Protein Sorbent Properties of Montmorillonite in Vitro and in Vivo Models

Study Overview
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Brief Summary
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Montmorillonite (MONT) is a phyllosilicate layered mineral with unique physicochemical properties, such as swelling and cation exchange capability. The aim of this project is to study, in healthy volunteers, the in vivo ability of MONT to reduce protein intestinal uptake. Furthermore, the study analyzed in vitro the MONT ability of immobilizing proteins.

Detailed Description
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Montmorillonite (MONT) is a phyllosilicate layered mineral; chemically it is hydrated sodium calcium aluminium magnesium silicate hydroxide. Potassium, iron, and other cations are common substitutes and the exact ratio of cations varies with source. Due to its unique physicochemical properties, such as swelling and cation exchange capability, as well its mechanical and thermal stability, MONT can be efficiently used as matrix for immobilization of proteins with high molecular weight (e.g. Albumin) and proteins with low molecular weight (e.g.Aflatoxines). Therefore, MONT is used for therapeutic purposes, in pharmaceutical formulations and in food preparations. In vitro assays made in our laboratory, showed that MONT absorbs, in stable manner, alimentary proteins: beta-lactoglobulins, alfa lactoalbumines and the ratio MONT-protein (w/w) 1:3, 1:5, 1:15, results in the protein capture of 50, 27, 0 % of proteins respectively. The aim of this project is to study, in healthy volunteers, the in vivo ability of MONT to reduce protein intestinal uptake evaluated with the blood MONT- protein ratio. Furthermore, the study analyzed in vitro the MONT ability of immobilizing proteins at different ratio (w/w) 1:3, 1:5, 1:15.

Official Title
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Protein Sorbent Properties of Montmorillonite in Vitro and in Vivo Models

Conditions
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Urea Cycle Disorders, Inborn, Other Metabolic Diseases

Intervention / Treatment
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* Dietary Supplement: montmorillonite 5 g
* Dietary Supplement: montmorillonite 3 g
* Dietary Supplement: Montmorillonite 1 g


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: healthy volunteers (12M, 13 F, 25-30 years). Fasting from morning Exclusion Criteria: not defined

Ages Eligible for Study
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Minimum Age: 25 Years
Maximum Age: 30 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: Diagnostic
Allocation: Non-Randomized
Intervention Model: Parallel Assignment
Masking: None (Open Label)


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: montmorillonite 5 g<br>5g Montmorillonite + 15 g protein (ratio 1:3) | Dietary Supplement: montmorillonite 5 g<br>* 5g montmorillonite + 15 g protein (ratio 1:3)<br>|
| Experimental: Montmorillonite 3 g<br>3g Montmorillonite + 15 g protein (ratio 1:5) | Dietary Supplement: montmorillonite 3 g<br>* 3g MONT+ 15 g protein (ratio 1:5)<br>|
| Experimental: Montmorillonite 1 g<br>1g Montmorillonite + 15 g protein (ratio 1:15) | Dietary Supplement: Montmorillonite 1 g<br>* 1g MONT+ 15 g protein (ratio 1:15)<br>|
| No Intervention: Control<br>A control goup will intake 15 g of protein alone | |

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Protein uptake evaluated with the blood MONT-protein ratio | A solution of MONT and whey protein dissolved in 200 ml of water at the three different ratios will be administered. The control group will intake 15g of protein alone. Blood samples will be centrifuged and analyzed by mass spectrometry (egilent HP5973 mass spectrometry). | before administration and 30, 60, 90, 120 ,180 minutes after | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Protein absorption by MONT in vitro | MONT and protein (whey protein) at three different ratio w/w (1:3, 1:5, 1:15) will be dissolved in acetate buffer 0.1M pH5 as well as a sample without MONT. Samples will incubate at RT for 1, 10, 30, 60 minutes and then centrifugate at 13000g. The surnatant will be analyzed by LOWRY method. Protein concentration will be expressed in mg/ml. The MONT ability of immobilization will be calculated respect to the protein control sample | at time of the trial | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Montmorillonite, proteic diet, in vitro and in vivo models