The Effects of

Hydrogen
Peroxide and
Catalase
October 4, 2010
Periods 6 & 7
Introduction
In order to obtain energy and building blocks from food, the digestive system must break
down proteins, fats, and carbohydrates. In this process, specific enzymes catalyze hydrolysis
reactions in which food polymers are broken up into monomers. Human cells are always making
poisonous chemicals. They do not die because they use enzymes to break down these chemicals
into harmless substances. Enzymes are proteins that speed up the rate of reactions that would
otherwise happen ore slowly. The enzyme is not altered by the reaction. Liver and other living
tissues contain the enzyme catalase. This enzyme breaks down hydrogen peroxide, which is a
harmful by-product of the process of cellular respiration if it builds up in concentration in the
cells. The reaction between hydrogen peroxide and catalase is as follows: 2H2O2  2H2O + O2.
This reaction is important to cells because hydrogen peroxide is produced as a byproduct of
many normal cellular reactions. If the cells did not break down the hydrogen peroxide, they
would be poisoned and die.
The purpose of this lab is the study the catalase found in liver cells, and the effects of
adding hydrogen peroxide to it under various conditions. Also, to test various other cells, such as
apple, ham and potato cells, for the presence of the enzyme catalase.

Hypothesis
Part A: If the liver is placed in 2 ml of 3% hydrogen peroxide, then the result will be a
high rate of reaction because catalase reacts with hydrogen peroxide to form hydrogen and
oxygen.
Part B: If pieces of potato, chicken, and apple are placed into 2 ml of hydrogen peroxide
separately, then only the chicken will have a reaction.
Part C: If the liver is placed in 2 ml of hydrogen peroxide at room temperature, then there
will be a high rate of reaction, otherwise the enzyme will become denatured at too high or too
low a temperature.
Part D: If the pH is neutral, then the enzyme will react properly with the hydrogen
peroxide.

Materials & Method

14 clean test tubes, one of which must be glass. A test tube holder. Scissors or a razor
blade to cut up the meat and other foods. A hot plate to boil and warm the water. One 10-ml
graduated cylinder to measure the hydrogen peroxide and 40 ml of a 3% hydrogen peroxide
solution. A thermometer and a stirring rod. pH paper. Fresh cow liver, chicken, apples and
potatoes. A small container to dump the wastes from the experiment. Half a cup of distilled water
to cover the liver when boiling. 3 clean glass cylinders to hold the water that will either be heated
or iced. A bag of ice for the ice bath. A stop watch to time the baths.
Part A
First, place 2 ml of the 3% hydrogen peroxide solution into a clean test tube. Using the
forceps and scissors, cut a small piece of liver and add it to the test tube. Push the liver slice into
the hydrogen peroxide with the stirring rod and observe the reaction. Place 2 ml of the hydrogen
peroxide solution into a clean test tube and again add a small piece of liver and observe the
reaction. Then pour off the liquid into a second clean test tube. Add a piece of liver to this used
liquid and observe what happens. Then, add another 2 ml of hydrogen peroxide to the liver
remaining in the first test tube and observe what happens.
Part B
Place 2 ml of hydrogen peroxide into a clean test tube. Then cut a small piece of potato
and add it to the solution and record what happens. In another test tube add 2 ml of hydrogen
peroxide and a small piece of chicken. To a third test tube, add a small piece of apple and 2 ml of
hydrogen peroxide. For each, observe and record the reaction.
Part C
Put a piece of cow liver into the bottom of the clean glass test tube and cover it with a
small amount of the distilled water. Fill one of the glass cylinders with water and put it on the hot
plate to boil. When it begins boiling, place the test tube into the boiling water bath for 5 minutes.
Using the test tube holder, remove the test tube from the boiling water and allow it to air cool.
Then pour out the water into the waste container and add 2 ml of hydrogen peroxide to the boiled
liver. Observe and record what happens.
Then, put equal amounts of liver into 2 clean test tubes and in 2 other test tubes, add 1 ml
of hydrogen peroxide to each. Put on of the liver test tubes and one of the hydrogen peroxide test
tubes into an ice bath (a cylinder filled with ice water) and the other 2 test tubes of liver and
hydrogen peroxide into a warm water bath (a cylinder filled with water and set on the hot plate
that is set to low with water temperature at 40°C). Time for 3 minutes, then pour the cold tube of
H2O2 (hydrogen peroxide) into the cold liver test tube and the warm tube of H2O2 into the warm
liver test tube and observe and record the reactions.
Part D
Prepare 3 tubes as follows: one tube with a pH of 3, so it is acidic, one tube with pH of 10
(basic), and one tube with a pH of 7 (neutral). Use the pH paper to ensure the pH of each test
tube. Use distilled water for the tube with pH 7. To each of the three tubes add 2 ml of hydrogen
peroxide. Add a small piece of liver, each about the same size, to each test tube. Observe and
record what happens.
Results
Here is a data table with the rates of reaction (with 0 being no reaction and 5 being a great
reaction) and observations recorded during the experiment.

Rate of Reaction
Observations
(0-5)

-Has the temperature of the test tube

Normal Liver 4 gotten warmer or colder? The tube got
warmer, so the reaction is exothermic.
-Explain your results (what is the
Part A Liver added to used liquid composed of?) The liquid after
0
peroxide the reaction is just H2O, so nothing
happens when a new piec of liver is
Reused Catalase 4 added to the used hydrogen peroxide.
-Is catalase reusable? Yes, it is.
-Which tissues contained catalase? The
Potato 1 potato contained catalase
-Do some contain more catalase than
Chicken 0 others? Yes. The potato contained less
Part B
catalase than the liver.
-How can you tell? The rate of reaction
Apple 0 tells how much catalase is contained in
each tissue.
-What will boiling do to an enzyme?
Boiled Liver 0 Boiling an enzyme will denature it.
-What is the “optimum” temp for
catalase? Warm (40°C)
-Why did the reaction proceed slowly
Part C Ice Bath Liver 3 at 0°C? Because the enzyme moved
slower because of the freezing temp
-Why did the reaction not proceed at
Warm Liver 5 all at 100°C? The enzymes were
denatured
Basic Solution (pH -Does there appear to be a pH
2 “optimum? If so, at what pH? Yes, at
10)
pH 10
-What is the effect of low or high pH
Part D Acidic Solution (pH 3) 1 on enzyme activity? At a low pH, the
liver got cooked, but there was little
Neutral Solution (pH reaction. At high pH, the reaction was
3
7) close to the rate of the neutral solution.
As shown to the left, the liver had the highest rate of reaction, so it must contain the most
catalase.

To the right, the effect of temperature on catalase activity is shown. When the liver is boiled, the
enzyme is denatured, so there is no reaction with hydrogen peroxide. However, when the liver is
put in freezing water, the enzyme slows down, so there is a reduced reaction occurring. The
optimum reaction occurs at 40°C.

Catalase seems to work best at a pH of 7. When the pH is too high or low, it becomes inactive, or
even denatured.

Error Analysis
Because there were many groups conducting the same experiment at once, and each
group had four people to decide what the rate of reaction was, the rate of reaction recorded may
not be accurate. The rate of reaction also may not be accurate because each step was not repeated
multiple times to ensure precision. There were not enough test tubes to use a completely clean
one every time, so some used ones were rinsed out and reused. They might not have been rinsed
completely clean, which may have affected some of the reaction rates. There is catalase
contained in chicken, just not in the part that was used in the experiment, which messed up the
hypothesis and results.

Discussion and Conclusion

From the data collected, one can conclude that the liver contains significant amounts of
catalase, which can be reused. Out of the chicken, potato, and apple, only the potato contained
even trace amounts of catalase, which caused a minimal reaction with the hydrogen peroxide.
The hypothesis that the chicken would have catalase was correct, although that was not
confirmed by the experiment because there is a minimal amount of catalase in chicken in
general, but most of it is in the liver, which was not the part tested. The experiment proved that
after a reaction with catalase, hydrogen peroxide really does turn into hydrogen and oxygen,
making it harmless to cells. Catalase has both an optimal pH and temperature in which it can
work in; the temperature being around 40°C and the pH being 7. Anything out a range of those
two will cause the enzyme to become denatured or inactive, signifying that there is a specific
environment in which the enzyme must have to work properly.
Further experimentation is needed to fully determine the exact temperature and pH range
in which catalase can be fully utilized. Additional experimentation is also needed to determine
how much catalase a potato contains, and the effects of temperature and pH on it.