Digestion Experiment Pre-Lab:

1. List the products or monomers formed in the digestion of each of the following substances:

a. lactose

b. sucrose

c. maltose

d. triglyceride

e. protein

2. What is a the difference between a qualitative and quantitative analysis?

3. What qualitative tests are there for:

a. proteins

b. carbohydrates

c. lipids

d. Proteins monomers

e. Carbohydrate monomers

f. lipid monomers
 Digestion Experiment
Purpose:
• In this experiment you will digest various fats, proteins and carbohydrates and then use
some general tests to detect the products formed in the digestion process.

In the digestive tract, large dietary molecules are converted into smaller molecules that can be
absorbed and used by the cells of the body. This process is called digestion.

Experimental Procedure:
This experiment will allow you to observe several reactions that occur during the digestion of
food in the mouth, stomach and small intestine. In each set of tests, control samples will be
used. The control samples contain everything but the enzyme that catalyzes the digestive
process. A control sample allows you to observe the importance of the enzyme. Some tests
require longer heating periods than others. Start those that require longer heating times first.

Hydrolysis of Starch Salivary Amylase:

• If you have eaten within the last two hours, rinse your mouth thoroughly with a cup of water.
In a 50 mL beaker, collect a volume of saliva equal to one third the volume of a 10 cm test
tube. This will be the source of salivary amylase.
• Fill a 250 mL beaker about two thirds full of tap water and begin warming the water to about
37°C. This will be used to incubate samples during the experiment.
• Label four test tubes A, B. C and D. Add 2.00 mL of 2% starch solution to each one.
• Add 4.00 mL of deionized water to test tubes A and B; these will be the controls.
• Add 4.00 mL of the saliva sample to test tubes C and D.
• Place the four test tubes into a 250 mL water bath set near 37°C.
• Then, using very clean glass stirring rods, stir the contents of tubes C and D for 3 minutes.
• Allow the four test tubes to remain in the water bath for a total of 30 minutes. During this
time, begin the next part.
• After 30 minutes, remove the test tubes and place them in a test tube rack.
• Add 2 drops of iodine solution to test tubes B and D.
• Record your observations.
• The solutions in test tubes A and C will be used in the following section for the Benedict test.

Hydrolysis of Starch Pancreatic Amylase:

• A boiling water bath will also be needed for this part of the lab.
• Label four test tubes E, F. G and H. Prepare each test tube as described below:
o Test tube E: 4 mL of deionized water (the control)
o Test tube F: 2 mL of 5% pancreatin solution + 2 mL of deionized water
o Test tube G: 2 mL of 5% pancreatin solution + 2 mL of 0.5% Na2CO3 solution (The pH
of this solution is near that of the small intestine.)
o Test tube H: 2 mL of 5% pancreatin solution + 2 mL of 0.2 M HCl solution (The pH of
this solution is near that of the stomach.)
• To each test tube, add 2 mL of 2% starch solution.
• Seal each test tube with a cork or Para-film and gently shake each one to mix the contents.
Remove the corks or film, place all four test tubes in the 37°C water bath, and hold them at
that temperature for 30 minutes.
• After 30 minutes, pour half the contents of each test tube into another test tube of the same
size (labeled E2, F2, G2 and H2) and add 2 drops of iodine solution to each of these second
tubes and mix.
• Save the remaining half of each solution (tubes E, F. G and H) for the Benedict test that
follows. Record your observations.
• Now test for the presence of reducing sugars (glucose and/or maltose).
• Add 5 mL of Benedict reagent to test tubes A and C and to the remaining solutions in test
tubes E, F. G and H. Place the six test tubes into a boiling water bath for 7 to 10 minutes.
• Record the colors you observe and indicate whether a precipitate forms. A positive test,
indicating the presence of maltose and/or glucose, is a red precipitate.

Digestion of Protein:
• Label and prepare three test tubes as follows:
o Test tube P1: 5 mL of deionized water + 1 mL of 0.1 M HCl (the control)
o Test tube P2: 5 mL of 2% pepsin (an enzyme) + 2 mL 0.1 M HCl (near stomach pH)
o Test tube P3: 5 mL of 2% pepsin + 1 mL deionized water (near neutral pH)
• Add a small piece of boiled egg white to each test tube and place each one in a water bath
at 37°C for one hour.
• Record any changes observed in the egg white in each test tube.
• Any differences observed between the control and the other two samples can be attributed
to the enzyme, pepsin, in acidic and near neutral environments.

Digestion of Triglycerides:
• Label three test tubes 1, 2 and 3.
• Add the following materials to the test tubes, using the supplied pipets as necessary.
o Test tube 1: 2 mL of 5% sodium choleate solution, a bile salt, 2 mL of 5% pancreatin
solution, an enzyme, and 10 drops of water.
o Test tube 2: 2 mL of 5% sodium choleate solution and 10 drops of water.
o Test tube 3: 2 mL of 5% pancreatin solution and 10 drops of water.
• To tubes 2 and 3 add 0.1 M NaOH solution drop by drop until the pH paper indicator turns
the color that indicates a pH of 7.
• Then add deionized water drop wise until the volumes of test tubes 2 and 3 have the same
volume as test tube 1.
• Add 5 drops of corn oil (or other triglyceride) to each test tube.
• Place the test tubes in a 37°C water bath. After 10 minutes, test the pH with the pH paper to
determine which samples have become more acidic, showing that digestion of the
triglyceride has taken place.
Concluding Questions:
Hydrolysis of Starch by Salivary Amylase:
Which sample(s) (B or D) gave a positive starch iodine test? Did digestion occur in either of
these starch samples?

Which sample(s) (A or C) gave a positive Benedict test? In terms of digestion, what is indicated
by a positive Benedict test?

Hydrolysis of Starch by Pancreatic Amylase:

Which sample(s) (E2, F2, G2 or H2) gave a positive starch iodine test? Did digestion occur in
any of these samples?

Which sample(s) (E, F. G or H) gave a positive Benedict test?

Indicate whether a precipitate formed and if one did what was its color:
E______________ G______________
F______________ H______________

At which pH was the digestion of starch the most complete? Why?

Digestion of Protein:
In which of the samples (P1, P2 or P3) did a visible change occur? What effect does 0.2 M HCl
have on the digestive process of protein?

Digestion of Triglycerides:
What is the color of universal indicator at pH 7?

What colors were observed in each sample after healing at 37°C for 10 minutes?

1 _____________ 2______________ 3________________

Which samples became more acidic during the heating?

Which samples did digestion occur?