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Different methodologies for the measurement of peptide amino acid (PAA) in blood and plasma were compared in sheep. Preparation of blood and plasma samples consisted of a deproteinization, either chemical with sulfosalicylic acid (0.04 g for 1 ml of sample) or physical by ultrafiltration (10,000-MW cut-off filters), with or without a subsequent ultrafiltration through a 3,000-MW cut-off filter. Peptide concentrations were determined by quantification of amino acid concentrations before and after acid hydrolysis of samples. Free amino acid concentrations were similar by all the method used (about 2.5 and 2.7 mM, for blood and plasma respectively). Peptide concentrations were higher with chemical deproteinization (10.6 and 4.2 mM, for blood and plasma respectively) than with physical deproteinization (5.7 and 3.3 mM, for blood and plasma respectively). When the deproteinized samples were further treated to remove material of molecular weight above than 3 kDa, peptide concentrations were significantly reduced, which indicates inefficiencies in the ability of the deproteinizing procedures in removing all the proteinaceous materials. Concentration of small PAA (< 3 kDa) in blood was about 1.5-fold that in plasma, mainly due to peptide Gly and Glu derived from the hydrolysis of the erythrocyte glutathione. The choice of a methodology for quantifying circulating peptides is discussed.

Digestion and portal net flux of nutrients were studied in sheep fed twice daily with fresh orchard-grass. Digestive flows were measured in six fistulated sheep using the double-marker technique. Three sheep were fitted with catheters and blood-flow probes, allowing nutrient net flux measurements across the portal-drained viscera (PDV), the mesenteric-drained viscera (MDV) and the rumen. Total tract apparent digestion of N was similar to portal net appearance of N, calculated as the sum of free amino acids (FAA), peptide amino acids (PAA), NH3, and urea net fluxes. PAA accounted for 25 % of non-protein amino acid net release across the PDV. With the exception of glycine and glutamate, the small intestine was the main contributor to this PAA net release. The essential amino acid (EAA) apparent disappearance between the duodenum and the ileum was lower than the net appearance of EAA (FAA + PAA) across the MDV. The value of PDV:MDV flux of free EAA was, on average, 78 %. The rumen accounted for 30 % of the net uptake of EAA by the PDV tissues not drained by the mesenteric vein. Rumen net release of acetate, propionate, butyrate, 3-hydroxybutyrate, and lactate accounted for 70, 55, 46, 77 and 52 %, respectively, of their portal net releases. Conversely, the small intestine was a net consumer of arterial acetate and 3-hydroxybutyrate. Dynamic study of nutrient net fluxes across the PDV showed that throughout a feeding cycle, the liver faced a constant flux of amino acids (AA), whereas volatile fatty acid and NH3 net fluxes varied in response to the meal. The present study specified, in forage-fed sheep, the partitioning of nutrient net fluxes across the PDV and the role of peptides in portal net release of AA. [ABSTRACT FROM PUBLISHER]

Different methodologies for the measurement of peptide amino acid (PAA) in blood and plasma were compared in sheep. Preparation of blood and plasma samples consisted of a deproteinization, either chemical with sulfosalicylic acid (0.04g for l ml of sample) or physical by ultrafiltration (10,000-MW cut-off filters), with or without a subsequent ultrafiltration through a 3,000-MW cut-off filter. Peptide concentrations were determined by quantification of amino acid concentrations before and after acid hydrolysis of samples. Free amino acid concentrations were similar by all the method used (about 2.5 and 2.7 mM, for blood and plasma respectively). Peptide concentrations were higher with chemical deproteinization (10.6 and 4.2 mM, for blood and plasma respectively) than with physical deproteinization (5.7 and 3.3mM, for blood and plasma respectively). When the deproteinized samples were further treated to remove material of molecular weight above than 3 kDa, peptide concentrations were significantly reduced, which indicates in,efficiencies in the ability of the deproteinizing procedures in removing all the proteinaceous materials. Concentration of small PAA (< 3 kDa) in blood was about 1.5-fold that in plasma, mainly due to peptide Gly and Glu derived from the hydrolysis of the erythrocyte glutathione. The choice of a methodology for quantifying circulating peptides is discussed. [ABSTRACT FROM AUTHOR]