1. Antioxidant Theory and Application
Mark Lange, PhD
Director, Quality Control
Market America

2. In this Presentation…. How antioxidants work Example antioxidants
Classic Quenching Theory
Antioxidants Protecting Cell Function
Example antioxidants
Coenzyme Q10 (CoQ10)
From fruits & vegetables
Resveravine ®, VitaBlue ®, BioVin ® ,
Cranberry

3. The Classic Quenching Theory
Free radical damage (oxidative stress) can cause degenerative disease.
Antioxidants neutralize (quench) free radicals.

4. Free Radical Formation is Normal to Life
All chemical reactions involve the transfer of unpaired electrons (free radicals).
Free radicals can cause damage to DNA and the cell structure.

5. Free Radicals Damage DNA
Reactive oxygen species and reactive nitrogen species are highly reactive compounds with the potential to damage DNA, proteins and lipids in cell membranes. Both LA and DHLA can directly scavenge and neutralize free radicals.

6. Reactive Oxygen Species are Important Signals for the Body

7. Metabolism
Metabolism is the dominant biological process in our bodies.
Metabolism creates almost all the free radicals in our bodies.

8. Antioxidants’ Big Impact
The ratio of antioxidants to food eaten is tiny.
How can this small amount of antioxidants consumed have such a big impact on our health?

9. Just when I thought I knew all the answers, they changed the questions!

10. Some Antioxidants Act Like Catalysts
Catalysts promote chemical reactions but are not consumed by them.
Extremely small amounts can have very large effects.

11. A New Take on Antioxidants
Antioxidants support and normalize the cell’s metabolism and structure when cells are under stress.
This promotes cell health, maintains good cell energy level, and normalizes cellular functions.

13. After seeing the chart, it’s pretty safe to say…..
The body is terribly complex.
In order to keep functioning, it has to be highly organized and controlled.

14. It Takes Energy to Keep Things Organized

15. Oxidative Stress Leads to Loss of Cellular Function
Diminished cellular energy leads to disorder.
Cellular reactions become less controlled and start to malfunction.
One cause of disease is linked to cell mechanisms malfunctioning.
Uncontrolled oxidative stress can cause a decrease in cellular energy.

16. Alpha Lipoic Acid & CoQ10 recommended.
MnSOD Antioxidant
Found in cell’s mitochondria
Stress creates excess free radicals
These attack mito and cell structure
AO’s neutralize free radicals and stabilize energy production
Alpha Lipoic Acid & CoQ10 recommended.

17. Although the body’s chemistry is controlled by genetics, we can optimize cellular bioenergetics through proper nutrition and exercise.

18. No One Shoe Fits All. No single antioxidant will do everything.
Depends upon genetics.
Depends upon overall health and cellular health.

19. What are antioxidants?
Chemicals that scavenge and neutralize free radicals and other pro-oxidants including: hydroxyl radical, singlet oxygen, superoxide anion, hydrogen peroxide, fatty acid hydroperoxides.
Generally donate electrons to stabilize radicals.
Many types of antioxidants

20. FATTY ACIDS (L-CARNITINE TRANSPORTS) CARBON DIOXIDE + FREE RADICALS
ATP in Mitochondrion
FATTY ACIDS (L-CARNITINE TRANSPORTS)
GLUCOSE
Mitochondrion (CoQ10)
As long as the cell is supplied with fuel and oxygen, the cycle of energy utilization and supply goes on unimpeded millions of times per second in our cells.
Keep in mind that this is a supply-and-demand cycle and if the cell is deprived of either food or oxygen, cellular energy metabolism suffers and cell function is compromised.
OXYGEN
Energy
CARBON DIOXIDE + FREE RADICALS

21. Dietary Sources of CoQ10 (per serving)
Beef: 2.6 mg
Peanuts: 0.8 mg
Egg: 0.1 mg
Fish: 0.9 mg
Cauliflower: 0.4 mg
Strawberries: 0.1 mg
Canola oil: 1.0 mg
Average Intake < 10 mg/d
Most people probably have a dietary intake of less than 10 mg/day of coenzyme Q10. Most likely in the 3-5 mg/day range. Rich sources of dietary CoQ10 include meat, poultry, and fish. Other rich sources include soybean and canola oils, and nuts.
When you fry food, about 25% of the CoQ10 is lost. Boiling foods does not change the CoQ10 content.

22. CoQ10 as an Antioxidant
In its reduced form, CoQ10H2 is an effective fat-soluble antioxidant. CoQ10H2 has been found to inhibit lipid peroxidation. Studies have shown when LDL is oxidized, CoQ10H2 is the first antioxidant consumed.
CoQ10H2 is not found in supplements because of it is an unstable molecule.

23. CoQ10 May Ease Fatigue after Exercise
Double-blinded, placebo-controlled, three crossover design
300 mg/d CoQ10 for 8 days to 17 people,
average age 37.5 years old
“Administration of CoQ10 may attentuate physical fatigue…”
"Antifatigue effects of coenzyme Q10 during physical fatigue." Kei Mizuno, et al, Nutrition Apr;24(4):
In this study 17 healthy volunteers were randomized and took oral coenzyme Q10 (300 mg/d) or placebo. To induce fatigue, subjects exercised on a stationary bicycle for 2 hours, rested for 4, and then did 2 more hours on the bike. After riding the bike for 30 minutes, they did an all out sprint for 10 seconds and another sprint 30 minutes before the end of the ride.
The results showed those participants taking CoQ10 could sprint faster on the bike than those on placebo. Also, the CoQ10 group complained less about fatigue.

24. CoQ10 Protects Against Effects of Aging
Yellow arrows: deformed backbone
Blue arrows: irritation
This study uses mice genetically bred to age quickly.
The top photo shows a twelve-month-old mouse that received no supplemental CoQ10.
Immobile and unresponse
Lesions in and around the eyes
Spinal and limb deformities
Coat was patchy and discolored
These are classic signs of degenerative aging suffered by elderly humans.
The group given conventional CoQ10 faired better, (Middle photo) but visible signs of pathological agiing were still clearly seen.
On the other hand, the mouse supplemented with one of the highly soluble form of CoQ10 was
Alert, responsive and energetic
No physical lesions or deformities
Glossy coast
The mouse at the bottom was shown to age 40% slower than the group receiving conventional coenzyme Q And 51% slower than the mouse taking no CoQ10
Yan J, Fuji K, Yao J, et al. Reduced coenzyme Q10 supplementation decelerates senescence in SAMP1 mice. Exp Gerontol Feb;21(2):130-40

25. ORAC of Selected Foods 2007 U.S. Department of Agriculture
Cranberry Blueberry Plum Blackberry Raspberry Strawberry Cherry Apple Pear 2941

26. Found in Tea, Cocoa, and Red Wine.
Catechin
Found in Tea, Cocoa, and Red Wine.

27. Found in Bilberry, Blueberries, and Cocoa
Cyanidin Structure
Found in Bilberry, Blueberries, and Cocoa

28. Cyanin Structure (Glucoside)
Elderberry.

29. Proanthocyanidin Structure
Found in Grape Seeds and Pine Bark.

30. Why are these such good antioxidants?
They donate an electron from the –OH group on Ring B.
This stabilizes (inactivates) the radical.
The polyphenol forms an aroxyl radical that will not react with tissue.
Vitamin C recycles polyphenols.

31. The Cell Membrane
cytoplasm

33. Berry Extracts
Resveravine® – whole grape extract that contains resveratrol,  -viniferin, and other polymers of resveratrol
BioVin® Advanced – grape skin extract that contains OPCs, anthocyanins, and resveratrol
Vita Blue® – wild blueberry extract that contains anthocyanins and pterostilbene
The trademarked, standardized extracts that will be discussed are shown here.-- Read Slide

34. What is Resveravine®?
A natural extract from grape vine combining the synergistic biological effects of trans-resveratrol and -viniferin
A standardized extract with 20% total trans-resveratrol and  -viniferin
The most concentrated source of Resveratrol from Vitis vinifera
What is Resveravine? Read Slide Please note that resveratrol is not a bioflavonoid like anthocyanins and OPCs that you are more familiar with, but is nonetheless a unique polyphenolic compound that has a number of beneficial biological effects.

35. infection and oxidative stress due to external attacks.
Resveratrol is naturally produced by plants as a defense system against
infection and oxidative stress due to external attacks. UV
Fungi
Copper
Ozone
time
Resveratrol level
Resveratrol is produced in response to environmental insults including ultraviolet light, fungal invasion, copper, and ozone. Red grapes contain substantial amounts of resveratrol and resveratrol polymers, but there are other plants such as the Japanese knotweed that also contain these compounds.

36. Sources of Resveratrol
Resveratrol is found in grapes, wine, grape juice, peanuts, and berries of Vaccinum species, including blueberries, bilberries, and cranberries. In grapes, resveratrol is found only in the skins.

37. Resveravine protects LDL from oxidation better than synthetic resveratrol, catechin and vitamin E
This slide shows that Resveravine is far more potent scavenger of oxygen radicals than vitamin E, catechin (a bioflavonoid), and even resveratrol.

38. Resveratrol is a Superior Antioxidant
Oxidative stress is implicated in numerous disease processes
A lead resveratrol investigator, Dr. Milos Sovak commented, “There is no question that resveratrol is one of the best free-radical scavengers and that it has many effects whose ramifications might affect not only longevity but also general health.”
If what I have shown you so far isn’t enough, then consider that resveratrol is also a potent antioxidant. READ SLIDE.

39. Protection Against Oxidative Stress
Influence of three related polyphenols on the oxidative stress of lymphocytes infected by the Epstein-Barr virus (EBV)
Peroxidation rate (%)
Here is one such experiment of resveratrol’s antioxidant activity. Human lymphocytes are infected with Epstein-Barr virus which creates oxidative stress. Various concentrations of resveratrol and resveravine are added to the cell culture medium. Note that at 2.5 micrograms per milliliter, Resveravine eliminated virtually all of the oxygen radicals.

40. Advantages of Resveravine®
Extremely potent : average ORAC value of 12,700.
Validated by in vitro tests.
Biological effectiveness unique due to synergistic composition of oligo-stilbenes (trans-resveratrol, -viniferin, and other resveratrol polymers).
Natural and safe extract from process using only food solvents and guaranteeing a natural, safe and controlled product from French vines.
Let’s look at some facts and figures about Resveratrol. READ SLIDE.

41. As mentioned, Resveravine also provides  -Viniferin, a dimer of Resveratrol :
Naturally synthesized by the plant, it is involved in the defense system against various fungi such as Botrytis cinerea .
It is naturally present in the grapes and wine
READ SLIDE. There are additional polymers (trimers, tetramers) in Resveravine that all act synergistically with one another.
Like Resveratrol,  -Viniferin has also very interesting biological properties:
anti-inflammatory
antioxidant effects

42. BioVin® Advanced
BioVin Advanced is a French red wine extract, offering a full 5% of pure resveratrol
Made exclusively from French red wine grapes, naturally high in resveratrol
Contains red wine polyphenols and trans-resveratrol
Let’s move on to another berrry extract—this one called BioVin® Advanced. READ SLIDE. It’s still being isolated from French red grapes like Resveravine®, but in this extract contains oligomeric proanthocyanidins (OPCs), anthocyanins, other phenolic acids and 5% naturally occurring resveratrol. Here we are looking at the possibility of more beneficial interactions among these compounds.

43. BioVin® Advanced vs. Wine
Amount of Resveratrol in 200mg BioVin® Advanced
Amount required to be equivalent to 200 mg BioVin® Advanced
Red Wine
White Wine
10 mg
10 glasses (1500 ml)
40 glasses (6000 ml)
This slide gives you a comparison of the amount of resveratrol in 200 mg of BioVin Advanced---that is, 10 mg---versus the amounts in 10 glasses of red wine or 40 glasses of white wine.

44. Benefits of BioVin® Advanced
Due to the synergistic blend of OPCs and resveratrol,
BioVin Advanced offers numerous health advantages
Cardiovascular health
Improves blood circulation
Strengthens blood vessels
Decreases platelet aggregation
Vision health
Excellent antioxidant capabilities
Skin health
Drawing on the OPC, anthocyanin, and resveratrol literature, BioVin Advanced offers numerous health benefits. READ SLIDE.

45. VitaBlue ® Wild Blueberry Extract
Through a unique proprietary process, VitaBlue has concentrated the phytochemicals of the blueberry into an extract of unmatched purity, anthocyanin content and antioxidant potency.
Only the finest, ripest North American wild blueberries qualify to be used in this patented extract.
12.5% Anthocyanins, 40% Polyphenolics, 6500 ORAC units, including pterostilbene.
We move on to another potent berry extract, namely Vita Blue™ Blueberry Extract , which concentrates anthocyanins, other polyphenolic acids, and another interesting compound pterostilbene—a close chemical cousin of resveratrol.

46. Benefits of Blueberries
Blueberries have been shown to be beneficial in several applications
Powerful antioxidant
Inhibits cellular proliferation and induces apoptosis (programmed cell death in unhealthy cells)
Promotes cardiovascular health
Contains COX-2 inhibitors
Promotes healthy vision
READ SLIDE Note the common thread to the previous two berry extracts — induction of apoptosis and inhibition of cell proliferation

47. Cranberry
Native Americans ate cranberries sweetened with maple syrup, in pemmican, and mixed with seal oil for gallbladder attacks.
Traditionally used as a diuretic to prevent kidney stones and treating bladder complaints.

48. Cranberry for Health
Proanthocyanidins prevent bacteria from clinging to the lining of the urinary tract, preventing infection.
Sobota AE. Inhibition of bacterial adherence by cranberry juice: potential use for the treatment of urinary tract infections. J Urol 1984;131:1013-6
Might help with peptic ulcers.
Burger O, Weiss E, Sharon N, Tabak M, Neeman I, and Ofek I. Inhibition of Helicobacter pylori adhesion to human gastric mucus by a high-molecular-weight constituent of cranberry juice. Critical Reviews in Food Science & Nutrition, (Suppl.).
Good oral health in preventing P. gingivalas build-up and resultant inflammation.
Y, Liu, R.H. Cranberries inhibit LDL oxidation and induce LDL
receptor expression in hepatocytes. Life Sci Aug 26;77(15):

49. Conclusions
Antioxidants quench free radicals and stabilize the functioning of stressed cells.
According to the U.S. government’s “5 A Day” plan to increase fruit and vegetable consumption, an estimated 1500 ORAC units is considered beneficial.
Some scientists in anti-aging research suggest that total daily intake should be increased to ORAC units.