The Quest for Equilibrium: Osmosis 1

The Quest for Equilibrium: Osmosis

Aaron Ventresca
Mrs. Carly Steininger
Honors Biology, Period IV
Cardinal Wuerl North Catholic High School
30 April 2018
The Quest for Equilibrium: Osmosis 2

Introduction
In order for the natural world to function, all organisms and interrelated systems must

live in harmony to maintain the delicate balance that sustains life. The delicate balance achieved

by various organisms, climates, and systems takes on many different forms. From the great cloud

forests of the Amazon Rainforest to the humble existence of the desert skink, all living things

rely on one another to create equilibrium in the natural world.

On a smaller level, every cell in existence constantly strives to maintain equilibrium to

sustain life. Cellular transport, the process in which the cell controls what enters and exits,

facilitates the maintenance of equilibrium (The Editors of the Encyclopaedia Britannica, 2018). In

general, there are two types of cell transport: passive and active. In passive transport, the cell

allows substances to flow from a high concentration to a low concentration. Moving along the

concentration gradient, substances that undergo passive transport are typically small molecules

that fit through the cell membrane. The cell membrane is selectively permeable; the cell membrane

allows only certain materials to pass through it. For example, the cell membrane allows water to

flow through it, but does not allow salt to do the same. Thus, the cell membrane is selectively

permeable. (University of Illinois, 2016)

The cell constantly strives to achieve equilibrium because it is constantly placed in different

environments that upset its typical resting state. Osmosis, the process in which water moves across

the cell membrane from a high concentration to a low concentration, allows the cell to adapt to

three potential osmotic solutions/situations (The Editors of the Encyclopaedia Britannica, 2018).

If the cell is placed in a hypotonic solution, there is a higher concentration of pure water outside

of the cell. Water will flow into the cell causing it to swell to reach equilibrium. If too much water

enters the cell, the cell may burst, undergoing cytolysis. Conversely, if the cell is placed in a

hypertonic environment, there is a higher concentration of water inside the cell. Water will flow
The Quest for Equilibrium: Osmosis 3

out of the cell causing it to shrink to reach equilibrium. If too much water leaves the cell, the cell

may shrivel, undergoing plasmolysis. If the cell is placed in an isotonic environment, however,

there is an equal concentration of water both inside and outside the cell. Thus, the cell is at

equilibrium. (University of Washington Department of Chemsitry, 2018)

Understanding the three types of osmotic solutions in addition to the applications of

osmosis is crucial in running various businesses. In grocery stores, hydration systems are

engineered to spray fresh produce with water, causing it to swell (hypotonic solution) to make it

appear appetizing to customers. On golf courses, horticultural engineers work to use reverse-

osmosis to water grass while performing conservation efforts at the same time (Advameg, 2018).

In addition, nurses and hospital technicians rely heavily on a strong understanding of osmosis

when helping patients recover from dehydration and injury through intravenous medical

treatments (IVs). In IVs, both salt and water are given to a patient so as to prevent cells from

bursting (cytolysis).

Using these previous examples as inspiration, a study was conducted to examine how the

rate of osmosis changes depending on the substances used and the strength of the concentration

gradient. Part I was performed to (1) observe the different effects of osmosis in different osmotic

environments, (2) model the permeability of the cell membrane, (3) describe how the rate of

osmosis differs because of different concentration gradients, and (4) assess how the rate of

osmosis changes as a cell approaches equilibrium. Part II was performed to model how dialysis

tubing is semi-permeable.

In this experiment, dialysis tubing was used to represent “model cells”. Dialysis tubing

was utilized because of its uniquely semi-permeable characteristics (University of Washington

Department of Chemsitry, 2018). Dialysis tubing is only permeable to certain fluids; thus,
The Quest for Equilibrium: Osmosis 4

dialysis tubing is the perfect model for the cell membrane. In this lab, six beakers were filled

with 200 ml of pure water in addition to one model cell. Each of the model cells created

represented a different osmotic environment. Beaker I (water in water) represented an isotonic

environment. Beaker II (20% glucose in water) represented a hypotonic environment. Beaker III

(40% glucose in water) represented a hypotonic environment. Beaker IV (60% glucose in water)

represented a hypotonic environment. Beaker V (80% glucose in 60% glucose, and water in 60%

glucose) represented a hypotonic environment and a hypertonic environment, respectively.

Beaker VI (glucose in water) was a less precise, yet still indicative data point regarding the

permeability of the cell membrane.

Part I of the experiment consisted of the first five beakers. In Part I, the constants were:

200 ml of solution in each beaker, the same temperature of water in each beaker, roughly the

same size piece of dialysis tubing in each beaker, the same time intervals between tests, the same

folding and tying pattern of dialysis tubing, the same drying procedures before measurements,

and roughly the same size of each beaker. The independent variable for Part I was the

concentration gradient and the amount of glucose in each baggie. The dependent variable was the

change in mass of the baggies. In Part I, the control group was Beaker I (water in water). Thus,

the experimental groups were Beakers II (20% in water), III (40% in water), IV (60% in water),

and V (80% in 60%, water in 60%). For Beaker I, the hypothesis would be: “If a pure water cell

is placed in an isotonic environment, then the cell will stay around the same mass.” For Beaker

II, the hypothesis would be: “If a 20% glucose cell is placed in a hypotonic environment, then

the cell will gain mass.” For Beaker III, the hypothesis would be: “If a 40% glucose cell is placed

in a hypotonic environment, then the cell will gain mass.” For Beaker IV, the hypothesis would

be: “If a 60% glucose cell is placed in a hypotonic environment, then the cell will gain mass.”
The Quest for Equilibrium: Osmosis 5

For Beaker V, the hypothesis would be: “If an 80% glucose cell is placed in a 60% glucose

environment, then the cell will gain mass, and if a purely aqueous cell is placed in a 60%

environment, then the cell will lose mass.”

Part II of the experiment consisted of a bag of starch in iodine-dyed water. In Part II, the

constants were: the same dialysis tubing, the same size beaker, and the same amount of water.

The independent variable for Part II was the location of the starch (in the dialysis tubing). The

dependent variable for Part II was the color change of the beaker and the baggie. The control

group for Part II was the “before” state of the beaker containing the iodine and starch baggie.

The experimental group for Part II was the “after” state of the beaker containing the selectively

permeable tubing with the iodine. The hypothesis for Part II was: “If cells truly have selectively

permeable membranes, then the iodine will diffuse into the model cell, causing its contents to

turn blue.”

In total, the following experiment details how osmotic tendencies occur in a controlled

environment. This report will provide critical details to understanding and applying osmotic

principles in the real world.

Materials
 Six 500 ml beakers  Two graduated cylinders
 Three L of pure water  Eight plastic spoons
 Seven pieces of dialysis tubing  Electronic scale
 Ribbon  Paper towels
 Scissors  Iodine
 Starch  Stopwatch/Clock
 20% glucose solution  Lab sheet
 40% glucose solution  Pipettes
 60% glucose solution  Writing utensil(s)
 80% glucose solution
The Quest for Equilibrium: Osmosis 6

Procedures
Part I:
1) Soak six pieces of dialysis tubing in one L of pure water.
2) After allowing dialysis tubing to become sufficiently wet and malleable, take one piece of
tubing, and fold one end horizontally, then vertically, then horizontally.
3) Tie a knot with ribbon around the end of the tubing. Cut off excess ends.
4) Fill the tubing with 5 ml of water.
5) Affix the other end of the tubing by folding the end horizontally, then vertically, then
horizontally.
6) Tie the other end of the tubing with ribbon. Cut off excess ends.
7) Repeat steps 2-6, using water, 20% glucose solution, 40% glucose solution, 60% glucose
solution, and 80% glucose solution in place of water.
8) Obtain and fill four beakers with 200 ml of pure water.
9) Obtain and fill the fifth beaker with 200 ml of 60% glucose solution
10) Mass each of the six baggies created. Record data on lab sheet.
11) The setups should be like the following:
a. Beaker I: water in water
b. Beaker II; 20% in water
c. Beaker III: 40% in water
d. Beaker IV: 60% in water
e. Beaker V: water in 60%, 80% in 60%
12) Simultaneously, place the baggies their respective beakers: water in water, 20% in water,
40% in water, 60% in water, water in 60%, and 80% in 60%.
13) After three minutes, simultaneously remove the baggies from the beakers.
14) Roll the baggies gently on paper towels to dry.
15) Mass each of the baggies. Record data on lab sheet.
16) Repeat steps 12-15 two more times.
17) Empty beakers down the sink and dispose of the model cell baggies. Wipe down lab
surfaces thoroughly. Wash hands after all else is completed.
18) Calculate the change in mass based on the mass measurements collected at regular three-
minute intervals. Record data on lab sheet.
Part II:
1) Soak one piece of dialysis tubing in 200 ml of pure water.
2) After allowing dialysis tubing to become sufficiently wet and malleable, take one piece of
tubing, and fold one end horizontally, then vertically, then horizontally.
3) Tie a knot with ribbon around the end of the tubing. Cut off excess ends.
4) Fill the tubing with about one teaspoon of starch.
5) Affix the other end of the tubing by folding the end horizontally, then vertically, then
horizontally.
6) Tie the other end of the tubing with ribbon. Cut off excess ends.
7) Obtain and fill a beaker with 200 ml of water.
The Quest for Equilibrium: Osmosis 7

8) Add eight drops of iodine to the water.

9) Place the model cell baggie into the water.
10) Observe the color of the beaker and the color of the contents of the dialysis tubing. Record
data on lab sheet.
11) One day later, observe the color of the beaker and the color of the contents of the dialysis
tubing. Record data on lab sheet.
12) Dispose of cell baggie and dump contents of beaker down sink. Wash hands after all else
is completed. (Diffusion Through Cell Membranes, 2018)
Results
Table 1 displays the data collected. The data shown represent the class averages for each

of the cumulative mass measurements after three, six, and nine-minute intervals. To acquire this

data, data was shared and then inputted into an EXCEL spreadsheet. After running some

arithmetic mean calculations, the averages for each of the measurement values were obtained.

Table 1 displays the cumulative mass changes for three, six, and nine-minute intervals. It is

important to note that cumulative mass does not represent mass or change in mass. Rather, it

represents a summation of change in mass values that create a cumulative trend (displayed on the

ogive graph in Figure 1).

Based off the data in Table 1, when a water-filled bag was placed in a purely aqueous

environment, the baggie gained 208 mg of mass after three minutes. After another three minutes,

the baggie had a total net mass again of 291 mg. After yet another three minutes, the baggie had

a total net mass gain of 249 mg. When a 20% glucose baggie was placed in water, the baggie

gained 317 mg after three minutes. From three to six minutes, the baggie ended with a total gain

of 534 mg. After one more measurement, the baggie ended up with a total net mass gain of 701

mg. When a 40% glucose baggie was placed it water, it gained 408 mg after just three minutes.

After a total of six minutes, the baggie had gained a total of 800 mg of mass. After a final total of

nine minutes, the baggie had gained a total of 1408 mg. When a 60% glucose baggie was placed

in water, it gained 567 after just three minutes. After six minutes, the baggie had a total mass
The Quest for Equilibrium: Osmosis 8

change of 1009 mg. After one measurement at a three-minute interval, the baggie ended up with

a net gain of 1409 mg. When a pure water baggie was placed in the 60% glucose solution, the

baggie lost 15 mg. After six minutes, the baggie lost 533 mg of mass. After a total of nine

minutes, the baggie lost 783 mg. When an 80% glucose baggie was placed in the 60% glucose

solution, the baggie gained 241 mg after three minutes. After six minutes, the baggie had a net

mass gain of 316 mg. After nine minutes, the baggie gained a net total of 399 mg.
Table 1: Changes in Mass Over Time*
Time Water in 20 % in 40% in 60% in Water in 80% in
Water Water Water Water 60% 60%
0 0 0 0 0 0 0
3 208 317 408 567 -15 241
6 291 534 800 1009 -533 316
9 249 701 1108 1409 -783 399
*All units are milligrams (mg)
Description: Table 1 details the class averages for the cumulative mass changes of each of the
baggies in the experiment. The data provided were collected at regular three-minute intervals.
To ensure that the data provided are correct, a specific delineation was made between change
in mass, net mass gain, and simply mass. Originally, mass values were collected from the lab.
The data values initially recorded on the lab sheet were thus the mass (in grams) of the
baggies between sessions in the beaker. To calculate change in mass, the formula “change in
mass=new mass-old mass” was used. Thus, the values for change in mass were then acquired
through calculations. The data provided above represent cumulative mass changes. Whenever
cumulative masses are calculated, the mass changes add on to one another to display how the
mass of the cell increases/decreases over time. It is important to note that the data provided
are uniform. The different mass changes were interpreted to form uniform information that
represents cumulative mass changes. Thus, just like cumulative frequencies sum to one,
cumulative masses keep adding onto one another.
The Quest for Equilibrium: Osmosis 9

Cumulative Mass Change vs. Time

2000
CUMULATIVE MASS GAIN (MG)

1500
1000
500
0
0 3 6 9
-500
-1000
TIME (MIN)
Water in Water 20% in Water 40% in Water
60% in Water Water in 60% 80% in 60%

Figure 1: Cumulative Mass Change vs. Time

Description: The above graph is a modified ogive. Ogive graphs display cumulative values- in this
case, mass. The x-axis depicts the time intervals at which the data values were collected: three
minutes, six minutes, and nine minutes. The y-axis depicts the cumulative mass gain of each of the
baggies in the experiment. These values were originally measured in grams but were converted to
milligrams to facilitate calculations and graphical analysis. Each of the colored series on the graph
represents a different baggie. The dark blue series represents water in water, the orange series
represents 20% in water, the gray series represents 40% in water, the yellow series represents 60%
in water, the light blue series represents water in 60%, and the green series represents 80% in 60%.

The data shown in Figure 1 represent the cumulative masses of each of the individual

cells tested. The graph shows how all the model cells tested have no net mass gain at zero

minutes. After three minutes, the series begin to diverge. The 60% in water gains the most mass,

approximately 567 mg. The 40% in water gains the second most mass, 408 mg. After six

minutes, the series begin to diverge even more. The 60% in water leads the way with 1009 mg of

cumulative mass, followed by the 40%, which has 800 mg of cumulative mass. After nine

minutes the series differentiate even further. The 60% in water has 1409 mg of cumulative mass,

and the 40% in water has 1108 mg of cumulative mass. It is important to also note that the water

in 60% loses a cumulative total of 783 mg of mass over the course of the experiment.
The Quest for Equilibrium: Osmosis 10

Table 2 sums up Part II of this experiment. Before the beaker was left to sit, the contents

of the baggie were white. The water in the beaker, having just been exposed to eight drops of

iodine, was of a yellow hue. After the beaker was left to sit overnight, the final results were

recorded. The beaker showed a clear color while the dialysis bag showed a bluish-purple color.

Table 2: Color Change with Addition of Iodine

Starting Color Color After One Day

Solution in Dialysis Bag White Bluish-Purple
Solution in Beaker Yellow Clear

Description: Table 2 is a qualitative summary of Part II of this experiment. In Part II, a starch-
filled baggie was placed in a beaker full of iodine and water. The beaker was left to sit
overnight. The next day, the results were recorded.

Discussion
The results in this lab were highly suggestive. Each of the simulated cells in Part I of the

experiment attempted to reach equilibrium, the aim of osmosis. Thus, all of the bags gained or

lost weight during Part I of the lab to reach equilibrium. As previously stated in the introduction,

all living things must exist in harmony and achieve a balanced equilibrium. The cells placed in

hypotonic environments (20% in water, 40% in water, 60% in water, 80% in 60%) all gained

weight to balance the concentration of solute. Each of these cells had a lower concentration of

pure water inside the cell because of the glucose solution; water flowed into the cell to help it

achieve equilibrium. The cell placed in a hypertonic environment (water in 60%) lost weight. In

this cell, there was a higher concentration of water inside the cell, causing the cell to expel water

out to balance the solute concentration. The cell placed in an isotonic environment (water in

water) gained weight due to the fluctuation in water flow in and out of the cell. Aquaporin

channel proteins are always open, allowing water to flow in and out freely (University of Illinois,
The Quest for Equilibrium: Osmosis 11

2016). Though it was already at equilibrium, the isotonic cell gained weight due to the properties

of selectively permeable membrane fluctuate tendencies.

Another trend identified by the data was the rate of osmosis. (Advameg, 2018) Figure 1

displays the cumulative masses of each series. The slope of each series is the rate of osmosis. It

is essential to notice how each series “levels off” as time goes on. Thus, as cells get closer to

equilibrium as time passes, the rate of osmosis decreases (The Editors of the Encyclopaedia

Britannica, 2018). As cells get closer to equilibrium, they do not need to work as hard to move

substances to reach a sustainable equilibrium.

Further, concentration gradient plays a large part in the rate of osmosis (The Editors of

the Encyclopaedia Britannica, 2018). The concentration gradient is the difference of solute

concentration inside and outside the cell. The biggest concentration gradients in this experiment

were water in 60%, and 60% in water. Compared to the other simulated cells, these two high-

concentration gradient scenarios experienced the greatest changes in mass. Thus, as the

concentration gradient increases, so does the rate of osmosis (University of Illinois, 2016). In a

mildly hypotonic environment (20% in water), the cell does not have to work hard to allow

equilibrium to be reached. When there is a strong concentration gradient, however, the cell

speeds up the osmotic process to maximize cellular function. Concentration gradient is integral

in determining the rate of osmosis; however, the data may suggest otherwise.

In this experiment, the 80% in 60% cell and the 20% in water cell both had the same

concentration gradient yet did not experience the same change in mass. This is due to the fact

that the beaker containing the 80% in 60% cell also contained the water in 60% cell. As the 20%

in water cell reached equilibrium, water from the external environment permeated the dialysis

tubing to balance the solute concentrations. During this process, the external environment
The Quest for Equilibrium: Osmosis 12

remained 100% pure water. Conversely, in the beaker holding the 80% in 60% cell and the water

in 60% cell, the external environment changed during the experiment. The water in 60% cell

represented a hypertonic environment; a significant amount of water came flowing out of the cell

into the environment to balance solute concentrations. Meanwhile, the 80% in 60% cell

represented a hypotonic environment. Thus, the water from the external environment began

entering the 80% cell. Having the water in 60% cell altered the external environment, as more

water was continually being added, diluting the 60% glucose concentration. Therefore, the

external environment in Beaker V was constantly changing due to the two cells present. In

Beaker V, all three environments would have eventually reached equilibrium; this phenomenon

takes time. The 80% in 60% cell did not gain as much weight in three minutes as the 20% in

water cell due to the changing external environment and the amount of time the experiment was

conducted.

Transitioning to Part II of the lab, the results once again were highly accurate. The inside

of the simulated cell turned blue due to the selective permeability of the dialysis tubing. Osmosis

is aimed at reaching equilibrium. The concentration of solute inside the dialysis baggie was

greater than that in the beaker. The iodine permeated the dialysis tubing. When starch and iodine

come in contact, the resulting substance is a bluish-purple color; the iodine died the starch inside

of the dialysis bag blue. Thus, the cell turned blue because of the permeability of the dialysis

baggie. As stated, the dialysis baggie is permeable to water and iodine but not starch.

Though this lab was highly successful and yielded accurate results, there are a few

sources of error to be considered. Primarily, the experiment was not conducted for a long enough

period of time. In Part I of the experiment, the masses of the cells were measured at three-minute

intervals. In the original instructions, the cells were supposed to be measured at 10-minute
The Quest for Equilibrium: Osmosis 13

intervals. If the cells were left longer in each of the beakers, more accurate results may have been

acquired. Further, the beaker water in Part I of the lab was not exactly room temperature. Due to

time constraints and human error, the water placed in each of the beakers was rather hot. The hot

water may have sped up the rate of osmosis, yielding skewed results. To add, the dialysis baggies

may not have been completely sealed. Though the baggies were double-knotted and folded

intricately, some solute may have leaked out of the ends of the baggies. This “leakage” may have

yielded skewed results. Finally, the baggies might not have been completely dry when they were

massed at the regular intervals. Failure to dry the baggies off completely may have added extra

mass that could have provided skewed results.

In the future, it would be beneficial to review the lab setup and important details in class

before the lab to ensure that time constraints are not an issue. The setup should be reviewed and

documented days in advance so that on the day of the actual lab, more accurate results can be

obtained over a longer experiment duration.

In sum, the osmosis lab performed was a great research tool that allows for cross-

curricular connections and the publication of personal research. The data provided suggest that

cells strive to reach equilibrium, changing the rate of osmosis if necessary. Cells will expel and

take in water to reach a balanced concentration. The goal of osmosis is to create balance in the

cell; osmosis is an endless quest for equilibrium.

The Quest for Equilibrium: Osmosis 14

References
Advameg. (2018). Osmosis- Real-Life Applications. Retrieved from Science Clarified.
Diffusion Through Cell Membranes. (2018). 1-4.
The Editors of the Encyclopaedia Britannica. (2018). Osmosis. Retrieved from Encyclopaedia
Britannica.
University of Illinois. (2016, December 30). Theoretical and Computational Biophysics Group.
Retrieved from Structure, Dynamics, and Function of Aquaporins.
University of Washington Department of Chemsitry. (2018). University of Washington.
Retrieved from Osmosis.