Physiological Effects of New Polyphenol-enriched Foods in Humans

• Sponsor: Federico II University (Other)
• Overall Status: Completed
• CT.gov ID: NCT01288859
• Collaborator: (none)
• Enrollment: 10
• Location: 1
• Arms: 6
• Duration (Months): 7
• Patients Per Site Per Month: 1.4

Study Details

Study Description

Brief Summary

Polyphenolic compounds exert several health benefits depending on bioavailability. Encapsulation may improve bioavailability of these compounds.This study will evaluate bioavailability of some polyphenols (curcumin and cocoa polyphenols) from new enriched-foods. In particular bread and nut based creams will be used as food matrices to include free or encapsulated polyphenols.

Condition or Disease	Intervention/Treatment	Phase
Healthy	Other: free curcumin; Other: encapsulated curcumin; Other: encapsulated curcumin + PQG; Other: free cocoa polyphenol; Other: encapsulated cocoa polyphenols; Other: control nut cream	N/A

Detailed Description

Polyphenolic compounds are abundant in foods and have been suggested to exert several health benefits including prevention of cancer, cardiovascular disease, suppressing inflammation, diabetes, etc. These properties are mediated by bioavailability of individual polyphenols that is, in turn, influenced by food properties (food matrix, technological processing, etc.), interaction with other compounds, and host related factors. Some well-studied dietary polyphenols include catechins from tea, curcumin from turmeric, procyanidins and anthocyanidins in grapes, berries, and dark chocolate. These compounds have strong antioxidative activities in vitro and have been suggested to have several beneficial health effects.

Curcumin, a phenolic compound deriving from turmeric spice, shows many biological and pharmacological effects, and clinical studies in humans proved that it's extremely safe and well tolerated even at very high doses (8-12 g/day). However, curcumin has not yet been approved as a therapeutic agent, because of its very low bioavailability depending on a poor absorption, rapid metabolism and rapid systemic clearance. Different approaches have been investigated to improve curcumin bioavailability. Among them, the use of adjuvants that interferes with metabolic pathways of curcumin, like piperine, represents one of the main strategies used to enhance its bioavailability. A further approach consists on manufacturing of curcumin containing liposomes, phospholipids complexes or nanoparticles. For polymer-based nanoparticles, maintenance of biological activity, increased absorption and delayed delivery was reported.

Cocoa based products are widely consumed in many countries, and indications of health benefits by some cocoa constituents, mainly polyphenols, have also been reported. Cocoa contains very high levels of polyphenols, in particular flavanols and among these, mainly epicatechins. The bioavailability of cocoa polyphenols has been measured in several human studies from acute consumption of cocoa rich beverages or chocolate. Monomeric flavonoids as well as dimeric and trimeric procyanidins were detected in human plasma over 2-3 hours from consumption. Plasma concentration of cocoa polyphenols were often in the nanomolar or low micromolar range. Donovan and coworkers demonstrated that commercial available chocolate samples contain a predominance of the less bioavailable (-)-catechin enantiomer as compared to the (+)-catechin which is present in most other plant derived foods. This may explain the relatively low bioavailability of catechins from chocolate and cocoa-containing products. The food matrix seems to be an important factor that may affect the bioavailability of cocoa polyphenols. For instance, proteins in the food matrix are supposed to form highly polymerized complexes with procyanidins, which can reduce bioaccessibility of these phenolic compounds. However, the potential negative effect of protein content in foods was not confirmed in a study that evaluated the cocoa polyphenols bioavailability after the intake of a milk-powder cocoa beverage. However, concurrent carbohydrates consumption seems to increase significantly the uptake of flavonols in humans. Little is known whether and to what extent oligomeric procyanidins from cocoa are absorbed. However, the biological activity of procyanidins with high polymerization degree may be partly attributed to their colonic breakdown products, including phenolic acids.

Actually there is also a growing interest in other biological properties of phenolic compounds in addition to their antioxidant effects; particularly, some evidences suggest that certain dietary phenols may modulate metabolic homeostasis. This is the case of chlorogenic acids, that are reported to play a potential role in modifying the pattern of intestinal glucose uptake and of other flavonoids (quercetin, catechins) that may modulate activation of GLP-1 receptor, involved in modulation of insulin and glucagon secretion, gastric emptying, and appetite.

In this randomized, crossover trial, serum, urine and fecal concentrations of curcumin and cocoa polyphenols, their parental compounds, metabolites and phenolic acids, following a two day multi-dose administration with six food sources, will be measured. All volunteers will undergo to six interventions - bread enriched with three different forms of curcumin (free, encapsulated, and encapsulated plus piperine) and nut based creams enriched with cocoa polyphenols (free or encapsulated one) or control cream (without enrichment). A one-week wash-out period will be included between two sequential treatments. Blood, urine and feces will be collected at time 0 (baseline), 24, and 48 hours; further blood samples at 0.5, 1, 2, 4, 6 hours after consumption of the first meal, and urine samples at 2 hours time intervals up to 10 hours, will be collected too. In particular, area under the curve (AUC) of serum and urine concentrations of parental compounds and metabolites in the time interval 0-24 hours will be calculated as primary outcomes. In addition the amount of total polyphenols in fecal samples will be measured.

Study Design

• Study Type: Interventional
• Actual Enrollment: 10 participants
• Allocation: Randomized
• Intervention Model: Crossover Assignment
• Masking: Single (Participant)
• Primary Purpose: Basic Science
• Official Title: Physiological Effects of New Polyphenol-enriched Foods in Healthy Humans
• Study Start Date: Dec 1, 2010
• Actual Primary Completion Date: Feb 1, 2011
• Actual Study Completion Date: Jul 1, 2011

Arms and Interventions

Arm	Intervention/Treatment
Experimental: encapsulated curcumin	Other: encapsulated curcumin; encapsulated curcumin-enriched bread, 1g/100g bread, 200g bread/day
Experimental: encapsulated curcumin + PQG PQG means Piperine, Quercetin and Genistein	Other: encapsulated curcumin + PQG; bread enriched with encapsulated curcumin plus piperine, quercetin and genistein, 1g/100g bread, 200g bread/day
Active Comparator: free cocoa polyphenol	Other: free cocoa polyphenol; nut cream enriched with free cocoa polyphenols, 1,5 g/100g cream, 100g/day per 1 day
Placebo Comparator: control	Other: control nut cream; nut cream, cocoa polyphenols 0g/100g cream, 100 g/day per 1 day
Experimental: encapsulated cocoa polyphenols	Other: encapsulated cocoa polyphenols; nut cream enriched with encapsulated cocoa polyphenols, 1,5 g/100g cream, 100g/day per 1 day
Active Comparator: free curcumin Subjects will consume bread added with free curcumin	Other: free curcumin; free curcumin in bread, dosage 1g/100g, 200g/day per 1 day

Outcome Measures

Primary Outcome Measures

1. Serum Polyphenol Concentrations Over 24h From Food Consumption [0, 0.5, 1, 2, 4, 6, and 24 hours post-dose]

   Area Under the Curves (AUC) from 0 to 24h of parent polyphenols was calculated using a trapezoidal rule applied to the concentration-time curves of compounds.

2. Urinary Excretion of Total Polyphenols [Time intervals: 0-2, 2-4, 4-6, 6-8, 10-24 hours post-dose.]

   Area Under the Curve (AUC) from 0 to 24h of total polyphenols (sum of parent polyphenols and metabolites)was calculated using a trapezoidal rule applied to the urinary concentration-time curves of compounds.

3. Amount of Total Fecal Polyphenols [0 and 24 hours post-dose.]

   Amount of parent polyphenols and metabolites in feces was calculated by multiplying net concentrations by the amount of feces.

Eligibility Criteria

Criteria

• Ages Eligible for Study: 18 Years to 45 Years
• Sexes Eligible for Study: All
• Accepts Healthy Volunteers: Yes

Inclusion Criteria:

• 18 - 45 years old, male and female

• Healthy by medical assessment

• Normal weight: BMI 18 - 25

• Sign of a written informed consent

Exclusion Criteria:

• Age > 18 and < 45 years old

• Pregnancy or breastfeeding

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

• Previous abdominal/gastrointestinal surgery

• Regular consumption of medication

• Antibiotic therapy within 2 months previous the study

• Food allergies and intolerances (celiac disease, lactose intolerance, nut allergy etc)

• Unwilling to consume experimental foods

• Concurrent participation or having participated to another clinical trial during the last 3 weeks

Contacts and Locations

Locations

	Site	City	State	Country	Postal Code
1	Department of Food Science	Portici		Italy	80055

Sponsors and Collaborators

• Federico II University

Investigators

• Principal Investigator: Vincenzo Fogliano, Professor, University of Naples
• Study Director: Paola Vitaglione, Researcher, University of Naples

Study Documents (Full-Text)

None provided.

More Information

Publications

None provided.

• Responsible Party: Vincenzo Fogliano, Professor, Federico II University
• ClinicalTrials.gov Identifier: NCT01288859
• Other Study ID Numbers: DSA-FF-01
• First Posted: Feb 3, 2011
• Last Update Posted: Aug 1, 2012
• Last Verified: Jul 1, 2012

• Keywords provided by Vincenzo Fogliano, Professor, Federico II University: polyphenols; curcumin; bioavailability; cocoa
• Additional relevant MeSH terms: Curcumin

Study Results

Participant Flow

Recruitment Details	The recruitment was performed at the ambulatory of Department of Food Science (University of Naples) among students of Agriculture Faculty. It began on December 2010 up to February 2011.
Pre-assignment Detail	Subjects were enrolled after signing a Consent Form and were randomized to the treatments. All subjects enrolled completed the study.

Arm/Group Title	Encapsulated Curcumin	Encapsulated Curcumin + PQG	Free Cocoa Polyphenol	Control Cream	Encapsulated Cocoa Polyphenols	Free Curcumin
Arm/Group Description	Subjects consumed 2 bread portions/day of bread enriched with micro-encapsulated curcumin(1g/100g portion, i.e. 2g/day)	Subjects consumed 2 bread portions/day of bread enriched with micro-capsules containing curcumin at dosage of 1 g/100 g plus 0.1% of Piperine, Quercetin and Genistein (PQG) (2g curcumin + 0.2g PQG/day) for one day.	Subjects consumed 3 nut cream portions (33g each) enriched with cocoa polyphenols at dosage of 1.5g/day.	Subjects consumed 3 nut cream portions (33g each).	Subjects consumed 3 nut cream portions (33g each) enriched with encapsulated cocoa polyphenols at dosage of 1.5g/day.	Subjects consumed 2 portions of bread enriched with free curcumin (1g/100g portion, i.e. 2g/day)
Period Title: Free Then Encapsulated Curcumin (1 Day)
STARTED	5	0	0	0	0	5
COMPLETED	5	0	0	0	0	5
NOT COMPLETED	0	0	0	0	0	0
Period Title: Free Then Encapsulated Curcumin (1 Day)
STARTED	5	0	0	0	0	5
COMPLETED	5	0	0	0	0	5
NOT COMPLETED	0	0	0	0	0	0
Period Title: Free Then Encapsulated Curcumin (1 Day)
STARTED	5	0	0	0	0	5
COMPLETED	5	0	0	0	0	5
NOT COMPLETED	0	0	0	0	0	0
Period Title: Free Then Encapsulated Curcumin (1 Day)
STARTED	5	0	0	0	0	5
COMPLETED	5	0	0	0	0	5
NOT COMPLETED	0	0	0	0	0	0
Period Title: Free Then Encapsulated Curcumin (1 Day)
STARTED	0	5	5	0	0	0
COMPLETED	0	5	5	0	0	0
NOT COMPLETED	0	0	0	0	0	0
Period Title: Free Then Encapsulated Curcumin (1 Day)
STARTED	0	5	5	0	0	0
COMPLETED	0	5	5	0	0	0
NOT COMPLETED	0	0	0	0	0	0
Period Title: Free Then Encapsulated Curcumin (1 Day)
STARTED	0	5	5	0	0	0
COMPLETED	0	5	5	0	0	0
NOT COMPLETED	0	0	0	0	0	0
Period Title: Free Then Encapsulated Curcumin (1 Day)
STARTED	0	0	0	5	5	0
COMPLETED	0	0	0	5	5	0
NOT COMPLETED	0	0	0	0	0	0
Period Title: Free Then Encapsulated Curcumin (1 Day)
STARTED	0	0	0	5	5	0
COMPLETED	0	0	0	5	5	0
NOT COMPLETED	0	0	0	0	0	0
Period Title: Free Then Encapsulated Curcumin (1 Day)
STARTED	0	0	0	5	5	0
COMPLETED	0	0	0	5	5	0
NOT COMPLETED	0	0	0	0	0	0
Period Title: Free Then Encapsulated Curcumin (1 Day)
STARTED	0	0	0	5	5	0
COMPLETED	0	0	0	5	5	0
NOT COMPLETED	0	0	0	0	0	0

Baseline Characteristics

Arm/Group Title	All Study Participants
Arm/Group Description	All participants followed all the periods
Overall Participants	10
Age (Count of Participants)
<=18 years	0 0%
Between 18 and 65 years	10 100%
>=65 years	0 0%
Age (years) [Mean (Standard Deviation) ]
Mean (Standard Deviation) [years]	31 (2)
Sex: Female, Male (Count of Participants)
Female	5 50%
Male	5 50%
Region of Enrollment (participants) [Number]
Italy	10 100%

Outcome Measures

1. Primary Outcome

Title	Serum Polyphenol Concentrations Over 24h From Food Consumption
Description	Area Under the Curves (AUC) from 0 to 24h of parent polyphenols was calculated using a trapezoidal rule applied to the concentration-time curves of compounds.
Time Frame	0, 0.5, 1, 2, 4, 6, and 24 hours post-dose

Outcome Measure Data

Analysis Population Description

The number of subjects was based on power calculations derived from our previous study. We calculated that, at α = 0.05 with a power of 80%, 8 subjects would allow us to detect a 20% difference in serum and urinary concentrations of parental compounds, glucuronides and phenolic acids.

Arm/Group Title	Free Curcumin	Encapsulated Curcumin	Encapsulated Curcumin + PQG	Control Cream	Free Cocoa Polyphenols	Encapsulated Cocoa Polyphenols
Arm/Group Description	subjects consumed bread enriched with free curcumin	subjects consumed bread enriched with encapsulated curcumin	subjects consumed bread enriched with encapsulated curcumin plus piperine, quercetin and genistein	subjects consumed cocoa-nut cream	subjects consumed cocoa-nut cream enriched with cocoa polyphenols in the free form	subjects consumed cocoa-nut cream enriched with encapsulated cocoa polyphenols
Measure Participants	10	10	10	10	10	10
Mean (Standard Error) [nmol*h/L]	1.7 (0.1)	12.1 (1.1)	7.7 (1.8)	0.01 (0.00)	7.3 (0.9)	0.5 (0.1)

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	Free Curcumin, Encapsulated Curcumin, Encapsulated Curcumin + PQG, Control Cream, Free Cocoa Polyphenols, Encapsulated Cocoa Polyphenols
	Comments
	Type of Statistical Test	Non-Inferiority or Equivalence
	Comments	The results from LC-MS/MS analysis of parent polyphenols were analyzed and expressed as the absolute changes from the baseline to reduce possible effects of inter-subject fasting variability
Statistical Test of Hypothesis	p-Value	0.02
	Comments
	Method	ANOVA
	Comments
Method of Estimation	Estimation Parameter	Mean Difference (Final Values)
	Estimated Value	1
	Confidence Interval	(2-Sided) 95%; to
	Parameter Dispersion	Type: Standard Error of the Mean; Value: 0.1
	Estimation Comments

2. Primary Outcome

Title	Urinary Excretion of Total Polyphenols
Description	Area Under the Curve (AUC) from 0 to 24h of total polyphenols (sum of parent polyphenols and metabolites)was calculated using a trapezoidal rule applied to the urinary concentration-time curves of compounds.
Time Frame	Time intervals: 0-2, 2-4, 4-6, 6-8, 10-24 hours post-dose.

Outcome Measure Data

Analysis Population Description

Basing on power analysis calculation on a previous work

Arm/Group Title	Encapsulated Curcumin	Encapsulated Curcumin + PQG	Free Cocoa Polyphenol	Control	Encapsulated Cocoa Polyphenols	Free Curcumin
Arm/Group Description	subjects consumed bread containing encapsulated curcumin	subjects consumed bread containing encapsulated curcumin plus PQG (i.e. Piperine, Quercetin and Genistein)	subjects consumed nut creams added with cocoa polyphenols	Subjects consumed white bread or nut cream	subjects consumed nut creams added with encapsulate cocoa-polyphenols	Subjects consumed bread added with free curcumin
Measure Participants	10	10	10	10	10	10
Mean (Standard Error) [nmol•h/L]	2.0 (0.8)	2.21 (1.27)	9.67 (6.7)	19.1 (12.3)	4.92 (3.36)	1.31 (0.81)

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	Free Curcumin, Encapsulated Curcumin, Encapsulated Curcumin + PQG, Control Cream, Free Cocoa Polyphenols, Encapsulated Cocoa Polyphenols
	Comments	Statistical analysis was performed using the statistical package SPSS for Windows (version15). By the analysis of variance (ANOVA) for repeated measures the subjective time curves for all measured compounds were compared and tested for the effect of treatment and of time as factors. For all tests, following a significant main effect in the ANOVA, individual means were compared using the Bonferroni test (p < 0.05). Results were considered significant at p < 0.05.
	Type of Statistical Test	Superiority or Other
	Comments
Statistical Test of Hypothesis	p-Value	0.01
	Comments
	Method	ANOVA
	Comments
Method of Estimation	Estimation Parameter	Mean Difference (Final Values)
	Estimated Value	0.9
	Confidence Interval	(2-Sided) 95%; to
	Parameter Dispersion	Type: Standard Error of the Mean; Value: 0.01
	Estimation Comments

3. Primary Outcome

Title	Amount of Total Fecal Polyphenols
Description	Amount of parent polyphenols and metabolites in feces was calculated by multiplying net concentrations by the amount of feces.
Time Frame	0 and 24 hours post-dose.

Outcome Measure Data

Analysis Population Description

[Not Specified]

Arm/Group Title	Encapsulated Curcumin	Encapsulated Curcumin + PQG	Free Cocoa Polyphenol	Control	Encapsulated Cocoa Polyphenols	Free Curcumin
Arm/Group Description	subjects consumed bread containing encapsulated curcumin	subjects consumed bread containing encapsulated curcumin plus PQG (i.e. Piperine, Quercetin and Genistein)	subjects consumed nut creams added with cocoa polyphenols	Subjects consumed white bread or nut cream	subjects consumed nut creams added with encapsulate cocoa-polyphenols	Subjects consumed bread added with free curcumin
Measure Participants	10	10	10	10	10	10
Mean (Standard Error) [nmol]	3.49 (2.13)	0.49 (0.01)	27.98 (13.97)	4.27 (4.52)	150.97 (54.65)	0.59 (0.38)

Statistical Analysis 1

Statistical Analysis Overview	Comparison Group Selection	Free Curcumin, Encapsulated Curcumin, Encapsulated Curcumin + PQG, Control Cream, Free Cocoa Polyphenols
	Comments
	Type of Statistical Test	Superiority or Other
	Comments
Statistical Test of Hypothesis	p-Value	0.04
	Comments
	Method	ANOVA
	Comments
Method of Estimation	Estimation Parameter	Mean Difference (Final Values)
	Estimated Value	0.2
	Confidence Interval	(2-Sided) 95%; to
	Parameter Dispersion	Type: Standard Error of the Mean; Value: 0.01
	Estimation Comments

Adverse Events

Arm/Group Title	Encapsulated Curcumin		Encapsulated Curcumin + PQG		Free Cocoa Polyphenol		Control Cream		Encapsulated Cocoa Polyphenols		Free Curcumin
Arm/Group Description	Subjects consumed 2 bread portions/day of bread enriched with micro-encapsulated curcumin(1g/100g portion, i.e. 2g/day)		Subjects consumed 2 bread portions/day of bread enriched with micro-capsules containing curcumin at dosage of 1 g/100 g plus 0.1% of Piperine, Quercetin and Genistein (PQG) (2g curcumin + 0.2g PQG/day) for one day.		Subjects consumed 3 nut cream portions (33g each) enriched with cocoa polyphenols at dosage of 1.5g/day.		Subjects consumed 3 nut cream portions (33g each).		Subjects consumed 3 nut cream portions (33g each) enriched with encapsulated cocoa polyphenols at dosage of 1.5g/day.		Subjects consumed 2 portions of bread enriched with free curcumin (1g/100g portion, i.e. 2g/day)
All Cause Mortality
	Encapsulated Curcumin		Encapsulated Curcumin + PQG		Free Cocoa Polyphenol		Control Cream		Encapsulated Cocoa Polyphenols		Free Curcumin
	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events
Total	/ (NaN)		/ (NaN)		/ (NaN)		/ (NaN)		/ (NaN)		/ (NaN)
Serious Adverse Events
	Encapsulated Curcumin		Encapsulated Curcumin + PQG		Free Cocoa Polyphenol		Control Cream		Encapsulated Cocoa Polyphenols		Free Curcumin
	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events
Total	0/10 (0%)		0/10 (0%)		0/10 (0%)		0/10 (0%)		0/10 (0%)		0/10 (0%)
Other (Not Including Serious) Adverse Events
	Encapsulated Curcumin		Encapsulated Curcumin + PQG		Free Cocoa Polyphenol		Control Cream		Encapsulated Cocoa Polyphenols		Free Curcumin
	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events	Affected / at Risk (%)	# Events
Total	0/10 (0%)		0/10 (0%)		0/10 (0%)		0/10 (0%)		0/10 (0%)		0/10 (0%)

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.