Peeling Away Problems

The Antioxidating, Antimicrobial, and Antimutagenic Effects of Tree Bark

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Extracting Antioxidants

1.      Weigh 5g of each tree bark powder and put into a 50mL tube. 

2.      Add 30mL of water to each tube. 

3.      Shake overnight. 

4.      Heat tube in 100ºC or 50ºC water for 10 minutes. 

5.      Centrifuge at 3000rpm for 5 minutes. 

6.      Transfer supernatant (clear top aqueous part) in to a new tube. 

7.      Add 20mL of water to the first tubes. 

8.      Shake them on a shaker at 200rpm for 4 hours. 

9.      Heat tube in 100ºC or 50ºC water for 10 minutes. 

10.  Centrifuge at 3000rpm for 5 minutes. 

11.  Combine 2 supernatants. 

12.  Add 5mL of chloroform (CHCl3) to the second tubes.

13.  Mix. 

14.  Centrifuge at 3000rpm for 5 minutes. 

15.  Remove supernatant (extract, 40mL) into flasks. 

16.  Transfer 1mL of extract into tubes to be used in the FRAP Assay and the Folin-Ciocalteau Assay. 

17.  Dry other 39mL using a vacuum evaporator to about 1mL. 

18.  Transfer into a new tube for the bacterial susceptibility test and Ames Test.

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