The Bioavailability of Red Clover Isoflavones

Study Overview
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Brief Summary
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Previous literature has shown that the molecular form of isoflavones (as aglycones or glycosides) and food matrices can influence the bioavailability of these compounds in humans and hence their efficacy. To determine the effects of processing and food matrices on the bioavailability of active compounds derived from red clover the investigators will execute a 5 phase, cross-over design, open label, RCT using 20 healthy women aged between 18 to 40 years. Participants will receive 5 different formulations of red clover and bioavailability will be monitored in blood plasma using LC-MS.

Detailed Description
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The aim of this study is to elucidate the effects food matrices and processing on the bioavailability of active compounds on the bioavailability of red clover derived isoflavones. 20 healthy participants will receive red clover derived isoflavones in five different formulations. The five formulations are split up into five phases: Phase 1) Fermented extract in liquid form Phase 2) Fermented extract (freeze-dried) in capsule form Phase 3) Fermented extract (freeze-dried) in tablet form Phase 4) Fermented extract (concentrated) mixed in yogurt Phase 5) Un-fermented red clover in capsule or tablet form The trial will last 5-6 weeks for each participant. Plasma sample collection will take place both pre-and post- ingestion of a formulation, participants will receive blood tests at intervals -15 min, 0 min, 2hr, 4hr, 6hr, 8hr and 12hr (on the day of ingestion) and on the subsequent two mornings (24hr and 48hr) following the initiation of a phase. There will be a minimum. 5 day intervals between all phases (washout periods). Isoflavones biochanin A, formononetin, genistein and daidzein present in plasma will be detected using HPLC-MS for each phase, thus creating a bioavailability curve that can be compared to each of the other phases. The primary endpoint is to determine whether fermented extract is more bioavailable than unfermented extract.

Official Title
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The Effects of Processing and Matrices on Bioavailability of Red Clover Isoflavones in Healthy Women

Conditions
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Healthy

Intervention / Treatment
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* Dietary Supplement: Fermented red clover isoflavones in aglycone form
* Dietary Supplement: Unfermented glycosides (as aglycone equivalents)


Participation Criteria
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Eligibility Criteria
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Inclusion criteria: Healthy women Age: 18-40 years Exclusion Criteria: Habitual intake of soy products, chickpeas or other supplements with a high content of isoflavones Hormone therapy Pregnant or breastfeeding Taking drugs affecting uptake Participation in other clinical trials within the last 3 months Severe cardiovascular, psychiatric, neurological and / or kidney disease Alcohol or substance abuse Acute illness

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 40 Years

Sexes Eligible for Study
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Female

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: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Active Comparator: Capsule (aglycone)<br>single bolus 40mg isoflavone formulation of fermented extract in freeze-dried capsule. Fermented red clover isoflavones in aglycone form | Dietary Supplement: Fermented red clover isoflavones in aglycone form<br>* Fermented isoflavones in aglycone form<br>|
| Active Comparator: Tablet (aglycone)<br>single bolus 40mg isoflavone aglycone formulation of fermented extract in freeze-dried tablet. Fermented red clover isoflavones in aglycone form | Dietary Supplement: Fermented red clover isoflavones in aglycone form<br>* Fermented isoflavones in aglycone form<br>|
| Active Comparator: Yoghurt (aglycone)<br>single bolus 40mg isoflavone aglycone formulation of fermented extract mixed with yoghurt Fermented red clover isoflavones in aglycone form | Dietary Supplement: Fermented red clover isoflavones in aglycone form<br>* Fermented isoflavones in aglycone form<br>|
| Active Comparator: Liquid extract (aglycone)<br>single bolus 40mg isoflavone aglycone formulation of fermented extract in liquid Fermented red clover isoflavones in aglycone form | Dietary Supplement: Fermented red clover isoflavones in aglycone form<br>* Fermented isoflavones in aglycone form<br>|
| Active Comparator: Tablet unfermented (glycoside)<br>single bolus 40mg isoflavone aglycone equivalent tablet formulation of unfermented red clover isoflavones Unfermented glycosides (as aglycone equivalents) | Dietary Supplement: Unfermented glycosides (as aglycone equivalents)<br>* Unfermented aglycone equivalents in glycoside form<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Bioavailability of isoflavones in terms of fermented vs unfermented formulations | As measured by iAUC plasma concentrations of isoflavones over 48 hours | -15min, 0hr, 2hr, 4hr, 6hr, 8hr, 12hr, 24hr, 48hr | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Bioavailability of isoflavones in terms of matrices (tablet, capsule, yoghurt and liquid) | As measured by iAUC plasma concentrations of isoflavones over 48 hours | -15min, 0hr, 2hr, 4hr, 6hr, 8hr, 12hr, 24hr, 48hr | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Bioavailability, Isoflavones, Plasma, Human, Processing, Cross-over