This study aims to assess the effects of broad bean hull (BBH) consumption on blood glucose and gut health. Broad bean (Vicia faba) is widely cultivated in Scotland, with the UK being the most significant European producer. The seed coat (hull or testa) is removed during broad bean processing. This is a significant secondary product that is largely discarded. Preliminary work showed that this material is comparable to wheat bran and is rich in fibre (49%) and protein (18%). Additionally, it showed a rich phytochemical profile and lower fat and carbohydrate content than wheat bran. Experiments also showed that BBH inhibited the activity of alpha-amylase and alpha-glucosidase enzymes, suggesting anti-diabetic properties. Overall, these results showed that BBH is a secondary crop product having potential as a functional food for humans. Therefore, the objective of this study is to assess in vivo in humans the physiological and functional effects of BBH. Using an acute phase randomised controlled crossover design, the study will assess how consuming BBH fortified breads affects plasma glucose and gut health. The study will recruit 18 volunteers, normal-overweight, aged 18-75 years, who habitually consume low amounts of fruits and vegetables (≤3 portions/day). The volunteers will attend two identical stand-alone intervention sessions lasting three days each following the screening. The order of the intervention sessions will be randomised. On the day before each intervention session, the participants will provide a baseline faecal sample and have a continuous glucose monitoring sensor (CGMS) attached. They will be also be given a standardised dinner. On the next morning, following a 10-12 hr fast, an indwelling antecubital cannula will be inserted, and a blood sample will be taken for measuring baseline levels of metabolites. The volunteers will be given a standardised portion of the BBH or control bread to consume, and further blood samples taken for the subsequent four hours. Breath samples will also be taken at the same time points for measuring gastric emptying. The volunteers will be provided with all the meals for the rest of the day and the subsequent two days. These will include two portions per day of either the BBH or control bread. The meals will be standardised for energy and macronutrients. The volunteers will be instructed to return to the Human Nutrition Unit on the fourth morning and provide a second faecal sample and remove the CGMS. Blood samples will be analysed for systemic bioavailability and metabolism of test meal components, glucose regulatory hormones and breath samples for quantifying gastric emptying. The faecal samples will be analysed for gut bioavailability and metabolism of test meal components, microbial counts, composition, and water content.
The study aims to confirm in vivo the effects of consuming broad bean hull on blood glucose control and gut health. Although in vitro evidence is promising, its benefits remain confirmed in vivo in humans. No studies have previously attempted to assess the impact of broad bean hull consumption on glycaemic and gut health outcomes in humans. Therefore, this study will be novel and provide useful information on the potential of using BBH as a functional food for human health. From a health perspective, it is essential to establish the bioavailability and metabolism of dietary constituents in humans. Certain metabolites are absorbed early in the gastrointestinal tract, whereas others are transported to the colon, where they are extensively metabolised by the gut microbiota. This study will characterise the bioactive metabolites present in the plasma and faecal samples obtained. This will inform on their availability to the systemic circulation and their excretion profiles. Using targeted quantitative analysis (LC-MS and GC-MS), metabolites and their in vivo concentrations will be measured. The principal metabolites analysed will be those produced by the phenylpropanoid pathway and protein and carbohydrate metabolism products. These will include derivatives and metabolites of the simple phenols, benzoic acids, phenolic acids, phenylacetic acids, phenypropionic acids, phenylpyruvic acids, phenyllactic acids, mandellic acids, phenolic dimers, acetophenones, benzaldehydes, cinnamaldehydes, benzyl alcohols, cinnamyl alcohols, indoles, isoflavones, coumarins, chalcones, flavanones, flavones, flavonols, anthocyanidins. Furthermore, short-chain fatty acids will be analysed in the participants' faecal samples. It has been shown that many of these compounds are bioactive and exhibit anti-oxidant and anti-inflammatory activity at in vivo concentrations. Therefore, these studies will provide important information on bioavailability and metabolism and correlate the indices with health biomarkers. In addition, compounds such as the carcinogenic heterocyclic amines, genotoxic nitrosamines, indoles, polyamines, bile acids, and other protein metabolites considered detrimental to human health will also be measured. It is likely, the carbohydrate and phytochemical content of plant proteins may also protect against the formation of these toxic and potentially carcinogenic protein by-products.
Study Type
INTERVENTIONAL
Allocation
RANDOMIZED
Purpose
BASIC_SCIENCE
Masking
NONE
Enrollment
9
The bean hull bread will deliver 21.96 g of fibre. Volunteers will consume a roll of bean hull bread (155g/portion) served with 25g of raspberry jam.
The control bread will deliver 4.53 g of fibre. Volunteers will consume a roll of plain bread (122g/portion) served with 25g of raspberry jam.
University of Aberdeen, Rowett Institute
Aberdeen, Aberdeen City, United Kingdom
Plasma key bioactive metabolites following the control and bean hull bread consumption
The intervention visit will involve a test meal to be consumed by subjects attending the Human Nutrition Unit (HNU) in the morning, following an overnight fast. They will be provided with a meal, which will be consumed within 15 minutes and blood samples will be collected postprandial over 4 hours. For both the control and test diet, plasma markers of the principal metabolites produced by the phenylpropanoid pathway and products of protein and carbohydrate metabolism will be measured over 4-hour interval and compared with baseline.
Time frame: over 4 hours
Plasma key bioactive metabolites following the control and bean hull bread consumption
The intervention visit will involve a test meal to be consumed by subjects attending the Human Nutrition Unit (HNU) in the morning, following an overnight fast. They will be provided with a meal, which will be consumed within 15 minutes and blood samples will be collected on the day. The participants will be provided with plain/bean hull bread rolls to be consumed on day 1 (intervention day), days 2 and 3. Fasted blood samples will be measured on day 4. The principal metabolites produced by the phenylpropanoid pathway and products of protein and carbohydrate metabolism following the 3 days consumption of the test and control diet will be measured in fasted plasma of day 4. These metabolites will be compared with the baseline (day 1) and between diets (day 4 vs day 4).
Time frame: Day 4
Faecal key bioactive metabolites excretion
The intervention visit will involve a test meal to be consumed by subjects attending the Human Nutrition Unit (HNU) in the morning, following an overnight fast. They will be provided with a meal, consumed within 15 minutes. Participants will also consume the plain/bean hull bread for days 1, 2, 3. A second faecal sample will be collected on day 4 to assess participants' gut bioavailability of key nutrients and bioactive following the chronic consumption of the plain/bean hull bread rolls. The principal metabolites produced by the phenylpropanoid pathway and products of protein and carbohydrate metabolism following the 3 days consumption of the test and control diet will be measured in faecal samples of day 4. These metabolites will be compared with the baseline (day 1) and between diets (day 4 vs day 4).
Time frame: Day 4 (post intervention)
Determine the effects of consumption bean hull fortified bread on Ghrelin levels
To investigate the effect of bean hull bread on ghrelin. The intervention visit will involve a test meal to be consumed by subjects attending the Human Nutrition Unit (HNU) in the morning, following an overnight fast. They will be provided with a meal, which will be consumed within 15 minutes and blood samples will be collected on the day. Ghrelin levels will be measured over 4 hours following the consumption of the plain/bean hull bread rolls.
Time frame: over 4 hours
Determine the effects of consumption bean hull fortified bread on GLP-1 levels
To investigate the effect of bean hull bread on the hormone GLP-1. The intervention visit will involve a test meal to be consumed by subjects attending the Human Nutrition Unit (HNU) in the morning, following an overnight fast. They will be provided with a meal, which will be consumed within 15 minutes and blood samples will be collected on the day. GLP-1 levels will be measured over 4 hours following the consumption of the plain/bean hull bread rolls.
Time frame: over 4 hours
Determine the effects of consumption bean hull fortified bread on glucagon
To investigate the effect of bean hull bread on the hormone glucagon. The intervention visit will involve a test meal to be consumed by subjects attending the Human Nutrition Unit (HNU) in the morning, following an overnight fast. They will be provided with a meal, which will be consumed within 15 minutes and blood samples will be collected on the day. Glucagon levels will be over 4 hours following the consumption of the plain/bean hull bread rolls.
Time frame: over 4 hours
Determine the effects of consumption bean hull fortified bread on PYY
To investigate the effect of bean hull bread on the hormone PYY. The intervention visit will involve a test meal to be consumed by subjects attending the Human Nutrition Unit (HNU) in the morning, following an overnight fast. They will be provided with a meal, which will be consumed within 15 minutes and blood samples will be collected on the day. PYY levels will be measured over 4 hours following the consumption of the plain/bean hull bread rolls.
Time frame: over 4 hours
Postprandial blood glucose responses (recording iAUC and AUC)
Measuring postprandial glucose levels following the test meals
Time frame: Every 5 minutes for 4 hours using a continuous glucose monitor device
Postprandial insulin responses (recording iAUC and AUC)
Measuring postprandial insulin levels following the test meals
Time frame: 4 hours and Day 4
Microbiota composition change
Changes in the composition of the microbiota will be based on next-generation sequence analysis of 16S rRNA (ribosomal ribonucleic acid) genes extracted from faecal samples provided by all volunteers
Time frame: Day 1 and 4
Gastric emptying
Measuring gastric emptying rate following the consumption of the two test meals
Time frame: 4 hours
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