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4.1 Studying the effects of temperature on the activity of salivary amylase. Problem Statement: What are the effects of temperature on salivary amylase activity? Hypothesis: The rate of reaction catalysed by salivary amylase is highest at 37 C. Variables: Manipulated: Temperature of medium of reaction Responding: The rate of reaction catalysed by salivary amylase Constant: Volume of saliva, volume and concentration of starch suspension and pH of medium. Materials and apparatus: Beakers, test tubes, syringes, droppers, glass rods, white tiles with grooves, thermometer, a test tube rack, stopwatch, water bath, 1% starch suspension, saliva suspension, iodine solution, ice cubes and distilled water. Procedure:

Diagram 1 1. Prepare 10 test tubes label A, B, C, D, E, P, Q, R, S and T. (P1) 2. Rinse mouth with warm water and collect saliva. (P5) 3. Dilute saliva with equal amount of distilled water. (P1)

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4. Use syringe to put 5 ml of 1% starch suspension into each test tubes label A, B, C, D and E respectively. (P1, P2) 5. Immerse test tubes A and P, B and Q, C and R, D and S, E and T respectively into different water baths with temperature keep constant at 0oc, 28C, 37C, 45C and 60C as shown in Diagram 1. (P1, P4) 6. Meanwhile, prepare a dry piece of white tile with grooves and place a drop of iodine solution into each groove. (P1, P2) 7. After 5 minutes, pour the starch suspension in test tubes A into saliva in test tubes P. Stir and start stopwatch. (P1, P2) 8. Use dropper to remove a drop of mixture from test tube A and place it into the iodine in the first groove on the tile. (P1) 9. Repeat step 7-8 for test tubes B, C, D and E. (P4) 10. Measure and record the time taken using stopwatch for the iodine solution to remain brown. (P3) 11. Calculate and record rate of reaction by using formula: Rate of reaction = 1/t (minute-1) (P3) Result Test Temperature tub (oC) e A B C D E 0 28 37 65 60 Time taken for the hydrolysis of starch to be completed (minutes) Rate of enzymatic reaction 1/t (minute-1)

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