Experiment 2-3

BIOCHEMICAL PROCESSES

Diffusion is known as passive transport. It is the process whereby a substance moves from an
area of higher concentration to a region of lower concentration. The greater the difference in
concentration between the two areas or regions, the faster will be the rate of diffusion. No external
source of energy is required.

Osmosis is the movement (diffusion) of water (solvent) through a semi-permeable membrane

from a weaker solution, one containing less dissolved solute, to a stronger solution, one containing
more dissolved solute. Osmotic pressure is required during the osmosis process.

Dialysis is the separation of solute particles from colloidal particles by means of a semi-
permeable membrane. In the body, the membranes, however, do not allow proteins to pass through
since proteins are colloids.

Surface tension is defined as the force that causes the surface of a liquid to contract. It is
responsible for the formation of drops of water on a greasy surface. All liquids exhibit surface tension;
the surface tension of water is higher than that of most liquids.

OBJECTIVES

1. To relate the particle concept of matter to biochemical processes.

2. To describe molecular motion and interaction.
3. To describe the following processes: diffusion, osmosis, dialysis and lowering of surface
tension.
4. To determine the effects of hypotonic, hypertonic and isotonic solutions on the red blood
cells.

MATERIALS

50 mL graduated cylinder water bath

2 pcs 250 mL beakers wire gauze
500 mL beaker tripod
6 test tubes bunsen burner
test tube rack evaporating dish
test tube brush glass funnel
test tube holder filter paper
iron stand pipette
iron ring stirring rod
microscope medicine dropper

Reagents Other materials

25% trichloroacetic acid solution gastric lining (longganisa
Fehling's solutions A and B membrane)
chloroform milk

1
 25% soap solution string
sulfur powder bile
distilled water 1/2 bar colorless agar agar
1M and 5M CuSO4 solutions 5 pcs glass slides w/
NaCl solutions (0.9%, 0.1% cover slips
and 5%) lancet
cotton
70% alcohol
ruler
cotton cloth

PROCEDURES

A. Diffusion

1. Place 1/2 bar of colorless agar agar in 100 mL of water. Boil the mixture until the agar
agar is completely dissolved. Screen the mixture using a cotton cloth.
2. Half-fill 2 test tubes with the hot/warm mixture. Solidify by allowing the mixture to cool.
3. In the first test tube, add 5 drops of 1M CuSO4 solution. Add 5 drops of 5M CuSO4 solution
to the second. Set aside for 1 to 2 hours.
4. Measure the depths of penetration of the two reagents in the test tubes. Compare.

B. Osmosis

1. Clean 4 glass slides with a piece of cotton. Label as 1, 2, 3 and 4.

2. Blood extraction:

a. Disinfect your ring finger with a cotton soaked in 70% alcohol solution.
b. Prick your finger once using the lancet.
c. Extract blood by pressing your finger. Place a drop of blood in each of the four
glass slides.
d. Clean your cut using a piece of cotton.

3. Use the fifth glass slide to spread the blood thinly in each glass slide.
4. Add 0.9% NaCl, 0.1% NaCl and 5% NaCl in the second, third and fourth slide, respectively.
5. Cover the four glass slides with cover slips.
6. Examine the slides under the microscope.
7. Compare the appearance of the red blood cells in the three glass slides with NaCl solutions
with that of the first.
8. Draw and describe the appearance of the blood cells in the glass slides.

C. Dialysis

1. Tie a string around the lower portion to close one end of the gastric lining.
2. Pour 25 mL of milk in the membrane. Close the open end with a string.
3. Wash off with distilled water any milk from the outside surface of the pouch.
4. Suspend the pouch from an iron ring into a 500-mL beaker. Add just enough distilled
water in the beaker to immerse about three-fourths of the pouch.

2
 5. Allow the milk to dialyze for an hour with constant stirring of the water outside.
6. Test the dialysate for proteins and sugars.

a. Test for Proteins

Place 1 mL of the dialysate in a test tube. Add 1 mL of 25% trichloroacetic acid

solution. If protein is present in the dialysate, a white precipitate will appear. The
mixture may also appear turbid.

b. Test for Sugars

Mix 1 mL each of Fehling's solutions A and B in a test tube. Heat the mixture in
a boiling water bath. Add 1 mL of the dialysate and continue boiling for 1 to 2 minutes.
A yellow or a red precipitate indicates the presence of a reducing sugar in the dialysate.

D. Surface Tension

1. Lowering Surface Tension Using Soap Solution

a. Prepare two test tubes labeled 1 and 2.

b. In each test tube, mix together 1 mL of distilled water and 1
mL of chloroform.
c. Add 1 mL of soap solution to test tube no. 2.
d. Shake both test tubes and allow them to stand for a few minutes.
e. Compare.

2. Lowering Surface Tension Using Bile Solution

a. Place 5 mL of distilled water in a dry evaporating dish.

b. Sprinkle a pinch of sulfur powder on the surface of the liquid. Observe.
c. Repeat the test using diluted bile-water mixture instead of water.
d. Compare the results.
e. Record your observations and results in the report sheet.

3
 REPORT SHEET

Group No.: _______________________________ Score: _______________

Name: __________________________________ Date Performed: _______
C/Y/S: ___________________________________ Date Submitted: _______

Experiment 2-3
BIOCHEMICAL PROCESSES

DATA and RESULTS

A. Diffusion

Depth of
System Penetration Observations / Explanations
(in cm)
________________________________________________
Agar agar with ________________________________________________
1M CuSO4 _____ ________________________________________________
solution ________________________________________________

________________________________________________
Agar agar with ________________________________________________
5M CuSO4 _____ ________________________________________________
solution ________________________________________________

B. Osmosis

System Drawing Observation / Explanation

_________________________________
Slide No. 1 _________________________________
Blood _________________________________

_________________________________
Slide No. 2 _________________________________
Blood + 0.9% NaCl _________________________________
solution
_________________________________
Slide No. 3 _________________________________
Blood + 0.1% NaCl _________________________________
solution
_________________________________
Slide No. 4 _________________________________
Blood + 5% NaCl _________________________________
solution

4
C. Dialysis

Test Positive / Negative Explanation

________________________
For Proteins ________________________
________________________

________________________
For Sugars ________________________
________________________

D. Surface Tension

System Observation Explanation

____________________________ ______________________________
Test tube no. 1 ____________________________ ______________________________
distilled water + ____________________________ ______________________________
chloroform ____________________________ ______________________________

____________________________
Test tube no. 2 ____________________________
distilled water + ____________________________
chloroform + ____________________________
soap solution
____________________________
Distilled water + ____________________________
sulfur powder ____________________________
____________________________
______________________________
______________________________
______________________________
______________________________
______________________________
______________________________
______________________________
______________________________

____________________________ ______________________________
Bile solution + sulfur ____________________________ ______________________________
powder ____________________________ ______________________________
____________________________ ______________________________

CONCLUSION

_____________________________________________________________________________________
_____________________________________________________________________________________
____________________________________________________________________________________

5
CLINICAL APPLICATIONS

1. What is a physiologic saline solution? Why are patients given this solution?
_________________________________________________________________________
_________________________________________________________________________
_________________________________________________________________________

2. Give at least one biological importance of diffusion.

_________________________________________________________________________
_________________________________________________________________________
_________________________________________________________________________

3. What is hemodialysis?
_________________________________________________________________________
_________________________________________________________________________
_________________________________________________________________________

4. Explain why mercury is toxic in its gaseous phase and nontoxic in its liquid
state.
_________________________________________________________________________
_________________________________________________________________________
_________________________________________________________________________