ACTIVITY 7

SALIVARY DIGESTION

A.) Objectives

At the end of this activity, students are expected to:

a.) Explain the nature of saliva

b.) Discuss the principles/rationale of each test

B.) Procedure

a.) Reaction of Saliva

Place a few drops of resting saliva in

three test tubes.

Test the reaction with

Phenolphthalein, Blue Litmus and
Congo red.

From the colors produced, estimate

the approximate pH of your saliva.
Use a pH paper.

Repeat experiment after stimulating

the flow of the saliva by chewing
paraffin vigorously for five minutes.
 b.) Inorganic Matter

Place 5 ml of saliva in a test tube

Acidify with concentrated Nitric acid

and heat to boiling to remove all the
proteins.

Filter, divide into four test tubes and

test the filtrate for:

Chloride by Phosphates by Sulfates by adding 1 Calcium By

adding 1 drop of adding 1 drop of drop of BaCl3 solution adding one drop
AgNO3 solution to Ammonium to test tube 3 of Ammonium of
test tube 1 Molybdate to test Potassium
tube 2 oxalate

c.) Influence of Temperature on Ptyalin Activity

Place 1 ml of starch solution to

each of the four test tubes

Immerse one test tube in an Keep the second at room Place the third in a
ice-water mixture, temperature water bath at 38

Add to these test tubes 10 drops

of saliva and shake well.

To the contents of the fourth

tube, add 10 drops of boiled
saliva.

At 15 minutes interval, remove

a sample from each test tube,
test with iodine solution and
benedicts solution.

Allow the test tubes to stand for

an hour.
d.) Influence of Dilution of Ptyalin Activity

Place 9 ml of water to each of the 6

test tubes.

To the first, add 1 ml of saliva and

mix thoroughly

Transfer 1 ml of this mixture to the

second tube and mix

Take 1ml of the mixture from the

second tube and transfer to the
third tube.

Take 1 ml of the mixture from the

third tube and transfer to the fourth
tube

Take 1 ml of the mixture from the

fourth tube and transfer to the fifth
tube

Take 1 ml of the mixture from the

fifth tube and transfer to the sixth
tube.

To each of the test tubes add 1 ml

of 1% starch solution.

Shake well and place in water bath

at room temperature of about 40 C
for 20 minutes

Now and then test a portion with

Iodine test and another portion with
Benedicts reagents.
C.). Results and Observation

a.) Influence of Temperature to Ptyalin Activity

SAMPLE IODINE BENEDICTS

Test tube 1 All the samples for the Iodine test Sky blue; No color change
turned black. Ideal result would be
lightening of the samples. But due to
procedural

Test tube 2 All the samples for the Iodine test Sky blue; No color change
turned black. Ideal result would be
lightening of the samples. But due to
procedural

Test tube 3 All the samples for the Iodine test Sky blue; No color change
turned black. Ideal result would be
lightening of the samples. But due to
procedural

Test tube 4 All the samples for the Iodine test Sky blue; No color change
turned black. Ideal result would be
lightening of the samples. But due to
procedural
 b.) Influence of dilution on Ptyalin activity

SAMPLE IODINE TEST BENEDICTS TEST

First Test Tube From brownish/yellow color, it From blue to light brown turbid
turned light clear blue. yellow

Second Test Tube From brownish yellow to very Remained blue

pale yellow

Third Test Tube Brownish yellow to pale yellow Remained blue

Fourth Test Tube Light green Clear

Fifth Test Tube Light green Light blue/remained blue

Sixth Test Tube Light green Remained blue

D.) Analysis and Conclusion:

1.) How did you know that digestion has taken place in the mouth?

Digestion is the process by which food is broken down by enzymes through hydrolysis.
That is, water is added to a large molecule, which splits into smaller molecules that can be
absorbed into the bloodstream. For example, water is added until proteins are broken down into
amino acids, starch is broken down into glucose, and fates broken down into glycerol and fatty
acids. Enzymes are necessary for digestion, just as they are required for chemical reactions in
the body. Salivary amylase hydrolyzes starch.

Starch is digested by salivary amylase in the mouth, a process described by the

following reaction:

Starch + water (catalyzed by amylase (enzyme)) = maltose

So a positive result for Benedicts test, a test for the presence of sugars e.g. maltose,
indicates digestion.

2.) Explain how temperature influences the process of digestion.

Like most chemical reactions, the rate of an enzyme-catalyzed reaction, in this case,
digestion, increases as the temperature is raised. A ten degree Centigrade rise in temperature
will increase the activity of most enzymes by 50 to 100%. Variations in reaction temperature as
small as 1 or 2 degrees may introduce changes of 10 to 20% in the results. In the case of
enzymatic reactions, this is complicated by the fact that many enzymes are adversely affected
by high temperatures. As shown in Figure 13, the reaction rate increases with temperature to a
maximum level, then abruptly declines with further increase of temperature. Because most
animal enzymes rapidly become denatured at temperatures above 40C, most enzyme
determinations are carried out somewhat below that temperature.

Over a period of time, enzymes will be deactivated at even moderate temperatures.

Storage of enzymes at 5C or below is generally the most suitable. Some enzymes lose their
activity when frozen.

Influence of temperature on Ptyalin Activity

If digestion has not taken place, the iodine test for starch will be positive. If starch is
present, a blue-black color immediately appears after a few drops of iodine are added.

If digestion has taken place, a test for sugar (maltose) will be positive. A color change of
blue to green to yellow to orange to red indicates the presence of maltose. Boiling the test tube
is necessary for the Benedicts reagent to react. When there is an increase in temperature,
there is an increase in kinetic energy which then increases effective collision. When effective
collisions are increased, there is an increase in the velocity of molecules which then increase
the rate of chemical reaction in Ptyalin. But, the optimum temperature for reaction is 40 degrees
Celsius. Anything above or below the optimum temperature, the rate of reaction would decrease
due to the inactivation of Ptyalin. In the experiment, 4 measures were made for the four test
tubes; the ice water, the room temperature, the 40 degree warm bath and the boiling saliva. It is
expected that in the Benedicts test, the one dipped in the ice water would become negative and
in the Iodine test, it would show a positive outcome. This shows that the salivary amylase is
more reactive in the Iodine test because even at low temperatures, a reaction can still occur.
Possibly, a higher temperature is needed in order for a reaction to take place in the Benedicts
test. In the room temperature, it is expected that both the Benedicts test and Iodine test would
display positive results. As weve said before, even in low temperatures, Iodine is already
reactive. This means that it is even more reactive when temperature is increased because
increasing temperature also increases the rate of reactivity of a solution (as explained above in
the previous sentences). Now, since temperature is now increased compared to that of the cold
bath, Benedicts test is now more reactive. This is why we can expect a positive result.
Benedicts test would display a positive result in the 38 degree water bath while the Iodine test
will display a negative result. This is because the Benedicts test becomes more reactive when
temperature increases especially when it reaches the boiling point. Having said this, we could
expect that the Benedicts test would react positively to the boiling saliva.
 Influence of dilution on Ptyalin activity

With the same amount of substrate, ptyalin activity is directly proportional to enzyme
concentration, so as the amount of enzyme concentration increases, the rate of reaction also
enzyme increases.

If the enzyme concentration was kept constant and the substrate concentration was
varied, increasing the amount of substrate increases the velocity of the reaction until it
reaches the maximum activity of the enzyme.

E.) Pictures