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首页> 外文期刊>Chromatographia >Quantitative Analysis of l-Arginine, Dimethylated Arginine Derivatives, l-Citrulline, and Dimethylamine in Human Serum Using Liquid Chromatography–Mass Spectrometric Method
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Quantitative Analysis of l-Arginine, Dimethylated Arginine Derivatives, l-Citrulline, and Dimethylamine in Human Serum Using Liquid Chromatography–Mass Spectrometric Method

机译：<重点型=“小型”> L -ARIPOLINE，二甲基化精氨酸衍生物，<重点型=“小胶囊”> L-/ EMPHASEN> -Citrulline，以及使用液相色谱 - 质谱 - 质谱法的人血清中的二甲胺 方法

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摘要

Nitric oxide (NO) is a small molecule involved in the regulation of many physiological processes. It plays a crucial role in the regulation of nervous system, immune and inflammatory responses, and blood flow. NO is synthesized by nitric oxide synthase (NOS) during two-step oxidation of l -arginine to l -citrulline. Intermediates and derivatives of NO metabolism, such as l -arginine, l -citrulline, asymmetrical dimethylarginine (ADMA), symmetrical dimethylarginine (SDMA), and dimethylamine (DMA), are investigated as potential biomarkers. In this article, we present a novel analytical method that allowed for simultaneous analysis of l -arginine, ADMA, SDMA, l -citrulline, and DMA, in a single-step extraction and derivatization using benzoyl chloride. In brief, aliquots of serum were mixed with internal standard solution mixture (50?μM D6-DMA, 20?μM D7-ADMA, and 100?μM D7-arginine) and 0.025?M borate buffer, pH 9.2 (10:1:5). The derivatization process was performed at 25?°C for 5?min using 10% benzoyl chloride. A reverse phase column was used for chromatographic separation. Quantitation was performed using following ions ( m/z ): 279.1457, 286.1749, 307.1717, 314.2076, 280.1297, 150.0919, and 156.1113 for l -arginine, D7-arginine, ADMA, SDMA, D7-ADMA, l -citrulline, DMA, and D6-DMA, respectively. The method was validated, and its assay linearity, accuracy and precision, recovery, and limits of detection (1.7?μM l -arginine, 0.03?μM ADMA, 0.02?μM SDMA, 0.36?μM l -citrulline, 0.06?μM DMA) and quantification (3.2?μM? l -arginine, 0.08?μM ADMA, 0.05?μM SDMA, 1.08?μM l -citrulline, 0.19?μM DMA) were determined. The method is sensitive, reliable, repeatable, and reproducible. It can be applied in the routine clinical/diagnostic laboratory. Graphical abstract.
机译：一氧化氮（NO）是涉及许多生理过程调节的小分子。它在调节神经系统，免疫和炎症反应和血液中起着至关重要的作用。在L -Arpinine的两步氧化至L-酸酐期间，不通过一氧化氮合酶（NOS）合成。没有代谢的中间体和衍生物，例如L -Arpinine，L-硫脲，不对称二甲基碱（ADMA），对称二甲基喹（SDMA）和二甲胺（DMA），作为潜在的生物标志物。在本文中，我们提出了一种新的分析方法，其在使用苯甲酰氯的单步萃取和衍生化中允许同时分析L -Arpinine，ADMA，SDMA，L-丙氨酸和DMA。简而言之，将等分试样的血清与内标溶液混合物（50μmD6-DMA，20≤μmD7-ADMA和100​​≤μmD7-精氨酸）和0.025μm硼酸盐缓冲液，pH 9.2（10：1： 5）。使用10％苯甲酰氯，在25℃下以25℃进行衍生化方法。反相色谱柱用于色谱分离。使用以下离子（m / z）：279.1457,2076,280.1297,314.2076,280.1297,150.076,280.1297,150.0919和156.1113用于L -Arpinine，D7-精氨酸，Adma，SDMA，D7-ADMA，L-丙属，DMA和D6-DMA分别。该方法经过验证，其测定线性度，精度和精度，恢复和检测限制（1.7？μmL-Arpinine，0.03≤μm，0.02≤μmSDMA，0.36ΩμmL-Citulline，0.06？μmdma）并定量（3.2？μm≤L-Arpinine，0.08≤μmAdma，0.05？μmsdma，1.08？μml-citrulline，0.19≤μmdma）。该方法是敏感的，可靠，可重复的和可重复的。它可以应用于常规临床/诊断实验室。图形概要。

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