Pamantasan ng Lungsod ng Maynila

College of Nursing Department

1st Semester A.Y. 2020 – 2021

A Group Laboratory Report

In Partial Fulfillment of the Requirements
In Biochemistry (Laboratory)

  
Presented to:
Prof. Ma. Joanna A. Astorga

Submitted by:
Alkayde, Gian Robert
Labang, Daniela Mae
Mabulac, Ellen Mynelle C.

BSN 1-2
November 13, 2020
 ACTIVITY NO. 2
Experiment no. 1: Diffusion Rate
“Chemical Diffusion in Agar”
I. OBJECTIVES OF THE STUDY
 To investigate diffusion as it applies to movement of particles in a semi-solid material
called agar.

II. MATERIALS USED

 2 Pieces of Test Tube with Agar
 Pencil
 Eyedropper (as many as possible)
 Methylene Blue
 Timer
 Potassium Permanganate

III. FLOWCHART/DIAGRAM
 Obtain two test tubes with agar
 Mark the test tubes as #1 and #2
 Add a small amount of methylene blue to test tube #1
 Quickly click on the START button on the timer and record on your printed Data Sheet
every 15 minutes the distance the dye has moved through the agar
 Add a small amount of potassium permanganate to test tube #2
 Quickly click on the START button on the timer placed on the table and record every 15
minutes the distance the dye has moved through the agar

IV. DATA AND RESULTS

V. INTERPRETATION OF THE RESULTS

o Based on the result, methylene blue, which has a molecular weight of 320 M diffuses to
the test tube with agar by 2 milliliters for 15 minutes, 4 milliliters for 30 minutes, 6
milliliters for 45 minutes and 7.5 millimeters for 60 minutes. While potassium
permanganate, which has a molecular weight of 158 M diffuses to the test tube with agar
by 4 milliliters for 15 minutes, 7 milliliters for 30 minutes, 14 milliliters for 45 minutes
and 21 millimeters for 60 minutes.

VI. CONCLUSION

o •Upon observing the two dyes, such as methylene blue and potassium permanganate, the
diffusion between them showed different fulfillment in terms of movement of particles in
each of the agar. Hence, if we will recall the definition of diffusion, it is the process of
equalization which involves movement of molecules from an area of high concentration
to an area of low concentration. And as stated before the experiment, how quickly
diffusion occurs is dependent on several factors given as the kinetic energy, nature of of
the environment and especially, the size of the molecules. Speaking of size of the
molecules, methylene blue possesses quite larger particles such as salt making it to
diffuse a little harder. Moreover, high viscosity of the agar causes particles in the blue
dye to diffuse more slowly. While on the other hand, potassium permanganate is
approximately half the molecular weight of methylene blue and diffuses more rapidly in
the agar. This gives us the idea that the particles in liquids can move around each other
paving the way for them to be evenly mixed and reach equilibrium. By this, we must
remember that even the solution undergo diffusion, the particles continue to move about
randomly due to their kinetic energy, but their concentrations do not change.
 ACTIVITY NO. 2
EXPERIMENT NO. 2: Passive Transport in Red Blood Cells
“Observations of Passive Transport in RBC”
I. OBJECTIVES OF THE STUDY
 To examine the three possible tonicities of the red blood cells in a solution such as
isotonic, hypotonic and hypertonic.
 To analyze the structure and describe each of the three representation.
 To observe how red blood cells behave in a particular solution percentage.

II. MATERIALS USED

 Beaker of isotonic solution (0.9% saline)
 Beaker of hypotonic solution (0.7% saline)
 Beaker of hypertonic solution (0.11% saline)
 Eyedroppers
 Vial of Human blood
 Three microscope slides
 Light microscope
 Pencil

III. FLOWCHART/DIAGRAM
 Obtain the three beakers of solution (by clicking on each of the blue beakers which lie on
the shelf to your left)
 Obtain the vial of blood from the shelf on your left (by clicking on the red bottle)
 Obtain the three microscope slides (by clicking on them)
 Label each of the slides as 0.7 and 0.9 and 0.11. Use at least pencil or marker in labeling.
 Place one drop of blood on each of the slides (by clicking on the dropper of the vial
containing the blood)
 Now add one drop from each of the solutions to the blood samples on each of the
corresponding slides.
 We will now view each of the slides using a compound light microscope.
 Click on the slide which is marked as 0.9 (Isotonic) (click on the focusing knob to see the
magnified image at 450 X)
 Click on the slide which is marked as 0.7 (Hypotonic) (click on the focusing knob to see
the magnified image at 450 X)
 Click on the slide which is marked as 0.11 (Hypertonic) (click on the focusing knob to
see the magnified image at 450 X)
IV. DATA AND RESULTS

 Sketches of the Three Tonicities

ISOTONIC (1200X)

HYPOTONIC (1200X)

HYPERTONIC (1200X)
IV. INTERPRETATION OF THE RESULTS

o Based on the results, the red blood cells (RBCs) in an isotonic solution (0.9% saline
solution) shows a biconcave disc shape which is normal size and shape of the RBCs.

o On a hypotonic solution (0.7% saline solution), the red blood cells (RBCs) are swelling
which appeared like a balloon shape.

o On a hypertonic solution (0.11% saline solution), the red blood cells (RBCs) are
shrinking which appeared like a prune shape.

V. CONCLUSION

o In case of passive transport, a substance moves down its concentration or electrical

gradient to cross the membrane using only its own kinetic energy or simply the energy of
motion. Kinetic energy is intrinsic to the particles that are moving such that there is no
input of energy to occur. Because we have known that movement of water and dissolved
substances had to overcome the permeability of the membrane. Upon conducting the
experiment, we have learned that when body cells or the red blood cells are placed in a
solution having a different osmotic pressure, the shape and volume of the cells change.
As the water moves by osmosis into or out of the cells, their volume increases or
decreases. So technically, a solution's tonicity is the measure of the solution's ability to
change the volume of cells by altering water content. The contained red blood cell in an
isotonic solution gives the cell a maintained normal shape and volume. Thus, the water
molecules enter and exit at the same rate, allowing the RBCs to keep their normal shape
and volume. Then the RBCs in a hypotonic solution has a lower concentration of solutes
than inside of the cell. Thus water molecules enter the cells faster than they leave,
causing the RBCs to swell and eventually burst. The rupture of RBCs in this manner is
called “haemolysis” which is the rupture of other types of cells due to placement in a
hypotonic solution. It is simply referred to as lysis. Next up, is the cells in a hypertonic
solution wherein there is a high concentration of solutes than which is inside the RBCs.
In such a solution, water molecules move out of the cells faster than they enter, causing
the cells to shrink. Such shrinkage of cells is called crenation.
 Activity no. 2
EXPERIMENT NO. 3: Osmosis and Dialysis
Part I: Osmosis
I. OBJECTIVES OF THE STUDY
 To observe the characteristic of osmosis in a sucrose solution and distilled water using
thistle tube.
 To determine the movement of pressure in mm by using osmotic pressure.
 To know that action taken by osmosis when placed near a higher concentration gradient.

II. MATERIALS USED

 Thistle tube (with mm markings)
 Thistle tube stand
 Dialysis tubing (selectively permeable membrane)
 Rubber band
 Sugar solution (20 % sucrose)
 Beaker

III. FLOWCHART/DIAGRAM
 Click on the thistle tube stand in order to bring it to the table
 Now click on the thistle tube funnel to attach it to the tube stand.
 Next we will put the dialysis tubing on the bulb end of the thistle tube. You will need
only a small piece. Click on the dialysis tubing on the shelf.
 Put a rubber band on the tube and dialysis tubing to hold it in place. You can
accomplish this by clicking on the rubber bands on the shelf
 Put the bulb of the thistle tube into the beaker of water. The water is distilled which
means that it has no solutes in it (100 % water). Click on the beaker labeled as H2O.
 Now we will pour the sucrose solution into the thistle tube from the top. The sucrose
solution is a 20 % sugar solution (80 % water). Click on the beaker labeled as
Sucrose.
 Record the original millimeter measurement of the sucrose solution in the column of
the thistle tube on your data sheet. Next, record the movement of the sucrose
solution every 15 minutes for 90 minutes. Click on the start button of the clock to
begin measuring.
 From the data you have collected you will need to produce two graphs. One of the
graphs will plot Osmotic Potential vs Time. The other graph will plot Osmotic
Change vs Time.
 Compare the two graphs derived from the results.
IV. DATA AND RESULTS
V. INTERPRETATION OF THE RESULTS

o Based on the result, the distilled water diffuses to the thistle tube with 20% sucrose
solution through a semi-permeable dialysis tubing. This diffusion of water makes the
volume of the solution inside the thistle tube to rise from 3 milliliters to 33 milliliters for
90 minutes.

VI. CONCLUSION

o Osmosis is defined as the type of diffusion in which there is net movement of a solvent
through a selectively permeable membrane. It is a passive process wherein the solvent is
water by which it moves by osmosis across a membrane from an area of higher water
concentration to an area of lower water concentration. Within the experiment, we've used
a thistle tube and a dialysis tubing to represent the selectively permeable membrane in the
cell. This demonstrates osmosis. A thick sucrose solution is filled in the thistle funnel and
the dialysis tubing is bound at its lower end. It is lowered in the beaker with distilled
water. A concentrated (hypertonic) solution and a dilute (hypotonic) solution are
separated by a semi-permeable membrane. As observed through the experiment, thistle
funnel is filled with sucrose solution and its mouth is tied with a rubber band. Then,
thistle funnel is kept inverted in a glass beaker that contains distilled water. After some
time the sucrose solution rises in the stem of a funnel that indicates that osmosis has
occurred. It is evident that the level of water in thistle funnel increases. Some time,
movement of water through the thistle tube can be seen. This means that water moves
from the region of higher concentration to lower concentration and the water occurs
through a semi-permeable membrane. Therefore, the force due to which the solution in
the tube increases is due to osmotic pressure.Further, stability of water level in the tube
shows that water concentration in both the beakers as well as the tube is the same and so
osmosis stop.
 Activity no. 2
EXPERIMENT NO. 3: Osmosis and Dialysis
Part II: Dialysis

I. OBJECTIVE OF THE STUDY

 To measure diffusion of small molecules through dialysis tubing, an example of a semi
permeable membrane.
 To observe the movement of a solute through a semi permeable membrane called
dialysis.
 To determine substance can pass through the membrane, in regards to the size of the
minute pores in the dialysis tubing.
II. MATERIALS USED

 30 cm section of dialysis tubing

 String
 15 % glucose solution
 1 % starch solution
 Glucose test strips
 Lugol’s solution
 Distilled water
 2 Test tubes
 Large beaker

III. FLOWCHART/DIAGRAM
 Obtain a 30 cm piece of dialysis tubing (click on the tubing to obtain the dialysis tubing)
 We will now tie one end of the dialysis tubing with the string. (Click on the string to tie
the tubing)
 We can now place the dialysis tubing in distilled water to moisten it so that it will be
easier to open.
 While we let the dialysis tubing soak, we can access the glucose and starch solutions.
 Bring the two solutions to the table by clicking on either of them.
 We will need to develop a method to test for glucose in a solution as well as test a
solution for starch.
 Use a table to record your results.
 We will be using glucose test strips to test a solution for the presence of glucose.
 When the strip is exposed to glucose it will change color from yellow to blue.
 We will be using Lugol’s solution to test for the presence of starch.
 When Lugol’s solution is in the presence of starch the solution will change to a dark blue
color
 Click on the test strips to bring them to the table.
 Next we will test the Glucose solution and the distilled water where the dialysis tubing is
now soaking for the presence of glucose.
 Click on one of the glucose test strips.
 You will be testing the glucose solution for the presence of glucose.
 Click on another test strip to test the distilled water for the presence of glucose.
 Remember a positive test will turn the yellow end of the strip to blue.
 Next we will test for starch in the starch solution and the distilled water.
 You will be using Lugol’s solution to test for starch.
 If starch is present the solution will turn a dark blue color.
 Click on the blue bottle of Lugol’s solution on the shelf to bring it to the table.
 You will need a test tube to test a small portion of the starch solution.
 Next click on the test tube rack.
 Click on one of the eyedroppers on the shelf to add starch to the test tube.
 Next click on the eyedropper in the bottle of Lugol’s solution to add a few drops to the
starch solution in the test tube.
 Record your results on your data sheet.
 Next we will test the distilled water for the presence of starch.
 Click on the test tube rack to get a clean test tube
 Click on a fresh eyedropper from the shelf to add distilled water to the test tube.
 Next click on the eyedropper in the Lugol’s solution to add a few drops to test for starch.
 Record your results
 Next we will open the dialysis tubing and fill it with about ½ full of starch solution and
about ½ full of glucose solution.
 Leave enough of the tubing so that you can tie the end shut
 Click on the dialysis tubing in the distilled water beaker to remove it so that it can be
opened.
 Next click on the starch or glucose solution to fill the dialysis tubing.
 Next click on the yellow string to tie the top of the dialysis bag.
 Now click on the dialysis bag to insert it into the distilled water.
 Leave the dialysis bag in the distilled water for 30 minutes.
 Click the start button on the timer.
 Remove the dialysis bag by clicking on it.
 Open one end of the dialysis tubing by clicking on the yellow string holding one end
together
 Click on the test tube rack to obtain a test tube.
 We will add a small amount of solution from the dialysis tubing to the test tube to test for
glucose and starch.
 Click on one of the blue eyedroppers on the shelf to add solution from the dialysis tubing.
 Next click on the one of the glucose test tapes to test for glucose.
 Record your results
 Now we can test for starch by using a few drops of Lugol’s solution.
 Click on the eyedropper in the Lugol’s solution to add to the dialysis tube solution.
 Record your results.
 We will now perform the same tests for glucose and starch on the solution of distilled
water in which the dialysis tubing soaked for 30 minutes.
 We will need a clean test tube. Click on the test tube rack.
 Click on one of the blue eyedroppers on the shelf to add solution from the distilled water.
 Next click on the one of the glucose test tapes to test for glucose.
 Record your results
 Now we can test for starch by using a few drops of Lugol’s solution.
  Click on the eyedropper in the Lugol’s solution to add to the distilled water.
 Record your results

IV. DATA AND RESULTS

V. INTERPRETATION OF THE RESULTS

o After 30 minutes of the dialysis bag with 1/2 full of starch solution and 1/2 full of glucose
solution been submerged into the distilled water inside the beaker, the distilled water was
tested. The tests shows that the water turns the glucose test strip to colour blue which
means it is positive for the presences of glucose. Hence, when tested with Lugol's
solution, the water did not change its colour which means it is negative for the presences
of starch.

VI. CONCLUSION

o Dialysis is the process of separating molecules in solution by the difference in their rates
of diffusion through a semipermeable membrane, such as dialysis tubing. During the
experiment, it was concluded that the dialysis tubing doesn’t allow all kinds of substances
to pass readily through the pores of its membrane. This means that it is selective in its
permeability to substances. The dialysis tubing was not permeable to all the three
solutions- glucose, starch and iodine (Lugol’s Solution). Rather, the tubing was
permeable to glucose and iodine but not starch. This could be known from the colour
change in the solutions in the beaker and the bag. The tubing was permeable to iodine
and so the content of the bag turned blue-black in colour indicating the presence of
starch. Glucose also readily passed through the pores of the membrane. After performing
corresponding tests on the solutions, the bag’s solution as well as the beaker’s solution
turned almost dark in colour. This shows the presence of reducing sugar in both solutions,
meaning that glucose passed into the beaker from the bag. Nonetheless, this experiment
shows that the natural tendency of these solutions is to move to the outside of the tubing.
However, the selectively permeable membrane kept the starch molecules from moving
out of the tubing. This was supported by the absence of starch outside the membrane. The
salt molecules, however, were able to pass through the membrane. The reason the salt
molecules can pass through the membrane and the starch molecules cannot is the relative
size of the molecules. Salt molecules are much smaller than starch molecules and can
pass through more easily. Starch molecules are made of many glucose molecules attached
to each other. Thus, they are quite large molecules in contrast to the relatively small salt
molecules. The smaller salt molecules pass through the membrane easily, but the larger
starch molecules cannot pass through the membrane.