Metabolomic Approach to Evaluate the Effect of Orange Juice Intake in Healthy Humans (METABORANGE).

Study Overview
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Brief Summary
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This study evaluates the effect of intake of two orange juice varieties in healthy adults. The participants will receive Bahia orange juice, Cara-Cara orange juice or a isocaloric control drink in a cross-over study.

Detailed Description
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The consumption of fruits and vegetables is linked to a decrease in the incidence of chronic diseases. Citrus fruits represent the main source of bioactive compounds such as flavonoids, vitamin C and carotenoids in the brazilian diet. Unlike the Bahia, the Cara Cara variety which is a spontaneous mutation of Washington navel orange is characterized by a bright red pulp due to high levels of lycopene. This difference in the chemical composition may lead to a different biological activity and then a greater protective effect on health. Since the biological effects generated from a food intake are the result of the interaction of various nutrients with different metabolic pathways, the investigators will apply the metabolomic approach to investigate the complex relationships between fruit intake, bioactive compounds and human metabolism. The investigators will propose to describe the metabolic pathways affected by orange juice intake and to identify biomarkers that can be used in nutritional interventions.

Official Title
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Metabolomic Approach to Evaluate the Effect of Orange Juice Intake in Healthy Humans: a Comparative Study Between Bahia and Cara Cara Orange Varieties.

Conditions
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Healthy Volunteers

Intervention / Treatment
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* Dietary Supplement: Orange juice


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Subjects like to drink orange juice Exclusion Criteria: History of cardiovascular disease History of gastrointestinal, liver or kidney disease History of alcoholism Diabetes and metabolic diseases Pregnant, trying to became pregnant or breast feeding Smokers To make use of vitamin and mineral supplements, antibiotics, antiacids, any drugs

Ages Eligible for Study
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Minimum Age: 20 Years
Maximum Age: 45 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: Treatment
Allocation: Randomized
Intervention Model: Crossover Assignment
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Bahia orange juice<br>250 ml twice daily for 7 days | Dietary Supplement: Orange juice<br>* Three days before the start of the study, all participants will restrict their diet avoiding citrus fruits (orange, tangerine, lemon) and its derivatives (washout period), red fruits and vegetables (strawberry, red lettuce, red grape, tomate). The day before the sample collection will be provided a controlled dinner. A fasting for 8 hours will be required before blood and urine collection. All participants will divided into three groups: I (Cara Cara), II (Bahia), III (Isocaloric control drink). They will consume 250 ml of each drink twice daily for 7 consecutive days, with a washout period of 7 days among each intervention.<br>|
| Experimental: Cara Cara orange juice<br>250 ml twice daily for 7 days | Dietary Supplement: Orange juice<br>* Three days before the start of the study, all participants will restrict their diet avoiding citrus fruits (orange, tangerine, lemon) and its derivatives (washout period), red fruits and vegetables (strawberry, red lettuce, red grape, tomate). The day before the sample collection will be provided a controlled dinner. A fasting for 8 hours will be required before blood and urine collection. All participants will divided into three groups: I (Cara Cara), II (Bahia), III (Isocaloric control drink). They will consume 250 ml of each drink twice daily for 7 consecutive days, with a washout period of 7 days among each intervention.<br>|
| Placebo Comparator: Isocaloric control drink<br>250 ml twice daily for 7 days | Dietary Supplement: Orange juice<br>* Three days before the start of the study, all participants will restrict their diet avoiding citrus fruits (orange, tangerine, lemon) and its derivatives (washout period), red fruits and vegetables (strawberry, red lettuce, red grape, tomate). The day before the sample collection will be provided a controlled dinner. A fasting for 8 hours will be required before blood and urine collection. All participants will divided into three groups: I (Cara Cara), II (Bahia), III (Isocaloric control drink). They will consume 250 ml of each drink twice daily for 7 consecutive days, with a washout period of 7 days among each intervention.<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Qualitative changes in urinary metabolome will be evaluated by Gas Chromatography coupled to Mass Spectrometry (GC-MS) and Liquid Chromatography coupled to Mass Spectrometry (LC-MS). |  | Urine collection will be performed at time 0, after 3, 6 hours and 7 days of treatment. | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Changes in the antioxidant enzymes will be evaluated in serum samples in different time points. |  | Blood collection will be performed at time 0, after 3, 6 hours and 7 days of treatment | 
| Changes in the miRNAs expression will be evaluated in plasma samples in different time points. |  | Blood collection will be performed at time 0, after 3, 6 hours and 7 days of treatment | 
| Qualitative and quantitative changes in fecal metabolome will be evaluated by an untargeted metabolomic approach based on Nuclear Magnetic Resonance spectroscopy (NMR). |  | Feces collection will be performed at time 0 and after 7 days of treatment. |