1. The Sensational Science Behind Antioxidants: Testing Facts vs. Myths of popular antioxidant products

2. Yesterday, you broke up into groups and selected a popular “antioxidant-rich” product to endorse Today you will chemically test the drinks to see if there’s truth to their marketing. You are the Research & Development Team and the Marketing Team for your product.

3. Victims of Oxidation Part I: The science behind antioxidants Free radical with unpaired electron Free radicals cause damage by oxidation

4. O     O         Molecular Oxygen O 2 O     O          O     O          H H O   O           O       H  O       H  Add two H + Add 3 rd e -  Hydroxyl ion OH - Add 1 st e - Add 2 nd e - Superoxide radical  O 2 - Peroxide radical  O 2 -2 Hydrogen peroxide H 2 O 2 Hydroxyl radical  OH Add 4 th e - O       H   Hydroxyl ion OH - Add 2H + O      H   H  O      H   H  Water   

5. Oxidation occurs when one substance transfers one or more its electrons to another substance Catechol Colorless + ½O 2 Benzoquinone Yellow-Brown + H 2 O Electrons lost to “Catechol is oxidized by O 2 to become benzoquinone” “Catechol loses its electrons to O 2 to become benzoquinone” Free radicals cause damage by oxidation

6. Catechol Colorless + ½O 2 Benzoquinone Yellow-Brown + H 2 O Phenol Oxidase Phenol Oxidase catalyzes the reaction that changes catechol, in the presence of O 2, into benzoquinone

7. Catechol Colorless + ½O 2 Benzoquinone Yellow-Brown + H 2 O Phenol Oxidase Phenol Oxidase catalyzes the reaction that changes catechol, in the presence of O 2, into benzoquinone + Antioxidant! X

8. Browning reactions are due to oxidation

9. Catechol Colorless + ½O 2 Benzoquinone Yellow-Brown + H 2 O Phenol Oxidase Phenol Oxidase catalyzes the reaction that changes catechol, in the presence of O 2, into benzoquinone

10. Can any of these products inhibit the browning reaction in mushrooms? Buffer pH 7 100X Daily Dose of Vitamin C x 10 Controls:

11. 1. Weigh mushroom 2. Chop mushroom into pieces and put into Ziploc bag Multiply mass by 4 and add that amount of buffer to the bag 3. Close bag tightly and mash gently with fingers 4. Wet the coffee filter with buffer and place over cup. Use pipette to transfer mushroom extract to filter Purifying mushroom oxidase Perform the tests!

12. Spot test 1.Using the pipette, add two drops of mushroom oxidase to every square on your table. 2.Add the appropriate reactants to each square in the order they are listed 3.Once all the drops have been added to the 6 squares, start the timer. 4.Observe the reaction for 5 minutes, then answer the following questions. Hint: Use a steady hand so that each drop is the same amount of liquid. Make sure the paper is level, so the drops do not move. Time = 0 min Time = 5 min

13. http://science-edu.larc.nasa.gov/EDDOCS/Wavelengths_for_Colors.html Double bonds absorb light

14. Light absorption is like a fingerprint for compounds

15. Absorbance Readout 0.430 Light Prism to disperse light Wavelength selector How does a spectrophotometer work? Catechol Colorless + ½O 2 Benzoquinone Yellow-Brown + H 2 O 0.5200.240 At Abs 490 nm:

16. Spectrophotometer Test 1.Label your test tubes #2, #4, #6 and buffer blank 2.Add 4 ml of buffer to “buffer blank”. Place the tube into spectrophotometer holder and use the left knob to set the numbers in the absorbance window to 0.000. 3.Add the reactants for Tube #2—use correct pipettes for each reactant! Mix by tapping the test tube, and place tube into spectrophotometer. Start the timer and record absorbance at t=0 min. 4.Remove tube at t = 4:30 min and mix by tapping test tube. Record absorbance at t = 5 min. 5.Repeat steps 3-4 for each Tubes #4, #6.

17. There are many different ways to test for antioxidants Our mushroom oxidase test may not be sensitive enough to pick up on antioxidant levels

18. Catechol Colorless + ½O 2 Benzoquinone Yellow-Brown + H 2 O Phenol Oxidase + Antioxidant! X

19. Catechol Colorless + ½O 2 Benzoquinone Yellow-Brown + H 2 O Phenol Oxidase + Antioxidant! X Your drink has antioxidants, but it may be very low amounts compared to 100X Daily Value of Vitamin C

20. Part II: The marketing behind antioxidants

21. Present a 3 minute marketing campaign that showcases:  Your knowledge on the mechanism and health benefits of antioxidants defense against free radicals.  Your product’s antioxidant test results.  Your opinion why your classmates should buy your antioxidant drink.

22. Edited by Ms. Theresa (Instructor note: show a video clip of antioxidants in the media. In my presentation, I edited together a video of 9 different commercials, interviews on the topic of antioxidants)

23. 1.Do not make scientific claims that you cannot back with evidence 2.Find a different approach to market the product

24. Much research has to be performed before claims can be made

25. “I recommend pizza since it is a vegetable and therefore has *tons* of antioxidants.” I’m actually a real doctor! Google me!

26. The best marketing campaign was for: The best scientific argument was made by: Write-in: Based on the overall campaign and facts, I would consider buying: Sample Ballot Which product is the best antioxidant drink on the market? ☐ 7-Up Antioxidant ☐ Activate Antioxidants ☐ FUZE Triple Berry ☐ Lipton Green Tea ☐ POM Wonderful ☐ Vitamin Water XXX ☐ Other ___________ ☐ 7-Up Antioxidant ☐ Activate Antioxidants ☐ FUZE Triple Berry ☐ Lipton Green Tea ☐ POM Wonderful ☐ Vitamin Water XXX ☐ Other ___________