DIFFUSION & OSMOSIS

Aguilar AB, Murillo JD, Onofre AS, Ramirez MFD, and Taborada J

BIO108 Lab Activity 03 Group 03

Science Department, College of Natural Sciences and Mathematics
Mindanao State University, Brgy. Fatima, General Santos City 9500 Philippines
4 September 2023

ABSTRACT

This lab report describes the mechanisms of osmosis and diffusion, including how a
molecule's size—small, medium, or large—as well as how the temperature affects
different molecular transport reactions and exchange. Three components are included in
this study: simulations, a wet lab, and a simple experiment carried out by the students to
investigate how solvent and solute molecules move across a semipermeable membrane
and how temperature and molecular size impact osmosis and diffusion. The
effectiveness of the diffusion rate is directly influenced by temperature and molecular
size, according to the results displayed in the tables. It also emphasizes the fact that
molecules, due to their size and charge, are unable to pass by diffusion and osmosis. The
outcomes of this lab exercise improved the students' comprehension of these procedures,
which will aid in their development of a thorough grasp of cellular respiration.

INTRODUCTION
The cell’s outer surface of the plasma membrane is in contact with the external environment, which
is a water solution that includes nutrients, wastes, gases, salts, and other substances that surround the cell.
On the other hand, the inner surface has contact with the cytoplasm, therefore the plasma membrane
regulates what enters and leaves the cell (Libretexts, 2020). The cell membrane allows the passage of some
materials but not all, thus it is said to be selectively permeable. Passive transport happens when there is no
energy requirement for substances to pass through the membrane. Two examples of this passive transport
include diffusion and osmosis (Libretext, 2020).
According to Helmenstine (2020), Osmosis refers to the movement of solvent particles across a
semipermeable membrane from a dilute solution, going into a concentrated solution. The solvent moves to
dilute the concentrated solution and equalize both the internal and external environment of the membrane
while diffusion is the movement of particles from higher concentration area to a lower concentration area
whose overall effect is to stabilize concentration throughout the medium. Both diffusion and osmosis
equalize the concentration of two solutions, however, their huge difference is that both solvent and solute
particles are free to move in diffusion while only the solvent or water molecules in osmosis cross the
membrane (Helmenstine, 2020).
Reinke et al. (2021) states that it is very important to understand how molecules move across the
cell membrane since it is a crucial concept in biology for the students to be able to understand more
advanced and broader concepts. A deep knowledge of diffusion and osmosis is needed to understand
cellular functions, may it be normal or abnormal, as well as life processes like water movement into the
roots of the plant, photosynthesis, cellular respiration, and many other biological phenomena (Reinke et al.,
2021). This activity aims to study the process of diffusion and osmosis through a simulation, and to observe
Page 1 of 9
BIO108 • Laboratory Activity 3• Group 03

how solute and solvent molecules move across a semipermeable membrane. Furthermore, this study aims
to analyze the factors that influence these processes such as concentration gradients and temperature.

MATERIALS AND METHODS

The students used a laptop and internet access (mobile data) in order to answer and acquire data for
the worksheet that was given which had two parts, the simulation wherein students were tasked to visit the
website “biology simulations” in which a simulation of osmosis and diffusion can be performed and
observed. Secondly, the making of their simple experiment based on the simulation. Materials included pen
and paper as well in order to draft the making of the simple experiment and to answer the worksheet that
was given.
In the first (1) part of the activity, the students were tasked to define the terms: solution, solvent,
solute, diffuse, dynamic equilibrium, and semipermeable membrane. Then they needed to access the
website https://www.biologysimulations.com/diffusion-osmosis with the use of a laptop with any browser
application. Here, the students can manipulate which molecules can be included in the observation. The
first run of the simulation should include the large molecules, medium molecules and small molecules. The
students were tasked to answer the question with a yes or no if the membrane is permeable based on the
size of the molecule. The next step was to set the large molecule on both sides to 2% and other molecules
to 0% and determine the percentage of water. Then, run the simulation for 60 seconds and observe how
much the solution concentration fluctuated. After that, record the greatest change in % over the 60 seconds.
The students needed to hover the mouse cursor over the graph to be able to see the values that were being
asked. The following step was to reset the simulation and set the large molecule concentration on side A to
5% and side B to 0 and set all other molecules to 0. Run the simulation again for 60 seconds and determine
the ending concentration of the large molecules on side A. Repeat the previous step for medium molecules
and small molecules. The students should answer the questions on the worksheet based on the data that was
being given and observed in the simulation.
For the second (2) part of the activity, the students were tasked to make a simple experiment where
they needed to test the effect of molecular size and temperature on the diffusion of molecules across a
semipermeable membrane. In this part, the students needed to state their hypothesis, identify their
manipulated, measured, and controlled variables, and test their hypothesis using the simulation. They were
tasked to record their observations and present the data in tables or graphs, whichever would be the most
appropriate and applicable.

RESULTS

1. Definition of terms correlated to the Diffusion and Osmosis Simulation:

Solution A mixture composed of two or more substances.

Solvent A substance that has the capacity to dissolve solutes to form a solution.

Solute It is a substance that is being dissolved in a solvent to form a

homogeneous solution.
BIO108 • Laboratory Activity 3• Group 03

Diffuse A process where particles move from higher to lower concentration.

Dynamic Equilibrium A balance between two opposing processes that are occurring
simultaneously.

Semipermeable It allows specific or certain molecules to pass through it.

Membrane

Table 1. Definition of terms

2. Simulation Background

In the solution, water is the solvent and the solvents are the small, medium, and large molecules.

3. Testing the permeability of the membrane from side A to side B based on the size of molecules

Molecule Is the membrane

permeable?

Large No

Medium Yes

Small Yes

Table 2. Permeability of the membrane to each molecule

4. Setting the large molecule on both sides to 2%, and the other molecules to 0%

The 98% of the solution is the water which can move easily between side A and B. The solution
fluctuated to 1.75% from its original concentration of 2%.

5. Setting the large molecule on side A to 5% and side B to 0 and also the other molecules.

The solution has 95% water content. On the 60 second mark, the 5% concentration of large
molecule in side A decreased to 3.89% and the ending concentration of water is 96.21%

6. Setting the medium molecule on side A to 5% and side B to 0 and also the other molecules.

The water in the solution is 95%. After running the simulation for 60 seconds, the 5%
concentration of medium molecules became 4% inside A and 0.84% inside B.

7. Setting the small molecule on side A to 5% and side B to 0 and also the other molecules.
BIO108 • Laboratory Activity 3• Group 03

The solution has 95% water content. After 60 seconds, the 5% original concentration of small
molecule became 3.09% and 1.79% in side B

8. Defining osmosis based on the simulation

It is a chemical and biological process wherein solvent molecules move from an area of lower
to higher solute concentration. The movements equalized the solute concentrations on both sides of the
membrane which is an important mechanism to maintain homeostasis.

9. Simple experiment

The effects of molecular size and temperature on the diffusion of molecules across a
semipermeable membrane.

Table 3. Result of the simple experiment. It shows that the molecule size and temperature directly affect
the efficiency of diffusion rate. The smaller the molecule size, the higher the diffusion rate. Same as the
higher the temperature, the higher the diffusion rate of molecules.

Temperature Concentration Level at 90 seconds

(°C)
Small Molecule at 5% Medium Molecule at Large Molecule at 5%
5%

A B A B A B
10 3.06 0.34 2.93 0.69 3.18 0
20 2.42 2.15 2.49 1.99 3.46 0
30 1.92 3.17 2.75 1.44 3.34 0
40 1.98 2.92 2.06 2.79 3.67 0
50 1.9 3.08 2.34 2.31 3.65 0

DISCUSSION

This laboratory activity consists of simulations, a wet laboratory, and a simple experiment. This
significantly helped the students understand the importance of osmosis and diffusion in various molecule
transport reactions and exchange. It will also cover the different factors, such as size of molecules (small,
medium, and large). As progress passes through the activity, each finished work is given as tables and
graphs, as seen from the results above. As such, they will be discussed accordingly.

1. Osmosis and diffusion background

In osmosis, which is a natural process, there are several important terms to understand.
First, we have the "solvent," which is usually water; it's the liquid in which other substances can
dissolve. Then, there's the "solute," which are the substances that get dissolved in the solvent
(Campbell and Reece 2008). Osmosis is all about the movement of solvent molecules from an area
BIO108 • Laboratory Activity 3• Group 03

with fewer solutes to an area with more solutes, through a "semipermeable membrane," like a
barrier with tiny holes. This movement continues until a "dynamic equilibrium" is reached, where
the solvent moves back and forth, but there's no net change in concentration. This balance is crucial
for many biological processes.

2. Simulation Background

Large molecules, such as proteins and polysaccharides, cannot pass through osmosis and
diffusion due to their size and charge. Osmosis and diffusion are passive processes driven by the
random movement of molecules from an area of high concentration to an area of low concentration.
Large molecules have a limited ability to pass through selectively permeable membranes, like the
lipid bilayer of a cell membrane, as their size hinders their movement. Additionally, the charge on
these molecules can further impede their diffusion, as they may be repelled by the charges on the
membrane. This is in accordance with Fick's law of diffusion, which states that diffusion rate is
inversely proportional to molecular size (Lodish B, etal., 2020).

3. Testing the permeability of the membrane from side A to side B based on the size of molecules

In the scenario described in statement number 4, where large molecules are initially present
at 2% concentration on both sides (A and B) of a selectively permeable membrane, and the other
molecules are at 0%, we can observe a change in the concentration of the large molecules over time
due to osmosis and diffusion. At the start, the large molecules cannot easily cross the membrane
due to their size, while water can readily move through the membrane. The 98% water on both
sides allows for the free movement of water molecules. This will result in a net flow of water from
the side with 98% water to the side with 1.75% solute concentration. This process continues until
an equilibrium is reached, where the concentrations on both sides stabilize. This concentration of
1.75% represents the point at which the movement of water molecules due to osmosis and the
diffusion of solute molecules reach a dynamic balance. The phenomenon can be explained by Fick's
laws of diffusion and the principles of osmosis. Fick's laws describe the net movement of particles
from areas of high concentration to areas of lower concentration. Osmosis, a specific type of
diffusion, applies these principles to the movement of water across a selectively permeable
membrane (Alber J, etal, 2018).

4. Setting the large molecule on side A to 5% and side B to 0 and also the other molecules.

The scenario you describe, where side A has a 5% concentration of large molecules and
side B contains 0% of both large and other molecules, can be explained by the principles of osmosis
and diffusion. According to Fick's laws of diffusion, molecules move from areas of higher
concentration to lower concentration, seeking equilibrium. In this case, water molecules on side B
will move into side A to dilute the higher concentration of the large molecules. This is in accordance
with the first law of thermodynamics, which states that energy (in the form of molecules) tends to
spread out or disperse to achieve a state of lower potential energy, which is often characterized by
a higher degree of randomness or entropy (Lehninger A, etal., 2017).
BIO108 • Laboratory Activity 3• Group 03

As time progresses, the concentration of the large molecules inside A decreases to 3.89%
while the water concentration inside A increases. This dynamic change is a result of the large
molecules moving out of side A into side B and the movement of water molecules in the opposite
direction, ultimately reaching a new equilibrium (Lehninger A, etal., 2017). This is in line with the
second law of thermodynamics, which states that in a closed system, entropy, or disorder, tends to
increase over time. In this case, the movement of molecules and the changes in concentrations
represent the system's tendency to achieve a higher level of randomness and equilibrium over time.

5. Setting the large molecule on side A to 5% and side B to 0 and also the other molecules.

The initial setup of setting the large molecule concentration on side A to 5% and side B to
0% results in a solution with 95% water content. After 60 seconds, the concentration of the large
molecule on side A decreases to 3.89% and the water concentration on side A increases to 96.21%.
This phenomenon can be explained by the principles of diffusion and the Law of Mass Action.
Diffusion is the process by which molecules move from an area of higher concentration to an area
of lower concentration. In this case, the concentration gradient between side A and side B drives
the diffusion of the large molecule from side A to side B. As a result, the concentration of the large
molecule on side A decreases over time. The Law of Mass Action states that the rate of a chemical
reaction is proportional to the product of the reactant concentrations (Berj J, etal., 2015). This
principle can be applied to the movement of water molecules from side B to side A. As the
concentration of the large molecule decreases on side A, the concentration of water molecules on
side B increases. This increase in water concentration on side B drives the movement of water
molecules to side A, leading to an increase in the water concentration on side A.

These principles provide a theoretical framework for understanding the dynamics of the
diffusion process and the changes in concentration observed in the given scenario.

6. Setting the medium molecule on side A to 5% and side B to 0 and also the other molecules.

In this diffusion simulation, there are two sides labeled A and B that contain a solution of
water molecules and "medium" molecules. Initially, side A contained a 5% concentration of the
medium molecules, while side B contained 0% medium molecules. The remaining 95% on both
sides was water. Diffusion is the process where molecules spread from regions of higher
concentration to regions of lower concentration until equilibrium is reached. This occurs because
molecules exhibit random thermal motion, so they will naturally disperse over time. Since side B
initially had no medium molecules, there was a concentration gradient driving the diffusion of the
medium molecules from side A to side B. After running for 60 seconds, the concentration of
medium molecules inside A decreased to 4% as some diffused to side B. The concentration in side
B increased to 0.84% as medium molecules moved across. However, equilibrium was not yet
reached, as the concentrations were still unequal after 60 seconds. This indicates the diffusion rate
was relatively slow for the experimental conditions. The diffusion rate depends on factors like the
temperature of the solution, molecular weight of the substances, and viscosity (Berj J, et al., 2015).
Smaller, lighter molecules diffuse faster. Higher temperatures also increase kinetic energy and
diffusion rates. The water and medium molecules likely had differences in these properties that
impacted the partial diffusion observed over 60 seconds. Overall, this simulation demonstrated how
BIO108 • Laboratory Activity 3• Group 03

concentration gradients are the driving force for diffusion according to Fick's laws of diffusion
(Berj J, et al., 2015). The system progressed toward equilibrium as molecules moved down their
concentration gradient, but full equalization of concentrations did not occur in just 60 seconds.

7. Setting the small molecule on side A to 5% and side B to 0 and also the other molecules.

This diffusion simulation started with an asymmetrical concentration of a "small" molecule

between sides A and B. Side A originally contained a 5% concentration of the small molecule,
while side B had 0%. The remaining 95% of the solution on both sides was water. Diffusion is the
spontaneous movement of molecules from regions of higher concentration to regions of lower
concentration, driven by the random thermal motion of molecules. Since side B initially lacked any
small molecules, there was a strong concentration gradient favoring the net diffusion of this
molecule from side A to side B. After simulating for 60 seconds, the concentration of small
molecules decreased to 3.09% on side A as some diffused across to side B. The concentration in
side B increased to 1.79% as small molecules moved down their concentration gradient.

However, full equilibrium was not reached within 60 seconds, as some concentration
difference remained. This indicates the diffusion rate was relatively slow for the specific
experimental conditions. Factors like molecular weight, temperature, and viscosity affect diffusion
rates, with smaller, lighter molecules diffusing faster (Berj J, al., 2015). The properties of the small
molecule likely slowed its diffusion compared to the water solvent. Additional simulation time
would be needed to eventually reach equal concentrations on sides A and B. Overall, this
demonstrates the process of diffusion down a concentration gradient according to Fick's laws (Berj
J, et al., 2015).

8. Defining osmosis based on the simulation

Diffusion is the spontaneous movement of molecules from a region of higher concentration

to a region of lower concentration, driven by the random temperature-mediated motion of
molecules (Berj J, et al., 2015). In this process, solvent molecules move through a permeable
membrane from an area of lower solute concentration to an area of higher solute concentration.
This net movement of solvent equalizes the solute concentrations on both sides of the membrane
over time. An important example is the diffusion of water across a biological membrane, which
helps maintain homeostasis in living organisms (Sheerwood L, al., 2016). Water will diffuse
through a membrane to dilute areas of higher solute concentration and concentrated areas of lower
solute concentration. This helps maintain optimal internal conditions within cells and in bodily
fluids. Imbalances in solute concentrations can disrupt important biological processes, so diffusion
of solvent is a key mechanism for regulating these gradients and avoiding extreme shifts in tonicity
or osmolarity (Sheerwood L, et al., 2016). Overall, the passive movement of solvent molecules
down their concentration gradient provides an elegant and efficient means of maintaining
homeostasis across membrane boundaries in biological systems.

9. Simple experiment
BIO108 • Laboratory Activity 3• Group 03

The Table 3 shows that. This diffusion simulation demonstrates how temperature and
molecule size impact diffusion rates. Initially, a 5 mol/L concentration of small, medium and large
molecules was set up on side A only. Side B had zero concentration. Higher temperatures generally
speed up diffusion by providing more kinetic energy to molecules (Berj J, etal., 2015). For the small
molecules, increasing temperature from 10°C to 50°C caused the concentrations on sides A and B
to become more equalized over the same time period. At 10°C, side A remained at 3.6 mol/L while
at 50°C it dropped to 1.9 mol/L, indicating faster diffusion. The medium molecules showed a
similar trend but diffused slower than the small molecules at all temperatures. Finally, the large
molecules diffused extremely slowly, with side B staying at 0 mol/L even at higher temperatures
(Biology Simulation, 2023). Molecular weight is a key determinant of diffusion rate, with smaller
molecules diffusing faster. The small molecules could diffuse quickly across the membrane due to
low molecular weight. The large molecules were much heavier and could not diffuse well. The
medium molecules exhibited intermediate behavior. This simulation showed how temperature
increases kinetic energy and accelerates diffusion, while larger molecular size slows diffusion. The
combined effects of these two factors were evident, with small molecules diffusing rapidly at high
temperatures but large molecules diffusing extremely slowly even with added thermal energy
(Biology Simulation, 2023). This demonstrates the molecular underpinnings of Fick's laws of
diffusion in action (Berj J, et al., 2015).

CONCLUSION

Osmosis is the process by which solvent particles move from a diluted solution into a concentrated
solution by passing through a semipermeable membrane. Diffusion is the movement of particles from a
higher concentration area to a lower concentration area, which has the overall effect of sustaining
concentration throughout the medium. The solvent moves to dilute the concentrated solution and equalize
both the internal and external environment of the membrane. According to our data, we can confirm that
the process of osmosis and diffusion had occurred. This experiment shows us that temperature and molecule
size have a direct impact on the diffusion rate's efficiency. It also highlights the fact that such large
molecules cannot pass through osmosis and diffusion because of their size and charge. The smaller the
molecule size, the higher the diffusion rate. Same as the higher the temperature, the higher the diffusion
rate of molecules. By conducting this experiment, we're able to get a comprehensive understanding of the
concept of how the process of osmosis and diffusion works in relation to a selectively permeable membrane.

LITERATURE CITED

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of the Cell. Garland Science.

Berg, J. M., Tymoczko, J. L., & Gatto, G. J. (2015). Biochemistry (8th ed.). W. H. Freeman and
Company.

Campbell, N. A., & Reece, J. B. (2008). Biology (8th ed.). Pearson.

BIO108 • Laboratory Activity 3• Group 03

Carolinastaff, & Carolinastaff. (2023, March 23). Diffusion and osmosis. Carolina Knowledge
Center.https://knowledge.carolina.com/discipline/life-science/anatomy-and-
physiology/diffusion-and-osmosis/

Helmenstine, A. M., PhD. (2020, February 6). Differences between osmosis and diffusion.
ThoughtCo.https://www.thoughtco.com/difference-between-osmosis-and-diffusion-
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Lehninger, A. L., Nelson, D. L., & Cox, M. M. (2017). Principles of Biochemistry. W.H. Freeman
and Company.

Libretexts. (2020, July 14). Diffusion and osmosis. Biology LibreTexts.

https://bio.libretexts.org/Learning_Objects/Worksheets/Biology_Tutorials/Diffusion_and
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Lodish, H., Berk, A., Zipursky, S. L., Matsudaira, P., Baltimore, D., & Darnell, J. (2020). Molecular
Cell Biology. W. H. Freeman.

Simulations (2023) Biology Simulations. Available at:

https://www.biologysimulations.com/simulations (Accessed: 07 November 2023).

Reinke, N. B., Kynn, M., & Parkinson, A. L. (2021). Immersive 3D experience of osmosis improves
learning outcomes of First-Year cell Biology students. CBE- Life Sciences Education,
20(1), ar1. https://doi.org/10.1187/cbe.19-11-0254