Mayboroda Oleg A, Neusüss Christian, Pelzing Matthias, Zurek Gabriela, Derks Rico, Meulenbelt Ingrid, Kloppenburg Margreet, Slagboom Eline P, Deelder Andre M
Biomolecular Mass Spectrometry Unit, Department of Parasitology, LUMC, Leiden, The Netherlands.
J Chromatogr A. 2007 Aug 3;1159(1-2):149-53. doi: 10.1016/j.chroma.2007.04.055. Epub 2007 Apr 29.
Analysis of amino acid profiles in urine and plasma is an essential part of modern clinical diagnostic routine. Here we present an approach for the analysis of amino acids in urine by capillary electrophoresis/time-of-flight (TOF) mass spectrometry. At first a method combining improved separation, high dynamic range, and high sensitivity is presented. Detection limits in the mid nM-range are achieved through the use of pH-mediated stacking injection in combination with modern TOF detection technology. The method can be easily applied to detect differences in the amino acid profile in urine in a clinical context. Moreover, beside amino acids low molecular weight amines, peptides and related metabolites can be profiled. As a proof of concept, urine samples from patients suffering from osteoarthritis have been analyzed. Finally, the introduction of multivariate data analysis in the work flow was evaluated on spiked urine samples and real clinical material.
尿液和血浆中氨基酸谱的分析是现代临床诊断常规的重要组成部分。在此,我们介绍一种通过毛细管电泳/飞行时间(TOF)质谱法分析尿液中氨基酸的方法。首先,提出了一种结合改进的分离、高动态范围和高灵敏度的方法。通过使用pH介导的堆积进样结合现代TOF检测技术,实现了中纳摩尔范围的检测限。该方法可轻松应用于临床环境中检测尿液中氨基酸谱的差异。此外,除了氨基酸外,还可以对低分子量胺、肽和相关代谢物进行分析。作为概念验证,对骨关节炎患者的尿液样本进行了分析。最后,在加标尿液样本和真实临床材料上评估了工作流程中多元数据分析的引入情况。