Analysis of organic acid markers relevant to inherited metabolic diseases by ultra-performance liquid chromatography/tandem mass spectrometry as benzofurazan derivatives
Analysis of organic acid markers relevant to inherited metabolic diseases by ultra-performance liquid chromatography/tandem mass spectrometry as benzofurazan derivatives
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DOI：
10.1002/rcm.3053
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发表时间：
2007-01-01
影响因子：
2
通讯作者：
Rashed, Mohamed S.
中科院分区：
文献类型：
--
作者：
Al-Dirbashi, Osama Y.;Santa, Tomofumi;Rashed, Mohamed S.
我们描述了一种新的方法，适用于确定有机酸，作为诊断标志物的几个遗传性代谢紊乱。我们利用液相色谱/串联质谱分析最近描述的苯并呋咱试剂的有机酸衍生物。衍生化步骤是必要的，以获得有机酸衍生物，适用于分析的反相液相色谱与高电离效率的质谱在正离子模式。在这项工作中，分析了一组含有5个或6个碳原子的相关二羧酸标记物，并对1型谷氨酸血症的特异性标记物谷氨酸和3-羟基谷氨酸进行了验证。通过在温和条件下使未处理的尿液与衍生化试剂反应来实现衍生化。在C18超高效液相色谱（UPLC）柱上分析反应混合物（50 x 2.1 mm，1.7 μ m），并在多反应监测模式下在5分钟内检测到。校准曲线线性高达至少1000 μ M，检测限为0.025和0.02 μ M的谷氨酸和3-羟基谷氨酸，（信噪比为3）。日内（n =11）和日间（n = 6）的变异系数优于11.2%。该检测方法成功应用于对照（n = 134）和1型谷氨酸血症（n = 55）尿液样本。版权所有（C）2007约翰威利父子有限公司
We describe a new approach applicable to the determination of organic acids that serve as diagnostic markers for several inherited metabolic disorders. We utilized liquid chromatography/tandem mass spectrometry for analysis of organic acid derivatives of a recently described benzofurazan reagent. The derivatization step was necessary to obtain organic acid derivatives suitable for analysis by reversed-phase liquid chromatography with high ionization efficiency for mass spectrometry in the positive-ion mode. In this work, a group of related dicarboxylic acid markers containing five or six carbon atoms were analyzed and validation was performed for glutaric and 3-hydroxyglutaric acids, the specific markers for glutaric acidemia type 1. Derivatization was achieved by reacting untreated urine with the derivatization reagent under mild conditions. The reaction mixture was analyzed on a C18 ultra-performance liquid chromatography (UPLC) column (50 x 2.1 mm, 1.7 mu m) and detected in the multiple reaction monitoring mode in 5 min. Calibration curves were linear up to at least 1000 mu M with detection limits for glutaric and 3-hydroxyglutaric acids of 0.025 and 0.02 mu M, respectively (signal-to-noise ratio of 3). Intra-day (n =11) and inter-day (n = 6) coefficients of variation were better than 11.2%. The assay was successfully applied to control (n = 134) and glutaric acidemia type 1 (n = 55) urine samples. Copyright (C) 2007 John Wiley & Sons, Ltd.