Bioavailability of Vitamin B12 in Bread Using Fermented Faba Bean as B12 Source in Healthy Volunteers (BeanBread)

Study Overview
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Brief Summary
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Vitamin B12 is naturally present in animal products, but it could be added to plant-based products through fermentation using the Propionibacterium freudenreichii bacterium. This study aims to investigate the absorption of vitamin B12 synthesized by Propionibacterium freudenreichii from products made with fermented faba bean. The study products are wheat breads of which 30 % of the dry weight has been replaced with fermented faba bean flour. One of the study breads contains vitamin B12 produced by P. freudenreichii bacteria, and the other bread (control bread) is fermented using L. brevis bacteria (does not produce vitamin B12). The participants will eat the study bread for two consecutive days during the study weeks, with a washout period of 12 days. Fasting blood samples are collected on day 1 before administration of the first dose of vitamin B12 and on day 3. Nutrient intake and food consumption will be analyzed from 3-day food records at the baseline and on the bread eating days. The study is carried out in a double-blinded crossover setting.

Detailed Description
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A human intervention study with healthy volunteers will be carried out to investigate the absorption of vitamin B12 synthesized by Propionibacterium freudenreichii from bread made with fermented faba bean. The study is conducted on healthy, 18-50-year-old volunteers. The study is carried out in a randomized, double-blinded, placebo-controlled crossover design. The study products are wheat breads of which 30% of the dry weight has been replaced with fermented faba bean flour. One of the study breads contains fermented faba bean flour containing vitamin B12 produced by P. freudenreichii bacteria, and the other bread (control bread) contains faba bean flour fermented with L. brevis bacteria (does not produce vitamin B12). In addition to the study breads, the study participants are given a vitamin supplement containing cyanocobalamin as a positive control during one study week and a placebo supplement as a negative control for two weeks. During each study period, the subjects eat study breads and supplements for two consecutive days, three times a day. The study participants are otherwise allowed to follow their usual diet. On the morning of the first and third study day of each study period, a blood sample is taken from the subjects, from which the vitamin B12 bound to holotranscobalamin II and the total vitamin B12 concentration in the serum are determined. At the baseline of the study, the blood samples are also analyzed for folate, methylmalonate, homocysteine and creatinine concentrations. Nutrient intake and food consumption will be analyzed from 3-day food records during the run-in week and each study week (total 9 days). The participants will report their gastrointestinal symptoms during the study periods.

Official Title
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Bioavailability of Vitamin B12 in Bread Using Fermented Faba Bean as B12 Source in Healthy Volunteers (BeanBread)

Conditions
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Vitamin B12 Absorption

Intervention / Treatment
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* Other: Vitamin B12 bread
* Other: Control bread


Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: Healthy Body mass index between 18.5 and 35 kg/m2 No exclusion criteria listed below Exclusion Criteria: Food allergies against ingredients in foods supplemented in the study pregnancy and/or breastfeeding Regular use of food supplements containg B12 Blood donation within 3 months Alcohol consumption more than 24 measures per week Smoking, using snuff Iron, B12 or folate deficiency Anemia small intestinal bacterial overgrowth (SIBO) Gastritis Helicobacter pylori infection Acid reflux Chron's disease Celiac disease Pancreatitis Abdominal surgery Eating disorder Use of supplements containing vitamin B12 in the last year Vitamin B12 injection treatment in the last year Use of metformin, protein pump inhibitors or H2-receptor blockers

Ages Eligible for Study
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Minimum Age: 18 Years
Maximum Age: 50 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: Prevention
Allocation: Randomized
Intervention Model: Crossover Assignment
Masking: Double


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Active Comparator: Vitamin B12 bread<br>Daily dose of bread containing B12 (15 µg of vitamin B12/d) and placebo tablet (0 µg of vitamin B12/d) for 2 consecutive days. | Other: Vitamin B12 bread<br>* Wheat bread with 30 % of wheat flour replaced with fermented faba bean flour containing vitamin B12 produced by P. freudenreichii bacteria.<br>|
| Active Comparator: Positive control<br>Daily dose of control bread (0 µg of vitamin B12/d and B12 tablet (15 µg of vitamin B12/d) for 2 consecutive days. | Other: Control bread<br>* Wheat bread with 30 % of wheat flour replaced with fermented faba bean flour.<br>|
| Placebo Comparator: Negative control<br>Daily dose of control bread (0 µg of vitamin B12/d and placebo tablet (0 µg of vitamin B12/d) for 2 consecutive days. | Other: Control bread<br>* Wheat bread with 30 % of wheat flour replaced with fermented faba bean flour.<br>|

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Vitamin B12 bound to transcobalamin II (holoTc) | Changes in serum holotranscobalamin concentrations after intake of vitamin B12 | 2 days | 
| Total B12 | Changes in serum total B12 concentrations after intake of vitamin B12 | 2 days | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| The intensity of gastrointestinal symptoms measured by a visual analogue scale | Study participants report the intensity of experienced stomach pain, cramps, bloating, flatulence, stomach noise, nausea, heartburn, unpleasant feeling and sudden need to defecate on a scale of zero (no pain at all) to 100 (very intense pain) | 4 weeks | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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cobalamin, holotranscobalamin, vitamin B12, absorption, propionibacterium freudenreichii, fermented faba bean