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Lab Report |5

Once all the H₂O₂ had reacted, any more KMnO₄ added would be in excess and therefore did

not decompose. The addition of excess KMnO₄ caused the solution to have a permanent pink or

brown tint. After this had been obtained, no more KMnO₄ was added. The amount of KMnO₄

added was proportional to the measure of the amount of H₂O₂ remaining, therefore allowing us

to calculate how much H₂O₂ had been remaining, and how much had been decomposed. This,

however, is only a general overview of the detailed experiment. The following explains in further

detail.

To observe a general reaction of catalase, 10 mL of 1.5% (0.44 M) H₂O₂ was added into

a 50-ml glass beaker. Into there, a 1 mL extract of fresh catalase was added. To demonstrate

factors of enzymatic activity, 5 mL of catalase was boiled and then cooled. About 1 mL of this

was added to another 10 mL of H₂O₂. Furthermore, catalase was exposed to both an acid (HCl)

and basic (NaOH) environment to see how it functioned. This part of the lab, Exercise 2A, gave

an insight on how catalase functioned.

Exercise 2B involved determining the amount of H₂O₂ initially present in a 1.5%

solution. This amount is known as the base line and is the basis for future procedures. Taking a

1.5% solution, 10 mL of H₂O₂ was placed in a clean glass beaker. Instead of enzyme solution, 1

mL of distilled water was added. Then, 10 mL of H₂SO₄ was added, forming a 21 mL solution.

After mixing well, a 5 mL sample was extracted into another beaker for titration. A magnetic

stirring was placed into the beaker, which was in turn placed on a hot plate. Using a burette,

KMnO₄ was added one drop at a time. After measuring the KMnO₄ used, the amount of H₂O₂

was calculated. Another trial of this procedure was conducted to avoid errors.

The following part of the lab called for examining how H₂O₂ decomposed spontaneously.

About 15 mL of H₂O₂ was placed in a beaker. It was stored uncovered at room temperature for

approximately 24 hours. From the remaining solution, a 5 mL extract was titrated to find the

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Lab Report |6

amount of H₂O₂ spontaneously decomposed. This was also repeated several times to ensure

accuracy.

The final portion of the lab observed the rate of enzyme-catalyzed H₂O₂ decomposition.

To observe the rate of reaction, the amount of substrate disappearing was measure over certain

periods of time. For this procedure, 7 clean cups were needed. In each of these cups, 10 mL of

1.5% H₂O₂ was added by a syringe. Along with that, 1 mL of catalase was added into each cup

one at a time, using another clean syringe. For example, one cup was left alone with catalase for

10 seconds. After 10 seconds, all enzymatic activity was halted with the addition of 10 mL of

sulfuric acid, which was added by yet another clean syringe. Another cup had the 1 mL catalase

extracted for 30 seconds. This procedure was consistent for 0, 10, 30, 60, 90, 120, and 180

seconds of catalase activity. From each cup, a 5 mL sample was removed to be titrated. This

allowed us to find out how much H₂O₂ had been remaining after that set time. After this

procedure was completed for all 7 times, it was repeated to ensure maintaining proper results.

RESULTS

For Exercise 2A, an enzyme-catalyzed reaction was observed. The enzyme in the reaction

was catalase. The substrate acted upon was hydrogen peroxide, which was broken down into

water and oxygen gas. The release of the gas was proven as oxygen by a glowing splint test.

When boiling catalase, the reaction does not occur as quickly, since the function of the enzyme

has been altered. Potatoes also have catalase stored in them, as observed when bubbles were

formed as pieces of potato were added to hydrogen peroxide.

The initial amount of H₂O₂ in a 1.5% solution had to be calculated, which acted as the

basis for the rest of the experiment. In order to find this, the molarity of 2% KMnO₄, which was

used as the titrant, had to be measured as well. The following calculations were made:

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Lab Report |7

2% KMnO₄ = 2 grams KMnO 100


₄ grams H O ₂ = 2 grams KMnO 100
₄ mL H O₂ = 20 grams

KMnO ₄1000 mL H O₂

20 grams KMnO4 1 mole KMnO 1 ₄ L 158.04 grams KMnO ₄ = 0.127 moles KMnO 1 ₄L

0.127 M (moles/liter) = 2% KMnO₄

The baseline measurement was calculated as follows:

3.70 mL KMnO₄ used to titrate 1.5% H₂O₂

3.70 mL KMnO ₄0.127 moles 1000 mL KMnO ₄ = 0.000470 moles KMnO₄

.000470 moles KMnO 5₄ moles H O ₂ 2₂ moles KMnO ₄ = 0.00118 moles of HO


₂ ₂ or 1180

µmoles

Baseline = 1180 µmoles

For Exercise 2C, the spontaneous conversion of H₂O₂ into H₂O and O₂ was observed in

an uncatalyzed reaction. The following calculations were made:

0.100 mL KMnO₄ used to titrate the solution

0.100 mL KMnO ₄0.127 moles 1000 mL KMnO ₄ = 0.0000127 moles KMnO₄

0.0000127 moles KMnO 5₄ moles H O ₂ 2₂ moles KMnO ₄ = 0.0000318 moles or 31.8 µmoles of

H₂O₂ remains

Amount of H₂O₂ spontaneously decomposed = Baseline - H₂O₂ remaining

1148 µmoles = 1180 µmoles – 31.8 µmoles

After leaving H₂O₂ uncovered overnight, approximately 1148 µmoles or 97% of the initial

amount was spontaneously decomposed without the aid of catalase.

For Exercise 2D, data was obtained by manipulating the time for catalase activity to see

how time would correlate to H₂O₂ decomposition. The following calculations were used to

determine the moles of H₂O₂ remaining from the catalyzed reaction, and how much had been

decomposed by the catalase:

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Lab Report |8

0 seconds – 3.70 mL of KMnO₄ was used as titrant

3.70 mL KMnO ₄0.127 moles KmnO 5 ₄moles H O ₂1000


₂ mL KMnO 2 moles
₄ KMnO ₄ =

0.00118 moles H₂O₂ remaining

= 1180 µmoles remaining after 0 seconds of catalase activity

Baseline - H₂O₂ remaining = H₂O₂ decomposed

1180 µmoles - 1180 µmoles = 0 µmoles

0 µmoles H₂O₂ decomposed after 0 seconds of catalase activity

10 seconds – 3.00 mL of KMnO₄ used as titrant

3.00 mL KMnO ₄0.127 moles KmnO 5 ₄moles H O ₂1000


₂ mL KMnO 2 moles
₄ KMnO ₄ =

0.000953 moles H₂O₂ remaining

= 953 µmoles remaining after 10 seconds of catalase activity

Baseline - H₂O₂ remaining = H₂O₂ decomposed

1180 µmoles - 953 µmoles = 227 µmoles

227 µmoles H₂O₂ decomposed after 10 seconds of catalase activity

30 seconds – 3.10 mL KMnO₄ used as titrant

3.10 mL KMnO ₄0.127 moles KmnO 5 ₄moles H O ₂1000


₂ mL KMnO 2 moles
₄ KMnO ₄ =

0.000985 moles H₂O₂ remaining

= 985 µmoles remaining after 30 seconds of catalase activity

Baseline - H₂O₂ remaining = H₂O₂ decomposed

1180 µmoles - 985 µmoles = 195 µmoles

195 µmoles H₂O₂ decomposed after 30 seconds of catalase activity

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Lab Report |9

60 seconds – 2.50 mL KMnO₄ used as titrant

2.50 mL KMnO ₄0.127 moles KmnO 5 ₄moles H O ₂1000


₂ mL KMnO 2 moles
₄ KMnO ₄ =

0.000795 moles H₂O₂ remaining

= 795 µmoles remaining after 60 seconds of catalase activity

Baseline - H₂O₂ remaining = H₂O₂ decomposed

1180 µmoles - 795 µmoles = 385 µmoles

385 µmoles H₂O₂ decomposed after 60 seconds of catalase activity

90 seconds – 2.50 mL KMnO₄ used as titrant

2.50 mL KMnO ₄0.127 moles KmnO 5 ₄moles H O ₂1000


₂ mL KMnO 2 moles
₄ KMnO ₄ =

0.000795 moles H₂O₂ remaining

= 795 µmoles remaining after 90 seconds of catalase activity

Baseline - H₂O₂ remaining = H₂O₂ decomposed

1180 µmoles - 795 µmoles = 385 µmoles

385 µmoles H₂O₂ decomposed after 90 seconds of catalase activity

120 seconds – 2.00 mL KMnO₄ used as titrant

2.00 mL KMnO ₄0.127 moles KmnO 5 ₄moles H O ₂1000


₂ mL KMnO 2 moles
₄ KMnO ₄ =

0.000635 moles H₂O₂ remaining

= 635 µmoles remaining after 120 seconds of catalase activity

Baseline - H₂O₂ remaining = H₂O₂ decomposed

1180 µmoles - 635 µmoles = 545 µmoles

545 µmoles H₂O₂ decomposed after 120 seconds of catalase activity

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L a b R e p o r t | 10

180 seconds -- 2.20 mL KMnO₄ used as titrant

2.20 mL KMnO ₄0.127 moles KmnO 5 ₄moles H O ₂1000


₂ mL KMnO 2 moles
₄ KMnO ₄ =

0.000698 moles H₂O₂ remaining

= 698 µmoles remaining after 180 seconds of catalase activity

Baseline - H₂O₂ remaining = H₂O₂ decomposed

1180 µmoles - 635 µmoles = 482 µmoles

10 30 60 90 120 180
0 sec sec sec sec sec sec sec
Baseline (mL) 3.70 3.70 3.70 3.70 3.70 3.70 3.70

Baseline (µmoles)* 1180 1180 1180 1180 1180 1180 1180


18.5
Final Reading (mL) 12.90 37.00 16.00 9.20 0 37.00 24.80
16.0
Initial Reading (mL) 9.20 34.00 12.90 6.70 0 35.00 22.60
Amount of KMnO₄ Consumed
(mL) 3.70 3.00 3.10 2.50 2.50 2.00 2.20
Amount of H₂O₂ Decomposed
(mL) 0.00 0.70 0.60 1.20 1.20 1.70 1.50
Amount of H₂O₂ Decomposed
(µmoles)** 0 227 195 385 385 545 482

482 µmoles H₂O₂ decomposed after 180 seconds of catalase activity

Table 1. This table shows an overview for all of the results for this part of the lab. The manipulated variable was the time
that the catalase was allowed to catalyze. The responding variable would be the amount of H₂O₂ remaining and
decomposed from the reactions. *Calculations for this measurement are shown in the results for 2B (see Lab Report 7)
**Calculations for these measurements are shown above (see Lab Report 8, 9, 10)

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CoolAsianDude left a comment

Hey. Feel free to use data from mine or leave comments on my lab reports that are
incorrect or whatever. I'm kinda rusty so I can't really help improve them.

01 / 31 / 2010
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xing ru lee replied:

can u help me do something about application of knowledge in genetic? It is one of the


chapter which is very difficult..!!! Thank you

02 / 01 / 2010

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do you want to publish more like this? i bet it would be of great help to students...

11 / 18 / 2009
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