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In this experiment we will be looking at the effects of pH on the enzyme Amylase

Which pH does Amylase work best at?

Place tube 6 and tube A into the bath for 2 minutes, while they are warming, place 1 drop of iodine into each well of the spotting tile - DON'T WRITE ON THE SPOTTING TILE.

Step 03

Using a measuring cylinder, add 5ml of starch solution to each of the tubes 6, 7, 8 and 9.Using a pipette, place 2ml of amylase into tube A

Step 02

Label 5 boiling tubes 6, 7, 8, 9 and AIdentify all equipment to be used, and ensure your workspace is tidy, set-up and organised!

Step 01


Start the stop watch and empty the remainder of the pipette solution back into tube 6. After 30 seconds extract some more solution and put one drop in the next spotting well, empty the rest back into tube 6. Repeat until the iodine no longer chnages colour

Step 06

Add the 5ml of pH Buffer to the pH 6 tube containing the starch solution.Add the amylase solution and immediately extract a drop using a clean pipette and place a drop into the FIRST spot of the spotting tile.

Step 05

In a clean measuring cylinder, measure out 5ml of the required pH buffer.Remove the 2 tubes from the water bath. Get ready with the stopwatch!

Step 04


  • Split into Pairs or Three's
  • Double-check you have everything you need - including paper, pen/pencil, ruler, goggles and gloves
Safety Precautions -
  • Wear goggles as some buffers are harmful
  • Avoid getting enzymes or iodine on skin or clothes as they can cause irritation and/or staining - wash off with water in case of contact



Clean up - your equipment, your workspace and wash your hands!

Step 09

Record your results in the relevant tables.Then draw a graph for the results of table 2 - with pH on the x-axis and time on the y-axis

Step 08

Repeat with the other pH buffers, 7 and 8, and if possible 9

Step 07



What caused the results observed?



What does "denaturing" mean?

What does this mean for enzymes in the body e.g. the stomach?

What effect do the other pH buffers have?

Which pH produced the fastest reaction?


Which pH?

Repeat the experiment with the other tubes!

Repeat every 30s until no colour change occursNote the time at which this happens

Immediately extract a small amount of the mixture andDrop it into the first well

Prepare 5ml of the required pH bufferAnd add to the starch solutionAnd then add the amylase.

Put 1 drop of iodine into Each well of the spotting tile

Add 2ml of amylase toTube labelled A

Place both tubes into a water bath for 2 minutes

Add 2ml of amylaseto tube labelled A

Add 5ml of starch solution toeach tube labelled 6,7,8 & 9

Label your boiling tubes

Method Summary
  • What is an enzyme?
  • Why are they important?
  • What is unique about them during a reaction?
  • What is the difference between a substrate and a product?
  • What does amylase react with?
  • What does Lipase reacts with?
  • What do proteases react with?
  • Name 2 things that can affect how effective an enzyme is?
  • Where in the body are enzymes produced?
  • What are enzymes made of?

What have you learned?