Name: JULIE ANN T. FULLANTE Date Performed: NOV.

9, 2020
ST
Year and Course: 1 YEAR: BSN Date Submitted: NOV. 10, 2020
Subject title: BIOCHEMISTRY__________ Code number: 1319
Name of Professor: MR. JOHN MICHAEL G. BALABA 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. Observe the process of diffusion in gelatin and water bath.

2. Test for the presence of substances albumin, NaCl and glucose in the dialyzing.
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

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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.

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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.

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V. RESULTS AND OBSERVATION

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

As I drop the dye on the surface of the colloid gelatin and leave it about 24 hours, I observe that the dye diffuse. The region of high
concentration where the molecules were highly concentrated and the area around that would be the area of low concentration where the
dye needed to diffuse would be as I produced to drop the dye into so it is to demonstrate that because the molecules in the blue dye were
much smaller than the molecules in the red dye they diffused at a faster rand and over a greater area compare to red dye.

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

These molecules, can travel through the gel in different directions or at differing speeds, allowing them to be separated from each
other. The dye molecules will move freely throughout the gel and they scatter.

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

Random motions of the molecules of dye and gel cause them to crash into one another and mix. The dye molecules then shift from an
area of greater concentration to a region of lower concentration. Eventually, in the solution, they are uniformly distributed. This indicates a
dynamic equilibrium has been achieved by the molecules. This process 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?

Like potassium permanganate, compounds shift from areas of higher concentration to areas of lower concentration as it diffuses
faster in purified water, which happens to have a lower concentration than tap water. The theory of chemical diffusion using potassium
permanganate in water is seen in this experiment. The purple solution spreads steadily across the beaker into the rest of the bath, producing
a less concentrated but uniformly colored purple solution.
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?

As what I observe in the result in the step 2 and 3, Diffusion times are temperature-dependent; the diffusion occurs rapidly in warm
agarose (or gelatin or Jello) they are faster than room temperature plates, and in cold plates they are slower.

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 and NaCl can diffused through the dialyzing membrane due to its smaller molecular weight solutes and
are tiny molecules which can move around easily.

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

In this experiment the colloidal solute albumin does not diffuse because it contains molecules of large molecular weight.

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

As I observed. Thus, smaller molecule the faster the net movement or they quicker diffuse. While the bigger molecule the slowly the
net diffusion from a region of higher concentration to lower concentration down the concentration gradient. The smaller molecule can
quickly travel throughout the surface area from an area of higher concentration to an area of lower concentration vice versa.

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

The different factors that cause to happen are affect that happening are: first, the TEMPERATURE and PRESSURE, increasing the
temperature or pressure increases the particles' kinetic energy, thereby increasing the diffusion rate. Second the SIZE and MOLECULAR
WEIGHT of the molecules, the smaller molecule the quicker. Lastly the SURFACE AREA, the greater the region over which diffusion will
occur, the higher the diffusion rate. These are some factors that affect the movement of materials into the cell through the cell membrane.
VI. CONCLUSION

It can be inferred that the rate of diffusion of a material is influenced by its concentration and molecular weight, based on the
outcome of the experiment. Like potassium permanganate, compounds shift from areas of higher concentration to areas of lower
concentration as it diffuses faster in purified water, which happens to have a lower concentration than tap water. A substance with a
lower molecular weight such as potassium permanganate has a higher diffusion rate when it comes to the molecular weight of substances
compared to a substance with a higher molecular weight like congo red. On the other hand, relative to potassium permanganate,
methylene blue, which happens to have a molecular weight of 319.85 g / mol, has a slightly lower rate of diffusion but faster than congo
red. As I observe, compared to gelatin, which is a colloid, the rate of diffusion in water is faster because water has more rooms to allow
the spread of molecules compared to colloids where space is small, which also restricts the movement of molecules, so the rate of
diffusion is slow. Diffusion is when molecules migrate from a region of high concentration to an area of low concentration, and the
various factors that allow it to occur influence the temperature of the molecules' size and weight and the distance that the molecules
have to disperse when the experiment happens. The diffusion of substances with smaller molecules happens faster in dialysis, which
refers to the diffusion of solutes through a selectively permeable membrane, much like what happened to chloride ions that began to
appear ahead of the calcium ions.
LIBRARY WORK:
 Define diffusion, colloid, net diffusion and dialysis

The transfer of a material from a region of high concentration to an area of low concentration is diffusion. In liquids and gases,
diffusion occurs because their particles pass spontaneously from place to place. For living organisms, diffusion is an essential process;
it's how substances pass in and out of cells. Moreover, colloid is a mixture in which very small particles of one substance are uniformly
distributed over another substance. In general, the particles are larger than the ones in a solution and smaller than the ones in a
suspension. Net diffusion is when the flow in one direction of the solvent particle is greater than the flow in the opposite direction of
the solvent particle. Furthermore, dialysis Isolation of suspended colloidal particles from dissolved ions or small-dimensional
(crystalloid) molecules by means of their unequal diffusion rates through semi-permeable membrane pores.

 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 Title Methods Results
published
 Author(s) Year Title Methods Results
published
A. Diffusion in water A. In Water Diffusion
In the test tube, half-filled with distilled
Two test tubes, one half- water, the KMnO4 crystals diffused more
filled with tap water and the othereasily. The diffusion was sluggish in the test
half-filled with distilled water, tube, half-filled with tap water, distinguished
were collected. The two test tubes by the development of a nebula-like structure
were then correctly numbered. suspended (it eventually disappeared) in the
Asaad, Two crystals of potassium center of the test tube. In the test tube with
Baldovino, permanganate (KMnO4) crystals dH2O, the nebula-like structure was not
Benuyo, were lowered using a pair of observed because the purple color rapidly
Dee, forceps on each of the two test dispersed across the entire tube.
Evangelista, 2014 Diffusion, tubes. In order to avoid any
Garcia, Dialysis and disruption, the test tubes were B. Diffusion in Colloid
Vicuña, Osmosis then placed on a shelf. The Results are shown through in a tabular
dissolution in each test tube of the format:
potassium permanganate crystals
was then observed. The dispersal Table 4.1 Distance travelled by Potassium
of a purplish, light color suggests permanganate, Methylene Blue and Congo
this breakup. The period was Red.
recorded when the color had Time Distance Travelled (cm)
completely dispersed in the water (min.) KMnO4 Methyle Congo
in both test tubes. ne Blue Red
5 1.0 0.3 0.2
10 1.4 0.5 0.3
15 1.5 0.6 0.3
 20 1.6 0.7 0.3
25 1.9 0.8 0.3
B. Diffusion in Colloid 30 2.1 0.9 0.4
35 2.1 0.9 0.4
Three test tubes were
collected, filled with a hardened 40 2.2 0.9 0.4
5% gelatin solution. The three On the basis of the above results, it can
test tubes were then applied be said that KMnO4 traveled the farthest
simultaneously with 1 mL of each distance, followed by blue methylene, and
of the following reagents: (1) finally red congo.
permanganate potassium, (2)
blue methylene, and (3) red C. Dialysis
congo. It reported the time that it Table 4.2 Results of Dialysis of Methylene
took for the stains to hit the Blue, Calcium and Chloride Ions
bottom of each test tube. The Time Chlori Calciu Methyle
distance traveled by each stain (min) de m Test ne Blue
through the gelatin was recorded Test
at five-minute intervals. For one 1.5
and a half hours, the same 2.5 ✔
procedure was performed. 3.5 ✔
4.5
C. Dialysis
5.5 ✔
There was a mixture of 3 6.5
mL of calcium chloride (CaCl2) After 2.5 minutes, the cloudy precipitate
solution and 2 mL of methylene started to emerge for the Chloride test, while it
blue in a small test tube. Then a took 5.5 minutes for the calcium test.
dried longganisa skin and a Methylene after 3.5 minutes. Through the
rubber band secured the mouth of beaker, blue spread.
the test tube. It made sure that the
seal did not have any leaks. In a E. Osmosis in Red Blood Cell
beaker filled with about 30 mL of Table 4.3 Size of Red blood Cells under
distilled water, half of the test different concentrations of NaCl
tube was then immersed, with the NaCl Concentration (M)
sealed mouth facing down. Two Time 0.07 M 0.15 M 0.30 M
separate test tubes were put in one (min.) Cell size in um
drop of it. The first test tube was 5 2 3 3
then inserted with a drop of silver 10 2 3 3
nitrate, while the second test tube 15 4 3 3
was added with a drop of 20 4 3 3
ammonium oxalate. This were 25 4 3 3
done to measure the presence of 30 4 3 3
chloride and calcium ions, 35 5 3 3
respectively, which a white,
40 4 3 4
cloudy precipitate would suggest.
Until the precipitate was clear in Based on the acquired data above, it can
both test tubes, the same process be said that under a NaCl concentration of 0.07
was repeated. In both test tubes, M, the size of the RBC increased, under the
the time when the precipitate NaCl concentration of 0.15 M, the size of the
became visible was registered. It RBC remained constant, whereas under the
was also observed if the blue NaCl concentration of 0.30 M, the size of the
methylene would disperse RBC increased.
through the beaker.
 Author(s) Year published Title Methods Results
In mouse and biophysical In silico modeling
studies combined with and in vivo research,
mathematical modeling, the deletion of aquaporin-1
study used experimental did not affect the
models of peritoneal icodextrin-induced
Johann Morelle, dialysis to evaluate the osmotic water transport,
Amadou Sow, mechanisms of colloid but did affect the
Charles-André Fustin, versus crystalloid osmosis crystalloid agent-induced
Catherine Fillée Mechanisms of Crystalloid versus across the peritoneal deletion. Icodextrin-
Elvia Garcia-Lopez, 2018 Colloid Osmosis across the membrane and to induced water flow was
Bengt Lindholm, Peritoneal Membrane investigate the water flow based on the presence of
Eric Goffin, mediating pathways large, colloidal fractions,
Fréderic Vandemaele, produced by the glucose with a close to unity
Bengt Rippe, polymer icodextrin. reflection coefficient, a
Carl M. Öberg low capacity for
Olivier Devuyst diffusion, and a minimal
impact on dialysate
osmolality. Strikingly
improved water and
sodium transport across
the peritoneal membrane
by combining crystalloid
and colloid osmotic
agents in the same
dialysis solution,
enhancing ultrafiltration
performance over that
obtained with either form
of agent alone.