Bioavailability of Hesperidine and Narirutin From Orange Juice to Identify Metabotypes in Hypertension

Study Overview
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Brief Summary
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Flavonoids are polyphenolic compound mainly found in fruits and vegetables with numerous beneficial health effects as protection against cardiovascular diseases by an antihypertensive effect. The intestinal microbiota plays a key role in the metabolization of these compounds, so that differences in the composition and activity of the microbiota between individuals can generate different metabotypes. Flavonoids are found mainly in their conjugated form linked to the monosaccharide rhamnose and need to be metabolized by the intestinal bacteria, releasing the rhamnose, to be absorbed and, thus, bioactive. The bacterial enzyme responsible of rhamnose hydrolysis is α-L-rhamnosidase, whose activity can vary considerably depending on the composition of the microbiota. In fact, a great interindividual variability has been observed in the ability to absorb flavonoids, which allows to classify individuals according to the corresponding metabotype. In a previous project, the investigators confirmed the interindividual differences in the bioavailability of hesperidin and narirutin, two flavonoids naturally present in orange juice. However, the role of the intestinal microbiota in the metabolism of hesperidin and narirutin needs to be elucidated. On this basis, the following hypothesis is presented: individuals with arterial hypertension can be classified into 3 different metabotypes that are the result of the ability to absorb hesperidin and narirutin, determined by the urinary excretion of their respective metabolites, and these metabotypes are associated with different microbiota enterotypes and with different fecal α-L-rhamnosidase activity.

Official Title
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Study of the Bioavailability of Hesperidine and Narirutin Provided by an Orange Juice to Identify Various Metabotypes in Prehypertensive and Hypertensive Grade 1 Individuals (FLAVOTIP)

Conditions
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Blood Pressure, Hypertension

Intervention / Treatment
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* Other: Orange juice rich in hesperidin and narirutin


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Men and women over 18 years of age Systolic blood pressure ≤159 mm Hg Sign the informed consent Exclusion Criteria: Body mass index (BMI) ≥ 35 kg / m2 Glucose > 126 mg / dL Systolic blood pressure ≥ 160 mm Hg and diastolic blood pressure > 100 mm Hg or taking antihypertensive drugs LDL cholesterol > 189 mg / dL Triglycerides > 350 mg / dL Anemia (hemoglobin ≤ 13 g / dL in men and ≤ 12 g / dL in women) Tobacco addiction Consumption of medicines, antioxidants or vitamin supplements 30 days before the study Use of antibiotics during the last 30 days prior to the study Consumption of prebiotics and / or probiotics during the 30 days prior to the study Clinical history of gastrointestinal disease or presence of intestinal disorders at the time of inclusion Chronic alcoholism Monitoring a vegetarian diet Pregnant or with intent to get pregnant Being in breastfeeding period Participation in a clinical trial or nutritional intervention study, in the last 30 days. Inability to follow the study guidelines

Ages Eligible for Study
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Minimum Age: 18 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
Intervention Model: Single Group Assignment
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Orange juice rich in hesperidin and narirutin<br>The consumption of the orange juice will be made in a single dose of 500 ml. The juice is presented in a concentrated and frozen format, packed in opaque cans of 500 mL, for which it must be thawed and diluted with mineral water up to 1.5 L before its ingestion. | Other: Orange juice rich in hesperidin and narirutin<br>* The product is a concentrated orange juice provided by the CITRUS Department of Florida (USA: www.FloridaCitrus.org).<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Percentage of absorption of hesperidin and narirutin | Quantification of the metabolites derived from hesperidin and narirutin in basal and 24-hour urine samples, using Liquid Chromatography coupled to Mass Spectrometry (LC-QqQ). From the profile of metabolites detected in urine, the percentage of absorption of hesperidin and narirutin with respect to the amount ingested will be quantified to determine the metabotypes, based on the urinary excretion corresponding to each volunteer: | After 24 hours of juice consumption | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Fecal α-L-rhamnosidase activity | Quantification of α-L-rhamnosidase activity in fecal samples by incubation of the samples with the substrate p-nitrophenyl α-Lrhamnopyranoside and the subsequent measurement of the aborsbance to a length wavelength of 405 nM in a spectrophotometer. | Basal | 
| Gut microbiota composition | Metagenomic analysis in fecal samples. The bacteria DNA will be extracted and massive sequenced by the Ion Torrent platform. The relative abundances of the different bacterial populations will be determined in fecal samples at different taxonomic levels, and microbial diversity will be analyzed. | Basal | 
| Enterotype classification | Metagenomic analysis in fecal samples. The bacteria DNA will be extracted and massive sequenced by the Ion Torrent platform. Enterotypes will be analyzed to classify volunteers in 3 enterotypes based on their intestinal microbiota composition. | Basal | 
| Creatinine levels | Quantification of creatinine concentration in basal urine and 24 hours urine samples by a colorimetric assay. | Basal and after 24 hours of juice consumption | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Hypertension, Flavonoids, Orange juice, Metabotypes