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Research question

How does the change of products containing enzymes affect the rate of the reaction of the
catalase in the products with H
2
O
2
?

Introduction
Catalase is an enzyme found in nearly all living organisms that are exposed to oxygen,
such as vegetables and fruit. Its main use in connected to the decomposition of hydrogen
peroxide (H2O2).



We had no trial runs before starting this experiment.

Aim
In this experiment we are going to investigate the rate of reactions for different products
containing catalase enzyme will react with a 4% H2O2 solution. We will do that by using
different products containing the enzyme (such as yeast, bananas, oranges etc.), making
them react with the acid, and measuring the amount of oxygen (O2) that comes out.

Hypothesis
My opinion is changing the products used (the independent variable) will give different
rates for their reaction with hydrogen peroxide (more specifically, the rate of reaction of
their catalase with H2O2). That is because every fruit or vegetable has a different
amount/concentration of catalase in it. We have had no trial preliminary data; therefore I
cannot find significance of different possible variables.

Variables
Independent variable - the fruit/vegetable/product used. This variable can be
manipulated by changing the product you use. Example: apple, potato, banana, etc.
However, the mass of the product used has to be the same for a fair test.
Dependent variable - the amount of O2 produced. This variable will depend on which
type of product is used, and will be different for every product used.
Controlled (fixed) variables:
The volume of H2O2 used should be kept the same - otherwise there will be more of the
chemical for the catalase to react with, makes it an unfair test
The concentration of H2O2 should be kept the same as well - that would change the rate
of the reaction in different cases, and that is not what we are investigating, makes it an
unfair test.
The amount of fruit/vegetables, their size, and the number of pieces - more products
used will provide more of the enzyme, which will speed up the reaction rate. Same
applies to their size. The number of pieces might affect the rate as well - as there is more
surface area to react with.
Temperature of H2O2 - higher temperature will increase the reaction rate.
Time for each reaction to elapse - more time will give the chemicals more time to react.
Time, when the reaction starts - as the transfer tube is put under water, it may get filled
with water. That water will require some time to be pushed out - therefore, we will see no
oxygen produced. We will start timing each reaction as soon as there is a tiny bubble on
the end of the tube (the end under water). That way, we can get more results (without half
of them being 0), and probably get more accurate results.
Apparatus
Conical flask (25-50 ml) x1
Test tube (medium size) x4 ! Scales x1
Test tube rack
Water bath or container filled with cold water
H
2
O
2
solution (4%) 400ml
4 products of choice (here yeast, orange, mandarin orange, plum)
Transfer tube for oxygen to go through with stopper on end
Measuring cylinders 10ml x1, 25ml x1, 100ml x1
Cutting board & knife ! Apron and goggles








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Method
1. Set up the apparatus, as shown in the image above.
2. Wear goggles and aprons for safety: H
2
O
2
solutions are corrosive.
3. Get 4 measures of H
2
O
2
(25 ml each) in test tubes, and put those into a test tube rack.
This will help you save time between each try. Remember to use the same beaker of
the solution, to keep the temperature of it relatively the same (fair test)
4. Cut up your product that is being used (in this experiment, plum, mandarin orange,
orange or yeast) into small pieces of 2.5 grams each (precision). Once again, get 4
measures of each fruit/vegetable. Preferably keep the number of pieces for each
measure the same, because the surface area can speed up the rate of the reaction (fair
test)
5. Put one of the measures of the product youre using in a flask, and get ready to start
the timer
6. Pour the H
2
O
2
solution into the flask and plug in the transfer tube
7. As soon as you see a bubble on the end of the tube that is inside the measuring
cylinder underwater, start the timer
8. Record the level of oxygen inside the measuring cylinder every 15 seconds.
9. Once you have measured the level of oxygen every 15 seconds for 2 minutes,
carefully pour out the contents of the flask through a sieve, rinse the equipment, re-fill
the measuring cylinder with water completely and insert the tube in it.
10. Repeat steps 3-9 4 times for each of the 4 products youre using
11. Once done, clean up the apparatus and make sure to wipe the table
12. When writing up the conclusion, do not forget to research on the experiment or ask
your friends with similar experiments to see if your experiment was accurate.



0 15 30 45 60 75 90 105 120
1 0 0,4 0,6 0,6 0,7 0,8 0,9 1,2 1,4 0,01167
2 0 0,3 0,6 0,8 1 1,1 1,2 1,6 1,8 0,01500
3 0 0,3 0,5 0,8 1 1,2 1,4 1,5 1,5 0,01250
4 0 0,2 0,5 0,7 1 1,2 1,3 1,5 1,7 0,01417 1,60
1 0 0 0 0 1 1,2 1,3 1,4 1,5 0,01250
2 0 0 2,5 3 3,5 4 4,3 4,5 5 0,04167
3 0 0 0,5 0,7 1,4 1,7 2 2,3 2,6 0,02167
4 0 1 1,7 1,8 2,1 2,5 2,6 2,8 6 0,05000 3,78
1 0,2 1,6 2 2,2 3 3,7 4 4,4 4,6 0,03833
2 0 0,5 1 1,5 2 2,4 3 3,8 4 0,03333
3 0 1 1,8 2 2,4 3 3,7 4 4,5 0,03750
4 0 1,2 1,8 2,4 3,2 3,8 4,4 4,9 6,6 0,05500 4,93
1 0 5 11 22 33 45 56 67 78 0,65000
2 0 5 15 25 35 47 55 65 70 0,58333
3 0 5 13 24 34 43 51 58 65 0,54167
4 0 5 18 32 45 60 70 78 82 0,68333 0,61
Table above: Raw data: amount of oxygen produced over time
Rate average (V
O2 produced/s)
Rate of reaction
(volume of oxygen
produced/s)
Amount of oxygen produced (cm3) 0.1
Orange
Yeast
Time of the reaction (s)
Product with
enzyme used
Try #
Plum
Mandarine Orange
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0,01167
0,01500
0,01250
0,01417 0,01
0,01250
0,04167
0,02167
0,05000 0,03
0,03833
0,03333
0,03750
0,05500 0,04
0,65000
0,58333
0,54167
0,68333 0,61
Table above: table of rates of reaction of plum, yeast,
orange, and mandarin with H2O2 solution
0,063964
Standard
deviation
Product used
Plum
Mandarine orange
Orange
Yeast
0,001521
0,017352
0,009559
Rate average (volume of O2/s) Rate (volume of O2/s)
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Conclusion
How does the change of products containing enzymes affect the rate of the reaction of the
catalase in the products with H
2
O
2
?
My hypothesis was correct from the bar chart we can clearly see, that the rate of
reaction changes if different products are used. However, we cannot state a clear pattern
in these results. We can say, that all of the products we used have one thing in common
they contain an enzyme called catalase. And the rate of reaction changes, because the
different products that we used have different amounts of catalase in them.
Rate can be calculated using the following formula:
Rate of reaction = amount of O
2
produced/time taken


Evaluation
I think our results are trustworthy. They are quite precise the standard deviation of the
average rates is less than 0.02 for all cases. That implies that the spread of our data
wasnt big at all, and that our experiment was precise.
After the raw results, I have graphed all results I got on 4 different line graphs. That does
not help me with my conclusion; however, it shows that the rate was more or less
consistent throughout the experiment for each product used.
After researching a bit on this experiment on the Internet, I found no experiments that
investigated different fruit/vegetables. And I couldnt compare the data from different
experiments, because the amount of chemicals used and their concentration was different
for all experiments. So, I asked a few of my classmates. And their results were similar to
this experiments. For example, Roy and Goncalo seem to have gotten the same trend in
their results with different reactants the rate of reaction changed. Therefore, our results
can be considered accurate.
Our results are reliable we had 4 repeats for every product used and used 4 different
products. We recorded the volume of oxygen produced 9 times for 2 minutes each.
Our results were also valid we kept the concentration of H
2
O
2
, amount of hydrogen
peroxide used, mass and number of slices of the products used the same throughout the
experiment.
My method was logical, valid and easy to follow nothing had to be changed or added to
the method during the course of the experiment.
Our experiment overall was pretty good apart from a few weaknesses. One of them was
that the products that we picked had a very different amount of catalase in them ranging
from almost none to immense amounts. Perhaps, some research should be done before
picking the products to have more visually appropriate results. Another weakness was
that our experiment was aimed at investigated something that cannot be described and
proved by scientific terms and reasoning the conclusion is very simple.
Therefore, this experiment could be modified to look at the rates of reaction of only one
catalase-containing product with different concentrations of hydrogen peroxide. Another
factor to investigate in this experiment would be if the temperature affects this reaction.

Problem Why was it a problem? Solution
Measurements taken may not
be precise.
Because the smallest
division on the smallest
measuring cylinder is 0.2;
therefore some data can
be not precise.
Use smaller measuring
cylinders with more
divisions, or use another way
of measuring (electronic?)
The timer had to be started
not as soon as the reaction
starts, and so some products
could have already been used
up => rate decreased mid-
reaction
There was still some
water to be pushed out of
the transfer tube, and no
oxygen would come out
until all that water was
pushed out
Plug the end of the tube
underwater with something,
so that no water can come in,
and as soon as the reaction
starts, the plug would come
out. Alternatively, use a one-
way tube (air and water can
come out, but not in)
Lost some time in the
experiment because we
underestimated some
reaction rates
For example, yeast: we
took a 50ml measuring
cylinder, thinking the
reaction cant be faster,
and we ran out of space
for air in less than a
minute
Have a trial run for every
product youre going to use
in the experiment before
starting the actual
experiment. (this would be a
good method improvement,
before all steps)

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