1. and add it to test tube

1.     Pour
all of the phosphate pH 7 buffer capsule into a 500 mL volumetric flask, add
about 200 mL of distilled water to the flask and stir the flask gently.

2.     Once
all the powder has dissolved, add the remaining volume of water to reach the
500 mL mark on the volumetric flask.  

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3.     Then,
cut cubes of 1 cm in width of peeled turnip, weigh it on a weighing balance and
cut and add turnip to achieve 50.000 g.

4.     Measure
300 mL of the pH 7 buffer and add it with the weighed turnip into a blender.

5.     Set
the blender for 1-3 minutes bursts at high intensity 3 times until the mixture
is homogenized.

6.     Leave
the mixture to rest for approximately 20 minutes.

7.     Once
the time was passed, filter the solution with filter paper into a beaker.

8.     Label
the beaker “turnip extract” and store it over ice or in the refrigerator.

 

Prepare the enzyme,
methylene blue-pH 5 buffer and substrate solutions 

1.     Obtain
and wear lab coat and goggles.

2.     Obtain
a test tube rack and two test tubes. Label the test tubes “E” for enzyme
extract and “MBB” for methylene blue-pH 5 buffer solution.

3.     Measure
20.00 mL of enzyme extract previously prepared with a 20.00 mL volumetric pipet
and add it to test tube “E”.

4.     Pour
all of the phosphate pH 5 buffer capsule into a 500 mL volumetric flask, add
about 200 mL of distilled water to the flask and stir the flask gently.

5.     Once
all the powder has dissolved, add the remaining volume of water to reach the
500 mL mark on the volumetric flask.  

6.     Measure
1 L of distilled water in a 1 L volumetric flask and add one drop of methylene
blue.

7.     Measure
10 mL of pH 5 buffer solution and 10 mL of diluted methylene blue solution
(each with a 10 mL volumetric pipet) and add them to test tube “MBB”

8.     Obtain
a second test tube rack and six test tubes labelled from “S6” to “S1”

9.     To
obtain the dilutions of hydrogen peroxide measure 1.00 mL of 30% hydrogen
peroxide (H2O2) solution with a 1.00 mL volumetric pipet
and mix it with the following volumes of distilled water in the corresponding
test tube

a.    
4.00 mL creating a 6.00% H2O2
solution

b.    
5.00 mL creating a 5.00% H2O2
solution

c.    
6.50 mL creating a 4.00% H2O2
solution

d.    
9.00 mL creating a 3.00% H2O2
solution

e.    
14.00 mL creating a 2.00% H2O2
solution

f.     
29.00 mL creating a 1.00% H2O2
solution

 

Set up the Colorimeter
and Logger Pro

1.     Prepare
a blank solution by adding 1 mL of 6% hydrogen peroxide solution, 1 mL of
methylene blue-pH 5 buffer solution and 1 mL of distilled water into a cuvette.

Measure these volumes using a 1 mL volumetric pipet.

2.     Connect
the Colorimeter to your Vernier interface, start the program and choose “New”
from the “File” menu.

3.     Place
the cuvette prepared in step 1 in the cuvette slot of the Colorimeter and close
the lid.

4.     Set
the wavelength to 635 nm by pressing the arrow buttons.

5.     Calibrate
the Colorimeter by pressing the “CAL” button. When the LED stops flashing, the
calibration is complete and you can remove the cuvette from the Colorimeter.

6.     Set
up Logger Pro with the following settings before collecting each trial

a.    
Choose “Data Collection” from the
Experiment menu

b.    
Change the data-collection duration to 3
minutes

c.    
Change the sampling rate to 120 samples/minute

d.    
Click “Done”

Collect data

1.     Measure
1.00 mL of solution from test tube “MBB” and 1.00 mL of the corresponding “S”
test tube, add them to a cuvette and place the cuvette in the Colorimeter.

2.     Measure
1.00 mL from test tube “E”.

3.     Press
“start” on Logger Pro

4.     With
the cuvette lid ready in the other hand, add the 1.00 mL from “E” quickly and to
the cuvette. Quickly add the cuvette lid and close the Colorimeter lid.  

5.     Press
“stop” on Logger Pro once

6.     For
each concentration of H2O2 solution, repeat the preceding
steps 1 to 5 thrice.

7.     Once
you have completed the 3 trials for each H2O2 concentration,
repeat the whole process but with Al3+ ions.

8.     Repeat
steps 3, 7 and 9 from the section titled “Prepare the enzyme, methylene blue-pH
5 buffer and substrate solutions”.   

10.  When
preparing the “MBB” test tube, weigh 0.250 g of aluminum sulfate
octadecahydrate (Al2(SO4)2 ·
18H2O, add it to the test tube since it has the highest volume of
water and mix until the salt has fully dissolved.