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Exercise no. 1
Movement of Substance through Cell Membrane
Experiment Demonstrating Diffusion
I. INTRODUCTION
If we put a teaspoon of instant ice tea on the surface of a glass of water, the
molecules soon spread throughout the solution. The molecules of both the solute (ice tea)
and the solvent (water) are propelled by random molecular motion. The initially
concentrated tea becomes more and more dilute. This process of the net movement of a
solute with the gradient (from an area of high concentration to an area of low
concentration) is called diffusion.
If the solute can pass through the membrane, diffusion will occur with net
transport of material from the region of initial high concentration to the region of initial
low concentration, and substance will equilibrate across the cell membrane. After a
while, the concentration of the substance will be the same on both sides of the membrane;
the system will be at equilibrium, and no more net change will occur.
II. OBJECTIVES
At the end of the experiment, each student will be able to:
1. Identify the process and understand the molecular basis of
diffusion.
2. Observe the process of diffusion and determine its significance.
III. MATERIALS:
2 – 250ml beaker, 1-500ml beaker, 1-1000ml beaker, 5 test tubes, test tube
rack, test tube brush, test tube holder, 3 x10 cellophane bag, 1-rubber
band, iron ring, 3 pipette, 3-medicine dropper, water bath, electric stove,
1-25ml graduated cylinder, 1m string, ruler
5g powder gelatin, 1ml methylene blue, 2g KMnO4, 2 g NaCl, 5% glucose
soln, 2g albumin powder, 2ml Nitric acid, 2ml silver nitrate, 2ml
Benedict’s soln.
IV. PROCEDURE:
Movement of dye through the gel
1. Preliminary preparation (a day or two before the laboratory period): add 5
gram (or 1 tsp) of gelatin to 25ml of cold water. Let stand for 5 minutes. Add 75
ml of boiling water and stir until dissolved. Pour about 15 ml of the solution into a
test tube. Fill the remaining solution to another test tube refrigerate until gelled.
2. Place a drop of methylene blue on the surface of the gel prepared ahead of time.
Set aside at room temperature. At the end of 1 hour, 2 hours, 6 hours,12 hours and
24 hours, observe whether the dye has move and if so in which direction and how
far.
Diffusion of KMnO4
3. Place a few crystal of potassium permanganate on the bottom of a beaker half
filled with water. Observe.
The dialyzing membrane
4. Prepare a bag of 3 x 10 inch of cellophane
5. Prepare a solution as follows in a large beaker (500ml): to 100ml of water add ½
tsp of NaCI plus 50ml of 5% glucose, and the uncooked albumin and place them
inside the cellophane bag. Both NaCI and glucose are crystalloids or true solutes,
whereas albumin is a colloidal solute. ( Better look at these terms up if you are not
sure what they mean)
6. Tie a string around the top of the bag and suspend it in a 1000ml beaker with
distilled water. You may use an iron ring and stand for this set-up. If the
cellophane bag is very thick you may pinch a very small hole in the bag and
suspend it in the distilled water. Let it stand for about an hour and then the test
indicated in steps a, b, and c.
a. Test for albumin: Using a pipette, pour about 5 ml of the fluid into the test tube.
Add a few drops of nitric acid. Note whether coagulation occurs. Nitric acid
coagulates albumin
In the test, the albumin is present. Testing of nitric acid and a urine sample made a
reaction in which it shows white ring that indicates the presence of the albumin in
the sample.
b. Test for the presence of NaCI: Using a pipette, pour about 5ml of solution into a
test tube and add a drop of silver nitrate. Note whether a precipitate forms. If
NaCI is present it will combine with silver nitrate to from a precipitate of silver
chloride.
During the test, a white particle or a white ring was visible on the solution which
indicates that the presence of the NaCl is visible. The silver nitrate solution is the
key to the chloride mixture.
c. Test for the presence of glucose: Using a pipette, pour about 5ml of solution into
a test tube and put 5ml of Benedict’s solution. Water bath the tube for 2 minutes
and let it cool slowly Note: whether a green, yellow, or red precipitate forms
indicates the presence of glucose/sugar.
The presence of glucose is positively visible. The change of the tint of the original
color which is aqua-blue turns into yellow because of the presence of the heat that
reduces the glucose or the sugar. The higher the concentration of heat shows the
decreasing number of sugar available.
V. RESULTS AND OBSERVATION
1. Did the dye diffuse through the colloid gelatin, in its gel state? Why?
Yes, the dye diffused because within the time period of observation. Because of
the random movement of the molecules that made diffusion take place.
2. In which direction did the dye move through the gel?
The dye slowly spread out from their position in the gel into the part where it has
low concentration.
3. What causes the movement of the dye through the gel? What do you call this
process?
The movement of the dye was because of the diffusion process in which the high
concentration part or the gel part moves to an area with low concentration which
is an area with less to no gel.
4. Did the result you observed in the 3 under the collection of data indicate that
potassium permanganate diffused through the water? Why?
Potassium permanganate diffused through the water faster than the other fluid
substance because its small molecule is soluble through the water and the room
temperature contributed on the progress of the diffusion. The higher the
temperature, the faster it diffuses.
5. Based on the result you observed in the step 2 and 3 do you postulate that
diffusion occurs rapidly through the gel, or a liquid, or both at the same rate?
Based on the result, the diffusion of a substances occurs rapidly depending on its
temperature, in other word, the gel or liquid – specifically water – are both at the
same rate.
6. Did the result in step 7, 8 and 9 indicate that the crystalloids glucose and NaCl
diffused through the dialyzing membrane? Why?
The crystalloids glucose did diffuse because it shows changes in color and its
small size can easily diffuse across the membrane. The solution process shows
that the substances did not dissolve. While the NaCl diffused because it has
smaller molecular weight glucose and it shows changes in color when it is mix in
silver nitrate and chloride ions.
7. Did the colloidal solute albumin diffuse through the membrane in this
experiment?
Yes, the colloidal solute albumin diffuses and can pass through the membrane
because its liquid substance can be mixed and shows a reaction.
8. From the experiments you performed in steps 2 and 3, explain the net diffusion of
solutes to its concentration.
The smaller the molecular weight the fastest it diffused. The process happened in
the experiments was that it started from a higher concentration to its low to no
concentration region.
9. Give at least three (3) factors that affect the movement of materials into the cell
through the cell membrane.
One factor that affect the movement of materials is the range of the concentration
gradient. The difference in concentration can affect the speed of the diffusion. The
higher the concentration the more rapid the diffusion. The temperature can also
affect the motion of the materials because the higher the temperature the faster it
will escalate the energy, therefore, the movements of the molecules speed up its
rate of diffusing. The mass of the molecule is also a factor for it can affect the
molecules rate in diffusing. The smaller the molecular weight, the faster it will
diffuse.
VI. CONCLUSION
In the overall review of diffusion, the simple diffusion is that the movement of particles
occurs along the direction of the concentration gradient. It is facilitated diffusion that the
movement of molecules can occur both in direction and it can be opposite of the
concentration gradient. And it can be the transport of substances across a biological
membrane from an area of higher concentration to a lower concentration with the help of
a transport molecule. Diffusion is defined by biology as the passive movement of
substances, such as biological molecules or ions.
LIBRARY WORK:
Define diffusion, colloid, net diffusion and dialysis
Diffusion- is when a molecule moves from a region of higher concentration to a region
of lower concentration.
Colloid - is when a certain substance is dispersed into another substance and its small
particles cannot be simply separated than those of suspension. Example of this are paint,
milk, and fog.
Net diffusion- is a particle of a solute in which the flow in one direction is bigger than the
particles of other solutes that travel in the opposite direction.
Dialysis- commonly done to people whose kidneys cannot naturally function well. This is
the process of removing toxins, solutes, and excess water in the blood.
