Department of Analytical Chemistry, Physical Chemistry and Chemical Engineering, University of Alcalá, Madrid, Spain.
Center for Proteomics and Metabolomics, Leiden University Medical Center, Leiden, The Netherlands.
Sci Rep. 2019 Jan 28;9(1):806. doi: 10.1038/s41598-018-37512-8.
Capillary electrophoresis-mass spectrometry (CE-MS) using a sheathless porous tip interface emerged as an attractive tool in metabolomics thanks to its numerous advantages. One of the main advantages compared to the classical co-axial sheath liquid interface is the increased sensitivity, while maintaining the inherent properties of CE, such as a high separation efficiency and low sample consumption. Specially, the ability to perform nanoliter-based injections from only a few microliters of material in the sample vial makes sheathless CE-MS a well-suited and unique approach for highly sensitive metabolic profiling of limited sample amounts. Therefore, in this work, we demonstrate the utility of sheathless CE-MS for metabolic profiling of biomass-restricted samples, namely for 20 µm-thick tissue sections of kidney from a mouse model of polycystic kidney disease (PKD). The extraction method was designed in such a way to keep a minimum sample-volume in the injection vial, thereby still allowing multiple nanoliter injections for repeatability studies. The developed strategy enabled to differentiate between different stages of PKD and as well changes in a variety of different metabolites could be annotated over experimental groups. These metabolites include carnitine, glutamine, creatine, betaine and creatinine. Overall, this study shows the utility of sheathless CE-MS for biomass-limited metabolomics studies.
无鞘毛细管电泳-质谱联用(CE-MS)采用无鞘多孔尖端接口,由于其众多优点,成为代谢组学中一种很有吸引力的工具。与经典的同轴鞘液界面相比,它的主要优点之一是灵敏度更高,同时保持 CE 的固有特性,如高分离效率和低样品消耗。特别是,能够从样品小瓶中的几微升材料中进行基于纳升的进样,使得无鞘 CE-MS 成为一种非常适合且独特的方法,适用于对有限量样品进行高度敏感的代谢物分析。因此,在这项工作中,我们展示了无鞘 CE-MS 在生物质受限样品代谢物分析中的应用,即用于多囊肾病(PKD)小鼠模型的 20 μm 厚肾脏组织切片的代谢物分析。该提取方法的设计方式使进样小瓶中的样品体积最小化,从而仍然允许进行多次纳升级重复进样以进行重复性研究。所开发的策略能够区分 PKD 的不同阶段,并且可以对实验组之间的各种不同代谢物的变化进行注释。这些代谢物包括肉碱、谷氨酰胺、肌酸、甜菜碱和肌酸酐。总的来说,这项研究表明无鞘 CE-MS 适用于生物质受限的代谢组学研究。