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采用液相色谱-串联质谱高分辨模式对人体汗液代谢组学分析的优化研究。

Optimization study for metabolomics analysis of human sweat by liquid chromatography-tandem mass spectrometry in high resolution mode.

机构信息

Department of Analytical Chemistry, Annex Marie Curie Building, Campus of Rabanales, University of Córdoba, E-14071 Córdoba, Spain; Institute of Biomedical Research Maimónides (IMIBIC), Reina Sofía Hospital, University of Córdoba, E-14071 Córdoba, Spain.

Department of Analytical Chemistry, Annex Marie Curie Building, Campus of Rabanales, University of Córdoba, E-14071 Córdoba, Spain; Institute of Biomedical Research Maimónides (IMIBIC), Reina Sofía Hospital, University of Córdoba, E-14071 Córdoba, Spain.

出版信息

J Chromatogr A. 2014 Mar 14;1333:70-8. doi: 10.1016/j.chroma.2014.01.071. Epub 2014 Feb 1.

Abstract

Sweat has recently gained popularity as a potential tool for diagnostics and biomarker monitoring as it is a non-invasive biofluid the composition of which could be modified by certain pathologies, as is the case with cystic fibrosis, which increases chloride levels in sweat. The aim of the present study was to develop an analytical method for analysis of human sweat by liquid chromatography-mass spectrometry (LC-Q-TOF MS/MS) in high resolution mode. Thus, different sample preparation strategies and different chromatographic modes (HILIC and C18 reverse modes) were compared to check their effect on the profile of sweat metabolites. Forty-one compounds were identified by the MS/MS information obtained with a mass tolerance window below 4 ppm. Amino acids, dicarboxylic acids and other interesting metabolites such as inosine, choline, uric acid and tyramine were identified. Among the tested protocols, direct analysis after dilution was a suited option to obtain a representative snapshot of sweat metabolome. In addition, sample clean up by C18 SpinColumn SPE cartridges improved the sensitivity of most identified compounds and reduced the number of interferents. As most of the identified metabolites are involved in key biochemical pathways, this study opens new possibilities to the use of sweat as a source of metabolite biomarkers of specific disorders.

摘要

汗水最近作为一种潜在的诊断工具和生物标志物监测手段受到关注,因为它是一种非侵入性的生物流体,其成分可以被某些病理改变,就像囊性纤维化一样,会增加汗液中的氯水平。本研究的目的是开发一种通过液相色谱-质谱(LC-Q-TOF MS/MS)在高分辨率模式下分析人汗的分析方法。因此,比较了不同的样品制备策略和不同的色谱模式(亲水作用色谱和 C18 反相模式),以检查它们对汗液代谢物谱的影响。通过获得的质量公差窗口低于 4 ppm 的 MS/MS 信息,鉴定了 41 种化合物。鉴定出的氨基酸、二羧酸和其他有趣的代谢物,如肌苷、胆碱、尿酸和酪胺。在测试的方案中,直接稀释后分析是获得汗液代谢组代表性快照的合适选择。此外,通过 C18 螺旋柱 SPE 小柱进行样品净化可提高大多数鉴定化合物的灵敏度,并减少干扰物的数量。由于大多数鉴定出的代谢物都参与了关键的生化途径,因此这项研究为将汗水作为特定疾病的代谢物生物标志物的来源开辟了新的可能性。

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