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The effect of concentration of catalase on the rate of


decomposition of hydrogen peroxide

Jungho Park
P.3
Jung Ho Park
IB Biology SL
Lab Instructor: Ms. Labudda Lab Date: November 11, 2015

The effect of concentration of catalase on the rate of decomposition of hydrogen peroxide

Research Question

How does different concentration of catalase change the rate of decomposition of hydrogen
peroxide into water and oxygen in a fixed time?

Background

Enzyme is proteins that act as biological catalysts in living organisms. Catalysts increase the
rate of metabolism in living cells without being consumed or altered permanently in a reaction,
and therefore they can be used over and over again. In catalytic reaction, substrates are bind to
the active site of the catalysts and broken down/built up into different molecules called products.

Catalase is an enzyme found in living cells which catalyzes the decomposition of hydrogen
peroxide into water and oxygen. Hydrogen peroxide is a cytotox-toxic in living cells-agent in high
level of concentration in animal and plant cells and it should be eliminated from living cells by
enzymes such as catalase.
The reaction of catalytic decomposition of hydrogen peroxide in living tissue by catalase is as
following:
2 H2O2 → 2 H2O + O2

In this experiment, we investigate the effect of concentration of catalase on the rate of


decomposition of hydrogen peroxide. We extract catalase from grinded potatoes and add
increasing amount of catalase to hydrogen peroxide in separate test tubes. Decomposition of
hydrogen peroxide into water and oxygen can be observed in form of foam/bubble on the
surface of hydrogen peroxide and grinded potato mixture. The concentration of catalase is
varied by adding different amount of potatoes and water with respect to the total volume of 10
mL of mixture of potatoes, water and hydrogen peroxide. The volume of hydrogen peroxide was
kept constant at 5 mL in all test tubes while the volume of water and grinded potatoes were
adjusted to manipulate the concentration of catalase in the mixture.

Hypothesis
The hypothesis for this experiment is that the rate of decomposition of hydrogen peroxide will
increase with the increase in the concentration of catalase. However, the rate of decomposition
of hydrogen peroxide will increase at slower rate with each increment of increase in catalase
concentration, and eventually stop increasing at a point in which the concentration of hydrogen
peroxide becomes a limiting factor. At high catalase (enzyme) concentration, all hydrogen
peroxide (substrate) have been broken down into water and oxygen (products) and there is no
hydrogen peroxide to decompose in the mixture, and no further increase in catalase
concentration will increase the rate of decomposition. The rate of decomposition will now
increase further with the increase in the concentration of hydrogen peroxide-increase in
substrates.

Justification for the research question

High level of hydrogen peroxide is toxic to living cells since hydrogen peroxide is a mild
oxidizing or mild reducing agent and can oxidise DNA, lipids and proteins. This experiment can
be used in medical industry to aid elimination of hydrogen peroxide in bodies lacking catalase
enzymes or having catalase that functions poorly. The decomposition reaction of hydrogen
peroxide can be boosted by injection of higher concentration of catalase into animal or plant
cells if it is determined in this experiment that higher concentration of catalase yields higher rate
of decomposition of hydrogen peroxide.

Variables

Independent variable: concentration of catalase extracted from grinded potatoes


- Concentration of catalase are varied by adding different amount of potato juice and
water to yield different catalase concentration-10%, 20%, 30%, and 40%-with respect to
total volume of 10 mL, with the volume of 3% hydrogen peroxide kept at 5 mL in all test
tubes.
- Limitations are the measurement of exact concentration of catalase extracted from
grinded potatoes since grinded potatoes were used as a primary source of catalase,
instead of pure catalase extract.
- The limitation could alter and add uncertainties on the catalase concentration in the
mixture since each potatoes contain different amount of catalase, and other substances
contained in potatoes could affect the decomposition of hydrogen peroxide. The errors
caused by this limitations were reduced by having five different test tubes for each
catalase concentrations-10%, 20%, 30%, and 40%- and by taking two minutes for the
duration of the reaction recorded instead of just one minute. The uncertainties in the
catalase concentration are +/- 5%.
Dependant variable: the height of foam, formed above liquid, measured with a ruler in
millimeters.
- The height of foam was measured from the surface of mixture to the top of foam.
- The limitation comes from the irregular shape and vulnerability of foam which makes it
difficult to determine the top of foam.
- The limitation caused by irregularity and vulnerability of foam can affect the
measurement of the height of foam.
- Measuring apparatus such as rulers and graduated cylinders also adds uncertainties of
+/- 0.5 mm and +/- 0.5 mL.

Constant variable: temperature, type of enzyme (catalase), pressure, reaction time recorded,
amount of H2O2 used in the mixture, total volume of the mixture, and measurement methods,
- The temperature and pressure were monitored and kept constant by conducting the
experiment in the same room with windows and doors closed. Limitations are individual
activity within the classroom which can change the temperature and pressure in the
room.
- The type of enzyme used, catalase, was kept constant by using grinded potato as the
primary source of enzyme throughout the experiment. Limitation is that potato might
possess other enzymes aside from catalase which also breaks down hydrogen peroxide.
- Reaction time was recorded for two minutes with stopwatch. Stopwatch was started as
soon as catalase was added in the test tube containing water and hydrogen peroxide,
and stopwatch was stopped after two minutes. Human error such as starting the
stopwatch a second or two late can affect the rate of reaction.
- The volume of hydrogen peroxide added in the test tube was kept constant at 5 mL in
five test tubes of all five catalase concentration. The limitation experienced in the
classroom was the method of distribution of hydrogen peroxide. 3% concentration
hydrogen peroxide was sitting in the beaker for a while and that might have caused
difference in actual concentration of hydrogen peroxide used in the experiment.
- The measuring method in which the foam was measured from on the surface of the
mixture in the test tube to the top of foam, was kept constant and used by all lab
partners. Since each lab members measured the height of foam in different
concentration of catalase, the measuring method could have been not consistent,
yielding higher chances of error and uncertainties of the data.
Apparatus

- Digital stopwatch, accurate to 0.01 seconds


- Ruler, accurate to mm
- 20 test tubes
- Test tube rack
- three 200 mL beakers

Materials
- 3% hydrogen peroxide from stockroom
- Potatoes
- Distilled water from the tap

Procedure

1. Obtained 20 test tubes and 4 test tube rack


2. Placed 5 test tubes in each test tube rack.
3. Labeled each test tubes as 10%, 20%, 30% and 40% respectively which indicate the different
concentration of catalase .
3. Obtained 3% hydrogen peroxide and distilled water placed each in separate 200 mL beaker.
4. Lab instructor grinded potatoes in a blender and filtered out the solid residue. Placed the
supernatant liquid in 200 mL beaker.
5. Mixture of distilled water, 3% hydrogen peroxide with four different concentration of
catalase-10%, 20%, 30%, and 40%- was made by adding the following volumes of each
solution shown in the tables below to add up total volume of 10 mL.

Raw Data
Amount of Catalase in Test Tube and the Height of the Foam

0%catalase into the test tube.


Test tube 3%H2O2 ml H2O ml Catalase ml Resulting foam
mm

A 5.0 5.0 0 0

B 5.0 5.0 0 0

C 5.0 5.0 0 0

D 5.0 5.0 0 0

E 5.0 5.0 0 0
(uncertainty is +/- 0.5 ml, +/- 0.5 mm)
1% catalase into the test tube
Test tube 3% H2O2 ml H2O ml Catalase ml Resulting foam
mm

A 5.0 4.0 1.0 5.0

B 5.0 4.0 1.0 7.0

C 5.0 4.0 1.0 4.0

D 5.0 4.0 1.0 6.0

E 5.0 4.0 1.0 7.0


(uncertainty is +/- 0.5 ml, +/- 0.5 mm)

2% catalase into the test tube


Test tube 3%H2O2 ml H2O ml Catalase ml Resulting foam
mm

A 5.0 3.0 2.0 11.0

B 5.0 3.0 2.0 10.0

C 5.0 3.0 2.0 11.0

D 5.0 3.0 2.0 11.0

E 5.0 3.0 2.0 11.0


(uncertainty is +/- 0.5 ml, +/- 0.5 mm)

3% catalase into the test tube


Test tube 3%H2O2 ml H2O ml Catalase ml Resulting Foam
mm

A 5.0 2.0 3.0 38.0

B 5.0 2.0 3.0 48.0

C 5.0 2.0 3.0 43.0

D 5.0 2.0 3.0 40.0

E 5.0 2.0 3.0 23.0


(uncertainty is +/- 0.5 ml, +/- 0.5 mm)

4% catalase into the test tube

Test tube 3% H2O2 ml H2O ml Catalase ml Resulting Foam


mm

A 5.0 1.0 4.0 62.0

B 5.0 1.0 4.0 65.0

C 5.0 1.0 4.0 56.0

D 5.0 1.0 4.0 63.0

E 5.0 1.0 4.0 65.0


(uncertainty is +/- 0.5 ml, +/- 0.5 mm)

6. Each lab members were in charge of conducting an experiment with different concentration of
catalase.
7. In all test tubes, catalase was added lastly. Stopwatch was started the moment catalase was
added in the test tubes, and stopped after two minutes.
8. After foam has appeared on the surface of the mixture, the height of foam was measured with
a ruler from the surface of the mixture to the top of the foam in mm.
9. The height of foam in each test tubes were recorded in google document.

Processed Data

The Average Height of the Foam Made in Each Solution


Concentration of Catalase(%) Height of the Foam (mm/min)

0% 0

1% 2.9

2% 5.4

3% 19.2

4% 31.1
(uncertainty of the height of the foam is +/- 0.5 mm/min)

The height of bubble formed in a minutes and the rate of decomposition of hydrogen peroxide.

Sample Calculations

1. Rate of reaction

The height of bubbles formed in 5 trials for each concentration of catalase was added then
divided by 5 to yield the average height of bubbles in each concentration. The average height of
bubbles in each concentration was then divided by 2 to get the rate of decomposition in mm/min
since the reaction of decomposition was carried out in two minutes.

The example calculation with the value obtained from 1% catalase concentration:

Average of the height of bubble formed:

(5mm+7mm+4mm+6mm+7mm) / 5 = 5.8mm

Rate of reaction (mm/s):

5.8 / 2 = 2.9mm/s

2. Uncertainty of the height of the foam

Since I have used a ruler with the smallest measurement of 1 mm, the uncertainty of the foam’s
height will be +/- 0.5 mm

Data Collection-qualitative data


When catalase (supernatant liquid extracted from potatoes) was added to the test tube, bubbles
foamed quickly throughout on the surface of the mixture.

Data presentation

Figure 1. The rate of decomposition of hydrogen peroxide (mm/min) against the concentration of
catalase (%). (used the standard deviation to show uncertainties)

Our table agreed to use standard deviation. As shown is raw data section, some solutions’ foam
heights are not constant. Therefore, we thought that the normal way of determining the
uncertainty (dividing the smallest unit measurement in the measuring device that was used by
two) would not represent our data well, but standard deviation will.

Conclusion

The rate of decomposition of hydrogen peroxide into oxygen and water, catalyzed by the
enzyme catalase from potato skin, increases with the concentration of catalase. Since the
catalysts boost up the speed of the reaction, the reaction rate will go up when there is more
catalysts, drawing exponential graph. The result supports the hypothesis in this experiment.
However, we also hypothesized that the rate of decomposition will increase with a slower rate
and eventually stop increasing at certain point in which the concentration of hydrogen peroxide
becomes the limiting factor of the rate of decomposition, which will be portrayed as a plateau
graph. Although it is officially known that catalyst had a limiting state where it can no longer
make the speed of reaction faster, this experiment does not support this part of hypothesis since
the graph in the result section is upward sloping without being stagnant. However, if we had
more trials with broader concentration of catalase, the graph will definitely going to be a form of
plateau graph.

Evaluation

The rate of decomposition of hydrogen peroxide is tested and observed with increasing
concentration of catalase from potato skin. The result we obtained agrees with the hypothesis
that the rate of decomposition of hydrogen peroxide increases with the concentration of catalase
in the mixture. Nevertheless, the other part of hypothesis that the rate of decomposition will stop
increasing with the catalase concentration at a point in which all hydrogen peroxide have been
used up has not been with an agreement with our experiment. This happened due to lack of
trials. I am very certain that if we had more trials with broader percentage of catalase
concentration, the results will agree with our second hypothesis as well.

A control trial was set up so that the mixture of distilled water and hydrogen peroxide contains
no potato skin, and thus no catalase enzyme.

30% catalase concentration had standard deviation of +/- 12.5 which makes the data
untrustworthy. This could have caused by the unevenly distributed potato skin in the mixture.
This can be prevented or improved if we use glass rod to mix the mixture thoroughly before
observing the reaction. Furthermore,I remember how the potato was already grinded for us, and
when it was time for me to get some catalase, I was able to see layers of potato bits separated
from the catalase. These might cause unequal distribution of catalase which will directly impact
the results.

Improvements
3 % concentration hydrogen peroxide solution was distributed in class to an open beaker. This
could have altered the actual concentration of hydrogen peroxide used in the experiment due to
errors coming from the environment such as evaporation of the solution. This will change the
height of the foam by reacting less/more with the catalase. (If the solution evaporates and make
the concentration higher, it means that there are more hydrogen peroxide compared to original
solution even if the mass is the same.)

In addition, the rate of decomposition did not stop increasing and did not stay constant with
increasing catalase concentration. One improvement to make to prove this part of hypothesis is
testing with higher concentration of catalase or increasing the testing catalase concentration
closer to 100% instead of stopping at 40 % catalase concentration.

Finally, our group should not forget about human error. Some of the teammates were not aware
of the fact that they should measure the foam right after 2 min. This must have affected the
heights of the foams directly. Next time, they should listen and read more precisely.