title: Global metabolic profiling using ultra-performance liquid chromatography/quadrupole time-of-flight mass spectrometry.
abstract: Currently, liquid chromatography-mass spectrometry (LC-MS) is one of the most important analytical technologies for detecting hundreds of metabolites in the field of metabolomics. A recent advance in LC that has impacted metabolomics is the development of UPLC (ultra-performance liquid chromatography). In this chapter, we describe the analytical methodologies for the global metabolic profiling of serum, urine, and tissue samples using UPLC-Q-TOF (quadrupole-time-of-flight)-MS. Aqueous metabolites are extracted after adding methanol/acetonitrile/acetone and then analyzed by UPLC-MS under positive and/or negative ionization mode. With the aid of multivariate statistical analysis, separation between various groups can be observed in the score plots, and biomarkers are screened in the loading/weight/VIP (variable importance in the projection) scatterplots. Furthermore, putative markers can be identified through comparison with the authentic standards based on tandem mass spectrometry (MS/MS) fragmentation pattern and LC retention. We expect that our protocol, with modifications if necessary, can be useful in many metabolomics studies and a wide range of research areas related to small molecules and LC-MS.
authors.affiliation: Department of Pharmaceutical Analysis, School of Pharmacy, Second Military Medical University, 325 Guohe Road, Shanghai, 200433, China.
authors.forename: Yunpeng
authors.initials: Y
authors.lastname: Qi
authors.forename: Yunlong
authors.initials: Y
authors.lastname: Song
authors.forename: Haiwei
authors.initials: H
authors.lastname: Gu
authors.forename: Guorong
authors.initials: G
authors.lastname: Fan
authors.forename: Yifeng
authors.initials: Y
authors.lastname: Chai
mesh_terms: D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D006801:Humans; D013058:Mass Spectrometry / Q000295:instrumentation / Q000379:methods*; D053858:Metabolic Networks and Pathways; D055442:Metabolome; D055432:Metabolomics / Q000295:instrumentation / Q000379:methods*; D012984:Software