| Literature DB >> 29392696 |
Jingjing Deng1, Guoan Zhang2, Thomas A Neubert3.
Abstract
Mass spectrometry (MS)-based techniques have been finding utility as sensitive, high throughput metabolite analysis tools for complex biological samples. We describe here a nanoflow liquid chromatography-tandem mass spectrometry (nano-LC-MS/MS) system we developed and applied to metabolic profiling of human cells. Metabolites are extracted from cells using methanol, and filtered through a C18 StageTip to remove large particles. Metabolite samples are separated by HPLC at a flow rate of 400-500 nl/min, then analyzed in both positive and negative ion modes in an LTQ-Orbitrap MS. Metabolite identification and differential analysis are performed using commercial or open source software. Protocols outlined in this chapter describe how nano-LC-MS can be applied to investigate metabolic profiling with limited biomass amount.Entities:
Keywords: Extraction; Liquid chromatography; Mass spectrometry; Metabolite
Mesh：
Year: 2018 PMID： 29392696 DOI： 10.1007/978-1-4939-7659-1_10
Source DB: PubMed Journal: Methods Mol Biol ISSN： 1064-3745