Name: Sherin M.

Blancada Date Performed: August 27, 2022

Year and Course: BS Nursing 1 Date Submitted: September 1, 2022
Subject title: Biochemistry Code number: 4977
Name of Professor: Zandra Mae Liston Score: ______________________

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. To illustrate and define net diffusion, dialysis, diffusion, and

colloid.

2. To establish a relationship between molecular weight and the

rate of diffusion through a cell membrane.

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

A cloudy turbid solution or precipitate appearing in the solution indicates the

presence of 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.

When a few drops of silver nitrate solution are added to a slightly acidic aqueous
solution containing chloride ions, a white precipitate of silver chloride occurs.

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.

As glucose are present, the solution hue turns reddish, and a yellow precipitate

builds over time. During heat process, lower glucose concentrations produce a red
to yellow precipitate.

V. RESULTS AND OBSERVATION

1. Did the dye diffuse through the colloid gelatin, in its gel state? Why?

The dye dispersed into the colloid gelatin, as we observed in this experiment. This
is due to the dye's ability to flow through the gelatin membrane from a high
concentration area to a low concentration area, or in other words, diffusion as the
dye spreads throughout the gelatin solution.

2. In which direction did the dye move through the gel?

The dye moves through the gel in an outward direction, allowing the dye to spread in the
gel.

3. What causes the movement of the dye through the gel? What do you call this
process?

Diffusion of particles or molecules occurs when heat, or thermal energy, excites

the particles or molecules and causes them to move. The cause of the movement
of the dye through the gel is because the gel is mainly composed of water and the
molecules of the dye move freely and randomly because of the water content.
Each molecule moves in a random direction, meaning that there will be a net
movement of dye molecules from areas of high concentration (the central band)
into areas of low concentration (the agar). This process is called Diffusion.
4. Did the result you observed in the 3 under the collection of data indicate that
potassium permanganate diffused through the water? Why?

Yes, because of the unequal mobility of the potassium permanganate particles, a

dense purple solution appears in the water at the bottom of the beaker. The
remaining water in the beaker will gradually absorb the purple solution, resulting
in a less dense but uniformly colored purple solution. The amount of potassium
permanganate will be relatively same since the potassium permanganate will
disperse throughout the liquid.

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?

Water has more spaces to allow the spread of molecules than gelatin, which is a
colloid, therefore the rate of diffusion is higher in water. Gel has limited space,
which also restricts the movement of molecules, therefore the rate of diffusion is
slower in gel.

6. Did the result in step 7, 8 and 9 indicate that the crystalloids glucose and NaCl
diffused through the dialyzing membrane? Why?

Yes. Both solutions demonstrate precipitation, according to the results. Because

of their lesser mass, smaller molecules can diffuse faster than larger molecules.

7. Did the colloidal solute albumin diffuse through the membrane in this
experiment?

No, the colloidal solute albumin was unable to diffuse across the membrane in
this experiment because it is a large molecule that cannot pass through the
dialyzing membrane.

8. From the experiments you performed in steps 2 and 3, explain the net diffusion of
solutes to its concentration.

The observations of the experiments in steps 2 and 3 demonstrate the relationship

between molecular weight and diffusion rate. Potassium permanganate, which has
a lower molecular weight than methylene blue, diffused faster into the gel
membrane than methylene blue. To sum up, a smaller molecular weight leads in a
faster diffusion rate.

9. Give at least three (3) factors that affect the movement of materials into the cell
through the cell membrane.

1) Membrane Thickness - For nutrients to diffuse into a cell they must traverse
the cell membrane. Cell membranes are extremely thin to allow for the
diffusion of materials across the cell membrane.
 2) Concentration Gradient - The greater the concentration gradient (the
difference in concentration either side of the membrane) the greater the rate of
diffusion.

3) Temperature and Pressure - As with any chemical reaction, increasing the

temperature or pressure increases the kinetic energy of the particles, thus
increasing the rate of diffusion.

VI. CONCLUSION

We can conclude from these experiments that the rate of diffusion of a substance
is affected by its concentration and molecular weight. Substances travel from
areas of higher concentration to areas of lower concentration, as potassium
permanganate did in water and methylene blue did in gelatin. Smaller molecules
diffuse quicker than larger molecules in dialysis, which refers to the diffusion of
solutes through a selectively permeable membrane. We observed  that the
concentration of solute particles has a clear effect on the rate of diffusion. In
addition, it has been determined that dialysis tubing does not allow all sorts of
substances to pass easily through the pores of its membrane. This indicates that it
is selective in its permeability to chemicals, which explains why albumin and
glucose could pass through dialysis tubing but not NaCl or glucose. We
discovered that a thermal temperature, can have a significant impact on how fast
the particles are moving throughout chemical reactions. Particles in a solid, liquid,
or gas move more rapidly as its temperature increases. The particles slow down as
the temperature drops. When a liquid is sufficiently cooled, it turns into a solid.

LIBRARY WORK:
● Define diffusion, colloid, net diffusion and dialysis

 Diffusion - The movement of molecules over a concentration gradient is referred

to as diffusion. It's a crucial phase that all living things go through. The
movement of chemicals into and out of cells is aided by diffusion. The molecules
shift from a higher to a lower concentration region until the concentration is
equalized.

 Colloid - A colloid is a heterogeneous combination of particles that are halfway

between those of a solution and those of a suspension. The particles are evenly
disseminated across the dispersion medium, which might be a solid, liquid, or gas.
A colloid's scattered particles are not as massive as those in a suspension,
therefore they do not settle out when left standing.
  Net diffusion - When the flow of solute particles going in one direction is greater
than the flow of solute particles moving in the other direction, this is referred to as
net diffusion.

 Dialysis - Dialysis is a separation method that uses selective and passive diffusion
via a semi-permeable membrane to remove tiny, undesirable chemicals from
macromolecules in solution.

● In an A4 bond paper research at least two (2) journals / articles about

diffusion, colloid, and dialysis by following the format (Times New Roman
font 13, landscape)
Author(s) Year published Title Methods Results
J Am Soc 2018 Mechanis They used In silico modeling
Nephrol ms of experimental and in vivo studies
Crystalloid models of showed that deletion
versus peritoneal dialysis of aquaporin-1 did
Colloid in mouse and not influence osmotic
Osmosis biophysical water transport
across the studies combined induced by icodextrin
Peritoneal with but did affect that
Membrane mathematical induced by
modeling to crystalloid agents.
evaluate the Water flow induced
mechanisms of by icodextrin was
colloid versus dependent upon the
crystalloid presence of large,
osmosis across colloidal fractions,
the peritoneal with a reflection
membrane and to coefficient close to
investigate the unity, a low diffusion
pathways capacity, and a
mediating water minimal effect on
flow generated by dialysate osmolality.
the glucose Combining
polymer crystalloid and
icodextrin colloid osmotic
agents in the same
dialysis solution
strikingly enhanced
water and sodium
 transport across the
peritoneal membrane,
improving
ultrafiltration
efficiency over that
obtained with either
type of agent alone.
Flynn MT, 2012 The The Forward The project was
Soler MP, Forward Osmosis Bag administered by the
Shull S, Osmosis (FOB) Kenya Water for
Broyan, Jr. Bag (FOB) investigation uses Health Organization
JL, investigati the osmotic (KWAHO) and was
Chambliss on pressure gradient closely observed by
JP, Howe across the UNICEF, USAF, and
AS, Gormly membrane to the Kenya Water
S, produce a total Minister. The results
Hammoudeh flow of water so far have been
M, Shaw H, through the outstanding. The
Howard K. membrane into people of Mudimbia
Forward the draw solution. found the HydroPack
Osmosis Driven by an had a great flavor and
Cargo osmotic pressure was easy to use. The
gradient, forward lab results showed
osmosis does not that hydration levels
require direct improved through the
energy input. test period and that
Forward osmosis there was no
is a low resource contamination in the
water treatment produced drink.
technology that
offers the
advantage of high
rejection of a
wide range of
contaminants than
traditional
membrane
processes.
 DOCUMENTATION

Preparation of Substances, Materials and Equipment

Movement
of dye through the
gel
Diffusion of KMnO4
 \

a. Test for albumin b. Test for presence of NACI c. Test for the presence of glucose

The dialyzing the membrane