scholarly journals Comparison of Dual Isotope and Ileal Sampling Methods for the Determination of the Amino Acid Digestibility of Pea Protein and Casein in Adult Humans

2021 ◽  
Vol 5 (Supplement_2) ◽  
pp. 873-873
Author(s):  
Florence Guillin ◽  
Claire Gaudichon ◽  
Laetitia Guerin-Deremaux ◽  
Catherine Lefranc-Millot ◽  
Gheorghe Airinei ◽  
...  
Keyword(s):  
Amino Acid ◽  
Sampling Method ◽  
Protein Quality ◽  
Plasma Samples ◽  
Tracer Method ◽  
Pea Protein ◽  
Dual Isotope ◽  
Reference Protein ◽  
Dual Tracer

Abstract Objectives The measurement of amino acid (AA) digestibility of protein through direct ileal sampling is highly invasive and inappropriate for vulnerable populations, such as children or elderly. The new dual tracer method relies on comparing meal and plasma isotopic ratios of 1/a test protein 2/a reference protein (or AA mix) of known digestibility, each one being labelled with a different isotope. The aim of this study was to compare this new indirect dual tracer method to standard ileal method, for the determination of AA digestibility of pea protein and milk casein. Methods Fifteen healthy adult volunteers completed the study and were equipped with a naso-ileal tube. They were given 9 portions of mashed potatoes containing either pea protein or casein isolates that were intrinsically labelled with 15N and 2H. A 13C algal free AA mix was added in the meals as the reference for dual tracer method. Plasma samples were collected regularly from before the first ingestion to 8-h later, while ileal digesta were collected continuously. For ileal sampling method, the AA digestibility (RIDAA) was determined using the recovery a non-absorbable marker (PEG-4000) perfused in the ileum, and the measurement of 15N enrichment of the digesta. For the dual tracer method, the amount of AA absorbed (AbAA) was calculated by the ratio of 2H/13C enrichments in plasma and in meals. The isotopic enrichments were evaluated in digesta, plasma samples and meals by GC-C-IRMS. The AA content was measured in digesta and meals by U-HPLC. Results Mean AbAA and RIDAA of pea protein were 102.2 ± 3.1% and 94.3 ± 1.5%, respectively. Mean AbAA and RIDAA of casein were 91.9 ± 2.0% and 97.1 ± 0.8%, respectively. The dual tracer method overestimated by 10% and 5% the AA digestibility of pea protein and casein, respectively, and the variability was high. The mean ileal AA digestibility of the 13C free AA mix was high (98.1 ± 1.1%), which validated our choice to use it as the reference ‘protein’ in the dual tracer method. Conclusions Several AA digestibilities obtained with dual tracer method were in the same range as the digestibilities from ileal sampling method. The variability was high and the effect of the protein source was inconsistent. After further research and validation, the dual tracer method could lead to notable advances in the determination of protein quality in humans. Funding Sources Roquette.

EVALUATION OF PROTEIN IN FOODS: III. A STUDY OF BACTERIOLOGICAL METHODS

1959 ◽  
Vol 37 (11) ◽  
pp. 1351-1360 ◽  
Author(s):  
C. G. Rogers ◽  
J. M. McLaughlan ◽  
D. G. Chapman
Keyword(s):  
Amino Acid ◽  
Protein Quality ◽  
Animal Origin ◽  
Rat Growth ◽  
Amino Acid Requirements ◽  
Growth Assay ◽  
Assay Methods ◽  
Limiting Amino Acid ◽  
Egg Powder

Bacteriological methods for the determination of protein quality were evaluated by comparison with protein efficiency ratio (P.E.R.) values determined by a standardized rat growth assay. Enzyme or acid hydrolyzates of foods were used as the source of amino acids with hydrolyzed whole egg powder as the reference standard. With Streptococcus faecalis A.T.C.C. 9790 autolysis occurred in media containing hydrolyzates of proteins deficient in lysine, and was largely responsible for results which did not agree with P.E.R. values. In methods employing Leuconostoc mesenteroides P-60 A.T.C.C. 8042, growth was influenced only by the most limiting amino acid relative to the requirements of the test organism.Results with enzyme hydrolyzates correlated poorly with P.E.R. values, whereas, with acid hydrolyzates, a good correlation was obtained for cereal proteins. A difference in amino acid requirements was largely responsible for the lack of agreement between the P.E.R. assay and methods employing L. mesenteroides, particularly for legumes and foods of animal origin. It was concluded that bacteriological assay methods which have been proposed for protein evaluation are unsatisfactory as screening procedures for the evaluation of protein in foods.

scholarly journals Challenging in Rats the Use of 13C Spirulina as Reference Protein for the Dual Isotope Method to Determine Amino Acid Bioavailability (P08-061-19)

2019 ◽  
Vol 3 (Supplement_1) ◽  
Author(s):  
Romain Tessier ◽  
Nadezda Khodorova ◽  
Juliane Calvez ◽  
Daniel Tomé ◽  
Claire Gaudichon
Keyword(s):  
Amino Acid ◽  
Goat Milk ◽  
Protein Digestibility ◽  
Male Rats ◽  
National Agency ◽  
Dual Isotope ◽  
Isotope Method ◽  
Reference Protein ◽  
Bonferroni Test

Abstract Objectives In order to establish DIAAS in humans, the FAO recommended to develop a new method to measure indispensable amino acid (IAA) digestibility. This method uses two isotopic labeling, one for the protein to test and one for a reference protein. Spirulina was chosen as the 13C reference protein due to its commercial availability and affordability. However, the real digestibility of spirulina protein has not been measured in vivo. This work aims to assess the digestibility of spirulina and its repeatability in different meal tests in rats. Methods 23 Wistar male rats were fed a test meal containing 0.5 g of 15 N protein from either spirulina (n = 7), sunflower n = 8) or goat milk isolate (n = 8) and 10 mg of 13C labeled spirulina. Rats were euthanized 6 h after the meal and their digestive luminal contents (stomach, small intestine, ileum, caecum, colon) were collected. Protein digestibility was determined for the test and the reference proteins by measuring 15 N and 13C enrichments in the digesta by EA-IRMS. Caecal IAA digestibility of 13C spirulina was determined by measuring the quantity of AA in the caecum by UPLC and the 13C enrichment in AA by GC-C-IRMS. Group effects were tested using one way ANOVA and differences between groups using Bonferroni test. Results Six hours after ingestion, most of the dietary 15 N and 13C were found in the caecum and colon. But there at least twice more 15 N nitrogen in the caecum and colon in the spirulina group than in the two other groups. Therefore, spirulina protein digestibility (86.0 ± 0.7%) was lower (P < 0.001) than sunflower (95.1 ± 0.5%) and goat milk digestibility (97.2 ± 0.2%). 13C spirulina digestibility tended to be different (P = 0.06) when mixed to spirulina (90.6 ± 0.6%), sunflower (88.8 ± 0.5%) or goat milk (89.0 ± 0.5%) isolates. The caecal IAA digestibility of 13C spirulina was lower in the spirulina group than in sunflower and goat milk groups for every IAA tested, and the mean was 91.6 ± 0.2% for sunflower, 91.4 ± 0.4% for goat milk and 85.4 ± 0.6% for spirulina. Conclusions Spirulina protein is of lower digestibility than other animal or plant proteins. Protein and amino digestibility of a tracer dose of 13C spirulina appears to vary depending on the protein component of the meal. These results question the use of spirulina as a reference protein for the dual isotope method. Funding Sources French Research National Agency (ANR), SOFIPROTEOL. Supporting Tables, Images and/or Graphs

scholarly journals Ileal digestibility of intrinsically labeled hen's egg and meat protein determined with the dual stable isotope tracer method in Indian adults

2018 ◽  
Vol 108 (5) ◽  
pp. 980-987 ◽  
Author(s):  
Sindhu Kashyap ◽  
Nirupama Shivakumar ◽  
Aneesia Varkey ◽  
Rajendran Duraisamy ◽  
Tinku Thomas ◽  
...  
Keyword(s):  
Amino Acid ◽  
Tracer Method ◽  
Dual Isotope ◽  
Ileal Digestibility ◽  
Egg White Protein ◽  
Meat Protein ◽  
Isotope Tracer ◽  
Hen’S Egg ◽  
The Mean ◽  
Cooked Meat

ABSTRACTBackgroundProtein quality assessment through the Digestible Indispensable Amino Acid Score requires accurate measurements of true ileal protein and amino acid digestibility, for which a dual isotope technique was recently developed. However, the ileal digestibility of indispensable amino acids (IAA) in humans from high-quality proteins is not well known.ObjectiveThe aim of this study was to intrinsically label hen's egg and meat protein by the use of uniformly 2H-labeled amino acids, and to measure their true ileal indispensable amino acid (IAA) digestibility via the dual isotope method in humans.Design2H-labeled lyophilized boiled egg white protein, whole boiled egg, and cooked meat were obtained from layer hens (BV-300) administered a uniformly 2H-labeled amino acid mix orally for 35 d with their daily feed. The ileal IAA digestibility of these proteins was determined with reference to digestibility of previously characterized [U-13C]spirulina in a dual tracer method in healthy Indian subjects whose intestinal health was measured by the plasma kynurenine-to-tryptophan (KT) ratio.ResultsAll subjects had normal KT ratios. The mean ± SD true ileal IAA digestibility of 2H-labeled egg white protein, whole boiled egg, and cooked meat was 86.3% ± 4.6%, 89.4% ± 4.5%, and 92.0% ± 2.8%, respectively. Leucine digestibility correlated with the KT ratio (r = −0.772; P = 0.009).ConclusionsUniformly 2H-labeled hen's egg and meat protein can be used to measure ileal IAA digestibility by the dual isotope tracer approach in humans. The mean IAA digestibility values for these high-quality proteins in the healthy Indians studied were similar to values obtained in earlier human and animal experiments. Leucine digestibility in these meal matrices correlated with the KT ratio, but this aspect needs further evaluation. This trial was registered at the Clinical Trials Registry of India (http://ctri.nic.in) as CTRI/2018/03/012265.

scholarly journals Meat protein quality of Dormitator latifrons (Pisces: Eleotridae): arguments for use by rural communities

2019 ◽  
Vol 7 (1) ◽  
Author(s):  
Mao Ernesto Rafael Basto-Rosales ◽  
Olimpia Carrillo-Farnés ◽  
Cynthia Eugenia Montoya-Martínez ◽  
Daniel Badillo-Zapata ◽  
Gustavo Gustavo Rodríguez-Montes de Oca ◽  
...  
Keyword(s):  
Amino Acid ◽  
Rural Communities ◽  
Essential Amino Acid ◽  
Protein Quality ◽  
Amino Acid Content ◽  
Protein Digestibility ◽  
Amino Acid Profile ◽  
Meat Protein ◽  
Reference Protein

The objective of this study was to evaluate the quality of meat protein of Dormitator latifrons for humans based on its Protein Digestibility Corrected Amino Acid Score (PDCAAS). Wild and cultured specimens were evaluated for amino acid content using HPLC equipment. The calculation of PDCAAS was performed as follows: milligrams of essential amino acid in 1 g of test protein per milligram of the same amino acid in 1 g of reference protein per true digestibility. To evaluate the protein of D. latifrons in relation to that of other fish, PDCAAS was calculated from the proteins of eight fish usually used in human nutrition. D. latifrons has a good essential amino acid profile, providing the same nutritional quality as those of other fish. Although the meat of wild D. latifrons contributes only 73% of human lysine requirements, it can be complemented with other lysine sources.

Multi-criteria assessment of pea protein quality in rats: a comparison between casein, gluten and pea protein alone or supplemented with methionine

2020 ◽  
pp. 1-9 ◽  
Author(s):  
Florence M. Guillin ◽  
Claire Gaudichon ◽  
Laetitia Guérin-Deremaux ◽  
Catherine Lefranc-Millot ◽  
Dalila Azzout-Marniche ◽  
...  
Keyword(s):  
Amino Acid ◽  
Wistar Rats ◽  
Protein Quality ◽  
Young Male ◽  
Diet Group ◽  
Experimental Conditions ◽  
Pea Protein ◽  
Male Wistar Rats ◽  
Amino Acid Score ◽  
The Impact

Abstract The objective of this study was to assess the nutritional quality of pea protein isolate in rats and to evaluate the impact of methionine (Met) supplementation. Several protein diets were studied: pea protein, casein, gluten, pea protein–gluten combination and pea protein supplemented with Met. Study 1: Young male Wistar rats (n 8/group) were fed the test diets ad libitum for 28 d. The protein efficiency ratio (PER) was measured. Study 2: Adult male Wistar rats (n 9/group) were fed the test diets for 10 d. A protein-free diet group was used to determine endogenous losses of N. The rats were placed in metabolism cages for 3 d to assess N balance, true faecal N digestibility and to calculate the Protein Digestible-Corrected Amino Acid Score (PDCAAS). They were then given a calibrated meal and euthanised 6 h later for collection of digestive contents. The true caecal amino acid (AA) digestibility was determined, and the Digestible Indispensable Amino Acid Score (DIAAS) was calculated. Met supplementation increased the PER of pea protein (2·52 v. 1·14, P < 0·001) up to the PER of casein (2·55). Mean true caecal AA digestibility was 94 % for pea protein. The DIAAS was 0·88 for pea protein and 1·10 with Met supplementation, 1·29 for casein and 0·25 for gluten. Pea protein was highly digestible in rats under our experimental conditions, and Met supplementation enabled generation of a mixture that had a protein quality that was not different from that of casein.

EVALUATION OF PROTEIN IN FOODS: III. A STUDY OF BACTERIOLOGICAL METHODS

1959 ◽  
Vol 37 (1) ◽  
pp. 1351-1360 ◽  
Author(s):  
C. G. Rogers ◽  
J. M. McLaughlan ◽  
D. G. Chapman
Keyword(s):  
Amino Acid ◽  
Protein Quality ◽  
Animal Origin ◽  
Rat Growth ◽  
Amino Acid Requirements ◽  
Growth Assay ◽  
Assay Methods ◽  
Limiting Amino Acid ◽  
Egg Powder

Bacteriological methods for the determination of protein quality were evaluated by comparison with protein efficiency ratio (P.E.R.) values determined by a standardized rat growth assay. Enzyme or acid hydrolyzates of foods were used as the source of amino acids with hydrolyzed whole egg powder as the reference standard. With Streptococcus faecalis A.T.C.C. 9790 autolysis occurred in media containing hydrolyzates of proteins deficient in lysine, and was largely responsible for results which did not agree with P.E.R. values. In methods employing Leuconostoc mesenteroides P-60 A.T.C.C. 8042, growth was influenced only by the most limiting amino acid relative to the requirements of the test organism.Results with enzyme hydrolyzates correlated poorly with P.E.R. values, whereas, with acid hydrolyzates, a good correlation was obtained for cereal proteins. A difference in amino acid requirements was largely responsible for the lack of agreement between the P.E.R. assay and methods employing L. mesenteroides, particularly for legumes and foods of animal origin. It was concluded that bacteriological assay methods which have been proposed for protein evaluation are unsatisfactory as screening procedures for the evaluation of protein in foods.

scholarly journals Amino acid digestibility and poultry feed formulation: expression, limitations and application

2010 ◽  
Vol 39 (suppl spe) ◽  
pp. 279-287 ◽  
Author(s):  
Wayne L. Bryden ◽  
Xiuhua Li
Keyword(s):  
Amino Acid ◽  
Protein Quality ◽  
Poultry Feed ◽  
Data Set ◽  
Ileal Digestibility ◽  
Feed Formulation ◽  
Amino Acid Availability ◽  
Sources Of Variation

The nutritional value or quality of dietary proteins used for poultry feed formulation varies: amino acid availability is an important measure of protein quality. Determination of ileal digestibility values has become the preferred method for estimating amino acid availability. This review discusses the different approaches to the expression of digestibility results, including correction for endogenous loss and the derivatisation of standardised values. Sources of variation in values include, the assay protocol, anti-nutritional factors in feedstuffs and feed milling. Feed formulating with ileal digestibility values should allow higher dietary inclusion levels of protein feedstuffs of lower quality provided that values of different feedstuffs are additive, the age of the bird and the use of feed enzymes are considered. An Australian data set of "ileal digestible amino acid values in feedstuffs for poultry" that has recently be published is described. This overview is intended to stimulate interest in the generation and application of ileal digestibility as a method for estimating amino acid availability in poultry nutrition.

scholarly journals Determination of True Ileal Amino Acid Digestibility in the Growing Pig for Calculation of Digestible Indispensable Amino Acid Score (DIAAS)

2020 ◽  
Vol 150 (10) ◽  
pp. 2621-2623
Author(s):  
Suzanne M Hodgkinson ◽  
Hans H Stein ◽  
Sonja de Vries ◽  
Wouter H Hendriks ◽  
Paul J Moughan
Keyword(s):  
Animal Model ◽  
Amino Acid ◽  
Protein Quality ◽  
Published Data ◽  
Adult Human ◽  
Indispensable Amino Acid ◽  
Growing Pig ◽  
Amino Acid Score

ABSTRACT Digestible indispensable amino acid score (DIAAS) has been recommended by the FAO for the evaluation of protein quality in human foods, but the application of DIAAS is currently limited because of a lack of published data on the true ileal amino acid (AA) digestibility of AAs in foods. The importance of DIAAS is highlighted. To calculate DIAAS, it is necessary to determine the true ileal AA digestibility of human foods using the growing pig as an animal model for the human based on previous FAO recommendations. A method is described in detail in Supplemental Methods to determine the true ileal AA digestibility of foods for humans using the pig as a model for the adult human. Adoption of the method will enable consistency in the development of databases on predicted true ileal AA digestibility in human foods for the calculation of DIAAS.

scholarly journals Need for Accurate and Standardized Determination of Amino Acids and Bioactive Peptides for Evaluating Protein Quality and Potential Health Effects of Foods and Dietary Supplements

2008 ◽  
Vol 91 (4) ◽  
pp. 894-900 ◽  
Author(s):  
G Sarwar Gilani ◽  
Chaowu Xiao ◽  
Nora Lee
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Ion Exchange ◽  
Bioactive Peptides ◽  
Protein Quality ◽  
The United States ◽  
Health Claims ◽  
World Health ◽  
Potential Health

Abstract Accurate standardized methods for the determination of amino acid in foods are required to assess the nutritional safety and compositional adequacy of sole source foods such as infant formulas and enteral nutritionals, and protein and amino acid supplements and their hydrolysates, and to assess protein claims of foods. Protein digestibility-corrected amino acid score (PDCAAS), which requires information on amino acid composition, is the official method for assessing protein claims of foods and supplements sold in the United States. PDCAAS has also been adopted internationally as the most suitable method for routine evaluation of protein quality of foods by the Food and Agriculture Organization/World Health Organization. Standardized methods for analysis of amino acids by ion-exchange chromatography have been developed. However, there is a need to develop validated methods of amino acid analysis in foods using liquid chromatographic techniques, which have replaced ion-exchange methods for quantifying amino acids in most laboratories. Bioactive peptides from animal and plant proteins have been found to potentially impact human health. A wide range of physiological effects, including blood pressure-lowering effects, cholesterol-lowering ability, antithrombotic effects, enhancement of mineral absorption, and immunomodulatory effects have been described for bioactive peptides. There is considerable commercial interest in developing functional foods containing bioactive peptides. There is also a need to develop accurate standardized methods for the characterization (amino acid sequencing) and quantification of bioactive peptides and to carry out dose-response studies in animal models and clinical trials to assess safety, potential allergenicity, potential intolerance, and efficacy of bioactive peptides. Information from these studies is needed for determining the upper safe levels of bioactive peptides and as the basis for developing potential health claims for bioactive peptides. This information is, in turn, needed by regulatory agencies for developing appropriate policy and regulations on adding these substances to foods and for determining if health claims are scientifically substantiated.

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