Barry Callebaut confirms

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Barry Callebaut confirms the power of
polyphenols in chocolate November 29, 2005
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Agenda

• Dirk Poelman, Barry Callebaut.
• Barry Callebauts research in polyphenols
• Dr. Alex Collie, PhD.
• The effects of polyphenols on cognitive
  functions
• Mr. Guillaume Tenca.
• The power of polyphenols - the positive effect
  on prostate cancer cells
• Questions Answers
• Drinks Chocolates

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CHOCOLATE, MORE THAN JUST INDULGENCE
Dirk Poelman FIE 29.11.2005
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Chocolate, Delicious and Healthy
Chocolate, More Than Just Indulgence
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Chocolate, Delicious and Healthy

• Healthy Components in Cocoa
• Vitamins Minerals
• Fibres
• Xanthines (Theo bromine)
• Anandamide
• Stearic Acid
• Polyphenols

Chocolate, More Than Just Indulgence
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Chocolate, Delicious and Healthy
Research Focus for Barry Callebaut
Strengthen the Natural Benefits of Cocoa and
Chocolate
Chocolate, More Than Just Indulgence
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ACTICOA Chocolate, The revitalizing Power of
Chocolate
Chocolate, More Than Just Indulgence
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Polyphenols, The active Substance of Chocolate

• What are Polyphenols ?
• Naturally occurring plant substances.
• Cocoa contains more polyphenols than red wine or
  tea.
• Cocoa polyphenols are more active as an
  anti-oxidant.
• Anti-oxidants protect against free radicals.
• Created via physical activity.
• Attack body cells ? impact on diseases and aging.

Polyphenols are very effective aniti-oxidants
that block free radical damage to the body.
Chocolate, More Than Just Indulgence
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Polyphenols, The active Substance of Chocolate

• Potential Health Benefits of Polyphenols
• Cardio vascular
• Cholesterol
• Inhibition of platelet function ? anti-thrombotic
• Cancer prevention
• Regulation of the immune system
• Improved brain function ? cognitive behaviour
• Activity level

Chocolate, More Than Just Indulgence
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Acticoa, a natural fortification Process

• During the normal cocoa processing from the
• cocoa tree up to the chocolate, there is a huge
• reduction in polyphenol content and
• anti-oxidant activity.
• Fermentation and drying
• Heat treatments during processing (mainly
  roasting and conching)
• Alkalisation treatment
• Polyphenol reduction from dry cocoa beans up to
  cocoa liquor

Chocolate, More Than Just Indulgence
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Acticoa, a natural fortification Process

• Because of the full vertical integration,
• Barry Callebaut can influence the whole chain
• and come to superior polyphenol levels.

The Acticoa process is designed to preserve the
naturally present high levels of active
polyphenols in cocoa, which are for a big part
lost during the normal processing.
Chocolate, More Than Just Indulgence
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Acticoa, a natural fortification Process
During fermentation polyphenols diffuse with cell
liquids from their storage cells and undergo
oxidation to condensed high molecular mostly
insoluble tannins
2 days of Sun drying of unfermented beans can
cause a 50 decrease in epicathechine content
Chocolate, More Than Just Indulgence
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Scientifically proven

• Research Results (HBe)
• 1) Test on animals
• Anti-oxidant effect reduction of free radicals
• Cognitive maintaining optimal brain function in
  24 months old rats
• Prevention of prostate cancer growth
• Prevention prostate enlargement
• 2) In vitro tests on human cells
• Prostate cancer high cell reduction of local
  and metastatic prostate cancer
  cells after polyphenol addition
• Cardiovascular positive effect on endothelium
  cells (endothelin-1 synthesis inhibition)
• 3) Tests on humans
• Cardiovascular decrease of the biomaker
  malondialdehyde
• Cognitive tests

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ACTICOATM Chocolate, with Polyphenols
Product Offering Barry Callebaut offers
chocolate with the highest Polyphenol content
available on the market Dark chocolate gt
minimum 3,2 polyphenol content Milk
chocolate gt minimum 1,1 polyphenol
content Available from liquid couverture to
blocks to finished products. Workable as standard
chocolate Available from February 2006
Chocolate, More Than Just Indulgence
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ACTICOATM chocolate is ....
Natural ACTICOATM chocolate preserves the
antioxidants that naturally occur in the
cocoa bean but are lost in chocolate
production Tasty ACTICOATM chocolate retains
the flavor of delightful true
chocolate Healthy ACTICOATM chocolate is
rich in active polyphenols
ACTICOATM chocolate makes you feel good in body
and mind
Chocolate, More Than Just Indulgence
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The effect of polyphenols on cognition

• Dr Alex Collie, PhD
• Senior Clinical Scientist, CogState Ltd, United
  Kingdom
• Senior Fellow, Centre for Neuroscience, The
  University of Melbourne, Australia.
• acollie_at_cogstate.com

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Cocoa polyphenols

• Cocoa is a rich source of polyphenols
• 2x red wine
• 4x tea
• 3 major types of flavanols in cocoa
• Catechin, Epicatechin, Procyanidin
• Anti-oxidant activity
• Rapidly absorbed by body
• Increase in plasma concentration after 2 weeks
  dietary supplementation

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Health benefits of polyphenols

• Improvement in cardiovascular health
• Lower cholesterol
• Decrease blood pressure
• Enhanced immune function
• Better response of immune cells
• Reduction in cancer risk
• Improvement in cognition ??
• Attention Vigilance
• Learning Memory
• Planning Problem solving

Engler et al (2004) J Am Coll Nutr Murphy et al
(2003) Am J Clin Nutr Wan et al (2001) Am J
Clin Nutr Rimm (2002) Am J Clin Nutr Keen et al
(2005) Am J Clin Nutr
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Polyphenols cognition - animal studies

• Dietary supplementation
• Minimizes aging-related cognitive deficits.
• Protects against memory impairment induced by
• Cerebral ischemia.
• Scopolamine administration.
• Heat shock.

Joseph et al (1999) J Neurosci Enno et al (2004)
Exp Gerontol Pu et al (2004) J Pharmaol Sci Kim
et al (2004) Biosci Biotech Biochem Joseph et al
(1998) J Neuorsci.
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Polyphenols cognition - animal studies

• Aged rates
• 8 weeks dietary supplementation.
• Polyphenol-rich foods.
• Morris water maze test before after.
• Positive effects on learning and memory.

Joseph et al (1999) J Neurosci
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Polyphenols cognition - human studies

• Population studies.
• Red wine consumption
• Rich source of polyphenols.
• PAQUID study in Bordeaux, France (1990s).
• 3-4 glasses red wine per day.
• Lower incidence of cognitive impairment.
• Lower rate of conversion to Alzheimers disease.
• Vitamin E
• Anti-oxidant properties.
• Higher intake associated with better cognition.
• Lower intake associated with worse cognition.

Letennaur et al (1993) Ann Intern Med Orgogozo
et al (1997) Rev Neuro Ortega et al (1997) Am J
Clin Nutr Perkins et al (1999) Am J Epidemiol.
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Polyphenols cognition - human studies

• Randomised, Controlled Trials (RCTs).
• Ginkgo Biloba
• Major active ingredient is flavanol polyphenols
  (24).
• Long-term consumption improves cognition
• Healthy young people.
• Healthy older people.
• Short-term consumption improves cognition
• Healthy young people.

Mix Crews (2002) Hum Psychopharm Kennedy et al
(2003) Psychopharm Stough et al (2001) Int J
Neuropsychopharm
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Short-term effects of ginkgo biloba

• Young adults
• Single-dose of ginkgo biloba or placebo.
• Placebo, 120mg, 240mg or 360mg.
• Computer cognitive tests before and after.
• Positive effects on attention at high doses.

Kennedy et al (2003) Psychopharm
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Polyphenols cognition - human studies

• Good evidence that polyphenols polyphenol-rich
  supplements can positively affect cognition
• Animals.
• Humans.
• To date, there have been no studies examining the
  effect of cocoa polyphenols.
• Barry-Callebaut studies
• Effect of a single dose of cocoa polyphenols on
  human cognition.
• Effect of 8 weeks dietary supplementation with
  cocoa polyphenols on human cognition.

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Measuring cognition in humans

• Maze Learning Test.
• Human analogue of Morris Water Maze.
• Administered via touch-sensitive screen.
• Task is to find your way through the hidden maze.
• Speed accuracy are recorded.
• Culture-neutral.
• Minimal language.
• Game-like.

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Study 1. Short-term effects

• Question Does a single-dose of polyphenol-rich
  chocolate result in short-term improvement in
  cognition?
• Randomised, controlled trial (RCT).
• Comparing polyphenol-rich chocolate to placebo.
• Test cognition before and after consumption.
• Cross-over design
• Group 1 chocolate then placebo
• Group 2 placebo then chocolate
• Healthy older people (50-65 years)

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Study 2. Long-term effects

• Question Does 8 weeks daily consumption of
  polyphenol-rich chocolate improve cognition in
  older people?
• Randomised, controlled trial (RCT).
• Comparing polyphenol-rich chocolate to placebo.
• Test cognition before, during and after
  consumption.
• Parallel groups design
• Group 1 Chocolate 1
• Group 2 Chocolate 2
• Group 3 Placebo
• Healthy older people (50-65 years)

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Summary

• Polyphenols positively effect numerous aspects of
  health.
• Good evidence from animal and human studies that
  polyphenols may enhance cognitive performance.
• However, there have been no cognitive studies of
  cocoa polyphenols.
• Barry-Callebaut studies are the first to address
  this important question.

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The positive effect of polyphenols

• Guillaume TENCA
• Business Development DirectorBIOalternatives
• France

www.bioalternatives.com
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Coffee, tea, wine or cocoa?
Comparison of the antioxidant activity of
commonly consumed polyphenolic beverages (coffee,
cocoa, and tea) prepared per cup serving. (J
Agric Food Chem. 2001) Nestle Research Center
Cocoa has more phenolic phytochemicals and a
higher antioxidant capacity than teas and red
wine. (J Agric Food Chem. 2003 Dec ) Seoul
National University
Cocoa gt red wine gt green tea gt black tea
BBC NEWS (2003/11/08) Cup of cocoa may keep the
doctor away !
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Food and prevention of cancer

• Connection between style of living and pathologic
  incidents.
• The importance of nutrition and cancer risk
  breast cancer, cancer of digestion and prostate
  cancer.

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Free radicals

• The oxygen molecule will take an electron and
  transform it into a free radical
• Free radical a result of detoxication of fat
  and chemical products medications, cigarette
  smoke, pollution, sunbeam, radiation stress

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Polyphenols

• natural exogenous antioxidant
• Find it in fruits, vegetables, cereals, chocolate
• and beverages such as tea, coffee or wine.

• Mechanisms
• Neutralise the free radicals (direct action)
• Stimulate the natural systems of detoxication
  (indirect action)
• Novel mechanisms of natural antioxidant compounds
  in
• biological systems involvement of glutathione
  and
• glutathione-related enzymes- Oct 2005 - Italia

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Polyphenols prevention of cancer

• Protection against mutagenicity and
  genotoxicity
• Inhibition of biochemicals markers of tumor
  initiation
• Inhibition of biochemical markers of tumor
  promotion
• Effects on detoxication enzymes
• Trapping of activated metabolites of
  carcinogens
• Antixocydant and free radical scavenging
  activity.

The American Journal of Clinical Nutrition
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In vitro effects of polyphenols from cocoa on the
growth of human prostate cancer and normal cells
RWEP1 (ATCC, CRL-11609) Human normal prostate
cell line Morphology epithelial
DU145 (ATCC, HTB-81) Human prostate cancer cell
line (metastasic site) Morphology epithelial
22Rv1 (ATCC, CRL-2505) Human prostate cancer cell
line (tumor site) Morphology epithelial
Cytotoxicity assays with 4 cocoa extracts at
several time of treatment (1H, 3H, 6H, 24H, 48H
et 72H) on each cells lines.
NEUTRAL RED TEST
The uptake of the NR vital dye into the
lysosomes/endosomes and vacuoles of living cells
is used as a quantitative indication of cell
number and viability after treatment with a
chemical test
SULFORHODAMINE B TEST
The SRB anionic dye quantifies viable cells by
measuring the protein content of cells.
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TIME DOSE DEPENDANT ASSAY ON 22Rv1 CELLS
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TIME DOSE DEPENDANT ASSAY ON DU145 CELLS
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Conclusion
In conclusion, for each cocoa extract 22Rv1 cells
were more sensitive to cocoa extract than DU145
cells. This results attempted to demonstrate
that the polyphenols of cocoa extract had a
specific anticancer activity against prostate
carcinoma and metastasis cells. Furthermore, our
results are the first to describe a potent
anti-proliferative effect of cocoa polyphenols on
human prostate cancer cells. t is interesting to
note that all cocoa extracts, at highest
concentrations, induce a complete inhibition of
tumor cells without effect on RWEP-1 cells.
Cocoa extract are more efficient on
non-metastasis cells than on metastasis tumors.
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Conclusion

• Polyphenols are the most abundant antioxidants
• Fruits, vegetables, cereals, tea, coffee, wine
  and cocoa
• Polyphenols can stimulate endogenous defense
  systems and detoxication
• Cocoa polyphenols present a strong interest for
  some disease prevention such as cancer.

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Questions Answers
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Drinks Chocolates