KENDRIYA VIDYALAYA ONGC CHANKHEDA

AISSE PRACTICAL EXAM 2021-2022

CHEMISTRY INVESTIGATORY PROJECT

TOPIC: Studying the action of salivary amylase

on starch and effect of pH and temperature on its
action

Name: Divya Yadav

Guided By: Mr. P.K. Patidar

Class:: 12th – B

Roll no.: 12218

Subject:: Chemistry

~CERTIFICATE~

Name: Divya Yadav

Class: 12th B

Roll no.: 12218

Institution: KV ONGC Chandkheda

This is certified to be bonafide work of the student in the K V ONGC

CHANDKHEDA ONGC, Laboratory during the academic year of 2021/2022.

No. of practicals certified 1 out of 1 in the subject of CHEMISTRY.

Examiner’s signature Teacher in charge

Principal

Date: Institution Stamp

 ~INDEX~
1. Certificate
2.Acknowledgement
3.Introduction
4.Aim
5. Requirements
6. Experiments
7. Observations
8.Conclusions
ACKNOWLEDGEMENT
I owe my debt to Mr. P.K. Patidar
for giving me the opportunity to carry out
this experiment in the Chemistry lab of KV
ONGC Chandkheda. I would like to extend
my heart full gratitude to our honourable
principal Mr. Ashok Rathi & my chemistry
teacher Mr. P.K. Patidar for being a constant
source of inspiration to me. At the end,
thanks to all who have given their direct or
indirect help in this project without which
it wouldn’t have been a success.

~INTRODUCTION~
Importance of saliva:
 Every health book insists on the chewing of food. The act
of chewing stimulates the release of saliva.
 Saliva helps in mastication of food and initiation of
digestion by hydrolysing almost 30% of the total starch
ingested by a human.
 However, the complete digestion of starch occurs only in
the small intestine by the action of pancreatic amylase.

Action of saliva:
 The enzyme ptyalin or salivary amylase, present in human
saliva hydrolyse the big molecules of food into many
molecules. For example, starch into mono-saccharides
maltose and glucose.
 Thus saliva not only helps in digestion of food but convert
it into energy generating substances.

Factors affecting enzyme activity:

 Enzymes and their activity are very sensitive to
temperature, pH and several other factors.
 Even a slight variation in these two factors, can disrupt the
action of enzymes. In other words, digestion of food by
salivary amylase is also effected by pH and temperature
and can be verified experimentally.
 There are certain favourable conditions for the activity of
enzyme called the optimum conditions.

Optimum Temperature: The temperature at

which the enzyme activity is the maximum and
decreases before or after that temperature is
called the optimum temperature for that enzyme.

Optimum pH: The pH at which the enzyme

activity is the maximum and decreases before or
after that pH is called the optimum pH for that
enzyme.
 ~AIM~
“To Study the digestion of starch by salivary amylase and
effect of temperature and pH on it”

 To study digestion of starch by saliva.

 To study the effect of temperature on the digestion of
starch by saliva.
 To study the effect of pH on the salivary digestion of
starch.
 REQUIREMENTS

 Beakers
 Test tubes
 Test tube holder
 Dropper
 Stopwatch
 Iodine solution
 Thermometer
 Dil. HCl (aq)
 Dil. NaOH (aq)
 Ice cubes
 Bunsen burner
Requirements
Experiments
 ~Experiment 01:
To Study the digestion of starch by salivary amylase.

 PROCEDURE:

1) Collection of Saliva - Rinse mouth throughly with cold

water and ensure that it does not contain any food particles. Now take
about 20ml of luke warm water in the mouth and gangle for about
three minutes so that saliva mixes up well with it. Spit this into a
beaker. Filter, if there is any suspended impurity clear filtrate is saliva
solution and contains enzyme ptyalin.

2) Preparation of starch solution - Take about 0.5g of

starch in a 100ml beaker and add enough water to make a paste. Dilute
the paste by adding 50ml water and boil for about 5 min.

3) Digestion of starch
(a) take 5ml of the starch solution in a test tube. Add 2 ml of saliva
solution into it. Mix the solutions well by shaking the tube carefully
and start a step watch.
(b) After one minute take out two drops of the mixture solution from
the test tube with the help of a dropper and transfer it into another test
tube containing about one ml of 1% iodine solution. Note the colour
produced, if any.
(c) Repeat this test after every one minute taking two drops of the
mixture solution and fresh 1% iodine solution continue until the test
shows no blue colour. Record the time and blue colour intensity.
  OBSERVATION TABLE

Time passed 1 min 2 min 3 min 4 min

Colour intensity Deep blue Blue Light No colour
blue

 OBSERVATION:
The blue colour of the starch solution fades down slowly and
turns colourless after a period of time.

 CONCLUSION:
Absence of blue colour on addition to iodine solution means
absence of starch in the mixture solution. That indicates whole
of the starch has got digested or hydrolysed by the action of
salivary amylase.

~Experiment 02:
To Study the effect of temperature on the activity
of salivary amylase.
 PROCEDURE:
1. Take beaker containing 15 ml of 1% starch solution + 3
ml of 1% NaCl solution.
2. Divide and pour this solution into three test tubes and
mark them as A, B and C.
3. Maintain the temperature of the beaker containing ice
cubes at 5°C.
4. Take beaker containing ice cubes and keep it on the
table.
5. Take another two beakers containing water and heat
over the Bunsen burner.
6. Now transfer experimental tube A into a beaker
containing ice.
7. Transfer the second experimental tube B into water
bath set at 37°C and third experimental tube C into the
beaker maintained at 50°C.
8. Using a dropper, take 1 ml saliva solution and transfer
the solution into test tube A.
9. Similarly, add 1 ml saliva solution into test tube B and
test tube C.
10. Immediately, using a dropper, take few drops from
experimental tube A and transfer this into first series
of test tubes having iodine solution.
11. Similarly, using fresh droppers, do the same procedure
for test tube B and test tube C and transfer the solution
into second and third series of test tubes having iodine
solution.
12. Note this time as zero minute reading.
13. After an interval of 2 minutes, again take a few drops
from each tube and add to the iodine tubes and note
the change in colour of iodine.
14. Keep on repeating the experiment at an interval of
every 2 minutes till colour of iodine does not change.

 OBSERVATION:
It takes less time to reach achromic point at 37°C, as
the enzyme is maximum active at this temperature,
while at higher and lower temperatures more time is
taken to reach the achromic point.

 CONCLUSION:
At a lower temperature, the enzyme salivary amylase is
deactivated and at the higher temperature, the enzyme is
denaturated. Therefore, more time will be taken by an
enzyme to digest the starch at lower and higher
temperatures.
Optimum temperature for the enzymatic activity of salivary
amylase ranges from 35°C to 37°C.

~Experiment 03:
To Study the effect of pH on the enzyme salivary amylase.

 PROCEDURE:
1. Take a beaker containing 15 ml of 1% starch solution +
3 ml of 1% NaCl solution.
2. Divide and pour this solution into three test tubes and
mark them as A, B and C.
3. Add pH tablet 5 into test tube A, pH tablet 6.8 into test
tube B and pH tablet 8 into test tube C.
4. Now transfer experimental tube A, B and C into a
beaker containing water and a thermometer for
recording temperature. Temperature of this beaker is
to be maintained at 37°C.
5. Using a dropper, take 3 ml saliva solution and add 1 ml
of solution to each of the three test tubes.
6. Immediately using a dropper, take few drops from
experimental tube A and transfer this into the first
series of test tubes having iodine solution.
7. Similarly, do the same procedure for test tube B and
test tube C and transfer the solution into second and
third series of test tubes having iodine solution.
8. Note this time as zero minute reading.
9. After an interval of 2 minutes, again take a drop from
each tube and add to the iodine tubes and note the
change in colour of iodine.
10. Keep on repeating the experiment at an interval of
every 2 minutes till colour of iodine does not change.

 OBSERVATION:
1. The blue colour of the solution remains intact in the case
of the solution with pH=05
 2. The blue colour of the solution remains intact in the case
of the solution with pH=08
3. The blue colour of the solution disappears after a while.

 INFERENCE:

1. pH is acidic (=05) in test tube 01, thus the

salivary amylase doesn’t show its activity.
2. pH is basic (=08) in test tube 01, thus the salivary
amylase doesn’t show its activity.
3. The test tube with slightly acidic pH (=6.8) shows
the action of salivary amylase and is favourable for
the digestion of the starch.

 CONCLUSION:
The enzyme salivary amylase is most active at pH 6.8.  Our
stomach has high level of acidity which causes the salivary
amylase to denature and change its shape. So the salivary
amylase does not function once it enters the stomach.