Biological Membranes: Dialysis, Osmosis, and Diffusion

Maravilla, Alexandra Louise M., Mercado, Antonette B.,

Manabat, Mariah Joy L. Ombrero, Karen O.Nunez, Gianne
Kianna A Nadonga, Nicole Angelyn T. Niez, Kimberly S.
Group 4 – BS Psychology 1PNASC2115, Department of Psychology, College of Social
Sciences and Development, Polytechnic University of the Philippines

ABSTRACT B

B\\B Biological membranes are thin boundary structures of molecular size on the surface of
cells and subcellular particles, as well as canaliculi and vesicles piercing the
protoplasm. Their most important function is to regulate the transport of ions, sugar and
amino acids, and other metabolic products. Biological membranes adapt their
composition such that the physical properties are maintained when the external
conditions change. This gives a role to change a composition, to temperature and
pressure. In this activity, different transport of activities and the different factors affecting
diffusion, osmosis and dialysis were observed. Potassium permanganate (KMnO4)
crystals was used for the diffusion in water. It was observed that the fastest rate of
diffusion of the solute KMnO4 crystals was on hot distilled water among the other types
and conditions of water which were tap water and distilled water. Therefore, the
presence of the large amount of kinetic energy in high temperature made solute
particles diffuse quickly. For the diffusion in colloid, gelatin was used to observed the
rate of diffusion. As the experiment result showed, the KMnO4 was the fastest among
the other test stains with the smallest particle. Congo red was next to KMnO4 and the
lowest was the methylene blue. It was the believed that the particle size affects the rate
of diffusion whereas the particle size was inversely proportional to the rate of the
diffusion. Solute concentration between two solutions is the factor that affect the rate of
osmosis. Based on the results, the increase in weight of the test tube is relatively
proportional to the period of time perform in the experiment. Moreover, the
concentration of solute is inversely proportional to the concentration of water. In dialysis,
the result showed that after two minutes, the methylene blue which has larger
JOURNAL OF LABORATORY REPORTS IN BIOCHEMISTRY
NASC 2115 – BIOCHEMISTRY LABORATORY

molecules, pass through the longganisa skin and change the color of the distilled water
in the beaker into blue. While chloride ions diffuse faster compared to calcium ions
because it has a smaller molecular size and both chloride and calcium ions reacted with
their reactants, the silver sulphate and oxalic acid. It was concluded that a dried
longganisa skin is a semipermeable membrane that permits the passage of solvent,
dissolved ions and small molecules but blocks the passage of larger molecules.

KEYWORDS: Biological membrane, osmosis, diffusion, dialysis

random motion which in turn triggers in

diffusion. Collisions between molecules
I. INTRODUCTION
are common: even in the air of
Diffusion and osmosis are both atmospheric pressure, a molecules
passive transport processes, meaning
collides with a neighbor every few
they require no energy input to move
nanoseconds.
substances. Both processes are
essential to the proper functioning of the Diffusion makes air a composition
biological process such the transport of uniform by redistributing chemical
species, such as oxygen in the air, until
water or nutrients between cells. equilibrium is reached: in other words
until the concentration gradient- the
Diffusion is the passive movement of the difference in concentration between two
molecules from an area of high areas- has been eliminated. if the
concentration of the species is not
concentration molecules to an area with
initially uniform, over time diffusion will
a lower concentration. Inside cells, cause a mass transfer in favor of a more
diffusion is the transport of small uniform concentration.

molecules across the cell membrane.

Molecules are always in motion.

Temperature, a physical quality people
commonly reference in their daily lives, is
directly related to molecular motion. It is II. MATERIALS
measure of the average of the kinetic Table 1.0: Materials needed for the
energy of the molecules in a material. Diffusion of solute in water
The energy of the molecules causes QUANTITY MATERIALS
JOURNAL OF LABORATORY REPORTS IN BIOCHEMISTRY
NASC 2115 – BIOCHEMISTRY LABORATORY

1 3 pcs. Test tubes 7 1 pack White Sugar

2 1 pc. Test tube rack 8 1 pc. 250 mL Beaker
3 1 pc. Stopwatch 9 500 mL Distilled Water
4 500 mL Distilled Water
5 1 pc. Thermometer
Table 3.0: Materials needed for Dialysis
Electric Stove
6 1 pc.
with Wire Gauze QUANTITY MATERIALS
Cold Distilled 1 4 pcs. Test tubes
7 500mL
Water Iron Clamp &
2 1 pc. Each
Iron Stand
Table 1.1: Materials needed for the 3 1 pc. Rubber band
Diffusion in a Colloid 4 500 mL Distilled Water
5 1 pc. Stopwatch
QUANTITY MATERIALS Longganisa
6 1 ply
1 3 pcs. Test tubes Casing
2 1 pc. Test tube rack Medicine
7 2 pc.
3 1 pc. Stopwatch Dropper
Medicine 8 1 pc. 250 mL Beaker
4 3 pcs.
Dropper Methylene Blue
9 Small amount
5 1 pack Gelatin Mixture Solution
6 1 pc. 500 mL Beaker Saturated
Electric Stove Calcium
7 1 pc. 10 Small amount
with Wire Gauze Chloride
8 500mL Distilled Water Solution
9 1 pc. Ruler 11 Small amount Silver sulphate
10 1 pc. Stirring Rod
Congo Red
11 Small amount III. METHODOLOGY
Solution
Methylene Blue A.1. DIFFUSION OF SOLUTES IN
12 Small amount
Solution
13 Small amount KMnO4 Solution WATER

The apparatuses prepared in this

Table 2.0: Materials needed for
experiment were a series of four test
Osmosis (Longanisa Casing Model)
tubes containing 10 mL of distilled water,
QUANTITY MATERIALS
1 1 pc Test tube tap water, hot distilled water of 70° (or
Iron Clamp & higher) and cold distilled water (on ice)
2 1 pc. each
Iron Stand
3 1 pc. Stopwatch Each test tubes were labeled
Longganisa accordingly. The experimenters, placed
4 2 plies
Casing
5 1 pc. Rubber band a “dot” amount of KMnO 4 crystals then
Triple beam took note of the time at which the crystals
6 1 pc.
balance
JOURNAL OF LABORATORY REPORTS IN BIOCHEMISTRY
NASC 2115 – BIOCHEMISTRY LABORATORY

were completely dissolved in the water
and repeat the determination.
Figure 1.0: Diffusion of solutes in various types
of Water (from left to right) –Hot distilled water,
tap water, cold distilled water, distilled water
and obtained the measurements for 120
minutes at10-minute intervals or until the
dye reaches the bottom of the tube.
Figure 2.0: Diffusion in a Colloid (from left to
right) – Potassium Permanganate [3], Methylene

A.2. DIFFUSION IN A COLLOID

The apparatuses used in this Blue [1], Congo Red [2]

experiment were sets of test tubes, B. OSMOSIS
medicine dropper, hot plate, wire gauze (Longanisa Casing Model)
and stirring rod. In addition, the reagents
Place 10 mL of saturated sugar
used were Potassium Permanganate
solution in a test tube and cover the
(KMnO4), Congo red (C32H22N6Na2O6S2),
mouth of the test tube with 2 plies of
and Methylene Blue (C16H18ClN3S)
Longanisa casing then secure it with a
The experimenters melted the gelatin
rubber band. Next, using the triple-beam
according to manufacturer’s procedures
balance, the experimenters obtained the
and poured 10 mL of the melted gelatin
weight of the covered test tube and
in the test tubes and allowed it solidify.
suspended it upside-down in a 250 mL
Then, overlaid 0.5 mL of the KMnO4 in
beaker containing distilled water using an
one of the gelatin-tube, and add the
iron clamp to fasten the test tube. Lastly,
same amount of Congo Red and
record the results obtained from the
Methylene Blue in a separate gelatin-test
weight of the test tube in a 10-minute
tubes. Next, measure the migration of the
interval for 120 minutes.
dye starting at the top level of the gelatin
JOURNAL OF LABORATORY REPORTS IN BIOCHEMISTRY
NASC 2115 – BIOCHEMISTRY LABORATORY

C.DIALYSIS Diffusion is affected by temperature, area

of interaction, steepness of the
Fill a test tube with 3.0 mL concentration gradient and particle size.
Methylene Blue solution and 5 mL Each of these factors, independently and
collectively can alter the rate and extent
Saturated CaCl2, cover the mouth of the
of diffusion.
test tube with 2 plies of Longanisa casing
and secure the cover with a rubber band. This experiment focuses about how the
temperature and type of the water affects
Then, suspend the test tubes upside- the rate of diffusion.
down in a 250mL beaker containing
distilled water and use an iron clamp to
fasten the test tube. Test the presence of
Methylene Blue, Calcium and Chloride
ion in the beaker-water after every
minute for the total duration of 30
minutes. Identify the presence of
Methylene blue by using visual
inspection, a faint blue color in the
beaker-water indicates the presence of
Methylene blue. To identify the presence Fig. 1.1: Diffusion of solute in different
temperature and condition of water (from
of Chloride, add a drop of Silver sulphate left to right): Hot distilled water, Distilled
test solution to 1-mL aliquot of the water, Tap water, and Cold distilled water.

beaker-water.Finally, tabulate the results

and identify which ion and/or molecule
passed through the longanisa skin first.
VI. Result and Discussion
1.1 Diffusion of solutes in water
Diffusion is the movement of a substance
from an area of high concentration to
an area of low concentration. Diffusion
happens in liquids and gases because
their particles move randomly from place
to place.
The experimenters used KMNO4
(Potassium Permanganate) as the solute
and H2O (Water) as the solvent. Four test
tubes were provided and each of the test
tubes was filled with 10ml of different
types of water. These are the cold
distilled water, hot distilled water, tap
water, and distilled water. The
experimenters then put a dot size amount
of the KMNO4 in each test tube to
determine the diffusion rate from fastest
to slowest.
JOURNAL OF LABORATORY REPORTS IN BIOCHEMISTRY
NASC 2115 – BIOCHEMISTRY LABORATORY
The experimenters did two trials of the
experiment because the first trial failed to
get the expected results. The following
results gathered in the first trial are the
following: For Hot distilled water the
solute took 48.14 seconds to diffuse. For
Distilled water the solute took 122
seconds to diffuse. For Tap water the
solute took 23.04 seconds to diffuse. For
Cold distilled water the solute took 132
seconds to diffuse. The experimenters
had a human error when they failed to
turn off the timer accurately after the
solute already diffused that resulted to
inaccuracy of results. However the
experimenters got the results in the cold
distilled water right thus they didn’t redo
it on the next trial.
The second trial results show that when
the KMNO4 (Potassium Permanganate)
was added in a hot distilled water the
solute diffused after 36.26 seconds. In
the second test tube when the KMNO4
(Potassium Permanganate) was added
in a distilled water the solute diffused
after 60.14 seconds. In the third test tube
when the KMNO4 (Potassium
Permanganate) was added in a tap water
the solute diffused after 44.02 seconds.
Lastly in the fourth test tube that has cold
distilled water the result of 132 seconds
remained.
With increase in temperature, the kinetic
energy of all particles in the system
increases. This increases the rate at
which solute and solvent molecules
move, and increases collisions. From the
results of the second trial this means that
it’s accurate that the Potassium
Permanganate diffused faster when put
in hot water mainly because its
molecules moved with greater energy
than in cold water.
However, coming from the results of the
experiment, it is found that two of the
results are inconsistent to the expected
rate of diffusion. Between the tap and the
distilled water, the Potassium
Permanganate should diffuse faster in
the distilled water because it is more pure
and has lesser impurities than the tap
water which contains more impurities that
impedes diffusion. For an instance if a
pinch of salt is added to water it will make
the diffusion slower because it will
contain other molecules which will
impedes diffusion. The experimenters in
the second trial failed to observe that the
distilled water diffused faster than the tap
water because the amount of Potassium
Permanganate put within the two test
tubes is not the same hence it caused
confusion in observing the results.
1.2 Diffusion in Colloid
Diffusion in colloid is slower than in
water. It is for a reason that they have
different states where in gelatin is solid
while water is liquid.
Gels are networks of cross-linked
molecules in water or other liquids. The
particles of gelatin move slower hence
they collide less frequently meaning that
they spread less and restricts diffusion.
JOURNAL OF LABORATORY REPORTS IN BIOCHEMISTRY
NASC 2115 – BIOCHEMISTRY LABORATORY
Fig. 1.2: Diffusion in Colloid (from left to
right): KMnO4 (Potassium Permanganate),
Methylene Blue, and Congo Red
The experimenters poured 10ml of
melted gelatin in each of the 3 test tubes
and waited a while for it to solidify. After
the gelatin is ready the KMnO4
(Potassium Permanganate), Methylene
blue, and Congo red were added in each
test tube with the amount of 0.5ml. The
experimenters proceeded in observing
the rate of diffusion in each solute and
collecting its measurements for every 10
minutes in a span of 2 hours.
In the first 10 minutes the diffusion of
KMnO4 measures 30 mm, while the
Methylene blue measures 28mm, and
the Congo red measures 38mm. On the
20 minute period it is observed that the
measurement of KMnO4 and Methylene
blue added 1mm while the Congo red
added 4mm in its measurement. The
KMnO4 maintained its measurement of
31mm until reaching the 100minutes
mark. Methylene blue on the other hand
maintained its measurement of 29mm
until its 60minutes mark. However,
Congo red’s measurement of 42mm
started from the 20minutes mark
remained only until reaching the 40
minutes mark. After finishing the 120
minutes the measurement of KMnO4 only
reached 34mm, while the Methylene blue
which initially diffused slower reached
42mm. Lastly, the Congo Red which
initially diffused a little bit faster than the
other two solutes reached until 60mm or
almost the bottom of the test tube at the
end of the 120 minutes.
Upon the data measurements collected,
it is observed that Congo red travelled
the farthest distance in a colloid followed
by the Methylene blue and the last is
KMnO4 (Potassium Permanganate).
The difference in diffusion rates of the
solutes varies in the size of their
molecules. Diffusion is faster for small
molecules than the larger ones.
However, the data gathered in the
experiment is not congruent to the
expected results. The differences in time
of pouring the gelatin in the test tube and
the inconsistency of its solidification are
the factors that affect the results of the
experiment.
2. Osmosis
Osmosis is a special example of
diffusion. It is the diffusion of a substance
through a semipermeable membrane
from a more dilute solution to a more
concentrated solution. A semipermeable
membrane is a barrier that permits the
passage of some substances but not
others.
JOURNAL OF LABORATORY REPORTS IN BIOCHEMISTRY
NASC 2115 – BIOCHEMISTRY LABORATORY

The osmotic pressure of a solution is the The experimenters obtained the weight
pressure difference needed to stop the of the test tube every 10 minutes in a
flow of solvent across a semipermeable span of 2 hours. The results shown as
membrane. The osmotic pressure of a follows:
solution is proportional to the molar
concentration of the solute particles in
solution. The more solute added to a Time in minutes Weight in grams
solvent, the higher the osmotic pressure.
10 31g

20 32g

30 32.5g

40 33g

50 33.5g

60 34g

70 34.5g

80 34.8g

90 35.1g

100 35.5g

110 35.5g

Fig. 2: Suspended test tube with saturated
sugar upside down in a beaker
In the experiment the longanisa casing
acted as the semipermiable membrane
to separate the two solutions. The high
concentrated solution is the saturated
sugar, while the low concentrated
solution is the distilled water. The test
tube with a 10ml saturated sugar was
covered with a 2 ply longanisa skin while
secured with a rubber band. It is then
suspended with a clamp in an upside
down setup in a beaker filled with 250ml
water.
120 35.7g
The results only show that the weight of
the test tube with the high concentrated
solution becomes heavier every 10
minutes within 2 hours which followed the
process of osmosis. The data gathered is
congruent to the expected results of the
experiment therefore it is considered as
a success.
2. Dialysis
JOURNAL OF LABORATORY REPORTS IN BIOCHEMISTRY
NASC 2115 – BIOCHEMISTRY LABORATORY

Fig. 3: Weight of covered test tube with 6 ✓ X

saturated sugar solution in grams for every 7 ✓ X
10-minutes interval for 120 minutes 8 ✓ X
9 ✓ X
10 ✓ X
11 ✓ X
12 ✓ X
13 ✓ X
14 ✓ X
15 ✓ X
16 ✓ X
17 ✓ X
18 ✓ X
19 ✓ X
20 ✓ Small
amount of
white color
21 ✓ Small
amount of
white color
22 ✓ Small
amount of
white color
23 Darker Small
Bluish amount of
white color
24 Darker Small
Bluish amount of
white color
25 Darker Large
Bluish amount of
white color
TABLE 1.0: Test for the presence of Methylene 26 Darker Large
blue (C16H18ClN3S) and Chloride in the breaker-
Bluish amount of
white color
water after every minute for thirty (30) minutes
27 Darker Large
PRESENCE
Bluish amount of
TIME OF PRESENCE white color
(MINUTES) METHYLENE OF CHLORIDE 28 Darker Large
BLUE Bluish amount of
1 ✓ X white color
2 ✓ X 29 Bluish Large
3 ✓ X amount of
4 ✓ X white color
5 ✓ X 30 Bluish Faint White
Precipitate
JOURNAL OF LABORATORY REPORTS IN BIOCHEMISTRY
NASC 2115 – BIOCHEMISTRY LABORATORY
Dialysis is a classic laboratory technique
that relies on selective diffusion of
molecules across a semi-permeable
membrane to separate molecules based
on size. Dialysis is used for a wide variety
of applications: desalting, removal of
labeling reagents, drug binding studies,
buffer exchange, cell growth and feeding,
virus purification, and blood treatment.
Typically, a sample and a buffer solution
(called the dialysate) are placed on
opposite sides of a dialysis membrane
which contains pores of a manufactured
size-range. Sample molecules that are
larger than the pores are retained on the
sample side of the membrane, but small
molecules pass through the membrane,
reducing the concentration of those
molecules in the sample. Alternatively,
desired components in the external
buffer solution can be slowly brought into
the sample.
The test tube was filled with 1.0 mL
Methylene blue (C16H18ClN3S) and 1.7
mL saturated Calcium chloride (CaCl2),
respectively.
It was monitored for a period of
thirty (30) minutes, and was tested for the
presence of Methylene blue and Chloride
ion for every passing minute. By the use
of visual inspection the presence of
Methylene blue was noted after the first
minute. It was indicated by the change in
color of the breaker-water from crystal
clear to a faint blue color, as the time
goes by, the color of the Methylene blue
became more obvious as gradient of the
solution turn into a much darker shade of
blue.
In the case of confirming the
presence of Chloride ion in the breaker-
water; an aliquot was taken then a few
drops of Silver nitrate (AgNO3) was
added. In the first twenty (20) minutes of
the experiment, after the Silver nitrate
was added; the only transformation
transpired was the color change. The
water in the aliquot became blurry white.
The emergence of white precipitate was
first observed after twenty five (25)
minutes were elapse indicating the
presence of Chloride. The succeeding
minutes made the white precipitate more
perceptible to the naked eye, and blurry
white color of the mixture still continues
to darken, thus making it much
recognizable than earlier.
JOURNAL OF LABORATORY REPORTS IN BIOCHEMISTRY
NASC 2115 – BIOCHEMISTRY LABORATORY
VII. CONCLUSION
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.
VIII. RECOMMENDATIONS
. The researchers then
recommend the repetition of the
procedure for the diffusion of a colloid
make sure that the gelatin is fully
disintegrated to not lead inconsistencies
in the results, where potassium
permanganate should be the fastest to
diffuse followed by Congo red and to
come in last by Methylene blue.
REFERENCES:

https://www.bbc.com/bitesize/articles/znqbcj6

https://biologydictionary.net/diffusion/
https://prezi.com/8vm9vq304bim/exercise-4-cell-transport/
https://www.biophysics.org/Portals/0/BPSAssets/Education/Documents/LessonPlanDiffu
sion_122115sm.pdf
http://leavingbio.net/osmosis-diffusion/
https://chem.libretexts.org/Bookshelves/Physical_and_Theoretical_Chemistry_Textbook_Maps/
Supplemental_Modules_(Physical_and_Theoretical_Chemistry)/Physical_Properties_of_Matter/
Solutions_and_Mixtures/Colligative_Properties/Osmotic_Pressure

https://www.physiologyweb.com/lecture_notes/membrane_transport/membrane_transpo
rt_processes_summary.html

https://www.scribd.com/presentation/99900245/Exercise-4-Metho
https://www.researchgate.net/publication/305493284_Diffusion_Dialysis_and_Osmosis

CONTRIBUTION/S: SIGNATURE

Abstract and Introduction – Maravilla, Alexandra Louise M. ______________

Mercado, Antonette B. _______________

Manabat, Mariah Joy L. _______________

Materials and Methodology – Ombrero, Karen O. ________________

Results and Discussion- Nunez, Gianne Kianna A. ________________

Nadonga, Nicole Angelyn T. _______________

Conclusion and Recommendation – Niez, Kimberly S. ________________

JOURNAL OF LABORATORY REPORTS IN BIOCHEMISTRY
NASC 2115 – BIOCHEMISTRY LABORATORY