Bioavailability of Protein and Amino Acids From Oilseeds in Healthy Volunteers
试验速览
- 阶段
- 不适用
- 入组人数
- 40
- 试验地点
- 2
- 主要终点
- Postprandial metabolism of glucose 2
研究概览
简要总结
The study aims to determine in healthy subjects the bioavailability of protein and amino acids of 4 protein sources: sunflower, rapeseed, lupin, flax. For this purpose, the investigators will compare two methods:
- the standard method consisting in measuring the ileal digestibility using ileal tubes
- an alternative method that has been proposed by an Food and Agriculture Organization (FAO) expert group: the dual isotope method
详细描述
4 groups of healthy volunteers are recruited (males and females from 18 to 65 y old), BMI from 18 to 30.
Each group tests a protein source (20g) incorporated in a biscuit that is divided in small portions to perform a repeated meal protocol. Protein sources (sunflower, rapeseed, flaxseed or lupin) are intrinsically labeled with 15N and deuterium (2H).
The day before the meal test, the volunteers are equipped with an intestinal tube that migrates until the ileum. On the day of the experiment, the position of the intestinal tube is checked by radiography in order to verify its location at the terminal ileum. A catheter is inserted in the forearm vein for blood sampling. A saline solution containing polyethylene glycol 4000 (PEG-4000, 20 g/L), used as a non-absorbable marker of the intestinal flow, is infused into the ileum in order to calculate the flow rate of the intestinal effluents. After a basal blood and urine sampling, as well as a basal collection of ileal effluents performed for 30 min, subject ingests at t=0 the first biscuit dose. The small biscuit portions are ingested every 30 min for 4h, to achieve an isotopic plateau. Tracer doses of carbon 13 (13C) amino acids are ingested concomitantly with the biscuit doses. Non absorbable markers are added to the test meal to correct for incomplete recovery at the ileal level.
The postprandial sampling period lasts for 8 h after the meal ingestion. The intestinal content is continuously collected over ice and pooled every 30 min. Blood is sampled every 30 min for 4 h, and hourly thereafter.Total urine is collected every 2 h.
Measurements:
研究设计
- 研究类型
- Interventional
- 分配方式
- Non Randomized
- 干预模型
- Parallel
- 主要目的
- Basic Science
- 盲法
- Single (Participant)
入排标准
- 年龄范围
- 18 Years 至 65 Years（Adult, Older Adult）
- 性别
- All
- 接受健康志愿者
- 否
入选标准
- •18<BMI<30
- •Insured under the French social security system
- •for women: use of birth control
- •signed informed consent
排除标准
- •Any dietary allergy
- •Latex allergy
- •Positive serology to HIV, hepatite C virus antibody, hepatite B virus surface antigen and core antibodies
- •Gluten intolerance
- •Abusive use of drugs or alcohol
- •Hypertension, diabetes, digestive disease, hepatic or renal disease, severe cardiac disease
- •Pregnancy
- •High sport practicing (>7h/wk)
- •Blood donation in the 3 months prior to the study
- •Participation in a clinical study in the 3 months prior to the study
- •No signed informed consent
研究组 & 干预措施
Sunflower
Biscuits containing sunflower isolate intrinsically labelled with 15N and 2H
干预措施: Sunflower (Dietary Supplement)
Rapeseed
Biscuits containing rapeseed isolate intrinsically labelled with 15N and 2H
干预措施: Rapeseed (Dietary Supplement)
Flaxseed
Biscuits containing flaxseed isolate intrinsically labelled with 15N and 2H
干预措施: Flaxseed (Dietary Supplement)
Lupin
Biscuits containing lupin flour intrinsically labelled with 15N and 2H
干预措施: Lupin (Dietary Supplement)
结局指标
主要结局
Postprandial metabolism of glucose 2
时间窗: time -30 minutes to 8 hours after the meal
Insulinemia (by ELISA) will be determined in plasma.
Percentage of ileal digestibility of plant proteins
时间窗: time -30 minutes to 8 hours after the meal
Total N and 15N will be measured by Elemental Analyzer coupled to an Isotopic Ratio Mass Spectrometer (EA-IRMS) and the percentage of dietary N reaching the terminal ileum will be then determined. This percentage will be corrected by the ileal flow rate which is estimated with measurement of PEG concentration in effluents by a turbidimetric methods. Ileal digestibility of plant proteins (i.e. N digestibility) will be calculated as 100 - corrected dietary N percentage.
Percentage of ileal digestibility of amino acids from plant proteins
时间窗: time -30 minutes to 8 hours after the meal
Amino acid concentrations will be measured by Ultra high performance liquid chromatography (UHPLC) and 15N, 2H and 13C enrichment in individual amino acids will be measured by Gas Chromatography-Combustion-Isotope Ratio Mass Spectrometry (GC-C-IRMS) in order to determine the percentage of dietary individual amino acids reaching the terminal ileum. This percentage will be corrected by the ileal flow rate which is estimated with measurement of PEG concentration in effluents by a turbidimetric methods. Ileal digestibility of plant amino acid will be calculated as 100 - corrected dietary amino acid percentage.
Percentage of net postprandial protein utilization of plant proteins
时间窗: time -30 minutes to 8 hours after the meal
Urea in plasma and urine (by enzymatic methods) will be determined and associated with 15N enrichment values (by EA IRMS) to determine the 15N transfer in plasma and urinary N pools and calculate the net postprandial protein utilization.
Postprandial metabolism of glucose 1
时间窗: time -30 minutes to 8 hours after the meal
Glycemia (by enzymatic methods) will be determined in plasma.
Postprandial plasma kinetics of dietary amino acids
时间窗: time -30 minutes to 8 hours after the meal
Amino acids in plasma will be determined by UHPLC and associated with 15N enrichment values in individual amino acids (by GC C IRMS) to determined the kinetics of amino acids from plant proteins.
次要结局
- Validation of the dual isotope method to assess the bioavailability of individual amino acids(time -30 minutes to 8 hours after the meal)
研究者
Robert Benamouzig
Professor
Institut National de Recherche pour l'Agriculture, l'Alimentation et l'Environnement