Healthy
Conditions
Brief summary
This study will evaluate the availability of phytonutrients in two blueberry varieties, chosen for their phytonutrient levels. This will be compared to phytonutrient-matched processed protein bar and a macronutrient-matched control meal, in healthy human volunteers. Blueberry phytonutrients will be analyzed in blood and urine over a four-day period, 48h prior to consumption and 48h after. The participants will consume each of the four meals over a 3-month period (4-way crossover design, 4 blocks of 4-day periods). The main objective of this study is to compare the proportions of blueberry phytonutrients recovered in the blood and urine after ingestion of the four treatments. We hypothesize that phytonutrient content will be predictive of human bioavailability and that a berry-enriched processed product will have similar phytonutrient bioavailability to unprocessed berries. The results of this study may establish if the nutritional value of a berry can be predicted or enhanced to provide elevated nutritional quality, with the ultimate goal of maximizing the health benefits of fruit consumption. As it is challenging for many to increase their fruit and vegetable intake to government recommended levels (5+ servings per day), the present proof-of-concept study explores a reasonable approach to help consumers achieve optimal health associated with high fruit and vegetable intakes, within the context of current consumption patterns, through enhancement of the nutritional density and bioavailability of common fruits and consumer products.
Detailed description
This study will evaluate the availability of phytonutrients in two blueberry varieties, chosen for their phytonutrient levels. This will be compared to phytonutrient-matched processed protein bar and a macronutrient-matched control meal, in healthy human volunteers. Blueberry phytonutrients will be analyzed in blood and urine over a four-day period, 48h prior to consumption and 48h after. The participants will consume each of the four meals over a 3-month period (4-way crossover design, 4 blocks of 4-day periods). The main objective of this study is to compare the proportions of blueberry phytonutrients recovered in the blood and urine after ingestion of the four treatments. After eligibility is confirmed, subjects will be randomly assigned to the four berry related interventions. The consumption of each intervention corresponds to one study period, which are separated by one-week washout. Blood will be collected at baseline and across 48h (1h, 3h, 6h, 9h, 24h, 48h) after intervention consumption while urine will be collected for 48h before and after intervention (-48h, -24h, 0-9h, 9-24h, 24-48h).
Interventions
OTHERa non-traditional (i.e., not typically available in the supermarket) blueberry cultivar bred using natural plant breeding techniques and established as having enhanced nutritive value
150 g of a non-traditional (i.e., not typically available in the supermarket) blueberry cultivar bred using natural plant breeding techniques and established as having enhanced nutritive value. Dietary restrictions will be observed (i.e. avoidance of food or supplements containing berry phytonutrients) for 7 days before each arm visit and throughout the study days.
OTHERa standard commercially available blueberry variety (i.e., cultivar)
150 g of a standard commercially available blueberry variety (i.e., cultivar). Dietary restrictions will be observed (i.e. avoidance of food or supplements containing berry phytonutrients) for 7 days before each arm visit and throughout the study days.
OTHERa "minimally processed" blueberry-rich protein bar
A "minimally processed" blueberry-rich protein bar matched to the phytonutrient content of the 150 g of the non-traditional blueberry. Dietary restrictions will be observed (i.e. avoidance of food or supplements containing berry phytonutrients) for 7 days before each arm visit and throughout the study days.
OTHERa control beverage of matched-nutritive content
The matched nutritive content of the blueberry-rich protein bar will be dissolved in whey protein Dietary restrictions will be observed (i.e. avoidance of food or supplements containing berry phytonutrients) for 7 days before each arm visit and throughout the study days.
Sponsors
North Carolina State University
Foundation for Food and Agriculture Research
Study design
Allocation
RANDOMIZED
Intervention model
CROSSOVER
Primary purpose
BASIC_SCIENCE
Masking
TRIPLE (Subject, Caregiver, Outcomes Assessor)
Eligibility
Sex/Gender
ALL
Age
25 Years to 65 Years
Healthy volunteers
Yes
Inclusion criteria
* male and female adults between 25-65 years; * non-smokers, non-tobacco users (no vaping nor deeping), or who ceased it ≥ 6 months ago; * who present no allergies to fruits or vegetables containing polyphenolic (e.g. anthocyanins, flavonoids) and phenolic acids such as blueberries, red apple, strawberry, red orange, purple onion and broccoli; * who present no allergies to dairy products, specifically whey protein, fructose or salicylates; * who are generally healthy and without chronic diseases including cancer, type 1 and 2 diabetes; * who are not prescribed thyroid or hypoglycemic medication or hormone replacement therapy (HRT) (due to the likely concomitant effects that these medications cause on the primary endpoint in the trial); * who has not been consuming any phytonutrient-containing supplements (e.g. with cocoa, coffee, berry, polyphenol, flavonoid, or anthocyanin extracts) for at least a month before the study and willing to not consume it during the study; * who lives within 40 miles from the North Carolina Research Campus (NCRC) campus; * those agreeing to restrict dietary intake of rich sources of phytonutrients targeted on the study during the wash-out and clinical sampling periods, agreeing to comply with a biological sampling protocol involving the collection of urine and blood samples, and to record their additional dietary intake over 2 days before each intervention, and two days after the intake of the intervention treatments; * who have BMI ≥18.5 and ≤ 30 (lbs/in2x703); * who have a successful (i.e., within normal range for healthy individuals) biochemical, hematological and urine analyses assessed by the clinical advisor as established during the screening period prior to final enrollment.
Exclusion criteria
* current smokers (vaping and deeping included), or ex-smokers ceasing \< 6 months before recruitment; * pregnant or breastfeeding; * subjects with existing or significant past medical history of vascular disease or medical conditions likely to affect the study measures i.e. vascular disease, circulatory (i.e. Reynaud's), diabetes, hepatic, renal, digestive, hematological, cancer, or thyroid disease; * fructose intolerant subjects or those with known allergy to salicylates, dairy products, specifically whey protein, or to berries; * those unprepared to adhere to dietary restrictions for 1 week preceding and during each intervention or unwilling to comply with the assessments per protocol; * who are in parallel participation in another research project involving dietary intervention and/or sampling of biological fluids/material; * those on therapeutic diets or having experienced substantial weight loss (to be judged by clinical advisor) within 3 months of screening; * those taking phytonutrient-containing supplements (e.g. with cocoa, coffee, berry, polyphenol, flavonoid, or anthocyanin extracts), unwilling to cease intake during, and 1 month preceding the trial, or unwilling to stop existing intake of other supplements or regular use of large-dose nutrient, herbal, and dietary supplements during the past one to two weeks, or planning to use them during the study; * prescribed thyroid, hypoglycemic medication or HRT medication -other medications will be assessed for suitability by the clinical advisor; * those having donated blood in the last month; * individuals that consume more than 1 and 2 drinks of alcohol per day for women and men, respectively, or more than 7 and 14 drinks per week for women and men, respectively (U.S. Department of Health and Human Services and U.S. Department of Agriculture Dietary guidelines 2015-2020); * currently on a weight-reducing plan or using weight-loss medications (e.g., selective serotonin reuptake inhibitors, steroids, Ritalin, appetite suppressants such as Diethylpropion or Amfepramone, and weight loss medications such as Alli, Xenical, Qsymia, Belviq, Contrave, and Saxenda), or planning to continue this treatment during the 10-week period of the study; * who has BMI\<18.5 and \>30 (lbs/in2x703); * who presents abnormal biochemical, hematological or urinary results, and measurements considered to be counter-indicative for the study, including: kidney and liver function, fasting glucose (especially if indicative of diabetes), lipid abnormalities, full blood count as established during the screening period prior to final enrollment.
Design outcomes
Primary
| Measure | Time frame | Description |
|---|---|---|
| Total Urinary Recovery of (Poly)Phenolic Metabolites Across Treatments | 4 days of urine collection per intervention; -48, -24, 0-9, 9-24 and 24-48 hours collections of urine. | Concentration of 185 (poly)phenolic metabolites in urine was measured after consumption of the treatments using broad spectrum metabolomic analysis via UPLC-MS/MS (ultra-performance liquid chromatography coupled with tandem mass spectrometry). Recovery of each metabolite was calculated by multiplying the concentration of the metabolite (ng/mL) by the volume of urine (mL) in each collection then converted to milligrams. The total urinary recovery of each (poly)phenolic metabolite was assessed as a measurement of their bioavailability, and it was established as the sum of the recovery from each urine collection (0 h to 48 h postintervention) minus the mean of the 48 h and 24 h baseline collections per participant and treatment. Then, it was averaged across participants per treatment and per metabolite, and further summed across metabolites. Therefore, data reported for each treatment represents the sum of the means of total urinary recovery across (poly)phenolic metabolites. |
| Serum Area Under the Curve (AUC) of (Poly)Phenolic Metabolites Across Treatments | 3 days of blood collection per intervention; 1 baseline collection, followed by 1, 3, 6, 9, 24 and 48 hours post-treatment collections of blood. | Concentration of 185 (poly)phenolic metabolites in blood was measured after consumption of the treatments using broad spectrum metabolomic analysis via UPLC-MS/MS (ultra-performance liquid chromatography coupled with tandem mass spectrometry). The area under the curve (AUC) of each (poly)phenolic metabolite in serum was assessed as a measurement of the bioavailability of these metabolites per participant and treatment. It was calculated using concentration and time data in the software Phoenix WinNonlin (version 8.3, Certara). Then, AUC was averaged across participants per treatment and per metabolite, and the mean AUCs were further summed across metabolites. Therefore, data reported for each treatment represents the sum of the means of AUC across (poly)phenolic metabolites. |
Secondary
| Measure | Time frame | Description |
|---|---|---|
| Maximum Serum Concentration [Cmax] of (Poly)Phenolic Metabolites Between the Treatments | 3 days of blood collection per intervention; 1 baseline collection, followed by 1, 3, 6, 9, 24 and 48 hours post-treatment collections of blood. | Concentration of 185 (poly)phenolic metabolites in serum was measured after consumption of the treatments using broad spectrum metabolomic analysis via UPLC-MS/MS (ultra-performance liquid chromatography coupled with tandem mass spectrometry). The maximum serum concentration \[Cmax\] of each (poly)phenolic metabolite in serum was calculated per participant and treatment using concentration and time data in the software Phoenix WinNonlin (version 8.3, Certara). Then, it was averaged across participants per treatment and per metabolite, and further averaged across metabolites. Therefore, data reported for each treatment represents the mean of the means of Cmax across (poly)phenolic metabolites. |
Countries
United States
Contacts
PRINCIPAL_INVESTIGATORColin D Kay, PhD
North Carolina State University
Participant flow
Recruitment details
75 participants were recruited and screened from the local community surrounding the North Carolina Research Campus, Kannapolis, North Carolina, between December 2019 and July 2021. Clinical screening and interventions were conducted at the Human Research Core at the Nutrition Research Institute (University of North Carolina-Chapel Hill). The trial was completed in October 2021.
Pre-assignment details
29 out of 75 screened participants were enrolled into the study and randomized to interventions. Of those not enrolled, 20 participants did not meet inclusion criteria, 17 declined to participate and 9 did not follow up.
Baseline characteristics
| Characteristic | — |
|---|---|
| Age, Continuous | 42.06 years STANDARD_DEVIATION 12.53 |
| BMI, kg/m2 | 24.75 kg/m2 STANDARD_DEVIATION 2.99 |
| Diastolic Blood pressure, mmHg | 71.54 mmHg STANDARD_DEVIATION 7.63 |
| Height (m) | 1.72 meter STANDARD_DEVIATION 0.1 |
| Region of Enrollment United States | 18 participants |
| Sex: Female, Male Female | 11 Participants |
| Sex: Female, Male Male | 7 Participants |
| Systolic Blood pressure, mmHg | 116.12 mmHg STANDARD_DEVIATION 13 |
| Weight (kg) | 73.8 kg STANDARD_DEVIATION 13.72 |
Adverse events
| Event type | EG000 affected / at risk | EG001 affected / at risk | EG002 affected / at risk | EG003 affected / at risk |
|---|---|---|---|---|
| deaths Total, all-cause mortality | 0 / 18 | 0 / 18 | 0 / 18 | 0 / 18 |
| other Total, other adverse events | 0 / 18 | 0 / 18 | 0 / 18 | 0 / 18 |
| serious Total, serious adverse events | 0 / 18 | 0 / 18 | 0 / 18 | 0 / 18 |
Outcome results
None listed