Annotation of the human serum metabolome by coupling three liquid chromatography methods to high-resolution mass spectrometry
Annotation of the human serum metabolome by coupling three liquid chromatography methods to high-resolution mass spectrometry
复制标题
DOI：
10.1016/j.jchromb.2014.04.025
复制
发表时间：
2014-09-01
影响因子：
3
通讯作者：
Junot, Christophe
中科院分区：
文献类型：
--
作者：
Boudah, Samia;Olivier, Marie-Francoise;Junot, Christophe
这项工作的目的是评估的相关性和多功能性的液相色谱耦合高分辨率质谱（LC/HRMS）进行定性和全面的研究人血清代谢组。为此，三种不同的色谱系统的基础上的反相（RP），亲水相互作用色谱（HILIC）和五氟苯基丙基（PFPP）固定相，与检测在正负电喷雾模式。首先评估了LC/HRMS平台检测、保留和分离657种代谢物标准品的能力，这些代谢物标准品代表了生物体液中出现的化学家族。在任一LC条件下有效保留超过75%，RP色谱柱仅保留不到5%。然后评价这三种LC/HRMS系统的血清代谢组覆盖率。基于RP、HILIC和PFPP的LC/HRMS方法的组合导致在负离子化模式下约1328个特征的注释和在正离子化模式下约1358个特征的注释，基于它们在至少一种色谱条件下的精确质量和精确保留时间。这些注释中只有不到12%被三个LC系统共享，这突出了它们的互补性。HILIC柱在负离子模式下确保了最大的代谢组覆盖率，而PFPP柱在正离子模式下最有效。总共，192注释确认使用我们的光谱数据库和74人通过执行MS/MS实验。这导致了266种代谢物的正式或推定鉴定，其中59种是首次在人血清中报告的。(C)2014爱思唯尔有限公司版权所有。
This work aims at evaluating the relevance and versatility of liquid chromatography coupled to high resolution mass spectrometry (LC/HRMS) for performing a qualitative and comprehensive study of the human serum metabolome. To this end, three different chromatographic systems based on a reversed phase (RP), hydrophilic interaction chromatography (HILIC) and a pentafluorophenylpropyl (PFPP) stationary phase were used, with detection in both positive and negative electrospray modes. LC/HRMS platforms were first assessed for their ability to detect, retain and separate 657 metabolite standards representative of the chemical families occurring in biological fluids. More than 75% were efficiently retained in either one LC-condition and less than 5% were exclusively retained by the RP column. These three LC/HRMS systems were then evaluated for their coverage of serum metabolome. The combination of RP, HILIC and PFPP based LC/HRMS methods resulted in the annotation of about 1328 features in the negative ionization mode, and 1358 in the positive ionization mode on the basis of their accurate mass and precise retention time in at least one chromatographic condition. Less than 12% of these annotations were shared by the three LC systems, which highlights their complementarity. HILIC column ensured the greatest metabolome coverage in the negative ionization mode, whereas PFPP column was the most effective in the positive ionization mode. Altogether, 192 annotations were confirmed using our spectral database and 74 others by performing MS/MS experiments. This resulted in the formal or putative identification of 266 metabolites, among which 59 are reported for the first time in human serum. (C) 2014 Elsevier B.V. All rights reserved.