Amino Acid Analysis

Amino acid analysis is used to determine the amino acid content of amino acid–, peptide- and protein-containing samples. With minor exceptions, proteins are long linear polymers of amino acids connected to each other via peptide bonds. The first step of amino acid analysis involves hydrolyzing these peptide bonds. The liberated amino acids are then separated, detected, and quantified. The method was first developed by Moore, Stein and coworkers in the 1950s using HCl acid hydrolysis, and, despite considerable effort by many workers, the basic methodology remains relatively unchanged. This unit provides an overview and strategic planning for amino acid analysis, discussing a range of methodologies and issues. In addition, several common methods used for analysis of l-amino acids are described in detail, including: HCl acid hydrolysis, performic acid oxidation for methionine and cysteine analysis, base hydrolysis for tryptophan analysis, analysis of free amino acids, and analysis of reactive lysine

True ileal amino acid digestibility of goat and cow milk infant formulas

Goat milk is used as an alternative to cow milk for the production of infant formulas. However, little is known about the protein quality and, specifically, about the digestible AA pattern of goat milk formulas compared with their cow milk counterparts. In this study, the true ileal AA digestibility of a goat milk infant formula was compared with a premium cow milk infant formula. The 3-wk-old piglet was used as a model for the 3-mo-old infant. Both milk formulas were prepared as described by the manufacturer, with titanium dioxide added as an indigestible marker. The formulas were fed to the piglets over a 2-wk trial period. Digesta from the terminal ileum were collected post euthanasia and analyzed for AA content, along with samples of the formulas. True AA digestibility was determined after correcting for endogenous AA loss at the terminal ileum of pigs fed an enzyme-hydrolyzed casein-based diet, followed by ultrafiltration (5,000 Da) of the digesta. Total urine and feces collection was also undertaken to determine the nitrogen retention from the diets. The true ileal AA digestibility was similar between the goat and cow milk infant formulas for all AA except Gly and Trp. There was no significant difference in the nitrogen retention of piglets fed the two different formulas. The goat milk infant formula and the premium cow milk infant formula were similar in terms of protein qualit

Mineral retention in three-week-old piglets fed goat and cow milk infant formulas

Goat milk and cow milk are commonly used in infant formula preparations and, as such, understanding the nutritional characteristics of infant formulas made from these milks is important. In this study, a goat milk infant formula was compared with an adapted (whey-enhanced) cow milk infant formula with respect to mineral absorption and deposition using the 3-wk-old piglet as a model for the 3-mo-old infant. Equal numbers of piglets (n = 8) were fed either the goat milk formula or the cow milk formula. The mineral composition of the prepared goat milk formula was higher than that of the prepared cow milk formula for most minerals, including calcium (75.1 vs. 56.7 mg/100 mL) but excluding iron, which was higher in the prepared cow milk formula (0.92 vs. 0.74 mg/100 mL). The amounts of calcium, phosphorus, and manganese absorbed by the piglets were significantly higher for the goat milk formula, whereas the amounts of zinc, iron, and magnesium absorbed were significantly higher for the cow milk formula. Apparent mineral absorption, relative to intake, was statistically higher in the cow milk formula for calcium and phosphorus, although the actual differences were very small ( less than 1.3%). For copper, zinc, iron, and magnesium there was no significant difference between treatments in apparent mineral absorption, whereas for manganese, absorption was higher for the goat milk infant formula. The absolute mineral deposition was higher in piglets fed the goat milk formula for calcium, phosphorus, and manganese, whereas iron deposition was higher in the piglets fed cow milk formula. For all other minerals tested, there were no significant differences between treatments. The goat milk infant formula provided a pattern of mineral retention in the 3-wk-old piglet very similar to that of the adapted cow milk infant formula. The minor differences observed between the 2 appeared to be due to the different mineral contents of the 2 formulas

Felinine stability in the presence of selected urine compounds

The stability of felinine, an amino acid present in feline urine, was investigated. Synthetic felinine was unstable in the urine of a selection of mammals. Felinine was found to stable in feline urine in which urea had been degraded. Synthetic felinine was found to react specifically with urea and did not react with urea analogues such as biuret or thiourea or other nucleophilic compounds such as ammonia which is more nucleophilic or acetamide and water which are less nucleophilic than urea. The reaction of urea and felinine was independent of pH over the range of 3¿10. Urea did not react with N-acetyl-felinine suggesting a felinine N-terminal interaction with urea. Mass spectral analysis of the reaction products showed the presence of carbamylated felinine and fragmentation ions derived from carbamyl-felinine. The physiological relevance of felinine carbamylation is yet to be determined

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How food protein structure can modulate the protein digestion behaviour: Evaluation by nutritional peptidomics

This study aimed to investigate the impact of protein aggregate structure on the nutritional valueof proteins, and especially on the extent of digestion, the nature and amount of peptides released.Ovalbumin was chosen as a model protein source. A range of four typical aggregate structures,from linear to spherical-agglomerated, were obtained after heating under different combinationsof pH and ionic strength. The non-aggregated and aggregated ovalbumins were digested usingan in vitro digestion model that simulated gastrointestinal digestion. The extent of digestion wasdetermined based on the disappearance of intact ovalbumin and the appearance of solublepeptides. The peptide profile of the digests was analyzed using LC-MS/MS.The extent of hydrolysis differed according to the aggregate structure with linear aggregatesbeing more extensively hydrolyzed than the spherical aggregates. The results suggest that thesurface area to volume ratio, and probably the degree of unfolding of its constituent proteinbefore ingestion are the major influencing criteria. Ovalbumin aggregation appeared to render anumber of peptide bonds accessible to digestive proteases, whereas these bonds were notaccessible in the native ovalbumin; moreover, cleavage sites appeared to be specific dependingon the structure of aggregates (fig.1). Nutritional peptidomics analysis by multivariate statisticalapproaches made it possible to connect the aggregate structure and the profile of generatedpeptides. For instance, it was possible to demonstrate that bioactive peptides were significantlypromoted after digestion of spherical and spherical-agglomerated aggregates.This work highlights the existing links between food structures resulting from technologicalprocesses and their breakdown during the digestive process. Such results imply that a fine tuningof unfolding and aggregation conditions can be used for targeted structuring that can lead to animprovement of the nutritional quality of food proteins