Effect of the methodology on peptide amino acid concentrations in blood and plasma of sheep

L. Bernard, B. Chauveau, D. Rémond
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引用次数: 3

Abstract

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 2.7mM, 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 (< 3kDa) 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.
方法对绵羊血液和血浆中多肽氨基酸浓度的影响
比较了绵羊血液和血浆中多肽氨基酸(PAA)的不同测定方法。血液和血浆样品的制备包括脱蛋白,或化学用磺基水杨酸(0.04 g / 1ml样品)或物理超滤(10,000 MW截止过滤器),随后通过3,000 MW截止过滤器进行超滤或不进行超滤。通过测定样品酸水解前后氨基酸浓度来测定肽浓度。所有方法的游离氨基酸浓度相似(血液和血浆分别约为2.5 mm和2.7mM)。化学脱蛋白组的肽浓度(血液和血浆分别为10.6和4.2 mM)高于物理脱蛋白组(血液和血浆分别为5.7和3.3 mM)。当脱蛋白样品进一步处理以去除分子量大于3 kDa的物质时,肽浓度显着降低,这表明脱蛋白过程去除所有蛋白质物质的能力效率低下。血液中的小PAA (< 3kDa)浓度约为血浆中的1.5倍,主要是由于红细胞谷胱甘肽水解产生的肽Gly和Glu。讨论了循环多肽定量方法的选择。
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