BABS1201 Team Enzyme Project

Team Protocol Form

Part 2 - Protocol Submission (Week 6, Practical 5)

Lab Day: Thursday 6th of April

Lab Time: 6-9pm

Are you in Lab G20 (furthest from BSB Student Office) or Lab G21 (closest to
BSB Student Office)?:

Demonstrator Name: Shin

Names of Team Members:

Name of Enzyme that Your Team is Investigating: Catalase

Equipment/Apparatus Required for Your Experiment:

21 x test tubes 4 x universal pH strips

1 x liver (approx. 500 grams) 2 x test tube racks

20 x pipettes 2 x razor blades

2 x stopwatches 2 x motor and pestle

4 x measuring cylinders 2 x thermometers

4 x beakers

Reagents/Materials Required for Your Experiment:

200 mL of Hydrogen Peroxide (3%)

Buffer solution of Sodium Phosphate (1 Mol) (.mL)

Buffer solution of . To make it acidic (.mL)

Method (outline your experimental protocol in words):

Effect of temperature:

1. Hot water baths were set up with the following temperatures: 50C and 100C. An
ice bath was made for one beaker to reach the temperature of 0C and a water
bath was set up at room temperature (37C).

2. 12 test tubes were filled with 5mL of Hydrogen Peroxide. Three repeat trials are
to be done for each temperature, as well as a control (liver and water) tested
with each trial.

3. 5mL of detergent and red dye was added to each test tube
 4. 10 grams of liver was cut with a razor and grinded with a motar and pestle, and
added to the test tubes of each of the four water bath.

5. The forth was measured with a ruler at 30 second increments until 15 minutes.

6. Results were recorded and tabulated.

Effect of pH:

1. Set up 6 test tubes and label them with appropriate pH. Also prepare and label one of them as a
CONTROL.

2. Add 10mL of correct pH solution. (HCl, Orange juice, Distilled water, Soapy water, NaOH). Do
not add anything into the CONTROL test tube as yet.

3. Check the pH of each solution with a pH strip and match the pH level with the colour chart
provided. Record pH in results table.

4. Add a 1x1cm cube of lung (containing enzyme catalase) to each test tube except the CONTROL
test tube and wait for 3 minutes.

5. Measure and record the height of the substance initially and then add 10mL of hydrogen
peroxide solution (the substrate) to each test tube and start timing.

6. Measure the maximum height of bubbles in each test tube after approximately 45 seconds.

7. Record results in a table and repeat experiment at least 3 times for accurate and reliable results.

Provide simple labelled diagrams of the procedures to be carried out and

apparatus to be used:
Provide a Risk Assessment for your experiment (see the risks described for the experiment
in you lab manual for examples, and adapt to specific those that apply to your experiment):
BABS Teaching Laboratory Enzyme:

Student Risk Management Plan catalase

Hazards Risks Controls

Razor blades You can slice you fingers or
hands
You can drop the razor
blade on your foot
Hot water baths Hot plates can burn
Care must be taken when cutting
Covered shoes must be worn
Lab coat and safety googles must
 hands/arms be worn to protect from splashes
Boiling water can splash Closed footwear
Chemical reagents: Hydrogen peroxide can Care must be taken when
Hydrogen peroxide cause skin and eye irritation transferring to test tube

Basic reagent. Lids must be placed back on after

.
Glass ware: such as test
tubes and beakers
Personal Protective Equipment
Acidic reagent use.
Bottles stored away from the
edge of the bench
Lab coat, goggles, shoes must be
worn
Test tubes or beaker can Enclosed shoes and safety
break googles must be worn
This can cut hands/ arms Gloves should be used when
and feet cleaning broken glass and it
should be placed in the correct
bin

Safety Glasses

Emergency Procedures
In the event of an alarm sounding, stop the experiment, turn off electrical equipment and wait for confirmation
from demonstrators to evacuate. Then pack up your bags and wash your hands. Follow the instructions of
the demonstrators regarding exits and assembly points. The BioScience assembly point is in the lawn in front
of the Chancellery. In the event of an injury inform the demonstrator. First aiders and contact details are on
display by the lifts.
Clean up and waste disposal
All used microfuge tubes should be placed in the biohazard bins. Used test tubes should be placed in the
appropriate container on the front bench of the lab. All solutions should be placed in the appropriate liquid
waste containers. Used pipette tips should be placed in approved biohazard sharps containers.
Declaration will be checked by demonstrator at the beginning of the practical class.
I have read and understand the safety requirements for this practical class and I will observe these
requirements.
Signature:Date:
End of Protocol

Please attach Part 1 of your protocol submission (independent research prepared by

individual team members, see p.25-32 of the Course Manual for details) and give both
Parts to your demonstrator at the beginning of your practical class in Week 6.

NB: There is no word/page limit to your protocol submission.

Table:

Temperature 0C 27C 50C 100C

Trial 1

Trial 2

Trial 3

Control