Beneficial Impact of Orange Juice Consumption on Risk Factors Associated With Cardiovascular Diseases (CITRUS)

Randomized, Parallel and Double Blind Placebo-controlled Study for the Evaluation of Both Acute and Chronic Role of Hesperidin Consumption in 100% Orange Juice

The primary aim is to examine both the acute and chronic effects of hesperidin consumption from 100% Florida orange juice in various doses on functional and systemic markers associated with cardiovascular disease (CVD) risks.

Study Overview

• Status: Completed
• Conditions: Cardiovascular Risk Factors
• Intervention / Treatment: Dietary supplement: Control; Dietary supplement: 100% Florida OJ; Dietary supplement: 100% Florida OJ-enriched

Detailed Description

The aim of this study is to compare the effect of different doses of hesperidin in 100% Florida orange juice (OJ) when regularly or postprandially consumed on cardiovascular risk markers; in addition, the plausible role and mechanism of the hesperidin will be investigated.

The sample size was calculated using a previously available bibliography using systolic blood pressure (SBP) as the primary outcome measure. A total of 84 subjects per study product group were needed, assuming variance components of approximately 20.0, to detect differences between the three groups (control, orange juice and hesperidin-enriched orange juice (10 mm Hg)) with a bilateral significance level of α=0.05 and a power of 80 %.

The sample size was computed to be sufficient to detect differences between treatment groups regarding the evolution in time of SBP levels. Justification of chosen sample size is based on the clinically meaningful difference assigned to δ=10.0 mm Hg, which is equivalent to a difference of approximately 7.4 % in patients with baseline SBP levels of approximately 135 mm Hg. Thus, a sample of 252 participants can be used for the chronic three arm parallel trial design (84 subjects/arm) and will allow us to detect small but clinically relevant differences between the three groups with statistical robustness and direct interpretation in terms of the chronic treatment effect.

To the acute postprandial tests, the investigators have chosen n=20 subjects per arm according to the most studies that have addressed the metabolic effects of a postprandial intervention have been performed using a very similar number of subjects with statistically good quality results.

The statistical analysis will follow the principles specified in the guidelines of the ICHE9 and CPMP/EWP/908/99 ICHE9 Points to Consider on Multiplicity Issues in Clinical Trials.

The continuous efficacy variables will be analyzed by an ANCOVA (analysis of covariance) with the baseline value as a covariate.

The efficacy outcomes will be determined using the absolute values and absolute differences from the baseline. The efficacy analysis will be performed using the Available Data Only approach. In addition, the analysis of the primary efficacy variable will be performed with the Baseline Observation Carried Forward approach.

A suitable hypothesis test will be applied to the rest of the variables according to the nature of each variable, such as the Fisher exact test for categorical variables, Student's T-test for continuous variables and Mann-Whitney U test for ordinal scale variables.

The statistical tests will be applied with an α=0.05 two-sided significance level. Post-hoc analyses and comparisons between pairs of groups will be done as for exploratory purposes.

In addition, the statistical plan will be transferred to the application form of the electronic data collection report (e-CDR), which allows the improvement of data management, diminishes human errors (according threshold values of each outcome) and, overall, guarantees the maximum exploitation of human data in the context of statistical analysis.

• Study Type: Interventional
• Enrollment (Actual): 159
• Phase: Phase 3

Contacts and Locations

Study Locations

• Spain
  • Tarragona
    • Reus, Tarragona, Spain, 43204
      • Technological Centre of Nutrition and Health (CTNS)

Participation Criteria

Eligibility Criteria

• Ages Eligible for Study: 18 years to 65 years (ADULT, OLDER_ADULT)
• Accepts Healthy Volunteers: Yes
• Genders Eligible for Study: All

Description

Inclusion criteria

• Men or women 18-65 years old
• No evidence of chronic disease
• No familial CVD history
• Written informed consent provided before the initial screening visit.
• Blood pressure (with no drug intervention) >120 mm Hg systolic blood pressure ≤ 159 mmHg

Exclusion criteria

• Body mass index (BMI) ≥ 35 kg/m2
• Glucose >125 mg/dl
• Systolic blood pressure ≥ 160 mm Hg and diastolic blood pressure >100 mm Hg or taking antihypertensive medications
• Total cholesterol >240 mg/dl
• LDL-cholesterol >160 md/dl
• TAG >350
• Smoking
• Pregnant or intending to become pregnant
• Use of medications, antioxidants, or vitamin supplements
• Chronic alcoholism
• Intense physical activity (5h/week)
• Intestinal disorders
• Following of a vegetarian diet
• Anemia (hemoglobin ≤13 g/dL in men and ≤12 g/dL in women)
• Current or past participation in a clinical trial or consumption of a research product in the 30 days prior to inclusion in the study
• Failure to follow the study guidelines.

Study Plan

How is the study designed?

Design Details

• Primary Purpose: PREVENTION
• Allocation: RANDOMIZED
• Interventional Model: PARALLEL
• Masking: QUADRUPLE

Number of Arms

3

Arms and Interventions

Participant Group / Arm	Intervention / Treatment
ACTIVE_COMPARATOR: Control; Control drink (placebo)	Dietary supplement: Control; 500 mL (250 mL; 2 times/day) of placebo drink for 12 weeks
EXPERIMENTAL: Natural Florida orange juice; 100% Florida orange juice (OJ) (natural content of hesperidin)	Dietary supplement: 100% Florida OJ; 500 mL (250 mL; 2 times/day) of 100% Florida OJ for 12 weeks
EXPERIMENTAL: Enriched Florida orange juice; 100% Florida orange juice (OJ) (enriched hesperidin content)	Dietary supplement: 100% Florida OJ-enriched; 500 mL (250 mL; 2 times/day) of 100% Florida OJ-enriched for 12 weeks

What is the study measuring?

Primary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Systolic Blood Pressure	During each visit, SBP will be measured after 5 min in a seated position in a comfortable room by the physician. The measurement will be taken in duplicate at 1-min intervals using an automatic sphygmomanometer (OMRON HEM-907; Peroxfarma, Barcelona, Spain), and the average of the two measurements will be calculated.	Every 2 weeks for a total of 12 weeks.

Secondary Outcome Measures

Outcome Measure	Measure Description	Time Frame
Diastolic Blood Pressure	During each visit, diastolic BP will be measured after 5 min in a seated position in a comfortable room by the physician. The measurement will be taken in duplicate at 1-min intervals using an automatic sphygmomanometer (OMRON HEM-907; Peroxfarma, Barcelona, Spain), and the average of the two measurements will be calculated.	Every 2 weeks for a total of 12 weeks.
Ischemic reactive hyperemia (IRH)	The endothelial-dependent vasomotor functions will be measured as IRH by a Laser-Doppler linear Periflux 5000 flowmeter (Perimed AB, Järfälla, Stockholm, Sweden)	Every 4 weeks for a total of 12 weeks
Platelet aggregation	Multiplate analyzer, Roche	Every 4 weeks for a total of 12 weeks
Homocysteine	Homocysteine concentrations will be measured by liquid chromatography-mass spectrometry (LC-MS/MS)	Every 4 weeks for a total of 12 weeks
C-reactive protein (inflammatory marker)	High sensitivity C-reactive protein (hsCRP) by standardized methods in a Cobas Mira Plus autoanalyzer (Roche Diagnostics Systems, Madrid, Spain)	Every 4 weeks for a total of 12 weeks
oxidized LDL (as oxidative stress biomarker)	Mercodia Oxidized LDL ELISA kit will be used to measure the oxidized LDL (mU/L).	Every 4 weeks for a total of 12 weeks

Other Outcome Measures

Outcome Measure	Measure Description	Time Frame
Transcriptomics	Plasma collected at 0 h (V1 and V7) of 20 samples each arm (volunteers in the postprandial study). These cells will be used to perform transcriptomics analysis to detect whole gene expression changes due to the chronic consumption of two doses of hesperidin in 100% Florida orange juice.	At week 0 (V1) and 12 week (V7).
Non-targeted Metabolomics	Plasma collected at 0 h (V1 and V7) of 20 samples each arm (volunteers in the postprandial study) will be used to perform non-targeted metabolomics by Nuclear Magnetic Response Spectroscopy (NMR) to detect metabolomic profile changes due to the chronic consumption of two doses of hesperidin in 100% Florida orange juice	At week 0 (V1) and 12 week (V7).

Collaborators and Investigators

• Sponsor: Technological Centre of Nutrition and Health, Spain
• Collaborators: University Rovira i Virgili; Hospital Universitari Sant Joan de Reus; Technological Centre of Nutrition and Health; Florida Department of Citrus
• Investigators: Principal Investigator: Rosa Solà Alberich, Prof, MD, • University Rovira i Virgili / Hospital Universitari Sant Joan de Reus

Publications and helpful links

Helpful Links

• Technological Centre of Nutrition and Health
• Florida Department of Citrus

Study record dates

Study Major Dates

• Study Start: January 12, 2016
• Primary Completion (ACTUAL): July 6, 2017
• Study Completion (ACTUAL): July 6, 2017

Study Registration Dates

• First Submitted: June 17, 2015
• First Submitted That Met QC Criteria: June 19, 2015
• First Posted (ESTIMATE): June 24, 2015

Study Record Updates

• Last Update Posted (ACTUAL): February 28, 2022
• Last Update Submitted That Met QC Criteria: February 25, 2022
• Last Verified: February 1, 2022

More Information

Terms related to this study

• Keywords: Cardiovascular disease; biomarkers; systolic blood pressure; orange juice; cardiovascular risk factor; metabolomics; transcriptomics; hesperidin
• Additional Relevant MeSH Terms: Cardiovascular Diseases
• Other Study ID Numbers: CITRUS 14-12