Basics of Antioxidants

March 12, 2009

1

Outline

1. Introduction

– Definitions

– Free Radicals

– Antioxidants (AOx)

– Oxidative stress

2. Types of AOx

– Different AOx: different mechanisms

– Dietary AOx

– Flavonoids

– Measuring AOx: ORAC

3. Health Effects

– Health effects for Vitamins E, A,C and Flavonoids

4. The AOx Trend: products and claims (Innova)

– Market for AOx

– AOx trend

– Products and claims for Confectionery and Bakery

5. Summary

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1. INTRODUCTION

3

Definitions

• Oxidation=

4

Losing electrons

• Reduction=

Gaining electrons

Free Radicals 1

• Oxygen→ required for life but it is highly

reactive

• During metabolism or infection reactive oxygen species (ROS) are formed→ Free

radicals (FR) – H₂O ₂ = Hydrogen peroxide

– HOCl = Hypochloric acid

– OH⁻ = Hydroxyl radical

– O₂⁻ = Superoxide anion

• Free radicals = unstable and highly reactive molecules (due to unpaired electron)

• The body can initiate the creation of free radicals to defend itself against viruses or bacteria.

5 Foster, 2007; EUFIC, 2009; IFIC, 2009

Other Free radicals formed during inflammation and disease: •Reactive Nitrogen Species (RNS)

FR can be formed by exposure to UV light, cigarette smoke, radiation, pollution etc.

Free Radicals 2

• Free radicals attack stable molecules and steal an electron- creating a new radical→ chain reaction.

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*CVD: cardiovascular disease

DNA

Mutations→

Cancer

Fatty acids

Lipid

peroxidation

→ CVD

Proteins

Enzyme inhibition

Denaturation &

degradation

Halliwell, 1997; Foster, 2007

Antioxidants

• Antioxidants (AOx) are part of the defense system of the body protecting against free radical damage.

• Most AOx donate an electron to FR without becoming FR themselves → chain reaction

stopped.

• AOx do not remove FR completely → keep them at an

optimum level.

• Dietary AOx not produced in the body→ Food

• Consumption of AOx is thought to provide protection

against oxidative stress → positive impact on health

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Image from www.welchs.com

Halliwell, 1997; Foster, 2007

Oxidative stress and disease

AOx

FR

Chronic Injury

Cancer, atherosclerosis, CVD, cataracts, immune

dysfunction, Alzheimer s disease, age-related macular

degeneration, rheumatoid arthritis, aging, etc.

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Oxidative Stress

2. TYPES OF ANTIOXIDANTS

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Types of Antioxidants

10

Different types of AOx →different AOx Mechanisms

Proper AOx: inhibitors interrupting the propagation

of the autoxidation chain reaction

Preventive AOx: inhibitors of FR oxidation reactions

Singlet Oxygen eliminators: e.g. carotenes

Reducing agents which convert hydroperoxides

into stable components in a non-radical way e.g. thiols

and sulfides.

Synergists of proper AOx: they increase the activity

of chain-breaking AOX in a mixture e.g. citric acid

Metal chelators which convert pro-oxidants

(especially iron or copper derivatives) into stable products (e.g. Quercetin,

tannins; phytates)

Inhibition of pro-oxidative enzymes

Pokorný, 2007

Different types of AOx →different measurement methods

FRAP ORAC TRAP

ORAC*

ORAC of Selected Foods- USDA database 2007

* all values presented are Total ORAC= H-ORAC and L-ORAC

Description Mean Unit

Spices, cinnamon,

ground 2675.4 µmol TE/1g

Sorghum, bran, hi-

tannin 2400.0 µmol TE/1g

Spices, Oregano, dried 2001.3 µmol TE/1g

Sorghum, bran, black 1008.0 µmol TE/1g

Cocoa, dry powder,

unsweetened 809.3 µmol TE/1g

Spices, basil, dried 675.5 µmol TE/1g

Sage, fresh 320.0 µmol TE/1g

Marjoram, fresh 273.0 µmol TE/1g

Rice bran, crude 242.9 µmol TE/1g

Nuts, pecans 179.4 µmol TE/1g

Ginger root, raw 148.4 µmol TE/1g

Elderberries, raw 147.0 µmol TE/1g

Peppermint, fresh 139.8 µmol TE/1g

Nuts, walnuts, english 135.4 µmol TE/1g

Nuts, hazelnuts 96.5 µmol TE/1g

Alchoholic beverage,

wine, table, red,

Cabernet Savignon 50.3 µmol TE/1g

Raspberries, raw 48.8 µmol TE/1g

Strawberries, raw 35.8 µmol TE/1g

Pears, raw 29.4 µmol TE/1g

Tea, green, brewed 12.5 µmol TE/1g

Tea, brewed, prepared

with tap water 11.2 µmol TE/1g

Lettuce, raw 2.73

µmol TE/1g

Cucumber, raw 1

µmol TE/1g

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*ORAC= Oxygen Radical Absorbance Capacity

Fruits

Spices/

herbs

Cocoa

Nuts

Wine

Tea

ORAC

12

New High

Antioxidant

Chocolate: Ox-Anti

ORAC method set up internally to measure AOx

capacity of fruits, chocolates, flours, etc.

Ox-Anti (Belcolade)

Acticoa (Callebaut)

Berries

Topfil Blueberry

Topfil Berrissimo

Rye flour

Wheat flour

Values measured internally and subject to a certain standard deviation. Please refer to the

lab for more information about these ORAC values.

Types of Antioxidants 2

13

Enzymes Other Low

molecular

weight AOx

Dietary AOx

Different types of AOx →different AOx Mechanisms, one defense system

SOD (Superoxide Dismutase)

Vitamin E, Vitamin C, β-Carotene,

Flavonoids Uric Acid

Glutathione

Polyphenols

Flavonoids Non- Flavonoids

Flavonols→ Quercetin

Flavones

Flavanols→ Catechin

Flavanones

Stilbenes → Resveratrol

Iso-Flavonoids

Anthocyanidins

Vitamins

A E C

ILSI Europe, 1996

Geissler & Powers, 2005

Phenolic alcohols

Hydroxybenzoic acids

Hydroxycinnamic acids

Lignans

Dietary Antioxidants

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Vitamin A & other Carotenoids

•Singlet oxygen quenchers; peroxyl radical scavengers

•Prevent singlet oxygen from generating FR

Vitamin E

•Proper AOx→chain breaking function

•Responsible for protecting the PUFA in cell membranes from oxidation

•Upregulates the activity of Aox enzymes

Vitamin C

• Proper AOx→chain- breaking function

• Powerful reducing agent -can directly scavenge superoxide, hydroxyl radicals, singlet oxygen and H₂O ₂

• Can re-generate Vitamin E from tocopheroxyl radical providing membrane protection

Geissler & Powers, 2005

Pokorný, 2007

Foster, 2007

Dietary Antioxidants

Zinc, Copper, Manganese & Selenium

•protect against oxidative stress indirectly serving as cofactors for AOX enzymes: SOD superoxide dismutase & GPx Glutathione peroxidase.

Polyphenols

•Effective FR scavengers; chain-breaking function

•Chelating agents : indirect AOx action

15 Geissler & Powers, 2005

Pokorný, 2007

Foster, 2007

Flavonoids

• Flavonols

• Flavanols

• Flavonones

• Anthocyanins

• Isoflavones

• Flavones

16

Flavonoids → most common group of

plant phenolics

→ +4000 identified in plants

→ Share a common structure (2 Benzene rings

and a central pyran ring) which determines their

AOx functioning

D Archivio, 2007

Flavonoids

Flavonols • Quercetin in Apples and Onions

Flavanols (Flavan-3-ols) • Epicatechin in cocoa and chocolate • Catechin in Red wine and Black tea • Epigallocatechin gallate found in Green tea

Anthocyanins • Cyanidin found in Orange juice • Malvidin found in Red wine

Isoflavones • Daidzein in Soy Milk

Flavanones • Hesperidin found in Orange Juice • Naringenin found in Grapefruit juice

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D Archivio, 2007

3. HEALTH EFFECTS

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Health Effects

• Epidemiologic evidence suggesting a protective effect of Fruits & Vegetables (F&V) against Cancer and Cardiovascular diseases (CVDs). – Meta-analysis by Gladys Block (2001)

– Studied relation between F& V and different types of Cancer

– 126/ 156 studies → beneficial effects

– Lung Cancer→24/25 studies showed beneficial effects.

• Conclusion: Increased consumption of F&V is protective against Cancer and CVDs

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Hypothesis: F&V contain compounds responsible for the health effects

ILSI Europe, 1996

Health Effects- Vitamin E V

itam

in E

• Ischemic heart disease (IHD)* • MONICA (1980s) higher plasma vitamin E correlated with lower risk

of IHD • CHD* • Nurses´Health Study and Male Health Profesionals Study – high

intakes of Vitamin E correlated with lower risk of CHD • Angina Pectoris • The Edinburg study – high levels of plasma Vitamin E associated with

lowest risk of Angina Pectoris • Prostate Cancer • SELECT study no effects on prostate cancer on treatment with

Vitamin E and Selenium, but a non-significant trend found of increased incidence of prostate cancer in the treatment group.

• ATBC study –significant reduction in the incidence of prostate cancer

20 *IHDis a disease characterized by reduced blood supply to the heart muscle, usually due to coronary artery disease (atherosclerosis of the coronary arteries) Angina Pectoris: severe chest pain.

*CHD is Coronary heart disease refers to the failure of coronary circulation to supply adequate circulation to cardiac muscle and surrounding tissue. *MONICA= Monitoring trends and determinants In Cardiovascular disease

*ATBC= Alpha Tocopherol, Beta-Carotene Cancer Prevention Trial.

Health Effects- Vitamin A

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Vitam

in A

(β

-C

aro

ten

e) • Cancer

• ATBC* study (1994)- heavy smokers • Found a statistically significant 18% higher incidence

of lung cancer in subjects supplied with β-Carotene. • CARET study(1990s)- heavy smokers • 28% increased risk for lung cancer in treatment group

and 17% increase in total mortality→trial terminated early

• Physicians Health Study • Supplementation with 50 mg β-Carotene every other day

showed no harm or benefit in smokers and non-smokers.

AOx – can potentially become

pro-oxidative!

*ATBC= Alpha Tocopherol, Beta-Carotene Cancer Prevention Trial. * CARET=Beta Carotene and Retinol Efficacy Trial

Possibility: Cancer process was

accelerated by β-Carotene in smokers

Health Effects- Vitamin C

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Vitam

in C

(A

scorb

ic a

cid

)

• CVD • Systematic review by Asplund, K (2002)- Cohort studies showed a

reduced risk of Cardiovascular events with high consumption of vitamin / high plasma levels of vitamin C.

• Randomized Clinical Trials did not show an effect of supplementation with Vitamin C.

• Vasodilation • Vasodilation improved in patients with endothelial dysfunction→

marker for onset of atherosclerosis and CVD • Cancer • EPIC- Norfolk study, Khaw et al (2001) and Loria et al (2000)- Both

found that high plasma vitamin C correlated with lower cancer mortality.

• Moertel et al (1985) RCT- High- dose vitamin C in patients with advanced Cancer→ no effect

• Cochrane Review (2008)- No convincing evidence for all AOx – including Vitamin C- for Gastrointestinal Cancer risk.

• Yeom et al (2007) -Quality of life improvement in Cancer patients with high dose Vitamin C supplementation.

EPIC= European Prospective Investigation of Cancer

Li and Schellhorn, 2007

Health Effects- Flavonoids

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Fla

vonoid

s

• Coronary Heart Disease • Zutphen Elderly Study; Finnish Mobile Clinic Health Survey; Ding et al 2006 (meta-analysis).

• Protective effects on non-fatal coronary heart disease • Vasodilation/ Endothelial function/ blood pressure • Duffy et al 2001; Taubert et al 2003 • Tea catechins improved short-term bloodflow and vasodilation

• Platelet function • Rein et al 2000 • Cocoa and wine flavonoids can inhibit platelet aggregation

• LDL Cholesterol • Osakabe et al 2001; Bearden et al 2000; Pearson et al 2001 • Cocoa flavonoids inhibit or retard LDL oxdiation- a marker for CVD

• Inflammation • Selmi et al 2006 • Cocoa Flavonoids modulate inflammation markers (eicosanoids)

• Cancer • Nurses´ Health Study II; Neuhouse et al 2004 • No overall association between intake of flavonols and the risk of breast cancer. • Quercetin may reduce the risk of lung cancer

Antioxidants and health effects

Antioxidants

CVD

-Vitamin E indications of reduced risk of

Ischemic Heart disease and Angina pectoris

-Flavonoids modulate inflammation markers

Cholesterol

- Flavonoids inhibit/retard LDL

Oxidation

Blood Pressure

- Flavonoids inhibit platelet aggregation

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Conclusions on Health Effects

• Good evidence for: – the role of FR in Cancer and CVD

– modulation of FR events by AOx

• Human studies suggest: – correlation between (some) AOx intakes and serum

concentration and incidence of mortality from cancer and CVD-

– But, results are not consistent!

• Optimal dosages are unknown: – the level of intake of AOx nutrients desirable for

optimal nutrition is still an open question.

– Genetics might play a role.

• Fundamental questions: – What causes the health effects? The food matrix as

a whole or the individual components ?

– F& V or supplements? 25

?

IFIC, 2006

Willet, 20001

Toner, 2004

4. THE AOX TREND: PRODUCTS AND CLAIMS

The Innova Database

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Market for AOx

27

10% of product

launches positioned as

“Antioxidant” were

tracked in the Bakery

and Cereals category

7% of product launches

positioned as

“Antioxidant” were

tracked in the

Confectionery category

5 year Data from 02/2004 – 02/ 2009

Antioxidant Trend for 2009 (Mellentin,J.)

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In 2008, AOx were listed as a

“Key Trend” for the year.

In 2009, AOx are predicted to

become a “Micro Trend”

Mellentin, 2008- 10 Key Trends in Food, Nutrition and Health.

The message has become a

category standard→ “all teas and

all dark chocolates contain AOx”

Regulatory problem→ what kind

of evidence needed to prove

health effects?

Industry option= add a point of

difference to the AOx message by

using specific and scientific

marketing (terms like Flavonoids

and ORAC)

Industry option= add a point of

difference to the AOx message by

combining different sources such

as Superfruits and Dark

Chocolate

Bakery & Cereals and Confectionery categories

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Bakery and Cereals category

Confectionery category 5 year Data from 02/2004 – 02/ 2009

Confectionery

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Brazil

“Natural source

of Antioxidant

Flavonoids”

“Natural

source of

Flavanol

Antioxidants

”

USA

“ With Grape pulp

extract, naturally-rich

in Antioxidants”

Netherlands

“ With Chilli, natural-

source of Antioxidants”

Germany

USA

“Natural source

of flavanol

antioxidants.

measured by an

ORAC (Oxygen

Radical

Absorbance

Capacity)

“ Extra

Antioxidants from

ACTICOA® ”

UK

“With Cacao

Flavanols for

good blood

circulation”

Bakery and Cereals

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USA

“Fiber plus

Antioxidants”

Sweden

“With two

sources of

antioxidants:

green tea extract

and vitamin E”

“A better protection

against oxidant stress,

thanks to addition of

natural antioxidants.”

India

Sweden

“A soft, sliced bread

with sunflower seeds.

Contains vitamins and

minerals, antioxidants,

and fiber”

France

“with precious

antioxidants in order to

protect your body from

aging”

“Purpur wheat, contains 10 times

more Anthocyanins than other

kinds of wheat. Anthocyanins have

an antioxidant effect”.

Hungary

“Natural

Antioxidants and

slow energy”

« Contains

natural

antioxidants ». Russia

Other products

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UK

“Contains

Antioxidants”

Cranberry and

Wine sauce

“Naturally rich in

Catechins”

Portugal

NL

« natural

antioxidants »

“Coffee Arabica

enriched with

Antioxidants”

France

India

“Source of

Antioxidants”

“With fibers,

Antioxidants,

Vitamins and

Minerals (from

berries)”

5. SUMMARY

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Summary

34

Free Radicals

•Needed for the defense mechanism of the body

•Unstable and reactive: an excess may damage cells

•Excess FR may damage DNA, proteins and fatty acids

Antioxidants

•Part of the defense system of our bodies

•AOX protect us from excess FR damage

•Keep FR at an optimum level to prevent oxidative stress and chronic disease

Types of Antioxidants

•Different types of AOX→different working mechanisms

• Major types: Enzymes, Dietary AOX, Other low molecular weight AOX

•Dietary AOX: Vitamins A, C, E and Polyphenols

•Flavonoids: most common group of polyphenols

Summary

Health Effects

•Good evidence for the role of AOX in modulation of FR

•Good evidence of the role of FR in the onset of some diseases (Cancer and CVD)

•No consistent results

•No optimal dosage yet

•Fundamental questions still to answer

AOX Trend

•AOX move from a key trend in 2008 to a microtrend in 2009

•AOX message has become a category standard

•Point of difference is searched by Industry

•Major category players: chocolate and tea

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Puratos wants to contribute

to the balanced diet of

consumers

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