Research question:

How do the different ph levels (5,6,7,8,9) affect the rate of reaction, as measured by the
rate of pressure increase (Pa.s-1), when hydrogen peroxide (H₂O₂) decomposes using
catalase from a potato?

Background research

What are enzymes:

Enzymes are biological catalysts that are used to speed up biochemical reactions by
lowering their activation energy (figure 1). Enzymes play an important role in cellular
metabolism. They split a substrate apart or they bind a substrate together. Most
enzymes are completely or partially made of proteins. There are different types of
enzymes, which include; catalase, diastase, pectinase and etcetera. Enzymes can be
used multiple times because they are not consumed when the reaction is happening.
Enzymes increase the rate of reaction but have no effect on the concentrations of
reactants and products at equilibrium.

(Figure 1)

https://www2.nau.edu/lrm22/lessons/enzymes/enzymes.html

Adding to this enzymes are extremely specific for their substrates in vivo, for example,
hydrogen peroxide only reacts with catalase. The active site of an enzyme is the
location on the enzyme where the substrate binds. Induced fit model explains that this
complementary binding where both enzyme and substrate change shape a little bit so
that they bind together at its maximum strength at the transition state of a catalytic
reaction.

Hypothesis

Variables

Control Variables - The length and volume of the

potato extract (3 cm) must be done
with the same corer to get the
same volume)
- The amount of hydrogen peroxide
(10 ml)
- The amount of ph buffer (10 ml)
- Duration of the reaction: The
conical flask has to be cleaned
every time we start a new
experiment because the flask has
to be clean every time
- Recording period of 60 seconds on
PASCO

Independent Variables pH buffer levels (5,6,7,8,9): this is the

variable that is changed in the
experiment. By adjusting the pH of the
reaction environment using different pH
buffer solutions, the effect of pH on the
rate of decomposition of hydrogen
peroxide can be researched. Each pH
level is used as a different experimental
condition, this allows for the comparison
of reaction rates with different acidity
levels.

Dependent Variables Rate of pressure change (±1 Pa/s): This

is the variable being measured in the
experiment. It shows the rate at which
gas is produced during the decomposition
of hydrogen peroxide catalyzed by the
catalase. The recordings of the pressure
sensor change in pressure over time, this
 allows the determination of the reaction
rate under various pH conditions to
happen.

Equipment list:

- PASCO relative pressure sensor: crucial for measuring the pressure changes that
happen throughout the decomposition reaction. It helps with accurate and
precise monitoring of the rate of pressure increase.

- Tube and bung that connect the pressure sensor to the conical flask: connect the
pressure sensor to the conical flask , making sure that the pressure changes
within the flask are transmitted accurately

- PASCO Airlink: a Bluetooth connection between the pressure sensor and the
laptop

- A laptop with PASCO Capstone software: necessary for collecting recording and
analyzing data from the experiment

- 25mL measuring cylinder: will be used to get exact measurements of hydrogen

peroxide and ph buffer before pouring it into the 250ml conical flask.

- Ph buffer following these pH levels (5,6,7,8,9): necessary to create different

reaction environments and investigate their effect on the reaction rate. The buffer
solutions ensure precise control of pH levels during the experiment.

- 250mL conical flask: used for preparing and storing solutions of hydrogen
peroxide and catalase before mixing them in the smaller conical flask.

- Knife & Corer: extracting catalase from the potato which is then added to the
reaction mixture. The corer helps with getting consistent potato samples, and the
knife helps with precise cutting

- Potato: will be used as the catalase during the experiment

- Cutting board: used to have a clean sterile board for when cutting the potato.

- Safety goggles: to protect the eyes from any foreign objects harmful to the eyes.

- Lab coat: also to protect the body from any foreign objects and also to keep
clothes uncontaminated.

- Timer
Method

1. Ensure you are wearing safety goggles throughout this experiment.

2. Using a corer to get a piece of potato and using the ruler measure 3 cm from the
potato that got cored out and place into the 250ml conical flask.

3. Use a 25mL measuring cylinder to measure 10ml of the pH buffer (5,6,7,8,9) and
pour all the 10ml of Ph buffer into the 250ml conical flask with the potato and
leave it for the potato to soak it in.
4. While the potato is absorbing the pH buffer in the 250ml conical flask, Connect a
PASCO relative pressure sensor to a bluetooth connector and turn on the
bluetooth connector. The bluetooth light on your Airlink device should now be
flashing red.

5. On a laptop, open PASCO Capstone, select “sensor data” and open a graph.

6. On the panel on the left called Available Wireless Devices, select the AirLink code
shown on your specific device. The bluetooth light on your Airlink device should
now be flashing green. PASCO Capstone should also recognise that you have a
relative pressure sensor connected. Close the panel on the left by clicking the
Hardware Setup button.

7. The y-axis on the graph should already show Relative Pressure. Change the units
from kPa to Pa. The x-axis of the graph should already show Time (s).

8. At the bottom of the screen, change the sample frequency from 20.00 Hz to 1.00
Hz. This means that the pressure will be measured once per second, rather than
20 times per second.

9. At the bottom of the screen, select Recording Conditions > Stop Condition >
Time Based. Change the record time to 60.000s and click OK. This will ensure the
recording period is 60 seconds.

10. Wash the 25 ml cylinder and measure 10 ml of hydrogen peroxide.

11. Pour the 10mL of hydrogen peroxide from the measuring cylinder into the 250mL
conical flask and swirl around to mix well with the pH buffer. Place the bung of
the pressure sensor onto the top of the test tube and twist it a bit to form a tight
seal. Press “start recording” in PASCO Capstone. You should see the data
forming a line with a positive slope stopping at 60s.
 12. Right click on the title of your data where it says “Run 1” in Capstone and rename
it to “Trial 1, (Conc)”

13. Repeat the procedure 4 more times, for a total of 5 repeated trials. To record a
new trial in Capstone, simply click the record button again. Rename each new
“run” to keep your data organised.

14. Before starting a new trial clean the conical flask

Data Processing Method

1. Use the line of best fit tool to select a Linear line of best fit. See Figure 1
below.

2. The value “m” given in the box is the slope of the line of best fit in Pa.s-1. Record
this value in your data table.

3. Click Delete Last Run to delete the data and reset the graph for your next trial.

Table used for recording data data

pH buffer Rate of pressure change (±1Pa/s)

levels
(20ml) 1 2 3 4 5 Mean Half range
5
6
7
8
9

Risk assessment
 Possible risk Precaution

Hydrogen peroxide Is a corrosive chemical and Wear gloves and safety

can cause irritation or goggles while handling
burns to the skin. As well hydrogen peroxide. As well
wear a lab coat to not ruin
caused damage to the
your clothes and protect
eyes. your body from it.

pH buffer solutions may contain chemicals that Avoid contact with skin and
could cause skin or eye eyes, and handle with
irritation. care.

Knife risk of cuts and/or puncture Use carefully and use

wounds if not handled well. appropriate safety
measures such as not
waving the knife around
and having a strong grip on
the handle.

Glassware may break and cause cut Wear gloves when

wounds and any contents necessary and wear a lab
in it can cause other bodily coat to protect yourself
from any shards of glass.
harm.

Electrical hazards Electrical equipment may Know how to turn off

get water or other liquids electrical devices and keep
spilled over it. cables away from liquids.