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.

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).

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.