Rice Anthocyanin Bioavailability in Humans

Study Overview
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Brief Summary
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Aims of the project To determine the rice anthocyanin content via high-performance liquid chromatographycoupled to mass spectrometry, HPLC-MS To study the oral bioavailability of anthocyanins in healthy volunteers after the intake of a serving (80 g) of black rice (Venere and Artemide varieties). To determine the plasma biokinetics parameters of rice anthocyanins via spectrophometric determination and high-performance liquid chromatographycoupled to mass spectrometry, HPLC-MS.

Detailed Description
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To limit the interindividual variability, The protocol will be designed as a prospective, randomized, cross-over pharmacokinetic study on healthy volunteers. Participants will be randomly divided into three groups (black rice Venere, black rice Artemide and white rice/control) by an allocation software ( http://graphpad.com/quickcalcs/randomise1.cfm ) which will generate different random sequences of the treatment assignment. Subjects will be deprived of anthocyanin-rich food sources 7 days before experimentation (wash-out period). Each volunteer will receive a complete list of anthocyanin-rich foods to be avoided, including berry fruits (i.e. blueberries, cranberries, raspberries, blackcurrants and elderberries), red/violet fruits/vegetables (i.e. grapes, cherries, pomegranates, red apples, plums, eggplants, tomatoes and peppers), red wine and other colored products (i.e. marmalade, jams and juices containing berries). Breakfast, lunch and dinner will be standardized 1 day before the experiment. Each group will randomly receive, during three different experimental days, one of the following treatments by different allocation sequences: 80g of black rice Venere (Group A) 80g of black rice Artemide (Group B) 80g of white rice (Group C) Therefore, each subject will be involved in three experimental days. The three treatments will be separated by a 7 day wash-out period. After an overnight fast, volunteers will consume the rice serving within 10-15 min, immediately after the first basal blood collection at 8:00 a.m. (baseline). Other blood withdrawals will be collected after 30 minutes, 1 hour, 2 hours, 3 hours. Chemical analyses. The extraction protocols of anthocyanins from body fluids will be developed, in order to reach the maximum yield of the analytes from the samples. Plasma anthocyanins will be measured by spectrophometric determination and high-performance liquid chromatographycoupled to mass spectrometry, HPLC-MS.

Official Title
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Anthocyanin Bioavailability and Plasma Antioxidant Capacity in Healthy Volunteers After Acute Rice Intake

Conditions
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Healthy

Intervention / Treatment
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* Other: Rice


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Healthy status Age: 18-45 Normal weight (BMI 18.5-24.9) who voluntarily accept to join the study, after informed consent. Exclusion Criteria: <18 Years; Pregnancy and lactation; Systemic disorders; Dietary supplements intake (vitamins, antioxidants, botanicals, phytochemicals) Abnormal hematological parameters; Heavy smoking and alcohol drinking; High-intensity physical activity.

Ages Eligible for Study
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Minimum Age: 20 Years
Maximum Age: 35 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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Accepts Healthy Volunteers

Study Plan
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How is the study designed?
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Design Details

Primary Purpose: Basic Science
Allocation: Randomized
Intervention Model: Crossover Assignment
Masking: Single


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Black rice Venere<br>Healthy volunteers will receive the black rice Venere in a cross-over randomized clinical trial (after 7 days wash-out). | Other: Rice<br>* Each group will randomly receive, during three different experimental days, one of the following treatments by different allocation sequences: 80g of black rice Venere (Group A) 80g of black rice Artemide (Group B) 80g of brown rice (Group C) The three treatments will be separated by a 7 day wash-out period. After an overnight fast, volunteers will consume the rice serving within 10-15 min, immediately after the first basal blood collection at 8:00 a.m. (baseline). Other blood withdrawals will be carried out a 0.5, 1.0, 2.0 and 3.0 h after rice administration.<br>|
| Experimental: Black rice Artemide<br>Healthy volunteers will receive the black rice Artemide in a cross-over randomized clinical trial (after 7 days wash-out). | Other: Rice<br>* Each group will randomly receive, during three different experimental days, one of the following treatments by different allocation sequences: 80g of black rice Venere (Group A) 80g of black rice Artemide (Group B) 80g of brown rice (Group C) The three treatments will be separated by a 7 day wash-out period. After an overnight fast, volunteers will consume the rice serving within 10-15 min, immediately after the first basal blood collection at 8:00 a.m. (baseline). Other blood withdrawals will be carried out a 0.5, 1.0, 2.0 and 3.0 h after rice administration.<br>|
| Sham Comparator: Complete white rice<br>Healthy volunteers will receive the complete white rice in a cross-over randomized clinical trial (after 7 days wash-out). | Other: Rice<br>* Each group will randomly receive, during three different experimental days, one of the following treatments by different allocation sequences: 80g of black rice Venere (Group A) 80g of black rice Artemide (Group B) 80g of brown rice (Group C) The three treatments will be separated by a 7 day wash-out period. After an overnight fast, volunteers will consume the rice serving within 10-15 min, immediately after the first basal blood collection at 8:00 a.m. (baseline). Other blood withdrawals will be carried out a 0.5, 1.0, 2.0 and 3.0 h after rice administration.<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Changes in plasma total anthocyanin detection | Total anthocyanins will be detected via spectrophometric determination; expressed as μg of catechin per mL of plasma | 0.0 (baseline, before starting the intervention), then after 0.5 hour, 1 hour, 2 hours and 3 hours | 
| Changes in plasma Cyanidin-3-O-glucoside detection | Cyanidin-3-O-glucoside will be detected via high-performance liquid chromatographycoupled to mass spectrometry, HPLC-MS) in plasma; expressed nM (nano molar) in plasma | 0.0 (baseline, before starting the intervention), then after 0.5 hour, 1 hour, 2 hours and 3 hours | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Changes in plasma antioxidant capacity ABTS | 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS+); the results are expressed as Trolox equivalent antioxidant capacity (TEAC, mmoleq Trolox/mL plasma) | 0.0 (baseline, before starting the intervention), then after 0.5 hour, 1 hour, 2 hours and 3 hours | 
| Changes in plasma antioxidant capacity DPPH | 2,2-diphenyl-1-picrylhydrazyl (DPPH); percentage of inhibition (I%) is calculated as [(ABScontrol 517 nm - ABSsample 517 nm/ABScontrol 517 nm)×100] | 0.0 (baseline, before starting the intervention), then after 0.5 hour, 1 hour, 2 hours and 3 hours |