Turn the Spectrophotometer on and let warm up 15-20 minutes.
Set the Spec 20's wave length to 500nm.
Make a blank test tube using 4mL of pH5 Buffer, 2mL .02% Hydrogen Peroxide, 1 mL .2% Guaiacol, and 2mL of the Phosphate Buffer. This should be put into one of the 13x100mm test tubes.
Next, we created three different sets of test tubes all containing different amounts of the enzyme extract.
In the first set Test tube S1 had 1ml of pH5 Buffer, 2mL of .02% Hydrogen Peroxide, and 1mL of .2% Guaiacol. Test Tube E1 had 2mL of pH5 Buffer, 1.5mL of the enzyme extract, and 1.5mL of the Phosphate Buffer (All of the test tubes have the same quantities as the first set of test tubes, the only change is the enzyme extract)
For test tube E2 the amount of enzyme extract is 2mL, and for test tube E3 the amount of enzyme extract is 1.75mL.
You want to time each test tube you put in the Spec 20 and record the Absorbancy every 20 seconds for between 240-400 seconds.
To test each tube pour test tube S1 into E1 then E1 back into S1. Start timing as soon as the liquid from S1 touches the liquid in E1.
Wipe the outside of the test tube with the lens paper so there are no smudges or marks and the glass is clear.
You have a period of 20-40 seconds before you should have the test tube in the Spec 20.
Repeat steps 8-10 with all remaining test tubes.
When finished dump solutions down the drain and clean all of your beakers and test tubes.