Chemistry HL IA

Comparing the different concentrations of a catalyst in the decomposition of hydrogen peroxide,

as measured by oxygen gas production.
Exploration
Introduction of background

Catalase is a common enzyme found in nearly all living organisms exposed to oxygen. This enzyme
catalyzes the decomposition of hydrogen peroxide to water and oxygen, which in high concentrations is
extremely toxic to a living organism. It is a very important enzyme in protecting the cell from oxidative
damage by reactive oxygen species..

Catalase is a common enzyme found in nearly all living organisms exposed to oxygen, this includes
bacteria, plants, and animals. The enzymes main purpose is to catalyze the decomposition of hydrogen
peroxide to water and oxygen.[1] Catalase protects the cell from oxidative damage by reactive oxygen
species (ROS), and turns the harmful compound into harmless molecules. The aim of this investigation is
to compare the amount of Catalase that is present in several different foods and determine which has the
highest.

Figure 1: Molecular structure of Catalase Figure 2: Visual representation of the enzyme

Hydrogen peroxide is a chemical compound with the formula H₂O₂. In the body it is produced primarily
in three places: lung, gut, and thyroid gland, as a result of normal metabolic processes. While having a
lower toxicity in smaller concentrations, Hydrogen peroxide can be fatal if ingested or acumulated in high
concentrations. In its purest form, it is a very pale blue liquid, being a slightly more viscous compound
water. However it’s rarely in a pure form, often distilled to a 3% concentration for household use,
however higher concentrations are made for more industrial uses. Hydrogen peroxide is classified as one
of the simplest peroxides, and has a wide range of common uses such as, an oxidizer, bleaching agent,
and antiseptic. Hydrogen peroxide is very unstable in nature, and decomposes readily to oxygen and
water with release of heat.

Catalase is a natural occurring enzyme that helps to catalyze the decomposition of hydrogen peroxide. It
has one of the highest turnover numbers of all enzymes. The high turnover number indicates that one

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catalase molecule can convert millions of hydrogen peroxide molecules to water and oxygen each second.
According to a study conducted by Britton Chance, a 1M solution of Hydrogen peroxide has a turnover
number of 3.5 x 10​7​ times a second.

Yeast is a living microscopic organism which converts sugar or starch into alcohol and carbon dioxide,
and contains high amounts of the enzyme catalase, and would be used to quikly decompose any remaining
hydrogen peroxide left over.

In nature different foods contain varying amounts of Catalase, and are often cited in articles and
supplements as sources of Catalase for different effects. Primarily marketed as a supplement to slow the
greying of hair. While unsure of the validity of those claims, they got me curious on whether or not the
claims of the existence of Catalase in different foods.

Variables

Independent The type of food used in creating each food puree mixture. Created in
Variable a 1:1 ratio of food item to water.

Dependent Variable The amount of gas produced in a set time, This will lead us to
compare the effectiveness, and estimated amount of catalase.

Controlled variables The amount of each The ratio of puree mixture to hydrogen
mixture/solution peroxide is kept the same at a 1:5 ratio. 5ml of
the puree mixture to 25 ml of Peroxide

Concentration The same concentration of peroxide was used

for each trial, as well as the same
concentration of puree mixture.

Temperature The mixtures were left to sit for 30 minutes to

reach room temperature before continuing in
the experiment

Time A time of 1 minute was used in all gas

collections.

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Materials

Solution Preparation Measuring Apparatus Experimental Materials

Sweet Potato Each item 250 mL Graduated Cylinder Rubber stopper with plastic
was from it's tube
naturally
White Onion grown state. 100 mL Graduated Cylinder 500 mL Erlynmyer flask

Banana Weighboats Yeast

Avocado Digital Thermometer 250 mL Beaker

Spinach Dropper

Broccoli Stopwatch

Distilled Water Electronic Balance

Stirring Rod

Immersion Blender

Large Plastic Container

Procedure
Creation of the Food Puree mixtures
1. Each food item is diced into roughly 1 cm pieces and set aside for 5 minutes to come to room
temperature.
2. Using a weighboat and electronic scale, 60 grams of the diced pieces are weighed and set into the
blender
3. 60 ml of distilled water are then added to the blender, and the mixture is blended until no visible
pieces are left, forming a homogeneous mixture.
a. The consistency varied from each food item, however the ratio of 60 grams of food to 60
ml of water was kept constant
4. The now pureed mixture is then transferred from the blender and into a larger container and sits
for 15 minutes to come to room temperature
a. An electric thermometer was used to ensure that each mixture had the same temperature
5. The Blender is washed out thoroughly under hot water and soap, rincing several times, before
repeating the steps with a different food item

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 a. This was repeated until all 6 food items were turned into pureed mixtures.

Creation of yeast mixture

1. 60 Grams of yeast are measured out using the electronic scale and weigh boats and poured into a
250 mL beaker
2. Using the graduated cylinder, 60 ml of water is measured out and poured into the beaker
containing the yeast.
3. The mixture is then stirred for about a minute before letting sit for 10 minutes.
4. This process was then repeated 6 more times per each food sample creating 6 100 ml mixtures in
total.
a. This was done to help keep the rate each mixture of yeast settled constant.

Figure 3, Representation of gas collection method

Experimentation
1. First gather all the necessary materials.
2. Then set up the gas collection. Filling a large container with water, before filling the 250
graduated cylinder with water and turning it upside down so the opening of the cylinder was was
submerged underwater, and the water stayed in the cylinder (figure 3)
3. The end of the stopper and tube was placed inside the cylinder, still keeping it submerged
underwater.
4. 25 ml of 3% Hydrogen Peroxide was measured out using the smaller graduated cylinder and
poured into the flask
5. Next 5 ml of the food mixture was measured out poured into the Hydrogen Peroxide flask quickly
placing the rubber stopper on
6. The flask was swirled around for about 5 seconds before letting the reaction take place.
a. The plastic tube would push out oxygen gas displacing the water in the upside down
graduated cylinder.
7. The reaction was timed for 1 minute before removing the tube from the cylinder and recording
the total gas
a. The end of the tube would be covered to prevent any gas from leaking out
8. The cylinder would be refilled and reset before replacing the end of the tube.
9. The stopper would be removed and the 100 ml yeast mixture would be poured into the flask
before resealing and swirling the flask for 10 seconds.
a. This was done in order to react to any remaining hydrogen peroxide leftover.

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 10. The reaction would let run until the rate of gas production slowed down to a significant amount.
11. The time and gas would be recorded before moving on to the next trial. Repeating each step for
each trial and each food item.

Experimental results

Initial gas collected

collected volume of gas per food item (± 0.5 mL)

Trials
Sweet White Spinach Broccoli Avocado Banana
Potato Onion

1 204 ml 1.00 ml 156 ml 97.0 ml 103 ml 73.0 ml

2 201 ml 2.00 ml 150 ml 96.0 ml 104 ml 77.0 ml

3 205 ml 0.00 ml 159 ml 95.0 ml 99.0 ml 78.0 ml

4 205 ml 2.00 ml 155 ml 97.0 ml 100 ml 79.0 ml

5 203 ml 1.00 ml 153 ml 93.0 ml 101 ml 77.0 ml

Average 203.6 ml 1.20 ml 154.6 ml 95.6 ml 101.4 ml 76.8 ml

204 ml 155 ml 101 ml

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As seen in the graph, there is a difference between each food and the amount of O2 gas collected
after the 1 minute period. A higher amount of gas would indicate a higher concentration of
Catalase present in the food due to it's high Turnover number.

It's seen that the Sweet Potato had the highest recorded gas output while the White Onion had virtually
none.

Gas collected after yeast was added after initial reaction

collected volume of gas per food item (± 0.5 mL)

Trials
Sweet White Spinach Broccoli Avocado Banana
Potato Onion

1 30.0 ml 230 ml 76.0 ml 135 ml 130 ml 154 ml

2 29.0 ml 228 ml 78.0 ml 137 ml 133 ml 150 ml

3 28.0 ml 236 ml 73.0 ml 139 ml 134 ml 153 ml

4 27.0 ml 235 ml 71.0 ml 136 ml 128 ml 152 ml

5 30.0 ml 233 ml 75.0 ml 139 ml 129 ml 154 ml

Average 28.8 ml 232.4 ml 74.6 ml 137.2 ml 130.8 ml 152.6 ml

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 232 ml 137 ml 131 ml 153 ml

By adding the data regarding the yeast results onto the previous bar graph, a better analysis of the trend
shown.

By combining the total gas collected from the same sample of hydrogen peroxide it becomes clear that
there is a limit to how much gas the decomposition of the peroxide can yield.

Calculations
First it’s important to note that by adding the initial gas collected from just the puree and then from the
yeast, the total amount of gas the sample released can be measured.
Sweet White Spinach Broccoli Avocado Banana Average
Potato Onion

Total 232.8 ml 233.2 ml 229.6 ml 232.6 ml 232 ml 229.8 ml 231.7 ml

collected
gas

Stoichiometry
Finding expected liters of Oxygen gas based on the strength of the peroxide.

Reaction taking place:

2H​2​O​2​ + Catalase -> 2H​2​O + O​2

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3% Hydrogen Peroxide implies that there are 30 g of peroxide in 1000 ml (1 liter), by divining 30 g/l by
the molar mass of hydrogen peroxide (34.0147 g/mol) the Molarity of 3% peroxide is able to be decided
to around 0.882 M.

M ol
By manipulating and filling in the information on the Molarity formula M = Liter the Mols of peroxide
can be deduced. In the samples 25 ml (0.025 L) of peroxide was used. From the mols of peroxide mols of
O​2​ can be calculated and from there using the relation 1 Mol O​2(g)​ = 22.4 L the total amount of oxygen gas
that when decomposed completely hydrogen peroxide can produce.

M ol 0.022 M ol H 2 O 2 1 mol O 2 22.4 l

0.882 = 0.025 mol = 0.022 H 2 O 2 * 2 mol H 2 O 2 * 1 mol O 2 = .2464 l (246.4 mL)

Qualitative Observations

Sweet Potato White Onion Spinach Broccoli Avocado Banana

-Created the - Faint - created a -created a -created a -created a

most foam buzzing thick green thin green dense foam dense foam
- inflated the sound foam foam -didn’t puff -didn’t puff
fastest -the mixture -inflated -inflated up much up much
- small fizz at had small relatively quickly -there was a -there was a
beginning, bubbles quickly -the flask temperature temperature
but quickie - no -the flask heated up change change
foamed up temperature heated up -a fizzing -audible -less audible
-the flask change -a fizzing sound fizzing fizzing
heated up sound - the foam - filled about - filled
- the flask - the foam filled less a third of the around a third
was almost nearly filled than half way flask of the flask
completely the flask
full of foam

Conclusion

For each of the foods chosen, the trials showed that certain foods had a higher concentration of
the enzyme catalase present. Some of the foods chosen like Broccoli and Avocado, showed a
very similar reaction, which indicates that they have similar levels of the enzyme, while the
potato was seen to have high concentrations of the enzyme. This investigation provided
significant results to prove that sweet potatoes have a high concentration of catalase.

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Errors
Systematic
Loss of O​2​ gas from preparing the next test: ​Precautions were taken to minimize the loss of gas
produced in the flask, but one potential error is when adding the yeast mixture which required the flask to
be opened, and for gas to potentially escape.

The reactions didn’t go to completion: ​As a reaction takes place, after a certain point the reaction
rate starts to decrease and slow down. This could explain why the calculated volume of O​2 gas
​ didn’t
match the average from the data.

Random
Errors in Solution Preparation: ​Due to the random error of the electronic balance used as well
as the graduated cylinder, the prepared solutions may not have been exactly 1%. This mostly
impacts the amounts for each substance, the puree and peroxide, which react with each other.

Errors in Environmental Conditions: ​Because of school closures due to Covid-19, this

experiment was not performed in an informal setting, a kitchen, rather than a school chemistry
lab. Apparatus were cleaned thoroughly before use and in between each test, but it's possible
there was food contamination in the saucepan as well as the immersion blender and large
containers.

Further Investigation

The impact temperature has on enzyme effectiveness: ​Many enzymes tend to behave
differently depending on the temperature at which they're kept or in. A potential future
investigation could involve further exploration involving changing the temperature each food is
kept at or prepared.

Preparation: ​Each food remained raw and un prepared. The biggest change that each item
underwent was the change into a puree. A future experiment could investigate the effects
cooking has on the presence of the enzyme

Different catalysts or substances: ​Hydrogen peroxide has many different catalysis, and it’s
possible to explore those venues more in depth to future investigate the effect catalysts have on
hydrogen peroxide, additionally it would be worth investigating on whether or not another
enzyme like Catalase exists, or if it has any other functions.

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Works cited

Chance, Britton. “THE COMPOSITION OF CATALASE-PEROXIDE COMPLEXES *.” ​Jbc.org,​

www.jbc.org/content/179/3/1311.full.pdf.

book, Chemical. “CATALASE.” ​CATALASE | 9001-05-2,​ 2015,

www.chemicalbook.com/ChemicalProductProperty_EN_CB9273573.htm.

Kaur, Kiranpreet. “Turnover Number.” ​Chemistry LibreTexts​, Libretexts, 11 Aug. 2020,

chem.libretexts.org/Bookshelves/Biological_Chemistry/Supplemental_Modules_(Biological_Chem
istry)/Enzymes/Enzymatic_Kinetics/Turnover_Number.

PC;, Chelikani P;Fita I;Loewen. “Diversity of Structures and Properties among Catalases.” ​Cellular
and Molecular Life Sciences : CMLS,​ U.S. National Library of Medicine,
pubmed.ncbi.nlm.nih.gov/14745498/.

Roskoski, Robert. “Turnover Number.” ​Turnover Number - an Overview | ScienceDirect Topics,​

2015,
www.sciencedirect.com/topics/biochemistry-genetics-and-molecular-biology/turnover-number.

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