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------The first experiment was unsuccessful and admitted no results as it

was found that the Hydrogen Peroxide used was faulty. Therefore the
first trial will not be recorded or graphed. The following table
records the results of the subsequent experiment in which the Hydrogen
Peroxide was effective.
Table 1: Reaction times collected from the experiment for varying pH
pH Level of Solution (in pH)
Reaction Time (min:ss)
Graph 1: Data from Table 1 [IMAGE]
As seen on the graph, reaction times were the fastest at an acid pH of

4. Reaction times rose dramatically for catalase in a strong acid pH

of 2. Reaction times also rose for reactions carried out in basic pH
The pH of liver was recorded as 5.5.

---------The purpose of this experiment was to investigate the effects of pH on
catalase activity. Since the enzyme was thought to usually be found in
a fairly neutral environment, it was hypothesized that the enzyme
activity would be slowed or stopped in pH solutions that were either
acidic or basic. It was expected that the reaction rate of the enzyme
would be the highest in a solution of pH 7. It was also expected that
the reaction rate would decrease as the pH deviated from 7. This was
due to the observation that the physiological environment in which
most cells are placed have a neutral pH.
The data indicated that the catalase worked most effectively in pH of
4, however it took only 11 seconds longer in the neutral solution.
This is only a small margin that could be explained by many reasons.
There is a possibility that the experiment carried out for a pH of 7
was compromised by slightly different conditions. When the filter
paper was dipped into the liquid liver solution, a fragment of liver
could have attached itself to the filter paper meaning that its weight
was greater and thus it could been more difficult to rise to the top
again. There could have been a slight miscalculation of the amount of
H2O2 put in the pH 7 solution.
Another possible explanation is that catalase works more efficiently
in slightly more acidic solutions. As liver was found to have a pH of
5.5, this being slightly acidic, this is a valid conclusion.
The data indicated that catalase works very inefficiently in a pH of 2
(strongly acidic). The amount of H+ in solution interfered greatly
with the structure of the enzyme causing denaturation.

Although the basic solutions permitted the catalase to carry out its
task eventually, it was not as efficient as the neutral or weak acidic
solutions taking approximately double the time to react.

Errors and Improvements

----------------------There are some ways in which this experiment could be improved. As the
first trial proved, it is essential to ensure that the chemicals used
are not stale. As measurements for the different amounts of chemicals
used, were judged by the human eye, there could have been
inaccuracies. Sometimes the same stirring rod was used for both the
acid, basic, and neutral cylinders, perhaps resulting in a 'neutral'
control solution being slightly acidic or basic.
To further this investigation, there are many potentially interesting
experiments. Essentially, repeating the temperature but with more
variation of pHs between 5 and 8 would lead to ascertaining the exact
optimal pH for catalase in the liver. Also, one could test the impact
of changes in temperature and enzyme concentration on reaction time as
well as testing other enzymes on their substrate to examine whether
comparable results were found. The most important thing is to repeat
the experiment to ensure that there were no problems such as
impurities or miscalculations.

The Decomposition of Hydrogen Peroxide into Oxygen and Water


Catalase is a specific enzyme. Enzymes act as a catalyst and are produced by living
cells. They speed up chemical reactions, an important process for all organisms
(Olson 2002). The enzyme catalase used in this experiment is found naturally in plants
and is known for prevention of oxidative stress from chemicals such as hydrogen
peroxide, H2O2. In aqueous solutions, the activity, specificity, and stability of
enzymes have a dependency on certain reaction conditions. Some of the important
factors affecting enzymes are temperature, pH and substrate concentration (Halling
2001). In the reaction of H2O2 with catalase, the temperature was an important factor
because we had to keep the catalase on ice. If the catalase were to heat up it would no
longer act as a catalyst, because it would be denatured. The sulfuric acid,(H2SO4),
that we added to the solution, lowered the pH and caused the catalase to no longer
When the plant Impatiens flannaganiae is exposed to too much light, an
overproduction of H2O2 results in the leaves. The plant in turn increases its
production of catalase to decompose H2O2. (Lall et al 1999). For our experiment, we
were measuring the amount of substrate, H2O2, which was decomposed into oxygen
and water over specific periods of time. Our hypotheses are:
H0 - The longer the catalase was allowed to react with H2O2, the amount of H2O2
remained the same, and therefore the drops of potassium permanganate, KMnO4, used
to quantify H2O2, would be the same.
HA- The longer the catalase was allowed to react, with H2O2, the amount of H2O2
decreased, and therefore the drops of potassium permanganate, KMnO4, used to
quantify the amount of H2O2, would decrease.
Materials and Methods
We began our experiment by obtaining a baseline measurement of how much H2O2
was in the solution. We put 2 ml of 1.5% H2O2 into a test tube. We added 4 drops of
distilled water, dH2O, which was kept in a plastic bottle, to this solution. We put on a
pair of safety goggles and then added 2 ml of sulfuric acid, H2SO4. We used a plastic
pipette to add the drops of KMnO4, which was contained in a glass bottle. We added
the drops one at a time, to the solution. We tapped the glass test tube each time to mix
the solution. The drops were added until a dark brown color remained uniform
throughout the test tube and the number of drops were recorded. After we did the
baseline, we then did timed reactions. We repeated the same sequence of steps, using a
clean test tube each time, except instead of adding 4 drops of dH2O, we added 4 drops
of catalase extract in a dropper bottle, which was kept on ice in between uses. We used
a timer and timed these reactions at 30 second, 1 minute, 2 minute, and 3 minute

intervals (Olson 2002). Microsoft Word was used to prepare this report. The data were
entered into Microsoft Excel, a spreadsheet program, to produce Table 1 and Graph 1.
Our results, as shown in Table 1, show a general trend that as the time increased, the
amount of drops of potassium permanganate decreased, with the exception of the 30
second timed reaction to the 1 minute timed reaction. The baseline measurement was
not timed and took a total of 90 drops before the solution retained a color. The 30
second timed reaction took 52 drops to retain a color, but the 1 minute reaction took
53 drops. The 2 minute reaction took 47 drops to retain a color. And the 3 minute
reaction took 34 drops to retain a color. The 2 minute and 3 minute reaction times
showed a decrease in the number of drops of potassium permanganate. There was no
data collected for a 6 minute trial. Graph 1 shows a general downward trend except
for the 30 second and 1 minute reaction times in which the number of drops of
KMnO4 increased from 52 drops to 53 drops of KMnO4. Discussion
The purpose of this experiment was to examine the effect of an enzyme, catalase, on
the decomposition of H2O2. The reaction examined was 2 H2O2 ----------------> 2
H2O + O2 catalase
A solution of H2O2 was prepared, the enzyme catalase was added and the reaction
above was allowed to proceed for varying lengths of time. The reaction was then
stopped by adding the acid H2SO4, which causes the denaturation of the catalase and
prevented the catalase from further reaction with the H2O2. The amount of H2O2 that
was decomposed in the presence of the catalase was quantified by adding drops of
potassium permanganate until no H2O2 remained. The amount of KMnO4 added is a
proportional measure of how much H2O2 remains (2 molecules of KMnO4 react with
5 molecules of H2O2). (Olson 2002)
Based on our results, I accept the HA hypothesis, that is, The longer the catalase was
allowed to react, with H2O2, the amount of H2O2 decreased, and therefore the drops
of potassium permanganate, KMnO4, used to quantify the amount of H2O2, would
decrease. However, there were a few problems that we had during our experiment.
First, the experiment was supposed to contain a 6 minute timed reaction but because
we ran out of time during the lab we were unable to complete it. Second, I believe that
an error was made on my part, because when I was dropping the drops of potassium
permanganate in the glass test tube with the pipette, the drops of potassium
permanganate got on the sides of the tube, therefore we aren't certain how many drops
actually went into the solution. This error may have caused an inaccurate count of the
number of drops of potassium permanganate that went into the solution because if we
added too few drops then there would still be H2O2 in the solution to react with

KMnO4 and no color would show. If I added too many drops of potassium
permaganate then I would have an inaccurate estimate of how many drops of KMnO4
it took to decompose the H2O2.
The results of the Impatiens flannaganiae experiment showed that cells in leaves
produce more catalase with increased exposure to light (Lall et al 1999). In our
experiment, instead of increasing the catalase, we allowed the experiment to continue
for a longer period of time. The amount of catalase did not change, but did allow
H2O2, to be decomposed. Possibly increasing the light exposure has the same effect
as allowing the reaction to occur for a longer period of time. I think that this
experiment should have had glass test tubes that were not so tall so that there would
be less of a chance of dripping the KMnO4 on the sides of the tube. Also, I think it
would be beneficial to have a plastic pipette that produced consistent size drops so
that there would be less chance of an error when we calculated the number of drops.
An experiment that might be done in the future would be possibly doing the same
experiment only at different temperatures to see if the temperature affects the rate of
the reaction. This experiment would demonstrate the importance of temperature which
affects enzymes. (Halling 2001) You could also do the same experiment but use a
different catalyst, or use more catalase.