Proteins can be classified as high or low biological value proteins depending on their composition in essential amino acids, which are those amino acids that the body cannot synthesize and must be provided through food. In recent years, there has been growing interest in reducing the consumption of proteins of animal origin, leading to the search for more sustainable protein options, such as vegetable proteins. However, vegetable options do not have a complete profile of essential amino acids. In this sense, soy protein is considered the reference vegetable protein because it has an adequate amino acid profile. However, the amount of methionine is considerably lower than that of an animal source, and it also has a considerable allergenic potential. The mixture of two types of complementary vegetable protein sources could serve as a strategy to achieve the profile of essential amino acids like that of an animal protein. The hypothesis of the present study is that the consumption of a mixture of vegetable proteins from legume and cereal sources will complement each other to achieve a bioavailability of essential amino acids equal to or greater than that observed when consuming soy protein.
An adequate protein intake is essential for the body's well-being and performance, because the body uses this nutrient in many cellular functions. Proteins are nitrogen-containing substances made up of amino acids (AA). Not only the quantity but also the quality of protein in the diet is important for the body's proper functioning. Protein quality depends on the AA profile, among other factors. 20 total AA (TAA) have been identified as necessary for human growth and metabolism. Of the 20 TAAs, nine of them are defined as essential AA (EAAs, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, threonine, tryptophan and valine), and the remaining eleven are considered non-essential AA (NEAAs, alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, proline, serine and tyrosine). In addition, branched-chain amino acids (BCAA, leucine, isoleucine, and valine) are found within the EAAs. The NEAAs can be synthesized by the body, while the EAAs must be ingested from the diet. In this sense, proteins can be classified into two groups: high or low biological value, depending on the presence or absence of all the EAAs in their composition. Today, this distinction is usually called animal or vegetable proteins, respectively, because proteins from meat, eggs, and dairy products have higher amounts of EAAs, including leucine, while proteins from legumes, seeds, or nuts have a deficit in some of them. In the diet, the consumption of a mixture of plant-based proteins (i.e., complementary proteins) is a common strategy to compensate for the fact that individual sources of plant-based proteins are often deficient in one or more EAAs. For example, the combination of rice protein (high in methionine but low in lysine) and pea protein (low in methionine but high in lysine) can produce a pea-rice protein blend with a balanced amino acid profile, free of deficiencies. Therefore, the formulation of a plant-based protein blend represents an opportunity to develop a high-quality protein option. The present study has as its main objective to evaluate the postprandial bioavailability of EAA from two plant-based protein sources present in a complete powdered meal, where one will have soy protein and the other product with a protein source from a mixture of rice and pea protein. The secondary objectives of the study are to compare the following kinetic parameters and the metabolic response of the two products with different plant-based protein sources. * Bioavailability of AAT (AAT-AUC0-240min), AANE (AANE-AUC0-240min), BCAA (BCAA-AUC0-240min) and of each of the EAA individually (EAA-AUC0-240min). * Time to reach maximum concentration of AAT (AAT-Tmax), AAE (AAE-Tmax), AANE (AANE-Tmax), BCAA (BCAA-Tmax) and of each EAA individually. * Maximum concentration of AAT (AAT-CMax), AAE (AAE-CMax), AANE (AANE-CMax), BCAA (BCAA-CMax) and of each EAA individually. * Insulinotropic response: insulin and blood glucose levels. * Objective hunger and satiety modulation: blood levels of hunger and satiety regulating hormones: glucagon-like peptide-1 (GLP-1), peptide YY (PPY), ghrelin. * Modulation of hunger and subjective satiety. * Changes in the expression of genes involved in oxidative and inflammatory metabolism. During the study there will be 3 visits: a preselection visit (V0), a visit for the first postprandial study (V1) and after one week washing period, a visit for the second postprandial study (V2).
Study Type
INTERVENTIONAL
Allocation
RANDOMIZED
Purpose
OTHER
Masking
DOUBLE
Enrollment
12
Eurecat
Reus, Spain, Spain
Bioavailability of essential aminoacids calculated by the Area Under The Curve (EAA-AUC 0-240min) of plasma essential aminoacids levels.
Fasting EAA levels in plasma will be determined before consuming the products until 4 hours postprandially at 6 points after consuming the product (30 min., 60min., 90min., 120 min., 180 min., 240 min.). Plasma EAA will be quantified by liquid chromatography coupled to triple quadrupole mass spectrometry (LC-MS/MS).
Time frame: At week 1 and week 2
Bioavailability of total amino acids (TAA-AUC0-240 min) levels in plasma.
Fasting TAA levels in plasma will be determined before consuming the product until 4 hours postprandially at 6 points after consuming the product ( 30 min., 60min., 90 min., 120 min., 180 min., 240 min.). Plasma TAA will be quantified by liquid chromatography coupled to triple quadrupole mass spectrometry (LC-MS/MS).
Time frame: At week 1 and week 2
Bioavailability of non-essential amino acids (NEAA-AUC0-240 min) levels in plasma.
Fasting NEAA levels in plasma will be determined before consuming the product until 4 hours postprandially at 6 points after consuming the product (30 min., 60min., 90 min., 120 min., 180 min., 240 min.). Plasma NEAA will be quantified by liquid chromatography coupled to triple quadrupole mass spectrometry (LC-MS/MS).
Time frame: At week 1 and week 2
Bioavailability of branched chain amino acids (BCAA-AUC0-240min) levels in plasma.
Fasting BCAA levels in plasma will be determined before consuming the product until 4 hours postprandially at 6 points after consuming the product ( 30 min., 60min., 90 min., 120 min., 180 min., 240 min.). Plasma BCAA will be quantified by liquid chromatography coupled to triple quadrupole mass spectrometry (LC-MS/MS).
Time frame: At week 1 and week 2
Bioavailability of each of the essential amino acids individually (AUC0-240min) levels in plasma.
Fasting each essential aminoacids levels in plasma will be determined before consuming the product until 4 hours postprandially at 6 points after consuming the product ( 30 min., 60min., 90 min., 120 min., 180 min., 240 min.). Plasma each essential aminoacids levels will be quantified by liquid chromatography coupled to triple quadrupole mass spectrometry (LC-MS/MS).
Time frame: At week 1 and week 2
Time for maximum plasma concentration (Tmax)
Time period for the maximum plasma concentration of TAA, NEAA, BCAA and each essential aminoacids
Time frame: At week 1 and week 2
Maximum plasma concentration (Cmax)
Maximum plasma concentration of TAA, NEAA, BCAA and each essential aminoacids.
Time frame: At week 1 and week 2
Insulinotropic response: Change in serum insulin levels (mU/L).
Fasting insulin levels in serum will be determined before consuming the product until 3 hours postprandially at 7 points after consuming the product (15min., 30min., 45min., 60min., 90min., 120min. and 180min.) Insulin levels will be measured by standardized chemiluminescence methods.
Time frame: At week 1 and week 2
Insulinotropic response: Change in serum glucose levels (mg/dL).
Fasting glucose levels in serum will be determined before consuming the product until 3 hours postprandially at 7 points after consuming the product (15min., 30min., 45min., 60min., 90min., 120min. and 180min.) Serum glucose levels will be determined by standardized spectrophotometry methods.
Time frame: At week 1 and week 2
Change of blood glucagon-like peptide-1 (GLP-1) levels (ng/mL).
Levels of blood GLP-1, will be measured by standardized chemiluminescence methods at extraction points: 0min., 30min., 60min., 120min., 180min. and 240 min.
Time frame: At week 1 and week 2
Change of blood peptide YY (PPY) levels (pg/mL).
Levels of blood PPY, will be measured by standardized chemiluminescence methods at extraction points: 0min., 30min., 60min., 120min., 180min. and 240 min.
Time frame: At week 1 and week 2
Change of blood ghrelin levels (pg/mL)
Levels of blood ghrelin will be measured by standardized chemiluminescence methods at extraction points: 0min., 30min., 60min., 120min., 180min. and 240 min.
Time frame: At week 1 and week 2
Modulation of subjective hunger
The evaluation of the sensation of subjective hunger will be assessed by using a 100 mm Visual Analogue Scale (VAS) at the following points: 0 min (baseline time), 15 min, 30 min, 45 min, 60 min, 90 min, 120 min, 180 and 240 min. The VAS is presented as a horizontal line 100 mm long, with two ends that represent the limits of the phenomenon to be evaluated: One end of the line is labeled "No hunger" (0), while the other end is marked as "extreme hunger" (100). The volunteer is asked to mark on the line the point that best represents the intensity of what he or she is feeling at that moment. The mark made is measured in millimeters from the bottom end (zero), and the value obtained is the quantitative representation of the phenomenon evaluated.
Time frame: At week 1 and week 2
Modulation of subjective satiety
The evaluation of the sensation of subjective satiety will be assessed by using a 100 mm Visual Analogue Scale (VAS) at the following points: 0 min (baseline time), 15 min, 30 min, 45 min, 60 min, 90 min, 120 min, 180 and 240 min. The VAS is presented as a horizontal line 100 mm long, with two ends that represent the limits of the phenomenon to be evaluated: One end of the line is labeled "not at all satisfied" (0), while the other end is marked as "totally satisfied" (100). The volunteer is asked to mark on the line the point that best represents the intensity of what he or she is feeling at that moment. The mark made is measured in millimeters from the bottom end (zero), and the value obtained is the quantitative representation of the phenomenon evaluated.
Time frame: At week 1 and week 2
Modulation of subjective fullness.
The evaluation of the sensation of subjective fullnes will be assessed by using a 100 mm Visual Analogue Scale (VAS) at the following points: 0 min (baseline time), 15 min, 30 min, 45 min, 60 min, 90 min, 120 min, 180 and 240 min. The VAS is presented as a horizontal line 100 mm long, with two ends that represent the limits of the phenomenon to be evaluated: One end of the line is labeled "no feeling of fullness" (0), while the other end is marked as "I have the greatest feeling of fullness I have ever had" (100). The volunteer is asked to mark on the line the point that best represents the intensity of what he or she is feeling at that moment. The mark made is measured in millimeters from the bottom end (zero), and the value obtained is the quantitative representation of the phenomenon evaluated.
Time frame: At week 1 and week 2
Modulation of subjective desire to eat food.
The evaluation of the sensation of subjective fullnes will be assessed by using a 100 mm Visual Analogue Scale (VAS) at the following points: 0 min (baseline time), 15 min, 30 min, 45 min, 60 min, 90 min, 120 min, 180 and 240 min. The VAS is presented as a horizontal line 100 mm long, with two ends that represent the limits of the phenomenon to be evaluated: One end of the line is labeled "I have no desire to eat food" (0), while the other end is marked as "I have the greatest desire to eat any food" (100). The volunteer is asked to mark on the line the point that best represents the intensity of what he or she is feeling at that moment. The mark made is measured in millimeters from the bottom end (zero), and the value obtained is the quantitative representation of the phenomenon evaluated.
Time frame: At week 1 and week 2
Modulation of subjective desire to eat anything fatty, salty, sweet or tasty.
The evaluation of the sensation of subjective desire to eat anything fatty, salty, sweet or tasty.will be assessed by using a 100 mm Visual Analogue Scale (VAS) at the following points: 0 min (baseline time), 15 min, 30 min, 45 min, 60 min, 90 min, 120 min, 180 and 240 min. The VAS is presented as a horizontal line 100 mm long, with two ends that represent the limits of the phenomenon to be evaluated: One end of the line is labeled "I have no desire to eat anything fatty, salty, sweet or tasty" (0), while the other end is marked as "I have the greatest desire to eat anything fatty, salty, sweet or tasty" (100). The volunteer is asked to mark on the line the point that best represents the intensity of what he or she is feeling at that moment. The mark made is measured in millimeters from the bottom end (zero), and the value obtained is the quantitative representation of the phenomenon evaluated.
Time frame: At week 1 and week 2
Expression of genes involved in inflammatory and oxidative metabolism
Changes in gene expression of genes involved in oxidative and inflammatory pathways will be quantified by transcriptomics of blood samples at baseline and at 240 min.
Time frame: At week 1 and week 2
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