Exemplar Chemistry IA

Score: 24/24

Relevant terms:
Rate of reaction, catalysis, activation energy, Arrhenius’s equation,
hydrogen peroxide.

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AN EMPIRICAL DETERMINATION OF THE ACTIVATION ENERGY OF DECOMPOSITION OF HYDROGEN PEROXIDE IN THE
PRESENCE OF CATALASE :

1. Introduction:
The study of catalysis in Chemical Kinetics remained with me as one of the great testaments for how
overwhelming practical significance can arise from theoretical discovery. At least from an industrial
perspective, catalysis, since the invention of the concept in 1794 by Scottish chemist Elizabeth Fulhame, has
been indispensable in industrial processes such as the Contact and Born-Haber cycle. However, through
deeper research I found that catalysts have been in use long before its discovery by mankind, existing as
enzymes which sustain reactions occurring within biological life. For example, the body’s defensive
mechanism against the unfavourable decomposition of hydrogen peroxide into hydroxyl radicals involves the
organic catalyst, ‘catalase’, which safely and quickly decomposes hydrogen peroxide into harmless products:
𝐶𝑎𝑡𝑎𝑙𝑎𝑠𝑒
(1) 2𝐻2 𝑂2 (𝑎𝑞) → 2𝐻2 𝑂(𝑙) + 𝑂2 (𝑔)
However, upon further exploration, I found that the catalysed reaction described in equation (1) is also part of
many industrial processes including food processing, pharmaceuticals, textiles and cosmetics (Trawczynska,
2020). Among these real word applications, catalase is used to remove hydrogen peroxide from milk prior to
cheese production (Roundy, 1958), and used to remove hydrogen peroxide from fabrics after bleaching
(Amorim, 2002). Application of catalase in industrial processes is highly dependent on the knowledge of its
kinetic parameters to maximise efficiency. Within this, the activation energy (Ea) of the catalysed reaction is
an important parameter for many industrial decisions (Trawczynska, 2020).

2. Research question and aim:

Extending from the exploration into the reaction’s kinetic parameters, the research question was formulated:
“What is the activation energy (kJ/mol) of the decomposition of hydrogen peroxide into water and oxygen
in the presence of catalase?”
The aim of this investigation is to derive the value for activation energy (Ea) for the decomposition of H2O2
in the presence of catalase using the experimentally determined relationship between the rate of reaction
(dependent variable), and the temperature under which it occurs (independent variable).

3. Background and hypothesis:

3.1 – Activation energy:
The activation energy (Ea) of a reaction is the magnitude of the minimum required kinetic energy of a
particle to allow the occurrence of the reaction (Brown and Ford, 2014). The known activation energy of the
decomposition of H2O2 in the absence of a catalyst has been confirmed empirically to be around 75 kJ/mol
(Cotton, 2011).
3.2 – The mechanism of catalysts:
Catalysts are substances that increase the rate of reaction by lowering the reaction’s required activation
energy without being consumed (Brown and Ford, 2014). Catalysts provide an alternative reaction pathway,
reacting with the substrate to form an intermediate product before forming the final product. The activation
energy of the steps involved within the alternative pathway are lower than when the catalyst is absent, thus
the rate of reaction is increased as more molecules have the required kinetic energy for a successful
collision. There are two types of catalysts, homogeneous and heterogeneous (Brown and Ford, 2014).
‘Catalase’ in the decomposition of H2O2 acts as a homogeneous catalyst, meaning that it exists in the same
phase as the reactant of the solution.
3.3 – Catalase: history, composition and mechanism:
The discovery of ‘catalase’ was a process of serendipity, when in 1818 French chemist Louis Jacques Thénard
observed the breakdown of hydrogen peroxide by an unknown substance (Glorieux, 2017). It was not until
the late 20th century that the amino acid sequence, molecular weight and three-dimensional structure of the
enzyme was determined. While the complete mechanism of catalase is not currently known, current
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speculations of the alternate pathway involve intermediate reaction with the iron-ion within the enzyme’s
coordination complex, in 2 stages (Andersson, 1991):
1. 𝐻2 𝑂2 + [𝐹𝑒(𝐼𝐼𝐼) − 𝐸𝑛𝑧𝑦𝑚𝑒] → 𝐻2 𝑂 + [𝑂 = 𝐹𝑒(𝐼𝑉) − 𝐸𝑛𝑧𝑦𝑚𝑒]
2. 𝐻2 𝑂2 + [𝑂 = 𝐹𝑒(𝐼𝑉) − 𝐸𝑛𝑧𝑦𝑚𝑒] → 𝐻2 𝑂 + 𝑂2 + [𝐹𝑒(𝐼𝐼𝐼) − 𝐸𝑛𝑧𝑦𝑚𝑒]
The ferric ion in the original enzyme reacts with hydrogen peroxide, breaking the O-O bond and forming [𝑂 =
𝐹𝑒(𝐼𝑉) − 𝐸𝑛𝑧𝑦𝑚𝑒] intermediate compound. In the second step, a second hydrogen peroxide molecule acts
as a reductant which regenerates the original enzyme [𝐹𝑒(𝐼𝐼𝐼) − 𝐸𝑛𝑧𝑦𝑚𝑒] while forming water and oxygen
products (Andersson, 1991). Both presumed steps have lower activation energy than the single-step
decomposition of H2O2. Thus, the effect of catalase could be visually signified on the Maxwell-Boltzmann
Distribution curve (figure 1), and Energy Profile diagram (figure 2), as shown in the diagrams below:
Figure 1: Maxwell-Boltzmann distribution displaying effect of catalyst on the Figure 2: Energy Profile diagram displaying effect of catalyst on
number of particles with required activation energy (Brown and Ford, 2011) activation energy (Brown and Ford, 2011)

= 75

Kinetic energy E (kJ/mol)

Ea(cat) Ea(uncatalyzed) = 75

3.4 – From the principle shown by these diagrams, a hypothesis of activation energy was generated:
“The activation energy (kJ/mol) of the decomposition of H2O2 in the presence of catalase will be lower than
75 kJ/mol.”
3.5 – Equations, Arrhenius’s graph and calculational approach:
The rate of the decomposition of H2O2 is affected by the concentration of reactants used. The relationship
between rate and concentration is given by the expression:
(1) 𝑅𝑎𝑡𝑒 = 𝑘[𝐻2 𝑂2 ]𝑛
Where ‘k’ is the rate constant and ‘n’ is the order of reaction. The decomposition of H2O2 in the presence of
catalase has been empirically determined by multiple studies to be a first order reaction (Greenfield and Price,
1954) (Ogura and Yamazaki, 1983). Thus the value of ‘n’ is 1. Equation 1 can be rewritten as:
𝑅𝑎𝑡𝑒
(2) 𝑘 = [𝐻 𝑂 ]
2 2

Equation 2 shows that the rate constant ‘k’ is dependent on the rate of decomposition and concentration of
H2O2. Adding a catalyst or changing the concentration of a catalyst will change the rate constant ‘k’ by
changing the rate of reaction. Similarly, changing the initial concentration of H2O2 will also change the rate
constant ‘k’. However, it is important to note that the activation energy of a reaction is only dependent on its
reaction mechanism. Changing the concentration of a catalyst, or initial concentration of H2O2 will not change
the alternate pathway that is already provided by the catalyst (Brown and Ford, 2014). As such, the activation
energy of the decomposition of H2O2 by catalase is a constant and will not be affected by the
concentration of H2O2 or concentration of catalase that is used.
To obtain ‘k’, the rate of reaction must first be empirically determined. The rate of reaction is, by definition,
the change in concentration of H2O2 over time. This value could be obtained by measuring the time it takes
for 10cm3 of gaseous product (O2) to be evolved. Using the ideal gas law (PV=nRT), the number of moles of
O2 gas evolved in this time can be calculated. Using the stoichiometric ratio between H2O2 and O2 of 2:1
(equation 1), one can then determine the number of moles of H2O2 consumed and ascertain the rate of its
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concentration change. Once the rate of reaction is known, using equation 3, we may then find the rate constant
k. The rate constant ‘k’ varies in relation to the temperature at which the reaction occurs. The empirically
obtained relationship between them could be used to calculate
the activation energy, using Arrhenius’s equation (Brown and
Ford, 2014):
𝐸
(3) ln 𝑘 = ln 𝐴 − 𝑅𝑇𝑎

Where A is the frequency of successful collisions, R is the gas

constant (8.3145JK-1mol-1) and T is temperature in Kelvin.

1
Equation 3 shows that the relationship between ln 𝑘 and is
𝑇
linear. By plotting these two variables, Ea may be derived
through the gradient (-Ea/R). (refer to figure 3, right) Figure 3: Arrhenius’s graph showing the relationship
in equation 3: (John, 2013)

3.6 – Choice of concentrations and quantity of substrate and catalyst:

As established previously in part 3.5, the choice of concentrations of H2O2 substrate and catalase catalyst will
not affect the value of the activation energy that will be calculated. However, this choice of concentrations
will effect the rate of reaction, and thus will also affect the ability to record the time of gas evolution with
minimal percentage uncertainty. For instance, a too high concentration of H2O2 and catalyst could mean that
10 cm3 of O2 gas evolves too fast to be accurately timed with a stopwatch, creating vast experimental
uncertainty. Furthermore, as the decomposition of H2O2 is an exothermic reaction, a too-high rate of reaction
could produce much heat, such that some of the H2O product also escapes as gas, making the measurement of
O2 evolution inaccurate. On the other hand, if too low concentration of H2O2 is used, or too little catalase is
used, the evolution of 10 cm3 of O2 gas would take too much time, limiting the number of repetitions that
could be achieved in the experiment.
Considering both extremes, a reasonable time frame for the collection of 10 cm3 of oxygen gas should be
targeted to be around 60 seconds (this, of course, is only an approximate target because the rate of evolution
will change at different temperatures). To express this is in moles per second, calculate the number of moles
𝑉𝑔𝑎𝑠
of gas in 10 cm3 using (𝑛 = ) and then divide by 60:
𝑉𝑚
0.01
(4) ÷ 60 = 7.44 × 10−6 moles per second.
22.4

Next, the concentrations of H2O2 and catalase needed to achieve this target should be determined. First, we
must establish the way in which catalase strength is measured. 1 ‘unit’ of catalase is internationally defined
as the “amount of catalase decomposing 1.0 µmoles of hydrogen peroxide per minute at pH 7.0 at 25oC, with
initial H2O2 concentration of 10.3 mM” (Beers, 1952, p. 6). Making a 10.3mM concentration of H2O2 from
provided stock solutions may be problematic due to the high uncertainties associated with multiple serial
dilutions. Thus, a 0.1M concentration was used to decrease the number of dilutions needed. Being a first order
reaction, a 10-fold increase in the concentration of H2O2 (10.3mM → 0.1M) will also increase the rate of
reaction by 10-fold.

Thus, at 0.1M concentration of H2O2, 1 unit of catalase decomposes 1 × 10−6 moles of H2O2 in 6 seconds,
meaning that 0.5× 10−6 moles of O2 is evolved (using the stoichiometric ratio of 2:1 between H2O2 and O2).
To express this in moles of O2 gas evolved per second, the number of moles of O2 evolved is divided by 6:
(5) 0.5 × 10−6 ÷ 6 = 8.33 × 10−8 moles per second.
1 unit of catalase evolves 8.33 × 10−8 moles of O2 per second (equation 5), but we need a target of
7.44 × 10−6 moles per second (equation 4). Thus, the number of units of catalase to achieve the target rate of
evolving 10 cm3 of O2 gas in 60 seconds is given by the calculation:
(6) (7.44 × 10−6 ) ÷ (8.33 × 10−8 ) ≈ 90 units of catalase
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 4. Variables:
4.1 – Table showing the independent variable and its uncertainty:
Independent: Method of variation and uncertainty:
(T) – Temperature The temperature under which the reaction takes place is varied through a water bath
of the reaction heating system. This apparatus consists of an in-built thermometer which digitally
mixture (Kelvins) measures the temperature of the water with an error of ±0.1𝐾. To covering a wide range
(±𝟎. 𝟏𝑲)
of data points, temperature was varied in 5K increments (288.0 K, 293.0K, 298.0 K, 303.0
K, 308.0 K, 313.0 K ±0.1𝐾). These values were chosen specifically because catalase
activity stays relatively constant under this temperature range (refer to figure 4, below).
Higher temperatures could not be used because Abuchowski shows that catalase
experiences decreasing activity at temperatures exceeding 313K due to ‘denaturation’,
the irreversible change in protein structure from the breaking of weak bonds in the
enzyme (Abuchowski, 1977).

Figure 4: Denaturation of catalase at

temperatures exceeding approximately 313K
(Abuchowski, 1977)

4.2 – Table showing the dependent variable and its uncertainty:

Dependent: Method of measurement and uncertainty:
(t) – Time for The time for the evolution of 10 cm3 of oxygen gas is measured by using a stopwatch to
the evolution time the displacement of 10 cm3 of gas in a pneumatic trough. As there is a certain degree
of 10 cm3 of
of human error in handling a stopwatch (random error), this is minimised by conducting 6
O2 gas
repeats for each temperature. An average will then be taken for all 6 recorded times to be
used in later calculations. The stopwatch has a measurement uncertainty of
±0.01𝑠. However, the experimental uncertainty – the differing measurements between
trials – is much greater than the measurement uncertainty, thus is used instead of it. The
experimental uncertainty of the dependent variable will be calculated by taking half the
range of the recorded times in all trials at each temperature.

4.3 – Table showing the controlled variables of the experiment:

Controlled: Rationale for control: Method to control:
Concentration The rate of reaction is affected by a change in the Each trial at each temperature will
of H2O2 concentration of the reactant used (Brown and Ford, use a constant initial concentration of
2014). This must be controlled to ensure temperature is hydrogen peroxide of 0.1 mol dm-3.
the only variable affecting the time 10cm3 of oxygen
gas is evolved.
Units of The number of units of catalase activity added to the The prepared catalase solution will
catalase hydrogen peroxide solution will affect the rate of its have 90 units of enzyme activity per
activity decomposition. If this variable is not controlled, the 5cm3 of solution. Only 5cm3 will be
derived rate constant ‘k’ will change relative to it, added to each trial.
consequently not being solely dependent on
temperature.
pH under Enzyme activity is impacted by changes in pH. At pH was controlled at 7 by using 5
which the extremely high or low pH levels, there can be an almost cm3 of pH 7 buffer solution, which
reaction complete lost in enzyme activity from denaturation of resists pH changes when an acid or
occurs:
catalase. The optimal pH level for catalytic activity for base is added.
catalase is approximately 7.0 (Williams, 1928).
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 Pressure: Pressure needs to be controlled at STP to ensure the This variable is automatically
valid use of the ideal gas equation (PV = nRT) to controlled, as the atmospheric
calculate the moles of O2 in 10cm3 of evolved gas pressure of Sydney is 1018 mb
(Brown and Ford, 2014). (WWO, 2020), which is
approximately 1 atm, with small
fluctuations in a laboratory setting.
Angle of Due to parallax error, the angle from which one Always observe the meniscus of the
observation observes the meniscus of the water in the gas collecting water in the gas collecting cylinder
for gas cylinder may influence the decision of whether 10 cm3 from directly horizontal level (eye
evolution:
of O2 gas has evolved. level).

5. Experimental Method
Modifications to method: The method underwent revision after its first trial. The original method of gas
collection involved the use of a gas syringe. Anticipating problems associated with ‘stiffness’ of the gas
syringe, the original method involved excessive lubrication of the syringe before use. However, in the first
trial the syringe nevertheless remained stiff despite clear effervescence observed in the test tube. I realised
that even if the syringe were to work, it would produce inaccurate results as the pressure required to move the
plunger would exceed STP. A modification of the original method was thus immediately conceived, involving
the ‘pneumatic trough’ system to collect O2 gas.
5.1 – Table showing the equipment list for the experiment:
Quantity Apparatus Rationale of use
1 Retort stand, boss-head Used to hold the pneumatic trough in vertical position for accurate gas
and clamp volume measurements.
1 Stopwatch (±0.01s)
1 10 cm3 graduated pipette
(±0.045 𝑐𝑚3 )
200 ml Catalase solution Stock solution provided by Southern Biological. Concentration (40
units/ml) information provided by company. A 1:1 dilution was
provided by staff for the experiment, lowering the concentration to 20
units/ml.
30 cm3 H2O2 aqueous solution Stock solution which will be diluted to form 0.1M required solution for
(6%) decomposition.
3
500 cm Deionised water Used for the dilution of hydrogen peroxide.
1 Wash bottle Used to transfer deionised water in the preparation of diluted solutions.
3
125 cm pH 7.0 Buffer solution Necessary to maintain pH at optimal level for catalase activity. 102.94
cm3 of Na2HPO4 was added to 22.06 cm3 of citric acid to create 125
cm3 of pH 7.0 buffer solution. The prepared solution was provided by
staff for the experiment.
1 Water bath Used to vary temperature from 288.0 K, 293.0 K, 298.0 K, 303.0 K,
308.0 K, 313.0 K (±0.1𝐾).
1 Water bath test tube rack
1 Side arm test tube This is where the reaction will take place, producing O2 gas which will
be measured in a pneumatic trough.
1 Test tube rubber stopper Prevent O2 from escaping into the air. All O2 must be collected in the
pneumatic trough.
1 Gas collecting cylinder Used to measure the quantity of evolved O2 gas.
(50 ml) (±0.1 cm3)
1 Delivery tube Used to connect the side-arm test tube to the pneumatic trough for
measuring the volume of gas evolution.
1 Volumetric flask (500ml) Used to store the prepared H2O2 solution.
1 Volumetric flask rubber Prevent escape/decomposition of stored hydrogen peroxide solution.
stopper
1 Safety glasses and gloves Safety equipment to avoid skin/eye contact with chemicals.

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5.2 – Method:
A. Preparation of Catalase solution (90 units)
The stock catalase solution contains approximately 40 units/ml, meaning in 5 cm3 of solution there are
approximately 200 units of catalase activity. Recall from section 3.6 that only 90 units of activity are needed in
each 5 cm3. This is approximated by a 1:1 dilution with water. A diluted solution of catalase was provided by staff.
B. Preparation of H2O2 solution (0.1M) from 6% (1.765M) stock solution
1. Using a measuring cylinder transfer 28 cm3 of 6% H2O2 solution to empty 500 mL volumetric flask.
2. Using a wash bottle containing deionised water, fill the volumetric flask up to the graduated line.
3. Immediately seal the volumetric flask to prevent instantaneous decomposition of H2O2 solution.
4. Gently shake the flask to ensure the prepared solution is homogeneous.
C. Preparation of apparatus (refer to figure 5, below)
1. Connect delivery tube to side-arm test tube. Ensure that the tube is tightly fitted, so no gas can escape.
This can be checked by filling the test tube to the rim with water and ensuring there is no leakage
between the side-arm and delivery tube.
2. Using a 10 cm3 graduated pipette (±0.045 𝑐𝑚3), measure exactly 5.000 cm3 of catalase solution, and
transfer to a side-arm test tube.
3. Using a graduated pipette, measure exactly 5.000 cm3 of pH 7 buffer solution (Na2HPO4 and citric
acid) and add it to the side-arm test tube containing the catalase solution.
4. Fill water bath, setting its temperature to 288.0 K (±0.1K).
5. Place the test tube containing the catalase and buffer solution into the water bath for 5 minutes (timed
using a stopwatch), ensuring that the temperature of the solution in the test tube has reached
equilibrium with that of the water bath.
6. Fill gas collecting cylinder with water and set up the pneumatic trough as per the diagram. Ensure that
the gas collecting cylinder is vertically held by the retort stand for accurate measurements (figure 5).
7. Ensure that the bottom of the gas collecting cylinder is held directly covering the opening of the
delivery tube such that all evolved gas will travel up into the cylinder.
D. Experimental procedure:
1. Using a graduated pipette, transfer 10cm3 of prepared H2O2 solution from the volumetric flask to the
test tube containing the catalase solution.
2. Immediately reseal the test tube with the rubber stopper and start a digital stopwatch.
3. Observe O2 gas gradually filling the gas collecting cylinder and record the time it takes for 10cm3 of
gas to evolve.
4. Repeat the experimental procedure 6 times at each temperature, 288.0 K, 293.0 K, 298.0 K, 303.0 K,
308.0 K, 313.0 K ±0.1 𝐾.
5.3 – Diagram of apparatus set-up:

Figure 5: Labelled Diagram of apparatus: Pneumatic

trough

Evolved 𝑂2 gas

Rubber
stopper

Retort stand

Delivery tube
Side-arm test tube

H2O2 catalase solution

Test tube rack
Water bath

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 6. Safety, ethical and environmental considerations:
6.1 – Safety considerations:
H2O2 “can be corrosive to the eyes, skin, and respiratory system. This chemical can cause burns to the skin
and tissue damage to the eyes.” (MSDS, 2015, p. 1) It is necessary to take caution by wearing gloves and
safety glasses. Avoid ingestion of H2O2, and inhalation of any evolved gas.
6.2 – Environmental considerations:
Safe disposal of leftover H2O2 solution is important, because H2O2 can cause environmental hazards. As a
strong oxidant and corrosive, “Upon interaction with some organic compounds hydrogen peroxide can
transform into an explosive ingredient” (ACS, 2016, p. 3). Ensure that the chemical is disposed in line with
the principles of green chemistry, achieved by employing evaporation methods.
6.3 – Ethical considerations:
There are no ethical considerations as no living organisms were involved in this investigation.
7. Raw Data Collection:
7.1 – Raw qualitative observations:
1. As temperature increased, the vigour of effervescence observed in test tube visibly increased.
2. The colour of the solution remained constant as the reaction progressed, the yellow/green colour of catalase.
3. The gas bubble can be observed ‘pushing through’ the delivery tube and escaping into the collection cylinder.
4. The rate of gas production was not constant. Effervescence is rapid in the initial seconds of the reaction.
After the initial ‘burst’, gas collection decreases to a slow and constant rate.
5. The temperature of the solution after the reaction was markedly higher than the initial temperature.
7.2 – Raw quantitative data:
Temperature Time taken to evolve 10 cm3 of O2 gas from the decomposition of H2O2 (t) (±𝟎. 𝟎𝟏𝒔)
(T) (±𝟎. 𝟏𝑲): Trial 1 Trial 2 Trial 3 Trial 4 Trial 5 Trial 6
288.0 68.23 67.54 67.82 68.36 67.95 68.57
293.0 64.37 64.01 64.23 63.89 64.44 64.22
298.0 60.93 60.22 61.01 60.53 60.66 60.87
303.0 60.98* 57.32 56.98 56.73 56.71 56.57
308.0 54.83 54.02 54.55 53.82 53.09 53.91
313.0 50.32 51.07 50.94 50.31 51.37 57.22*
*This value was discarded as an outlier and not included in calculations as it deviates greatly from the other
recorded values at this temperature.
7.3 – Sample calculation: Average time of 10 cm3 O2 evolution and its experimental uncertainty at 288K:
Average time (𝒕̅): Experimental uncertainty (∆𝒕):
6 trials were performed at each temperature, and an average The experimental uncertainty is
taken to minimise random human error. At 288K: determined by half the range of
̅
(𝑡) = (68.23 + 67.54 + 67.82 + 68.36 + 67.95 + 68.57) ÷ 6 recorded values (t) for trials at each
(𝑡̅) = 68.08 temperature. At 288K:
∆𝑡 = (68.57 − 67.54) ÷ 2
∆𝑡 = ±0.52
This calculational method was applied to all other temperatures (T), producing the table below:
7.4 – Table showing average time (𝒕̅) for the evolution of 10 cm3 of O2 evolution and its experimental
uncertainty (∆𝒕) at each temperature (T):
Temperature (T) (±𝟎. 𝟏 𝑲): Average time (𝒕̅): Experimental uncertainty (∆𝒕):
288.0 68.08 ±0.52
293.0 64.19 ±0.28
298.0 60.70 ±0.40
303.0 56.86 ±0.38
308.0 54.04 ±0.87
313.0 50.80 ±0.53
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 8. Data processing and uncertainty propagation:
The purpose of this section is to fulfil the aim of the report by determining the activation energy of the
decomposition of hydrogen peroxide by catalase. First, O2 evolution must be converted to H2O2 consumption:
8.1 – Calculation for number of moles of H2O2 consumed in the evolution of 10 cm3 of O2:
Step Calculation Uncertainty propagation
Moles of O2 𝑉𝑔𝑎𝑠 Volume of O2 has a measurement uncertainty of
𝑛=
evolved 𝑉𝑚 ±0.2 cm3, as 2 readings must be made (initial and
0.0100 final), each with an uncertainty of ±0.1 cm3.
= 4.41 × 10−4 𝑚𝑜𝑙 ±𝑉𝑔𝑎𝑠
22.7 ±∆𝑛 =
𝑉𝑚
±0.0002
= ±8.81 × 10−6 𝑚𝑜𝑙
22.7
Moles of H2O2 H2O2 and O2 are in a 2:1 ratio. The uncertainty on the number of moles of O2 is
−4 −4
consumed 2 × 4.41 × 10 = 8.81 × 10 doubled to find the uncertainty on moles of H2O2:
moles of H2O2 must be consumed. 2 × (±8.81 × 10−6 ) = ±1.76 × 10−5 𝑚𝑜𝑙
8.2 – Sample calculation for the average rate of reaction at 288K:
Step Calculation Uncertainty propagation
Change in ∆[𝑛] Total volume (V) was added using 3 pipettes, each
concentration ∆[𝑅𝑒𝑎𝑐𝑡𝑎𝑛𝑡𝑠] = 𝑉 with ±0.045 𝑐𝑚3 measurement uncertainty. The
−4
∆[𝑅𝑒𝑎𝑐𝑡𝑎𝑛𝑡𝑠] 8.81 × 10 uncertainty of solution volume (±𝑉) is thus:
at 288K ∆[𝑅𝑒𝑎𝑐𝑡𝑎𝑛𝑡𝑠] =
0.0200 ±0.045 × 3 = ±0.135 𝑐𝑚3
∆[𝑅𝑒𝑎𝑐𝑡𝑎𝑛𝑡𝑠] = 4.41 × 10−2
mol dm-3 Percentage uncertainty of ∆[𝑅𝑒𝑎𝑐𝑡𝑎𝑛𝑡𝑠] is sum of
percentage uncertainty on moles of H2O2 and
percentage uncertainty on volume (V):
1.76 × 10−5 0.000135
100% × ( + ) = 2.67%
8.81 × 10−4 0.0200
Average rate Rate of Reaction = ∆[𝑅𝑒𝑎𝑐𝑡𝑎𝑛𝑡𝑠] The experimental uncertainty on ∆𝑡 at 288K is ±0.52
of reaction at ∆𝑡
4.41×10−2 as established in 7.3. Percentage uncertainty of rate of
288K Rate of Reaction = 68.08
reaction is calculated by adding percentage uncertainty
Rate of Reaction = 6.47 × 10−4 of ∆[𝑅𝑒𝑎𝑐𝑡𝑎𝑛𝑡𝑠] and percentage uncertainty of ∆𝑡:
mol dm-3 s-1 0.52
2.67% + 100% × ( ) = 3.44%
68.08
8.3 – Sample calculation for K constant, and ln K at 288K:
Step Calculation Uncertainty propagation
K constant Recall section 3.5, the 𝐶1
𝐶2 = 𝑉1 ×
at 288K: decomposition of H2O2 𝑉2
in the presence of The absolute uncertainty of [𝐻 𝑂
2 2 ] is thus calculated by:
𝐶1
catalase is shown to be ±[𝐻2 𝑂2 ] = ±∆𝑉1 ×
𝑉2
a first order reaction: Where ±∆𝑉1 is the measurement uncertainty of volume of stock H2O2
𝑅𝑎𝑡𝑒 solution added to make the dilution (±0.0005). ±[𝐻2 𝑂2 ] thus equals:
𝑘= 1.765
[𝐻2 𝑂2 ] ±0.0005 × = ±1.77 × 10−3 𝑚𝑜𝑙𝑑𝑚−3
−4 0.500
6.47 × 10
𝑘= Percentage uncertainty of K is the sum of percentage uncertainty
0.1
𝑘 = 6.47 × 10−3 s-1 on rate of reaction and percentage uncertainty on [𝐻2 𝑂2 ] (absolute
uncertainty divided by 0.100 molar concentration):
1.77 × 10−3
3.44% + 100% × ( ) = 5.19%
0.100
ln K at ln 𝑘 = ln(6.47 × 10−3 ) The absolute uncertainty for ln x is equal to ± ∆𝑥.
𝑥
288K ln 𝑘 = −5.04 ∆𝑘
Thus, the absolute uncertainty for ln 𝑘 is 𝑘 , which is the same as
the relative uncertainty for K, which is ±0.0519.
9|Page
This calculational process was applied to all other temperatures. The table below summarises the processed
values for rate of reaction, K and ln K at each temperature:
8.4 – Table showing rate of reaction, K constant and ln K and the uncertainty of 𝐥𝐧 𝐊 at each
temperature for the decomposition of hydrogen peroxide by catalase:
Temperature (Temperature)-1 Rate of reaction K constant
−4 ln 𝑘 ∆ ln 𝐾 (±)
(K) -3
(10 K) (10 moldm s ) -3 -1
(10−3 s-1)
288.0 3.47 6.47 6.47 -5.04 0.0519
293.0 3.41 6.86 6.86 -4.98 0.0487
298.0 3.36 7.26 7.26 -4.93 0.0509
303.0 3.30 7.75 7.75 -4.86 0.0510
308.0 3.25 8.15 8.15 -4.81 0.0605
313.0 3.19 8.67 8.67 -4.75 0.0548

9. Analysis of results and determining the activation energy (𝑬𝒂 )

𝐸𝑎 1
Arrhenius’s equation (ln 𝑘 = ln 𝐴 − ) shows that (ln 𝑘) has a linear relationship with ( ). When
𝑅𝑇 𝑇𝑒𝑚𝑝𝑒𝑟𝑎𝑡𝑢𝑟𝑒
𝐸𝑎
expressed in the form ‘𝑦 = 𝑚𝑥 + 𝑏’ the gradient (m) of the equation is (− ). As ‘R’ equals 8.3145 J mol-1
𝑅
𝐸𝑎 1
K-1, the activation energy (𝐸𝑎 ) can be calculated from the gradient (− ). Accordingly, was
𝑅 𝑇𝑒𝑚𝑝𝑒𝑟𝑎𝑡𝑢𝑟𝑒
graphed against ln 𝑘 with vertical error bars showing the uncertainty, ∆ ln 𝐾 (table 8.4). Note that horizontal
error bars are negligible due to the extremely small percentage uncertainty of temperature.
9.1 – Arrhenius Graph showing the relationship between (Temperature)-1 (x-axis) and 𝐥𝐧 𝒌 (y-axis)
for the decomposition of hydrogen peroxide by catalase:
-4.65
Figure 6: Arrhenius graph showing
-4.7 the relationship between
-4.75
(Temperature)-1 (x-axis) and 𝑙𝑛 𝑘 (y- LoBF
axis)
-4.8 y = -1050x - 1.37
-4.85

-4.9 LoWF1
y = -1440x - 0.091
𝐥𝐧 𝒌

-4.95
𝑹𝟐 = 0.9989
-5

-5.05 LoWF2
-5.1
y = -671x - 2.66

-5.15
0.00315 0.0032 0.00325 0.0033 0.00335 0.0034 0.00345 0.0035
(Temperature)-1 / (K)-1
Figure 6 shows the linear relationship between the processed values of (Temperature)-1 and ln 𝑘 for the
decomposition of hydrogen peroxide by catalase. In particular, it shows the line of best fit (blue), as well as
the lines of worst fit (orange/grey) to demonstrate the range of error for the gradient. The equations of the
three lines indicate that the value of the gradient is (-1440), with a range of error from (-1440) to (-671).
This gradient information can be used to calculate the activation energy (𝐸𝑎 ) and its range of error:
9.2 – Calculation of activation energy and its range of error:
Step 𝐸𝑎 Max 𝐸𝑎 Min 𝐸𝑎
Gradient 𝐸𝑎 𝐸𝑎−𝑚𝑎𝑥 𝐸𝑎−𝑚𝑖𝑛
𝐸 − = −1050 − = −1440 − = −671
(− 𝑎) 𝑅 𝑅 𝑅
𝑅
Calculation 𝐸𝑎 = −1050 × (−8.3145) 𝐸𝑎−𝑚𝑎𝑥 = −1440 × (−8.3145) 𝐸𝑎−𝑚𝑖𝑛 = −671 × (−8.3145)
of activation 𝐸𝑎 = 8730 J mol-1 𝐸𝑎−𝑚𝑎𝑥 = 10200 J mol-1 𝐸𝑎−𝑚𝑖𝑛 = 7580 J mol-1
energy: 𝐸𝑎 = 8.73 kJ mol-1 𝐸𝑎−𝑚𝑎𝑥 = 10.2 kJ mol-1 𝐸𝑎−𝑚𝑖𝑛 = 7.58 kJ mol-1
10 | P a g e
Thus, we have satisfied the initial aim of our exploration. The experimental value for activation energy of
the decomposition of hydrogen peroxide by catalase is 𝟖. 𝟕𝟑 kJ mol-1 with a range of error from 𝟕. 𝟓𝟖 kJ
mol-1 to 𝟏𝟎. 𝟐 kJ mol-1.

9.3 – Analysing the impact of uncertainties and trend correlation:

Despite small measurement uncertainties, the impact of propagated experimental uncertainties is observably
greater on the result. The uncertainty of the final calculated activation energy is half the range of error,
10200−7580 ±1310
= ±1310 J mol-1. This is calculated to be a percentage error of 100% × = ±𝟏𝟓. 𝟏%.
2 8730
±15.1% is a fairly large final uncertainty because the initial experimental uncertainty is greatly magnified
during the course of its propagation, especially due to the multi-step process of the calculation in part 9.
Another reason for the large final uncertainty is because the experiment was done over a limited range of
temperatures. Considering graph 9.1, one may imagine that a larger range of temperatures will ‘stretch-out’
the y-axis, leading to a smaller range of error for gradient, and thus activation energy. However, due to the
effects of denaturation explained in part 4, it is impossible to increase the temperature range without
undermining the validity of the experiment.
Large percentage uncertainties undermine the validity of results. However, the precision (how strong the
correlation is) of the results is high. This is testified by the value of ‘Pearson’s Coefficient of Determination’
(R2), which was shown in graph 9.1 to be 0.9989. Pearson’s Coefficient of Determination measures the
strength of association between two variables. A high R2 value for this investigation means that data points
deviate minimally from the trend-line, and that the relationship between ln K and Temperature is strong. The
conclusion that this analysis yields is that we can be sure that ln K and Temperature follow a general
positive linear relationship, but we are not as sure of the value of the final calculated activation energy
(which could be any value within the large uncertainty range).

10. Evaluation:
Conclusion:
10.1 – Conclusion on research question
This investigation aimed to derive the value for the activation energy (Ea) of the decomposition of H2O2 in the
presence of catalase. My original hypothesis stated that the activation energy must be lower than 75 kilojoules
per mole, which is the activation energy of the decomposition of H2O2 without a catalyst. The investigation
concludes with an experimental value for the activation energy of hydrogen peroxide in the presence of
catalase, of 8.73 kJ/mol, with a percentage error of ±15.1%. The possible values within this range of error
are well below 75 kilojoules per mole, thus supporting the hypothesis. This satisfaction of the hypothesis is
a source of confirmation of the results, as a catalysed reaction must have a lower activation energy than the
uncatalyzed reaction.
10.2 – Conclusion in relation to scientific context:
In ‘Elements of Physical Chemistry’, written by English chemist Peter Atkins, the literature value for the
activation energy of the decomposition of H2O2 in catalase is quoted to be 8 KJ/mol (Atkins, Paula, 2013).
This literature value shows the experimental value of this paper, 8.73 KJ/mol to be extremely accurate, with
a percentage error of only 9% from this theoretical value. However, as activation energy is empirically
determined, a ‘literature value’ does not exist in the strict sense of the term, only many different experimental
values derived from a range of rigorous studies. For instance, another study using a similar approach to the
one in this paper, conducted in 2018, concluded 𝐸𝑎 to be 12.9 kJ/mol (Milek, 2018). Because there is no
absolute theoretical value for the activation energy of the reaction to compare against, it does not make sense
to calculate a percentage error. Rather, the accuracy of the experiment comes from its general agreement
with the range of values determined by other, more rigorous studies done in the scientific community.

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Furthermore, scientific justification can be provided for the qualitative data in part 7.1, which relate mainly to
the observation of effervescence within the test tube and gas collection cylinder. Effervescence is the
appearance of foaming in a solution that results from the release of gas. The vigour of effervescence is thus a
qualitative indication of the rate of reaction. There are 2 significant observations that relate to effervescence
and gas release mentioned in part 7.1:
1. Effervescence increases with temperature increase. This is means that at higher initial temperatures,
there is also a higher rate of reaction. Temperature raises the rate of reaction because temperature is
the average kinetic energy of the particles. A higher average kinetic energy means more particles will
have the necessary energy for a successful collision, thus raising the rate of reaction.
2. Effervescence decreases over time. This means that as time progresses, the rate of reaction decreases.
This is because as time progresses, reactants are being turned into products, meaning that the
concentration of H2O2 decreases over time. Fewer available particles for collision also means that the
likelihood of successful collisions decreases, thus lowering the rate of reaction.
Finally, another important observation was that the temperature of the solution increased markedly over time.
Although the specific increase in temperature was not measured, it was felt when handling the test tube after
the reaction took place. This indicates that the reaction is an exothermic reaction, meaning that there is a net
release of thermal energy from the forming of chemical bonds.

Strengths, weaknesses and further investigation:

10.3 – Strengths of the investigation:
Strength: Significance:
Low initial Because trials are in general agreement with each other, this indicates the reliability
experimental error of the values obtained for the times of oxygen evolution. Furthermore, the
of time to evolve synonymity of the raw data also testifies that there was probably a synonymity of
3
10 cm of oxygen conditions between trials, meaning that controlled variables were effectively kept.
(in table 7.2)
High accuracy and Precision is the measure of the strength of correlation in my results. Accuracy is the
precision of results measure of how ‘correct’ the final value is relative to a literature value. The strong
linear correlation between ln K and Temperature (9.1), and the general agreement of
the calculated activation energy range with the empirical values of other studies
show that the results I obtained were both precise and accurate.
Good choice of In section 3.6, I worked backwards to obtain the most optimal concentrations to use
concentrations of for catalase and hydrogen peroxide so that the evolution of 10 cm3 in each trial
catalase and H2O2 could be done in approximately 60 seconds. This meant that enough trials could be
done, as well as minimising the percentage uncertainty associated with the fast
reactions (of handling the stopwatch) of low time values.

10.4 – Weaknesses and potential improvements of the investigation:

Analysis of error: Possible solution:
Systematic error: Oxygen is soluble in water, The solubility of oxygen in water is negatively
thus the volume of gas measured will be lower correlated with temperature, meaning that at higher
than the actual volume of gas evolved. temperatures, the amount of oxygen dissolved in
water decreases (Tromans, 1998). Thus, the effect of
this source of error on the experiment can be greatly
minimised by increasing the temperature of water
used in the pneumatic trough system (not the water
bath system).

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 Systematic error: The pressure inside the Calculate the actual pressure in a pneumatic trough
pneumatic trough system is higher than STP as a system rather than using STP. The pressure of within
result of vapour pressure from the evaporation of a pneumatic trough system is determined by the sum
water. Because STP was used in calculations, the of STP and the vapour pressure of the water used in
number of moles of O2 gas calculated was lower the system (Miller, 1860). This is a more accurate
than the actual number of moles evolved value to determine the number of moles of O2 evolved
(PV=nRT). through PV = nRT.
Systematic error: The experiment is Record down the temperature of the solution inside
exothermic, thus the temperature was not the test tube at 10 second increments during the
constant during the course of each reaction. In course of the reaction. Rather than using the initial
the context of graph 9.1, this means that the temperature as the value for the independent variable,
actual data point for ln K occurs at a higher use the average temperature during the course of the
temperature (and thus lower T-1) than what was reaction to approximate a more accurate value which
recorded. Therefore all data points on the graph considers the error of heat release.
should be shifted to the left to a certain extent.
Whether the gradient remains constant depends
on whether the amount that each point is shifted
is the same.
Random error: Human error of timing the The impact of this source of random error is minimal
evolution of 10 ml of gas. because I deliberately chose concentrations that will
allow the collection of gas over a time of around 60
seconds. In consideration of this, a few more or few
less milliseconds due to human error will hardly be
significant.
Systematic error: Human error of time delay This can be solved by modifying the apparatus of the
between starting the reaction and placing stopper experiment. Rather than having to place a rubber
on test-tube. stopper on the opening of the test tube after adding the
H2O2 to the catalase, the H2O2 could be added using a
one-way valve instead, thus eliminating the systematic
time delay of placing a rubber stopper.

10.6 – Further investigation:

Further study into the effect of the catalase enzyme on the activation energy of decomposition of H 2O2 may
take the form of differentiating between different types of the enzyme. Indeed, catalase is divided into 3 main
classes: (i) “classic” monofunctional catalases, (ii) catalase-peroxidases, and (iii) nonheme manganese-
containing catalases. A further investigation may take the form of determining how the type of catalase affects
the activation energy of the reaction in its presence. Particularly, this may provide greater insight into which
specific type of catalase to use in different industrial applications. For instance, catalases are used in the
industrial applications such as food or textile processes to remove hydrogen peroxide from sterilization
(Zamocky, Gasselhuber, Furtmuller, & Obinger, 2012). Using a type of catalase with lower activation energy
may maximise the efficiency of this process.
Another extension of this paper may take the form of investigating the specific effects of denaturation on
catalase. While the empirical method of this paper involved only a limited range of temperatures to control
the variable of catalase activity, further investigation could increase temperatures beyond 313 K to observe
the effects of denaturation. One would predict that at higher temperatures, the activity of catalase would
become impaired, and the linear trend between ln K and Temperature would not be observed.

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