title: Metabolic profiling of body fluids and multivariate data analysis.
abstract: Metabolome analyses of body fluids are challenging due pre-analytical variations, such as pre-processing delay and temperature, and constant dynamical changes of biochemical processes within the samples. Therefore, proper sample handling starting from the time of collection up to the analysis is crucial to obtain high quality samples and reproducible results. A metabolomics analysis is divided into 4 main steps: 1) Sample collection, 2) Metabolite extraction, 3) Data acquisition and 4) Data analysis. Here, we describe a protocol for gas chromatography coupled to mass spectrometry (GC-MS) based metabolic analysis for biological matrices, especially body fluids. This protocol can be applied on blood serum/plasma, saliva and cerebrospinal fluid (CSF) samples of humans and other vertebrates. It covers sample collection, sample pre-processing, metabolite extraction, GC-MS measurement and guidelines for the subsequent data analysis. Advantages of this protocol include: •Robust and reproducible metabolomics results, taking into account pre-analytical variations that may occur during the sampling process•Small sample volume required•Rapid and cost-effective processing of biological samples•Logistic regression based determination of biomarker signatures for in-depth data analysis.
authors.affiliation: Luxembourg Centre for Systems Biomedicine, University of Luxembourg, Belvaux, Luxembourg; Integrated BioBank of Luxembourg, Strassen, Luxembourg.
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Trezzi
authors.affiliation: Luxembourg Centre for Systems Biomedicine, University of Luxembourg, Belvaux, Luxembourg.
authors.forename: Christian
authors.initials: C
authors.lastname: Jäger
authors.affiliation: Functional Proteomics, Medizinisches Proteom-Center, Ruhr-University Bochum, Germany.
authors.forename: Sara
authors.initials: S
authors.lastname: Galozzi
authors.affiliation: Functional Proteomics, Medizinisches Proteom-Center, Ruhr-University Bochum, Germany.
authors.forename: Katalin
authors.initials: K
authors.lastname: Barkovits
authors.affiliation: Functional Proteomics, Medizinisches Proteom-Center, Ruhr-University Bochum, Germany.
authors.forename: Katrin
authors.initials: K
authors.lastname: Marcus
authors.affiliation: Paracelsus-Elena Klinik, Kassel, Germany; University Medical Center Goettingen, Institute of Neuropathology and Department of Neurosurgery, Goettingen, Germany.
authors.forename: Brit
authors.initials: B
authors.lastname: Mollenhauer
authors.affiliation: Luxembourg Centre for Systems Biomedicine, University of Luxembourg, Belvaux, Luxembourg; Braunschweig Integrated Centre of Systems Biology, University of Braunschweig, Rebenring 56, Braunschweig, Germany; Department of Computational Biology of Infection Research, Helmholtz Centre for Infection Research, Braunschweig, Germany.
authors.forename: Karsten
authors.initials: K
authors.lastname: Hiller
keywords: Blood; Body fluids; Cerebrospinal fluid; Data analysis; GC–MS; Metabolic profiling of body fluids and multivariate data analysis; Metabolite extraction; Metabolomics; Saliva