title: From sample treatment to biomarker discovery: A tutorial for untargeted metabolomics based on GC-(EI)-Q-MS.
abstract: This tutorial provides a comprehensive description of the GC-MS-based untargeted metabolomics workflow including: ethical approval requirement, sample collection and storage, equipment maintenance and setup, sample treatment, monitoring of analytical variability, data pre-processing including deconvolution by free software such as AMDIS, data processing, statistical analysis and validation, detection of outliers and biological interpretation of the results. For each stage tricks will be suggested, pitfalls will be highlighted and advice will be provided on how to get the best from this methodology and technique. In addition, a step-by-step procedure and an example of our in-house library have been included in the supplementary material to lead the user through the concepts described herein. As a case study, an interesting example from one of our experiments at CEMBIO Research Centre is described, presenting an example of the use of this ready-to use protocol for identification of a metabolite that was not previously included in Fiehn commercial target library.
authors.affiliation: Centre for Metabolomics and Bioanalysis (CEMBIO), Faculty of Pharmacy, Universidad San Pablo CEU, Campus Monteprincipe, Boadilla del Monte, 28668, Madrid, Spain. Electronic address: annalaura.mastrangelo@ceu.es.
authors.forename: Annalaura
authors.initials: A
authors.lastname: Mastrangelo
authors.affiliation: Centre for Metabolomics and Bioanalysis (CEMBIO), Faculty of Pharmacy, Universidad San Pablo CEU, Campus Monteprincipe, Boadilla del Monte, 28668, Madrid, Spain. Electronic address: ferrarini.alessia@gmail.com.
authors.forename: Alessia
authors.initials: A
authors.lastname: Ferrarini
authors.affiliation: Centre for Metabolomics and Bioanalysis (CEMBIO), Faculty of Pharmacy, Universidad San Pablo CEU, Campus Monteprincipe, Boadilla del Monte, 28668, Madrid, Spain. Electronic address: frstolle@ceu.es.
authors.forename: Fernanda
authors.initials: F
authors.lastname: Rey-Stolle
authors.affiliation: Centre for Metabolomics and Bioanalysis (CEMBIO), Faculty of Pharmacy, Universidad San Pablo CEU, Campus Monteprincipe, Boadilla del Monte, 28668, Madrid, Spain. Electronic address: antogar@ceu.es.
authors.forename: Antonia
authors.initials: A
authors.lastname: García
authors.affiliation: Centre for Metabolomics and Bioanalysis (CEMBIO), Faculty of Pharmacy, Universidad San Pablo CEU, Campus Monteprincipe, Boadilla del Monte, 28668, Madrid, Spain. Electronic address: cbarbas@ceu.es.
authors.forename: Coral
authors.initials: C
authors.lastname: Barbas
mesh_terms: D015415:Biomarkers / Q000378:metabolism*; D008401:Gas Chromatography-Mass Spectrometry / Q000379:methods*; D055432:Metabolomics
chemical_list: D015415:Biomarkers
keywords: Database; Fingerprinting; GC-MS; Global profiling; Metabolomics; Methodology