Idborg Helena, Zamani Leila, Edlund Per-Olof, Schuppe-Koistinen Ina, Jacobsson Sven P
Stockholm University, Department of Analytical Chemistry, Svante Arrhenius väg 10-12, SE-106 91 Stockholm, Sweden.
J Chromatogr B Analyt Technol Biomed Life Sci. 2005 Dec 15;828(1-2):9-13. doi: 10.1016/j.jchromb.2005.07.031. Epub 2005 Sep 21.
Complex biological samples, such as urine, contain a very large number of endogenous metabolites reflecting the metabolic state of an organism. Metabolite patterns can provide a comprehensive signature of the physiological state of an organism as well as insights into specific biochemical processes. Although the metabolites excreted in urine are commonly highly polar, the samples are generally analyzed using reversed-phase liquid chromatography mass spectrometry (RP-LC/MS). In Part 1 of this work, a method for detecting highly polar metabolites by hydrophilic interaction liquid chromatography-electrospray ionization mass spectrometry (HILIC/ESI-MS) is described as a complement to RP-LC/ESI-MS. In addition, in an accompanying paper (Part 2), different multivariate approaches to extracting information from the resulting complex data are described to enable metabolic fingerprints to be obtained. The coverage of the method for the screening of as many metabolites as possible is highly improved by analyzing the urine samples using both a C(18) column and a ZIC-HILIC column. The latter was found to be a good alternative when analyzing highly polar compounds, e.g., hydroxyproline and creatinine, to columns typically used for reversed-phase liquid chromatography.
复杂的生物样本，如尿液，含有大量反映生物体代谢状态的内源性代谢物。代谢物模式可以提供生物体生理状态的全面特征，以及对特定生化过程的深入了解。尽管尿液中排泄的代谢物通常具有很高的极性，但这些样本一般采用反相液相色谱 - 质谱联用（RP-LC/MS）进行分析。在本研究的第一部分，描述了一种通过亲水作用液相色谱 - 电喷雾电离质谱联用（HILIC/ESI-MS）检测高极性代谢物的方法，作为RP-LC/ESI-MS的补充。此外，在一篇配套论文（第二部分）中，描述了从所得复杂数据中提取信息的不同多变量方法，以获得代谢指纹图谱。通过使用C(18)柱和ZIC-HILIC柱分析尿液样本，该方法筛选尽可能多代谢物的覆盖范围得到了极大提高。在分析高极性化合物（如羟脯氨酸和肌酐）时，发现后者是通常用于反相液相色谱的色谱柱的良好替代品。
