Diffusion, Dialysis and Osmosis

Group 2
Asaad, Baldovino, Benuyo, Dee, Evangelista, Garcia, Vicua

Biology 22 Lab 3B
August 22, 2014

Abstract
Cell is the basic unit of life and all living organisms are made up of cells. In order for the cell to sustain life, the cell
must maintain internal stability. To do this, the cell makes use of a semi permeable membrane that acts as a regulator for the
materials that pass through the cell. The materials may pass through the cell either passively by physical processes such as
diffusion or actively by physiological processes such as active transport. Five different activities were done to predict the
movement of the materials to the differentially permeable membranes; to discuss the importance and functions of the cell
membrane; and to know the factors that affecting the rate of diffusion of the materials to the cell membrane. Using the methods in
the activities, results were tabulated for each of the activities. Quantitative results show that the distilled water, the chloride test
and potassium permanganate has the fastest diffusion or dialysis in their respective groups. For the osmosis, the water goes to the
sucrose filled test tube. Also, different concentrations of NaCl solution affect the red blood cells differently. It was found out that
the rate of diffusion is affected by the concentration and molecular weight of the substance. Substances move from higher areas
of concentration to lower areas of concentration. Also, diffusion rate is lower if the substance has a higher molecular weight.

Introduction
According to the cell theory, all
organisms are made up of cells. It is the most
basic unit of life. In fact, there are various
single-celled life forms. More complex
organisms, such as plants and animals, are multicellular; they have specialized cells that work
together to maintain life. In response to
environmental changes, cells need to maintain
internal stability to sustain life. The cell
regulates its composition, using a limiting semipermeable membrane, in order to preserve
constant condition despite the changes in its
environment. This semi-permeable membrane is
referred to as cell or plasma membrane. The cell
membrane,
in
response
to
different
environmental changes, acts as a barrier to some
particular substances while at the same time,
allowing the passage of other materials. By
controlling the passage of different materials, the
cell can manipulate its own composition.
Passive movement of materials is possible
through different processes such as diffusion.
Osmosis and dialysis are the two forms of
diffusion processes. Physiological processes
such as active transport, phagocytosis, and
pinocytosis can possibly move the materials
actively.

Method
A. Diffusion in water

Two test tubes were obtained, one halffilled with tap water and the other half-filled
with distilled water. The two test tubes were
then labeled properly. In each of the two test
tubes, two crystals of potassium permanganate
(KMnO4) crystals were dropped using a pair of
forceps. The test tubes were then placed on a
rack so as to prevent any disturbance. The
dissolution of the potassium permanganate
crystals in each test tube was then observed.
This dissolution is indicated by the dispersal of
a purplish, light color. The time when the color
had completely dispersed in the water in both
test tubes were recorded.
B. Diffusion in Colloid
Three test tubes filled with hardened 5%
gelatin solution were obtained. The three test
tubes were then added with 1 mL of each of the
following
reagents
simultaneously:
(1)
potassium permanganate, (2) methylene blue,
and (3) congo red. The time it took for the
stains to reach the bottom of each test tube was
recorded. At five-minute intervals, the distance
travelled by each stain through the gelatin was
recorded. The same procedure was done for one
and a half hours.
C. Dialysis

In a small test tube, 3 mL of calcium

chloride (CaCl2) solution and 2 mL methylene
blue were combined. The mouth of the test tube
was then sealed with a dried longganisa skin and
a rubber band. It was made sure that there were
no leaks on the seal. Half of the test tube was
then immersed, with the sealed mouth facing
down, in a beaker filled with about 30 mL of
distilled water. The test tube was held vertically
and it was made sure that the sealed mouth does
not touch the bottom of the beaker. A test tube
holder was used to hold the test tube in place.
For every 30 seconds, 2-4 drops of water were
aspirated from the beaker. A drop of it was
placed in two separate test tubes. A drop of
silver nitrate was then added to the first test tube
whereas a drop of ammonium oxalate was added
to the second test tube. These were done to test
the presence of chloride and calcium ions
respectively, which would be indicated by a
white, cloudy precipitate. The same procedure
was repeated until the precipitate became
evident in both test tubes. The time when the
precipitate became evident in both test tubes was
recorded. It was also observed if the methylene
blue would diffuse into the beaker.
D. Osmosis
A small test tube was filled with 5 mL of
a 30% sucrose solution. The mouth of the test
tube was then sealed with longganisa skin and a
rubber band. It was made sure that there were
no leaks on the seal. The test tube was then
inverted and the level of the liquid inside the test
tube was noted. The test tube was then
immersed, with the sealed mouth facing down,
in a beaker filled with about 60 mL of distilled
water. The test tube was held vertically and it
was made sure that sealed mouth does not touch
the bottom of the beaker. A test tube holder was
used to hold the test tube in place. The test tube
was left immersed in this position for about two
and a half hours. Afterwards, it was observed
whether the level and weight of the liquid inside
the test tube increased.
E. Osmosis in Red Blood Cell
Three clean microscope slides were
prepared and labeled A, B, and C respectively.

Three drops of blood was extracted from a

female volunteer by pricking her index finger
with a sterile lancet. It was made sure the skin
of the volunteers index finger was swabbed first
with alcohol before doing the procedure. A drop
of blood was placed on each of the three slides.
The three slides were then added with a drop of
each of the following solutions: (1) 0.07 M NaCl
to the blood on slide A, (2) 0.15 M NaCl to the
blood on slide B, and (3) 0.30 M NaCl to the
blood on slide C. The blood and the NaCl
solutions were mixed thoroughly, and the
mixture was smeared evenly on the surface of
the slides. The slides were focused at 400X
using three different microscopes. With the aid
of an ocular micrometer, the diameter of the
cells in each slide was measured for every five
minutes for forty minutes. Possible changes in
the shape of the cells in each slide were
observed.

Results
A. Diffusion in Water
The KMnO4 crystals diffused faster in
the test tube half-filled with distilled water. In
the test tube half-filled with tap water, the
diffusion was slow, marked by the formation of
a nebula-like structure suspended at the middle
of the test tube (it disappeared eventually). The
nebula-like structure was not observed in the test
tube with dH2O since the purple color quickly
scattered throughout the entire tube.

B. Diffusion in Colloid
Results are shown through in a tabular format:
Table 4.1 Distance travelled by Potassium
permanganate, Methylene Blue and Congo Red.
Time (min.)

Distance Travelled (cm)

KMnO4

1.0

Methylene
Blue
0.3

Congo Red

10

1.4

0.5

0.3

15

1.5

0.6

0.3

0.2

20

1.6

0.7

0.3

25

1.9

0.8

0.3

30

2.1

0.9

0.4

35

2.1

0.9

0.4

40

2.2

0.9

0.4

Based on the data, it can be said that the

KMnO4 travelled the farthest distance, followed
by methylene blue, and lastly by congo red.

C. Dialysis
Table 4.2 Results of Dialysis of Methylene Blue,
Calcium and Chloride Ions
Time (min)
1.5
2.5
3.5

Chloride
Test

Calcium
Test

Methylene
Blue

4.5
5.5

6.5

For the chloride test, the cloudy

precipitate started to appear after 2.5 minutes,
while for the calcium test, it took 5.5 minutes.
The methylene blue diffused into the beaker
after 3.5 minutes.
D. Osmosis
Some of the water in the beaker set-up
was transferred to the test tube containing 30%
sucrose. Initially, the water in the beaker was
filled with 60 mL, then after 2 hours, 10 mL of
the water in the beaker shifted to the sucrosefilled test tube.

E. Osmosis in Red Blood Cell

Table 4.3 Size of Red blood Cells under

different concentrations of NaCl
NaCl Concentration (M)
Time (min.)

0.07 M

0.15 M
Cell size in um

0.30 M

10

15

20

25

30

35

40

Based on the acquired data above, it can

be said that under a NaCl concentration of 0.07
M, the size of the RBC increased, under the
NaCl concentration of 0.15 M, the size of the
RBC remained constant, whereas under the
NaCl concentration of 0.30 M, the size of the
RBC increased.

Discussion
Diffusion is the process in which
molecules move from a more concentrated
solution to a less concentrated solution. That is
why in experiments A and B, it was seen that the
molecules of the solutes in the different test
tubes diffused. However, it was observed in the
results that the different substances showed
different times of diffusion with one diffusion
faster than the others. For experiment A, the test
tube filled with distilled water diffused faster
than the test tube filled with tap water. The
reason for this is that distilled water is purer than
tap water because tap water has impurities such
as bacteria, minerals etc. This makes tap water
more concentrated than distilled water. Because
of this, KMnO4 was able to diffuse faster in
distilled water.
The concentration of a substance is a
major factor in the speed of diffusion. This can
be observed the experiment B in which diffusion
is observed in colloidal materials. Unlike water,

these substances have colloidal suspensions that

affect the rate of diffusion. In the experiment,
three reagents namely potassium permanganate
(KMnO4), methylene blue, and congo red were
simultaneously added into three separate test
tubes containing gelatin. After forty minutes, it
was observed that among the three reagents
potassium permanganate was the one to diffuse
the farthest.
Compared to the two other
reagents, potassium permanganate has the
smallest and lightest molecules which it faster
and easier for it to diffuse. When it comes to the
diffusion of colloids, the molecular weight of the
chemical is a primary factor. The smaller and
lighter the molecular weight of a chemical, the
faster it will diffuse.
Dialysis
involves
separation
of
substances in a solution by their unequal
diffusion
through
their
semipermeable
membranes. In the experiment, it was observed
that a cloudy precipitate formed. This happened
due to the reaction between the ions and their
reactants. The chloride ions diffused faster than
the calcium ions because chloride ions have
smaller molecules compared to the calcium ions.
Methylene blue was observed to have diffused
into the beaker. However, the molecules of
methylene blue are very large that the membrane
of the dried longganisa skin will not allow the
passage of the molecules. The methylene blue
should not have diffused into the beaker. The
dried longganisa skin is considered as a
semipermeable membrane because it allowed
smaller molecules such as the chloride ions and
calcium ions to pass through and prevented
larger molecules from passing.
Osmosis is the process in which water
permeates through a cell through its membrane.
Water is the one that diffuses in this process.
Water is transported from substances with higher
amounts of water to substances with lower
amounts of water. That is why in experiment D,
some of the water transferred to the test tube
filled with 30% sucrose since there is a lower
concentration of water in the test tube than in the
beaker filled with distilled water. Osmosis is
also observable in experiment E. In this
experiment, it can be determined that the setup
with 0.15 M NaCl is an isotonic solution

because there was no change in the size of the

cell. This means that the concentration between
the cells and the solution are the same 0.15
NaCl. On the other hand, the setup with 0.07 M
NaCl is a hypotonic solution because it caused
the cells to get bigger. This means that the
solution has greater amounts of water than the
cell. Lastly, the setup with 0.30 M is a
hypertonic solution because the solution has
lesser amounts of water than the cell. However,
results showed that the 0.30 M setup caused the
cells to become bigger when theoretically it
should have caused it to shrink.

Conclusion and Recommendation/s

Based on the result of the experiment, it
can be concluded that the rate of diffusion of a
substance is affected by its concentration and
molecular weight. Substances move from areas
of higher concentration to areas of lower
concentration just like what happened to
potassium permanganate when it diffused faster
in distilled water which happens to have a lower
concentration than tap water. When it comes to
the molecular weight of substances, a substance
with lower molecular weight like potassium
permanganate (158.03 g/mol) has a higher
diffusion rate compared to a substance with
higher molecular weight like Congo red with a
molecular weight (696.66 g/mol). On the other
hand, methylene blue which happens to have
molecular weight of 319.85 g/mol has a slightly
lower diffusion rate compared to potassium
permanganate but faster than congo red.
The rate of diffusion is faster in water
compared to gelatin which is a colloid because
water has more spaces to allow the spread of
molecules compared to colloids where space is
limited which also limits movement of
molecules therefore the rate of diffusion is slow.

In dialysis, which refers to the diffusion

of solutes across a selectively permeable
membrane, the diffusion of substances with
smaller molecules occurs faster just like what
happened to chloride ions which started to
appear ahead of the calcium ions.
In osmosis, which is defined as the
diffusion of water from a hypotonic solution or
low concentration of solute to a hypertonic
solution or high concentration of solute, there
should be a balance on the ionic concentrations
(isotonic solution) on both sides of the cell
membrane for the cells to survive. This was
shown in experiment E in the 0.15 M NaCl,
where the size of the RBC remained constant.
Therefore this is the isotonic solution which is
needed for the cell to survive. On the other
hand, results showed that the 0.30 M NaCl setup
caused the size of the RBC to become bigger
when theoretically it should have caused it to
shrink. Perhaps, there was an unknown mistake
in the conduction of the experiment in the 0.30
M NaCl setup. The researchers then recommend
the repetition of the procedure for the 0.30 M
NaCl setup to further validate the results.

Bibliography
Campbell, N., Reece J. (2008). The Cell.
Biology (8th ed.). San Francisco,
CA: Pearson Education, Inc. (pp. 94).
Campbell, N., Reece J. (2008). Membrane
Structure
Results
In
Selective
Permeability. Biology (8th ed.). San
Francisco, CA: Pearson Education, Inc.
(pp. 131).
Mendoza E. E., Religioso, T. F. (2008).

Chemistry. Quezon City, Philippines:

Phoenix Publishing House Inc.
Smith,

D. Osmosis, Diffusion and Cell

Transport. 2003. 25 August 2014
<http://teachers.henrico.k12.va.us/godwi
n/strauss_s/sscwebpage/tutorials/cell_tra
nsport_tutorial.pdf>.