Efficacy of Polyphenols From Milk and Dark Chocolate

• Sponsor: Nestlé (Industry)
• Overall Status: Completed
• CT.gov ID: NCT00513344
• Collaborator: (none)
• Enrollment: 6
• Location: 1
• Arms: 3
• Duration (Months): 46
• Patients Per Site Per Month: 0.1

Study Details

Study Description

Brief Summary

Dark chocolate is one of the richest sources of polyphenols though it has been hypothesised that the bioavailability and therefore probably the bioefficacy of epicatechin from milk chocolate was reduced compared to dark. This study is designed to compare milk and dark chocolate as a source of polyphenols with a control "chocolate" for improving a risk biomarker for vascular disease.

Condition or Disease	Intervention/Treatment	Phase
No Disease	Dietary Supplement: Dark Chocolate; Dietary Supplement: Milk Chocolate; Dietary Supplement: Control (polyphenol-free) "Chocolate"	N/A

Detailed Description

Dark chocolate is one of the richest sources of polyphenols, for example, a standard 40g portion of dark chocolate contains 400-800 mg of polyphenols, compared to red wine (170 mg /100ml) or an apple (200 mg/piece). Cocoa polyphenols, most notably the catechins, can exist in both lipid and water-based environments (amphipathic), meaning they can spare both lipophilic and hydrophilic vitamins. There have been a number of human trials conducted using chocolate or cocoa and measuring various endpoints. Most have been conducted with dark chocolate. An article in Nature found that the bioavailability of epicatechin from milk chocolate was substantially reduced compared to dark, and even dark taken with a glass of milk (Serafini et al 2003). The hypothesis was that the milk proteins bind to polyphenols, making them unavailable. Subsequent studies have not been able to reproduce this, but none have been conducted using solid chocolate as the first study, all have been done using a drink matrix, which may completely alter the binding interactions of the polyphenols and protein. To this end, this study is designed to compare solid chocolates as a source of polyphenols for improving a risk biomarker for vascular disease.

This study is designed as a blinded, three arm crossover trial. The primary outcome measure is to compare endothelial function after consumption of 3 chocolates (1 milk, 1 dark, 1 polyphenol-free control) with a secondary outcome of arterial stiffness. All volunteers will take all chocolate types in a crossover design. Subjects will undergo medical screening, anthropometry, physical activity and dietary assessments before randomization for the order of consumption.

Study Design

• Study Type: Interventional
• Actual Enrollment: 6 participants
• Allocation: Randomized
• Intervention Model: Crossover Assignment
• Masking: Single (Investigator)
• Primary Purpose: Basic Science
• Official Title: Efficacy of Polyphenols From Milk and Dark Chocolate
• Study Start Date: Jun 1, 2008
• Actual Primary Completion Date: Jun 1, 2009
• Actual Study Completion Date: Apr 1, 2012

Arms and Interventions

Arm	Intervention/Treatment
Experimental: dark chocolate containing polyphenols dark chocolate	Dietary Supplement: Dark Chocolate; 1 portion
Experimental: Milk chocolate containing polyphenols Bespoke milk chocolate	Dietary Supplement: Milk Chocolate; 1 portion
Active Comparator: Control chocolate with no polyphenols cocoa-free chocolate	Dietary Supplement: Control (polyphenol-free) "Chocolate"; one portion

Outcome Measures

Primary Outcome Measures

1. Change in Reactive Hyperemia Index (RHI) From Baseline (20 Min Before Product Intake) to 2 Hours Following Product Intake [Baseline and 2 hours]

   Value of RHI at 2 hours minus value at baseline. RHI reflects the endothelial function of a vessel at the distal phalanx of a finger, i.e. the capacity of the vessel to dilate after an ischemia. RHI is the increase of blood flow following the occlusion of the brachial artery during 5 minutes by the inflation of an armcuff. RHI was measured by peripheral arterial tonometry using a fingerprobe connected to an EndoPat analyser.

Secondary Outcome Measures

1. Change in Arterial Stiffness From Baseline (20 Min Before Product Intake) to Two Hours Following Product Intake [baseline and 2 hours]

   Value of arterial stiffness at 2 hours minus value at baseline. Arterial stiffness is also automatically calculated by peripheral arterial tonometry which consists in measuring the peripheral vessel endothelial response to an ischemia provoked by a 5-min occlusion of the humeral artery using an armcuff. An increase in arterial stiffness means an increase in the resistance of the vessel wall which reflects an impaired endothelial response to ischemia.

Eligibility Criteria

Criteria

• Ages Eligible for Study: 25 Years to 45 Years
• Sexes Eligible for Study: All
• Accepts Healthy Volunteers: Yes

Inclusion Criteria:

• 25- 45 years, male and female

• Healthy as determined by the medical questionnaire

• Normal weight: BMI 19 - 25

• Having given informed consent

Exclusion Criteria:

• Intestinal or metabolic diseases/disorders such as diabetic, renal, hepatic, hypertension, pancreatic or ulcer, including lacto-intolerance.

• Have had a major gastrointestinal surgery.

• Have a regular consumption of medication.

• Have an exceptionally high intake of chocolate or similarly high polyphenol foods.

• Have a high and regular intake of vitamin supplements

• Have an alcohol intake: > 2 units a day

• Patient who cannot be expected to comply with treatment.

• Smoker

• Having a nut allergy

• Unwilling to consume chocolate

• Currently participating or having participated in another clinical trial during the last 3 weeks.

• Having given blood in the past three weeks

• More than 3 x 45 min of exercise per week

Contacts and Locations

Locations

	Site	City	State	Country	Postal Code
1	Nestle Research Center	Lausanne	Vaud	Switzerland	1000

Sponsors and Collaborators

• Nestlé

Investigators

• Principal Investigator: Karen A Cooper, PhD, Nestlé
• Study Director: Gary Williamson, PhD, Nestlé

Study Documents (Full-Text)

None provided.

More Information

Publications

• Fisher ND, Hollenberg NK. Aging and vascular responses to flavanol-rich cocoa. J Hypertens. 2006 Aug;24(8):1575-80.
• Heiss C, Finis D, Kleinbongard P, Hoffmann A, Rassaf T, Kelm M, Sies H. Sustained increase in flow-mediated dilation after daily intake of high-flavanol cocoa drink over 1 week. J Cardiovasc Pharmacol. 2007 Feb;49(2):74-80.

• Responsible Party: Nestlé
• ClinicalTrials.gov Identifier: NCT00513344
• Other Study ID Numbers: 06.38.MET
• First Posted: Aug 8, 2007
• Last Update Posted: Aug 14, 2013
• Last Verified: Jul 1, 2013

• Keywords provided by Nestlé: Endothelial function; Arterial stiffness; Dark chocolate; Milk chocolate; Polyphenols

Study Results

Participant Flow

Recruitment Details	5 healthy subjects, 2 females and 3 males, were recruited within the staff of the Nestlé Research Center in June-July 2008.
Pre-assignment Detail	From 3 days before the first test session no foods containing epicatechin or catechin or procyanidins should be ingested. The subjects received a single dose of the 3 products in a randomised order with a minimum of 3 days washout in between (crossover design). The duration of the study was a minimum of 15 days.

Arm/Group Title	Each Subject Tested the Three Products Randomly Following
Arm/Group Description	Before the 1st intervention, no food with polyphenols was admitted Each subject was administered the three products randomly(one product per one-day intervention) according to the six possible sequencies: Either dark chocolate first, then milk chocolate then control or dark chocolate first, then control, then milk chocolate or control first, then dark chocolate, then milk chocolate or Control first, then milk chocolate, then dark chocolate or Milk chocolate first, then control, then dark chocolate or milk chocolate first, then dark chocolate, then control r dark chocolate was given once in the morning (1 day) Products were administered in the morning of the testing day. The testing days (interventions) were separated by a three-day wash-out period.
Period Title: Diet Restriction (3 Days)
STARTED	5
COMPLETED	5
NOT COMPLETED	0
Period Title: Diet Restriction (3 Days)
STARTED	5
COMPLETED	5
NOT COMPLETED	0
Period Title: Diet Restriction (3 Days)
STARTED	5
COMPLETED	5
NOT COMPLETED	0
Period Title: Diet Restriction (3 Days)
STARTED	5
COMPLETED	5
NOT COMPLETED	0
Period Title: Diet Restriction (3 Days)
STARTED	5
COMPLETED	5
NOT COMPLETED	0
Period Title: Diet Restriction (3 Days)
STARTED	5
COMPLETED	5
NOT COMPLETED	0

Baseline Characteristics

Arm/Group Title	Entire Study Population
Arm/Group Description	Includes groups randomized to receive control first, milk chocolate first, and dark chocolate first
Overall Participants	5
Age (years) [Mean (Standard Deviation) ]
Mean (Standard Deviation) [years]	34 (7)
Age, Customized (participants) [Number]
<= 28 years	0 0%
between 28 and 42 years	5 100%
>= 42 years	0 0%
Sex: Female, Male (Count of Participants)
Female	2 40%
Male	3 60%
Region of Enrollment (participants) [Number]
Switzerland	5 100%

Outcome Measures

1. Primary Outcome

Title	Change in Reactive Hyperemia Index (RHI) From Baseline (20 Min Before Product Intake) to 2 Hours Following Product Intake
Description	Value of RHI at 2 hours minus value at baseline. RHI reflects the endothelial function of a vessel at the distal phalanx of a finger, i.e. the capacity of the vessel to dilate after an ischemia. RHI is the increase of blood flow following the occlusion of the brachial artery during 5 minutes by the inflation of an armcuff. RHI was measured by peripheral arterial tonometry using a fingerprobe connected to an EndoPat analyser.
Time Frame	Baseline and 2 hours

Outcome Measure Data

Analysis Population Description

Taking into account the fact that there is no background data and that we need to have accurate evaluation of central tendency and dispersion, a minimum of five complete subjects are needed. Data from all randomized subjects were considered in the intention to treat model (for the primary outcome).

Arm/Group Title	Control Chocolate With no Polyphenols	Milk Chocolate Containing Polyphenols	Dark Chocolate Containing Polyphenols
Arm/Group Description	cocoa-free chocolate Control (polyphenol-free)	Bespoke milk chocolate Milk Chocolate : 1 portion	dark chocolate Dark Chocolate : 1 portion
Measure Participants	5	5	5
Mean (Standard Error) [percentage of the pulse wave amplitude]	2.12 (0.52)	2.30 (0.71)	2.26 (0.65)

2. Secondary Outcome

Title	Change in Arterial Stiffness From Baseline (20 Min Before Product Intake) to Two Hours Following Product Intake
Description	Value of arterial stiffness at 2 hours minus value at baseline. Arterial stiffness is also automatically calculated by peripheral arterial tonometry which consists in measuring the peripheral vessel endothelial response to an ischemia provoked by a 5-min occlusion of the humeral artery using an armcuff. An increase in arterial stiffness means an increase in the resistance of the vessel wall which reflects an impaired endothelial response to ischemia.
Time Frame	baseline and 2 hours

Outcome Measure Data

Analysis Population Description

[Not Specified]

Arm/Group Title	Control Chocolate With no Polyphenols	Milk Chocolate Containing Polyphenols	Dark Chocolate Containing Polyphenols
Arm/Group Description	cocoa-free chocolate Control (polyphenol-free)	Bespoke milk chocolate Milk Chocolate : 1 portion	dark chocolate Dark Chocolate : 1 portion
Measure Participants	5	5	5
Mean (Standard Deviation) [percentage of baseline]	-13.23 (11.16)	1.23 (12.87)	-9.49 (15.03)

Adverse Events

Adverse Event Reporting Description	Adverse Events Not Collected
Arm/Group Title	Dark Chocolate		Milk Chocolate		Control
Arm/Group Description
All Cause Mortality
	Dark Chocolate		Milk Chocolate		Control
	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events
Total	/ (NaN)		/ (NaN)		/ (NaN)
Serious Adverse Events
	Dark Chocolate		Milk Chocolate		Control
	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events
Total	0/0 (NaN)		0/0 (NaN)		0/0 (NaN)
Other (Not Including Serious) Adverse Events
	Dark Chocolate		Milk Chocolate		Control
	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events
Total	0/0 (NaN)		0/0 (NaN)		0/0 (NaN)

Limitations/Caveats

[Not Specified]

More Information

Certain Agreements

All Principal Investigators ARE employed by the organization sponsoring the study.

There is NOT an agreement between Principal Investigators and the Sponsor (or its agents) that restricts the PI's rights to discuss or publish trial results after the trial is completed.