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Development of Precolumn Derivatization–LC/MS for Amino-Acid-Focused Metabolomics

机译：氨基酸聚焦代谢物质的术前衍生化-LC / MS的开发

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Focused metabolomics facilitates the understanding of the current phenotype of living organisms, involves highly practical methods for investigation, and allows for the acquisition of highly quantitative data for important endogenous metabolites. The authors developed novel precolumn derivatization reagents for the analysis of amino acids, which are particularly important endogenous metabolites, through precolumn liquid chromatography/tandem mass spectrometry (LC/MS/MS). One of the reagents facilitated sub-femtomole-to-attomole-scale amino acid detection. We also developed an automated precolumn derivatization amino acid analyzer using another reagent for liquid chromatography/mass spectrometry (LC/MS). With this device, nearly 40 amino acids can be analyzed within a short time (<10 min), compared with the time (~2 h) taken by a conventional automated postcolumn derivatization amino acid analyzer. For highly accurate and precise analysis of free amino acid concentration in human plasma, we validated all steps, from blood collection to measurement, by using the proposed analyzer and developed a test procedure for studying free amino acid metabolomics in human plasma. While calculating the reference intervals of plasma amino acid concentrations in healthy individuals, it was found that the balance of amino acid concentrations in plasma is useful for risk screening of diseases including cancer, diabetes, and myocardial infarction. This functional-group-targeted analysis through LC/MS for amino acid was also effective for dipeptide, tripeptide, and D, L-amino acid analyses and amino acid detection in tissue by imaging mass spectrometry.
机译：聚焦的代谢组语有助于了解目前的生物体表型，涉及高度实用的调查方法，并允许获得重要内源代谢物的高度定量数据。作者开发了一种新的预漏衍生化试剂，用于分析氨基酸，其是突出液相色谱/串联质谱（LC / MS / MS）的特别重要的内源代谢物。其中一种试剂促进了亚毫小骨对术级级氨基酸检测。我们还开发了一种用于液相色谱/质谱（LC / MS）的另一种试剂的自动捕柱衍生氨基酸分析仪。通过该装置，与传统自动化后级衍生化氨基酸分析仪相比，可以在短时间（<10分钟）内分析近40个氨基酸。对于人血浆中的游离氨基酸浓度的高度准确和精确分析，我们通过使用所提出的分析仪从血液收集到测量中验证了所有步骤，并开发了用于研究人血浆中游离氨基酸代谢物的测试程序。在计算健康个体中血浆氨基酸浓度的参考间隔的同时，发现血浆中氨基酸浓度的平衡可用于风险筛查包括癌症，糖尿病和心肌梗死的疾病。通过对氨基酸的LC / MS进行氨基酸的这种官能组靶向分析对于通过成像质谱法通过成像质谱法对偶氮，三肽和D，L-氨基酸分析和氨基酸检测也是有效的。

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