16.8 -1.7 21.6 13.4 28.0
NaCl 5.2 -7.3 0.0 0.0 10.7
Glucose 10.9 -0.9 11.5 5.1 25.0
Protein 28.5 -0.9 0.0 -4.5 4.6




Table 2:

The change in mass of a potato with the concentration of the solution the
potato is submerged in.
Water 1 Sucrose 2 NaCl 3 Glucose 4 Protein 5
Molar concentration of
sucrose solution (in M)
Percent of
change in
mass
Percent of
change in
mass
Percent of
change in
mass
Percent of
change in
mass
Percent of
change in
mass
1 M -26.3 -25.9 -25.3 -24.4 -30.3
0.8 M -21.3 -25.9 -24.7 -24.4 -23.7
0.6 M -18.8 -18.2 -16.2 -18.6 -19.6
0.4 M -1.3 -3.8 3.9 -2.3 -8.8
0.2 M 6.3 1.3 6.4 10.8 10.9
0 M 7.5 6.5 12 14 13.04

-30.0
-20.0
-10.0
0.0
10.0
20.0
30.0
40.0
H20 Sucrose NaCl Glucose Protein
M
a
s
s

d
i
f
f
e
r
e
n
c
e

(
i
n

%
)

Surrounding Solution
Relating how the surrounding solution affects
the diffusion of the solution inside the dialysis
bags.
Water Sucrose NaCl Glucose Protein



Analysis:
The graphs above show the relationship between the type of solutions and the
molar concentration of a solution and how they affect diffusion and osmosis
occurring in a cell. If the solution is hypotonic compared to the hypertonic cell
inside the solution, then the cell will increase in mass because water will go
into the cell via osmosis. For example, if glucose is placed on a beaker with
surrounding water, the mass of the glucose dialysis bag will increase by a
10.9%. This means that water entered the dialysis bag containing glucose
increasing the mass of the bag. Also, in beaker number 1, when the potato
was placed in a hypotonic solution containing 0.2M of sucrose, the mass of
the potato increased by a 6.3%. This means that water diffused into the
potato. On the other hand if the solution is hypertonic compared to the
hypotonic cell, the mass of the bag will decrease because water will move out
of the cell. A clear example of this is a water bag placed in a sucrose solution.
The water dialysis bag lost water, which will leave the cell to the sucrose
solution. This causes the cell to lose mass by a -19.8%. In the potato
-35
-30
-25
-20
-15
-10
-5
0
5
10
15
20
0 0.2 0.4 0.6 0.8 1 1.2
M
a
s
s

d
i
f
f
e
r
e
n
c
e

(
i
n

%
)

Surrounding concentration (in M)
Investigating the relationship between molar
concentration and how it affects osmosis in a
solution.
Water
Sucrose
NaCl
Glucose
Protein
experiment, in beaker number 5, the potato was submerged in a 1M of
sucrose solution, which is hypertonic compared to the potato. This caused the
potatoes mass to decrease by a -30.3%, meaning water left the potato and
went to the sucrose solution.

Conclusions

The purpose of this experiment was to analyze the effect of having different
concentrations of solutions in comparison to the concentration of the cell
submerged into a solution. Also, to identify what role does water potential play
in the relationship between hypertonic and hypotonic solutions. During the
experiment, we closely observed how osmosis takes place when you change
the solution that surrounds a cell as well as changing the content of the cell.
Also, we managed to alter the molar concentration in which potato cells were
submerged and see the change in water potential and its relation with the
altering of the concentration both inside and outside the cell.

The major findings of this experiment revolve around water potential.
Hypotonic solutions have a high water potential, while hypertonic solutions
have a low water potential. Water will move from a high water potential to a
low water potential. A solution is hypertonic when it has a lower concentration
in comparison to another solution, which has a higher concentration or is a
hypotonic solution. Therefore water will move from a hypotonic solution to a
hypertonic solution. Solutions may increase in concentration in two ways,
increasing the molar concentration or having various solutions, which have
different solute concentrations. Both of these factors affect the rate of diffusion
of small nonpolar molecules as well as osmosis.

My hypothesis was supported by the experiment because solutions having a
lower concentration of solute decreased its water potential because water
diffused to hypertonic solutions, which have a lower water potential. If the
solution has a high molar concentration for example a molar concentration of
0.8, the hypotonic cell will decrease its water potential because water via
osmosis will go out of the cell. Again water moves from a hypotonic solution to
a hypertonic solution with the concentration gradient of water movement. In
the same way, if the solution in which a cell was submerged contains a higher
concentration than the cell, water will leave the cell into the hypertonic
solution.

The findings in this experiment are very similar to those established by other
researchers. Diffusion will always move particles from a high concentration
area to a low concentration area (Bowen, 2000). According to IHW, dialysis
tubing will work as a permeable membrane, which allows only small nonpolar
molecules to diffuse freely through the membrane. Both molecules and water
are constantly moving through the plasma membrane. Molecules and water
move with the concentration gradient respectively. This means that molecules
will move from a hypertonic solution to a hypotonic solution, while water
moves from a hypotonic solution to a hypertonic solution. (IHW, 2005)

The movement of water always moves from a high water potential, found in a
hypotonic solution to a low water potential found in hypertonic solutions. Also,
molecules move with the concentration gradient, this means from a hypertonic
solution to a hypotonic solutions. Both of these movements of molecules
occur in order to reach an isotonic solution. This means a balance between
solute and solution in comparison to the other solution. This is why molecules
are constantly moving through the membrane in order for osmoregulation to
take place. Therefore if molecules can diffuse through the membrane they will
go from a high concentration to a lower concentration, which can either be the
cell or the solution. If molecules cannot diffuse freely through the molecule,
water will diffuse from a high water potential to a low water potential in order
to reach an isotonic solution.

During this experiment various errors were introduced. One of the most
important of these was the final mass measurements of the dialysis bags of
group 5. These measurements altered the final percentage change, this
means that these results didnt make sense since as previously said when two
solutions are placed together and a permeable membrane is diving them,
molecules will move from a high concentration to a low concentration and
water will move from a low solute concentration to a high solute concentration.
This means that these data cant be used to support this experiment because
the results arent valid. Other errors such as s slight change in mass of the
potato cubes also altered the final results of the experiment.

In order to improve this experiment, it is very important to make accurate
calculations in order to have more precise data, which is very useful to
compare the different effects of diffusion and osmosis. Measurements such as
the potato cubes should be exact; volumes found in the beakers in which the
cells are submerged should be as accurate as possible. The timing is
essential in this experiment; therefore all 5 potatoes and dialysis bags must
remain in the beakers the same time in order to have the same time for
diffusion and osmosis to take place. Also, this experiment could improve by
having a wider range of molar concentrations in order to have more data to
analyze this objective.

In a further study, it would be interesting to experiment with dead cells and
live cells. How will diffusion change in a dead cell and in a living cell? Also, to
investigate with other variables that affect osmoregulation such as the
quantity of water placed in a beaker as well as the concentration inside the
cell. Testing different concentrations of solutes inside the cell placed in a
water solution and see how diffusion and osmosis are affected by these
change in solute concentration.











Bibliography
Bowen, R. (2000, july 1). Mechanisms for Transport Across Membranes. Retrieved
october 17, 2012, from
http://arbl.cvmbs.colostate.edu/hbooks/cmb/cells/pmemb/transport.htm
l
Gil, J. (2012, october). Transport of molecules.
IHW. (2005, march). Movement across membranes. Retrieved october 17, 2012,
from http://www.biologymad.com/resources/diffusionrevision.pdf