Experiment No.

18
Date: 12.10.2015

Aim: To study the effect of different pH on the activity of salivary amylase on starch.
Materials required: Test tubes, test tube stands, pipettes, beakers, cotton, funnel, water bath, spirit lamp, burner, measuring
cylinder, starch, sodium chloride, distilled water, iodine, pH tablets of different ranges, cavity blocks and thermometer.
Preparations:
A. 1% starch solution: Add 1g soluble starch to 10 mL distilled water to make thick paste. Boil 90 mL of distilled water and add
the 10 mL prepared starch paste into it. Stir well. Leave solution overnight. Then filter solution to get 1% starch solution.
B. 1% NaCl solution: Dissolve 1g NaCl in 100 mL of distilled water.
C. Collection of saliva: Make thin film of cotton and dip it in water. Spread the cotton over mouth of funnel to act as filter
after draining out excess water from it. Keep funnel over a clean test tube. Chew a piece of rubber and pour incoming saliva
into funnel to filter it. Take 1 mL of filtered saliva and add 19 mL of water to it to get 1:20 dilution of the enzyme.
Procedure:
1. Make three experimental tubes each having 5mL of 1% starch solution + 1mL of 1% NaCl solution. Add a tablet of different pH
into different test tubes i.e., pH=3, pH=7 and pH=10 respectively. Put the test tubes in water bath at 37 o C.
2. Add 1 mL diluted enzyme in all the three experimental tubes.
3. Take iodine in cavities of cavity block.
4. Take a drop of solution from each the three experimental tubes and put it into the different cavities of cavity block having
iodine. Note the time as zero minute and record the change in colour of iodine.
5. After an interval of 2 minutes again take a drop from each tube and pour into the iodine present in cavity of cavity block and
again observe the change in colour of iodine.
6. Keep on repeating the experiment after interval of every 2 minutes till the colour of iodine does not change.
7. Note the time taken by different experimental tubes till they do not give any colour with the iodine (reach achromic point).
Observation:
Sl. No. Time Reaction with iodine
(in minutes) Test tube with pH=3 Test tube with pH=7 Test tube with pH=10
1. 0 Blue colour Blue colour Blue colour
2. 2 Blue colour Blue colour Blue colour
3. 4 Blue colour Blue colour Blue colour
4. 6 Blue colour Blue colour Blue colour
5. 8 Blue colour Blue colour Blue colour
6. 10 Blue colour Blue colour Blue colour
7. 12 Blue colour No change in colour of iodine Blue colour
Result: The enzyme salivary amylase digested the starch in the tube having pH=7 (neutral), and gave achromic point at 12
minutes. It did not act in tubes having pH=3 (acidic) and pH=10 (alkaline) as colour of iodine always changed in them.
Precautions:
1. Filter the saliva through a wet cotton film and not through a filter paper.
2. All the weighing and measuring should be done accurately.
3. There should be no air bubbles while measuring saliva.
4. All the glassware used must thoroughly clean and dried.